Progress in clinical trials of cell transplantation for the treatment of spinal cord injury:how many questions remain unanswered?

Spinal cord injury can lead to severe motor,sensory and autonomic nervous dysfunctions.However,there is currently no effective treatment for spinal cord injury.Neural stem cells and progenitor cells,bone marrow mesenchymal stem cells,olfactory ensheathing cells,umbilical cord blood stem cells,adipose stem cells,hematopoietic stem cells,oligodendrocyte precursor cells,macrophages and Schwann cells have been studied as potential treatments for spinal cord injury.These treatments were mainly performed in animals.However,subtle changes in sensory function,nerve root movement and pain cannot be fully investigated with animal studies.Although these cell types have shown excellent safety and effectiveness in various animal models,sufficient evidence of efficacy for clinical translation is still lacking.Cell transplantation should be combined with tissue engineering scaffolds,local drug delivery systems,postoperative adjuvant therapy and physical rehabilitation training as part of a comprehensive treatment plan to provide the possibility for patients with SCI to return to normal life.This review summarizes and analyzes the clinical trials of cell transplantation therapy in spinal cord injury,with the aim of providing a rational foundation for the development of clinical treatments for spinal cord injury.     

The importance of olfactory ensheathing cells for spinal cord injury

<正>We have been conducting clinical trials on olfactory ensheathing cell(OEC)transplantation for incomplete chronic spinal cord injury(SCI)since 2005.Long-term follow-up results verified that sensory and motor functions improved in most patients to varying degrees.However,the main source of OECs     

Neural plasticity after spinal cord injury

Plasticity changes of uninjured nerves can result in a novel neural circuit after spinal cord injury, which can restore sensory and motor functions to different degrees. Although processes of neural plasticity have been studied, the mechanism and treatment to effectively improve neural plasticity changes remain controversial. The present study reviewed studies regarding plasticity of the central nervous system and methods for promoting plasticity to improve repair of injured central nerves. The results showed that synaptic reorganization, axonal sprouting, and neurogenesis are critical factors for neural circuit reconstruction. Directed functional exercise, neurotrophic factor and transplantation of nerve-derived and non-nerve-derived tissues and cells can effectively ameliorate functional disturbances caused by spinal cord injury and improve quality of life for patients.     

Rebuilding motor function of the spinal cord based on functional electrical stimulation

Rebuilding the damaged motor function caused by spinal cord injury is one of the most serious challenges in clinical neuroscience. The function of the neural pathway under the damaged sites can be rebuilt using functional electrical stimulation technology. In this study, the locations of motor function sites in the lumbosacral spinal cord were determined with functional electrical stimulation technology. A three-dimensional map of the lumbosacral spinal cord comprising the relationship between the motor function sites and the correspond-ing muscle was drawn. Based on the individual experimental parameters and normalized coordinates of the motor function sites, the motor function sites that control a certain muscle were calculated. Phasing pulse sequences were delivered to the determined motor function sites in the spinal cord and hip extension, hip lfexion, ankle plantarlfexion, and ankle dorsilfexion movements were successfully achieved. The results show that the map of the spinal cord motor function sites was valid. This map can provide guidance for the selection of electrical stimulation sites during the rebuilding of motor function after spinal cord injury.     

Edaravone combined with Schwann cell transplantation may repair spinal cord injury in rats

Edaravone has been shown to delay neuronal apoptosis, thereby improving nerve function and the microenvironment after spinal cord injury. Edaravone can provide a favorable environment for the treatment of spinal cord injury using Schwann cell transplantation. This study used rat models of complete spinal cord transection at T9. Six hours later, Schwann cells were transplanted in the head and tail ends of the injury site. Simultaneously, edaravone was injected through the caudal vein. Eight weeks later, the PKH-26-labeled Schwann cells had survived and migrated to the center of the spinal cord injury region in rats after combined treatment with edaravone and Schwann cells. Moreover, the number of PKH-26-labeled Schwann cells in the rat spinal cord was more than that in rats undergoing Schwann cell transplantation alone or rats without any treatment. Horseradish peroxidase retrograde tracing revealed that the number of horseradish peroxidase-positive nerve fibers was greater in rats treated with edaravone combined with Schwann cells than in rats with Schwann cell transplantation alone. The results demonstrated that lower extremity motor function and neurophysiological function were better in rats treated with edaravone and Schwann cells than in rats with Schwann cell transplantation only. These data confirmed that Schwann cell transplantation combined with edaravone injection promoted the regeneration of nerve fibers of rats with spinal cord injury and improved neurological function.     

Collagen scaffold combined with human umbilical cord-mesenchymal stem cells transplantation for acute complete spinal cord injury

Currently, there is no effective strategy to promote functional recovery after a spinal cord injury. Collagen scaffolds can not only provide support and guidance for axonal regeneration, but can also serve as a bridge for nerve regeneration at the injury site. They can additionally be used as carriers to retain mesenchymal stem cells at the injury site to enhance their effectiveness. Hence, we hypothesized that transplanting human umbilical cord-mesenchymal stem cells on collagen scaffolds would enhance healing following acute complete spinal cord injury. Here, we test this hypothesis through animal studies and a phase I clinical trial.(1) Animal experiments: Models of completely transected spinal cord injury were established in rats and canines by microsurgery. Mesenchymal stem cells derived from neonatal umbilical cord tissue were adsorbed onto collagen scaffolds and surgically implanted at the injury site in rats and canines;the animals were observed after 1 week–6 months. The transplantation resulted in increased motor scores, enhanced amplitude and shortened latency of the motor evoked potential, and reduced injury area as measured by magnetic resonance imaging.(2) Phase I clinical trial: Forty patients with acute complete cervical injuries were enrolled at the Characteristic Medical Center of Chinese People's Armed Police Force and divided into two groups. The treatment group(n = 20) received collagen scaffolds loaded with mesenchymal stem cells derived from neonatal umbilical cordtissues;the control group(n = 20) did not receive the stem-cell loaded collagen implant. All patients were followed for 12 months. In the treatment group, the American Spinal Injury Association scores and activities of daily life scores were increased, bowel and urinary functions were recovered, and residual urine volume was reduced compared with the pre-treatment baseline. Furthermore, magnetic resonance imaging showed that new nerve fiber connections were formed, and diffusion tensor imaging showed that electrophysiological activity was recovered after the treatment. No serious complication was observed during follow-up. In contrast, the neurological functions of the patients in the control group were not improved over the follow-up period. The above data preliminarily demonstrate that the transplantation of human umbilical cord-mesenchymal stem cells on a collagen scaffold can promote the recovery of neurological function after acute spinal cord injury. In the future, these results need to be confirmed in a multicenter, randomized controlled clinical trial with a larger sample size. The clinical trial was approved by the Ethics Committee of the Characteristic Medical Center of Chinese People's Armed Police Force on February 3, 2016(approval No. PJHEC-2016-A8). All animal experiments were approved by the Ethics Committee of the Characteristic Medical Center of Chinese People's Armed Police Force on May 20, 2015(approval No. PJHEC-2015-D5).     

A flexible electrode array for determining regions of motor function activated by epidural spinal cord stimulation in rats with spinal cord injury

Epidural stimulation of the spinal cord is a promising technique for the recovery of motor function after spinal cord injury.The key challenges within the reconstruction of motor function for paralyzed limbs are the precise control of sites and parameters of stimulation.To activate lower-limb muscles precisely by epidural spinal cord stimulation,we proposed a high-density,flexible electrode array.We determined the regions of motor function that were activated upon epidural stimulation of the spinal cord in a rat model with complete spinal cord,which was established by a transection method.For evaluating the effect of stimulation,the evoked potentials were recorded from bilateral lowerlimb muscles,including the vastus lateralis,semitendinosus,tibialis anterior,and medial gastrocnemius.To determine the appropriate stimulation sites and parameters of the lower muscles,the stimulation characteristics were studied within the regions in which motor function was activated upon spinal cord stimulation.In the vastus lateralis and medial gastrocnemius,these regions were symmetrically located at the lateral site of L1 and the medial site of L2 vertebrae segment,respectively.The tibialis anterior and semitendinosus only responded to stimulation simultaneously with other muscles.The minimum and maximum stimulation threshold currents of the vastus lateralis were higher than those of the medial gastrocnemius.Our results demonstrate the ability to identify specific stimulation sites of lower muscles using a high-density and flexible array.They also provide a reference for selecting the appropriate conditions for implantable stimulation for animal models of spinal cord injury.This study was approved by the Animal Research Committee of Southeast University,China(approval No.20190720001) on July 20,2019.     

Transplantation of placenta-derived mesenchymal stem cell-induced neural stem cells to treat spinal cord injury

Because of their strong proliferative capacity and multi-potency, placenta-derived mesenchymal stem cells have gained interest as a cell source in the field of nerve damage repair. In the present study, human placenta-derived mesenchymal stem ceils were induced to differentiate into neural stem cells, which were then transplanted into the spinal cord after local spinal cord injury in rats. The motor functional recovery and pathological changes in the injured spinal cord were observed for 3 successive weeks. The results showed that human placenta-derived mesenchymal stem cells can differentiate into neuron-like cells and that induced neural stem cells contribute to the restoration of injured spinal cord without causing transplant rejection. Thus, these cells promote the recovery of motor and sensory functions in a rat model of spinal cord injury. Therefore, human placenta-derived mesenchymal stem cells may be useful as seed cells during the repair of spinal cord injury.     

Early methylprednisolone impact treatment for sensory and motor function recovery in patients with acute spinal cord injury★ A self-control study

BACKGROUND： For the treatment of spinal cord injury, any pathological changes of the injured tissue should be primarily corrected or reversed. Any remaining fibrous function and neurons with intact structure should be retained, and the toxic substances caused by ischemia-hypoxia following spinal cord injury, should be eliminated to create a favorable environment that would promote neural functional recovery. OBJECTIVE： This study was designed to investigate the effects of the impact of early methylprednisolone-treatment on the sensory and motor function recovery in patients with acute spinal cord injury. DESIGN： A self-control observation. SETTING： Department of Spine Surgery, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China. PARTICIPANTS： Forty-three patients with acute spinal cord injury were admitted to the Department of Spine Surgery, First Affiliated Hospital of Nanjing Medical University, between October 2005 and September 2007. These patients were recruited for the present study. The patients comprised 33 males and 10 females, and all met with the inclusive criteria namely, the time between suffering from acute spinal cord injury and receiving treatment was less than or equal to eight hours. METHODS： According to the protocol determined by the State Second Conference of Acute Spinal Cord Injury of USA, all patients received the drop-wise administration of a 30-mg/kg dose of methylprednisolone （H200040339, 500 mg/bottle, Pharmacia N.V/S.A, Belgium） for 15 minutes within 8 hours post injury. After a 45-minute interval, methylprednisolone was administered at 5.4 mg/kg/h for 23 hours. MAIN OUTCOME MEASURES： Prior to and post treatment, acupuncture sense and light touch scoring were performed at 28 dermatomic area key points, including occipital tuberosity and supraclavicular fossa. At the same time, motor scoring of key muscles among 10 pairs of sarcomeres was also performed. RESULTS： All 43 patients participated in the final analysis. There was no s     

Combined transplantation of GDAsBMP and hr-decorin in spinal cord contusion repair****○

Following spinal cord injury, astrocyte proliferation and scar formation are the main factors inhibiting the regeneration and growth of spinal cord axons. Recombinant decorin suppresses inflammatory reactions, inhibits glial scar formation, and promotes axonal growth. Rat models of T8 spinal cord contusion were created with the NYU impactor and these models were subjected to combined transplantation of bone morphogenetic protein-4-induced glial-restricted precursor-derived astrocytes and human recombinant decorin transplantation. At 28 days after spinal cord contusion, double-immunofluorescent histochemistry revealed that combined transplantation inhibited the early inflammatory response in injured rats. Furthermore, brain-derived neurotrophic factor, which was secreted by transplanted cells, protected injured axons. The combined transplantation promoted axonal regeneration and growth of injured motor and sensory neurons by inhibiting astrocyte proliferation and glial scar formation, with astrocytes forming a linear arrangement in the contused spinal cord, thus providing axonal regeneration channels.     

Neuroprotective effect of melatonin against ischemia/reperfusion-induced neuronal apoptosis in mouse cerebellum

BACKGROUND: Some experiments have demonstrated that melatonin (N-aceyl-5-methoxytryptamine, Mel) has antioxidation. However, whether it has neuroprotective effect in the ischemia/reperfusion injury of central nervous system is unclear. OBJECTIVE: To observe the protective effect of Mel on ischemia/reperfusion-induced cerebellar neuronal apoptosis of rats, and the action mechanism. DESIGN: Controlled observation experiment. SETTING: Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: Eight Sprague-Dawley rats aged 7–8 days and weighing 10–12 g were provided by Medical Experimental Animal Center, Tongji Medical College,Huazhong University of Science and Technology. Anti-cytochrome C monoclonal antibody was purchased from R & D Company; 7-dichlorodihydrofluorescein diacetate(DCFH-DA), rhodamine 123 and Mel were purchased from Sigma Company (USA). Lactate dehydrogenase (LDH) kit was purchased from Nanjing Jiancheng Bioengineering Institute. METHODS: This experiment was carried out in the laboratory for Department of Biochemistry and Molecule Biology, Tongji Medical College between October 2002 and March 2004. Cerebellar neurons of rats were cultured in vitro. After oxygen-glucose deprivation (OGD) for 90 minutes, 1×10–4,1×10–6, 1×10–9 mol/L Mel was added, respectively, namely high-, middle-, and low-concentration Mel groups. Cells, which were cultured by OGD, served as model group, and control group, in which OGD intervention was omitted, was set. ①Cytochrome C level of mitochondrial cells in each group was detected by ELISA method. ②LDH activity in the cell culture fluid was measured, and cell membrane permeability change was analyzed. The cells in the Mel group with the lowest LDH activity served as Mel treatment group, i.e. cells were cultured with OGD, and then Mel was added; Meanwhile, Mel prevention group was set, i.e. Mel was added before OGD. Intervention was not changed in the model group and control group. ③ DNA level was analyzed and cell apoptosis was observed by agarose gel electrophoresis(AGE). ④Mitochondrial transmembrane potential of cells, and apoptotic way in each group were analyzed by confocal laser scanning microscopy. MAIN OUTCOME MEASURES: ①Mitochondrial cytochrome C level of cerebellar nerve cells. ②LDH activity of cerebellar nerve cells. ③ DNA AGE results. ④Mitochondrial transmembrane potential change. RESULTS: ①Mitochondrial cytochrome C level of cerebellar nerve cells: cytochrome C was obviously released at 6 hours of OGD-reperfusion. Mel inhibited the release of cytochrome C in dose-dependent manner. ②LDH activity of cerebellar nerve cells: LDH activity (A value) was significantly lower in the high- and middle-concentration Mel groups than in the model group (P < 0.05). LDH activity (A value) in the low-concentration Mel group was 0.415 0±0.012 9, indicating that Mel could decrease LDH activity of OGD-treated cell supernatant and promote membrane stablization in dose-dependent manner. ③AGE results of DNA: 1×10–9 mol/L was considered as the best concentration of melatonin. Cell DNA was extracted for AGE. Results presented typical ladder shape, indicating apoptosis appeared, while apoptosis was lessened in the Mel treatment group and Mel prevention group.④Mitochondrial transmembrane potential change: Experimental results showed that green fluorescein was evenly distributed in cerebellar granule cells cultured normally, and the axons of neurons were very clear. The body of neurons was condensed and the axons disappeared after cerebellar granule cells undergoing OGD injury. Mel could completely reverse the effect of OGD. CONCLUSION: Mel can enhance cerebellar neuronal membrane stabilization of rats in dose-dependent manner, and suppress OGD-induced apoptosis of cerebellar granule cells by preventing against mitochondrial apoptosis.     

Effect of intracranial hypertension on cerebral hemorrhage induced autonomic nerve imbalance

BACKGROUND: Cerebral hemorrhage can cause the imbalance of nerve function, whereas its mechanism and main impact factors are still not quite clear. OBJECTIVE: To explore the rules about the changes of intracranial pressure in brainstem hemorrhage and internal capsule hemorrhage, and analyze the role of intracranial hypertension in the changes of nerve function caused by cerebral hemorrhage. DESIGN: A self-controlled trial. SETTING: Department of Physiology, Tianjin Medical University. MATERIALS: Sixty-five healthy male Japanese white rabbits with long ears (1.5–1.8 kg) were supplied and fed by the Department of Animal Experiment of Tianjin Medical University. The RM6240B biological signal collecting and processing system was used. METHODS: The experiments were conducted in the Department of Physiology, Tianjin Medical University from August 2001 to May 2006. ① The rabbits were anesthetized, then fixed onto the brain stereotaxic apparatus, and afterwards fenestration on skull and intubation to lateral ventricle were performed.The dynamic changes of intracranial pressure were monitored continuously. Rabbits were infused with autologous arterial blood (0.3 mL) into midbrain corpora quadrigemina inferior colliculus to induce model of acute brainstem hemorrhage; models of internal capsule hemorrhage were established by infusing autologous arterial blood into internal capsule. ② The dynamic intracranial pressures under the above conditions were recorded continuously with the RM6240B biological signal collecting and processing system. ③ An animal model of persistent intracranial hypertension was established by infusion of physiologic saline into lateral ventricle. ④ The changes of the intensity of autonomic nerve discharge were analyzed, using the biological signal collecting and processing system before and after hemorrhage and under persistent intracranial hypertension. ⑤ Ten animal models of internal capsule hemorrhage and 10 of brainstem hemorrhage were selected respectively, then gross pathological samples were cut open, and the accuracy of hemorrhage models was affirmed. Histological sections in hemorrhage point and around this point were prepared for with hematoxylin and eosin staining, and the pathological changes were observed under light microscope. MAIN OUTCOME MEASURES: ① Changes of intracranial pressures before and after internal capsule hemorrhage and brainstem hemorrhage; ② Changes of the discharge intensity of cervical vagus nerve trunk in animal models of internal capsule hemorrhage, brainstem hemorrhage and persistent intracranial hypertension without hemorrhage; ③ Accuracy of location of internal capsule hemorrhage and brainstem hemorrhage confirmed by gross pathological samples and sections. RESULTS: Totally 65 rabbits were involved in the analysis of results. ① Dynamic state of intracranial pressure: Intracranial pressure increased obviously at 45 minutes after internal capsule hemorrhage and brainstem hemorrhage, the intracranial pressures were (1.31±0.30), (1.82±0.45) kPa, which were obviously higher than those before hemorrhage [(1.04±0.18), ( 1.05±0.19) kPa, P < 0.01]. ② Discharge of vagus nerve: Under intracranial hypertension, the discharge of cervical vagus nerve trunk was enhanced, and the discharge intensity of vagus nerve trunk was significantly different before and after persistent intracranial hypertension [(364.28±78.55), (1252.19±151.75) μV·s, P < 0.01]. The discharges of cervical vagus nerve trunk were significantly enhanced after internal capsule hemorrhage and brainstem hemorrhage (P < 0.01). ③ Validation of hemorrhage sites: The hemorrhage sites were internal capsule and brainstem on histopathological sections. CONCLUSION: Intracranial pressure may play an important role in the pathophysiological process of vagus nerve imbalance caused by cerebral hemorrhage.     

Spinal cord injury in patients with systemic lupus Erythematosus——Clinical manifestations, imaging characteristics and treatment

BACKGROUND： There are fewer reports on systemic lupus erythematosus （SLE） related myelitis, and definite and uniform therapeutic program is not available. OBJECTIVE： To observe the clinical manifestations, imaging characteristics, results of laboratory examination and treatment of SLE. DESIGN： A retrospective case analysis. SETTING： Department of Neurology, the Second Affiliated Hospital of Sun Yat-sen University. PARTICIPANTS： Totally 1 052 SLE inpatients were selected from the Second Affiliated Hospital of Sun Yat-sen University from January 1995 to May 2005, and they all accorded with the diagnostic standards for SLE set by American Rheumatism Association in 1982. 124 of them were diagnosed to have damage of central nervous system. Inclusive criteria： Patients with one of the focal physical signs, including mental and behavior disorders, headache, seizure and involvement of nervous system. Exclusive criteria： Patients with hypertensive encephalopathy, damage of nervous system due to uremia and infection of central nervous system. Spinal cord lesion occurred in 15 female cases of 23 - 51 years old. Informed consents were obtained from all the participants. METHODS： The physical signs, laboratory examinations, therapeutic program and prognosis were recorded in the 15 patients with symptoms of spinal cord lesions. All the patients underwent MRI scan of brain or lesioned segment of spinal cord, and 8 cases of them underwent lumbar puncture to determine intracranial pressure, routine and biochemical examinations were cerebrospinal fluid were performed. The disease activity of SLE in systems beyond central nervous system was evaluated with modified lupus activity criteria count （LACC）. MAIN OUTCOME MEASURES：① Incidence of SLE related myelitis, attack age distribution and its association with the activity of SLE; ② Comparisons of the clinical characteristics, cranial and spinal cord MRI manifestations, different therapeutic program and prognosis. RESULTS： All the 15 SLE patients were involved in the analysis of results. ① The incidence of SLE related myelitis was low （1%, 15/1 052）. ②SLE related myelitis occurred mostly when the SLE symptoms were active, and only a few occurred at the stable period. ③ Among the SLE patients, MRI displayed abnormal changes in 71% （10/14）, the typical changes appeared abnormal signals at corresponding spinal segments, manifested as prolonged T1 and T2 signals, thickened spinal segments. Lumbar segments were mostly involved. ④ Of the 9 patients treated with hormone impact, 7 cases （78%） had obvious improvements, and the effects were better in those treated with immunosuppressor combined with intravenous immunoglobulin of large dosage. CONCLUSION：① Myelitis is a rare complication of SLE.② MRI serves as a valuable supplementary approach in the diagnosis of SLE related myelitis without specificity. ③ Steroid pulse combined with immunosuppressor and intravenous immunoglobulin of large dosage is effective in the treatment.     

Hyperglycemia induces protein nonenzymeatic glycosylation in brain neurons of diabetic rats at early stage

BACKGROUND: Protein nonenzymatic glycosylation is supposed to be one of mechanisms for chronic complications development in diabetes mellitus, and therefore, might play an important role in the neuronal degeneration. OBJECTIVE: To study the protein nonenzymatic glycosylation in brain neurons of diabetic rats, and to analyze the pathway of neuronal degeneration at the early stage of hyperglymecia. DESIGN: Randomized controlled animal experiment. SETTING: Department of Endocrinology, First hospital Affiliated to General Hospital of Chinese PLA and Beijing Laboratory for Brain Aging, Xuanwu Hospital Affiliated to Capital Medical University. MATERIALS: Thirty-five male Wistar rats (grade Ⅱ), aged 3 months old, and 11 male purebred Kunming mice (grade Ⅲ) without special pathogen, aged 3 months old, were provided by the Animal Room of Capital Medical University. METHODS: This experiment was carried out in the Beijing Laboratory for Brain Aging, Xuanwu Hospital Affiliated to Capital Medical University in 1998. The rats in the diabetic model group were intraperitoneally injected into 10 g/L STZ according to 60 mg/kg to establish rat models of diabetes mellitus. The blood glucose and body mass of rats in each group were determined respectively at 1, 2 and 3 months after modeling. The antibodies of advanced glycosylation end products (AGEs) of bovine serum albumin (anti-BSA) were self-prepared: ①The antigen of AGEs-BSA was prepared.②Eleven male Kuming mice (grade Ⅱ) of 3 months old without special pathogen were selected to inoculate AGEs-BSA. ③ The animals were immunized. ④Primary purification and detection of poly-antibodies of AGEs: the AGEs were performed immunohistochemical examination at 1 month after diabetic modeling by ELISA method. MAIN OUTCOME MEASURES: ① Detection results of blood glucose and body mass of rats in two groups at different time points. ② Determination of polyclonal antibody titer of AGEs-BSA. ③ Changes in immunohistochemical image of AGEs in brain tissue of rats in two groups. RESULTS: Thirteen rats in the diabetic model group and fifteen rats in the normal model group entered the stage of final analysis. ①Changes of blood glucose and body mass: At 1, 2 and 3 months after modeling, the blood glucose of rats in the diabetic model group were respectively(28.8±2.8),(23.1±5.5),(25.4±5.1) mmol/L, which were significantly higher than those in the normal control group [(6.2±0.9),(6.1±0.8),(6.1±0.7) mmol/L,P < 0.01]; At 1, 2 and 3 months after modeling, the body mass of rats in the diabetic model group were respectively (250.1±52.2),(263.8±50.0),(261.5±42.6) g, which were significantly lower than those in the normal control group [(422.6±36.2),(462.6±39.0),(485.0±28.8) g,P < 0.01].②Determination of antibody titer of immune serum: The mice were treated by AGEs-BSA of different concentrations twice. After that, the titer of AGEs -BSA was determined, and the results of which indicated that a higher absorbance existed at 1∶1 000. ③Determination of antigen concentration: The final titer of antibody in the abdominal dropsy was determined, and the results of which suggested that there was a much higher absorbance in the AGEs-BSA at the concentration of 5–50 mg/L. ④Determination of antibody titer in abdominal dropsy: The antibody titer in abdominal dropsy was detected by ELISA method with antigen at 20 mg/L, which indicated that the maximum absorbance (1.265±0.039) existed at 1∶4 000, and very larger absorbance (0.982±0.067) at 1∶20 000. The polyclonal antibody of AGEs-BSA was successfully prepared. ⑤Immunohistochemical detection results: The immunohistochemical staining of AGEs showed there were positive neurons in the first month in the diabetic model group, whereas it was not significant in the normal control group. The positive substances were found mainly in the cytoplasm. CONCLUSION: Hyperglycemia at the early stage of diabetes mellitus (1 month after modeling) can lead to protein nonenzymeatic glycosylation in brain neurons, and no obvious reactions mentioned above are found in the normal control group. It suggests that the degenerative changes of tissue structure of central nervous system are related with protein nonenzymeatic glycosylation caused by hyperglycemia.     

Changes in neuronal apoptosis and apoptosis modulatory factors in cerebral ischemia/reperfusion

BACKGROUND: The high concentration of glutamate release is the main cause for neuronal cell death. The relationship between glutamate level and apoptosis during ischemia/reperfusion injury is still unclear. OBJECTIVE: To observe the neuronal apoptosis at 24 and 72 hours following cerebral ischemia/reperfusion in rats, and analyze the possible influencing factors. DESIGN: A randomized controlled animal experiment. SETTING: School of Medicine, Southern Yangtze University. MATERIALS: Totally 30 male adult Sprague Dawley (SD) rats of clean grade, weighing 240–290 g, were obtained from Shanghai Experimental Animal Center, Chinese Academy of Sciences. The rats were randomly divided into sham-operated group (n=10) and model group (n=20). Each group was observed at 24 and 72 hours after ischemia/reperfusion, 5 rats at each time point in the sham-operated group, whereas 12 at 24 hours and 8 at 72 hours in the model group. Kits for determining apoptosis and Bcl-2 were bought from Wuhan Boster Biological Technology, Co., Ltd.; Kit for calcineurin from Nanjing Jiancheng Bioengineering Institute. METHODS: The experiment was carried out in the Functional Scientific Research Room of Southern Yangtze University from June to October in 2006. ① Right middle cerebral artery was occluded by inserting a thread through internal carotid artery (ICA). The surgical process for the sham-operated rats was the same as that in the model group except a nylon suture inserted the ICA. According to Longa five-degree standard, the neurological deficit evaluation of rats was evaluated after surgery, and grades 1–3 were taken as successful model establishment. The blood was recirculated by withdrawing the nylon filament under anesthesia at 2 hours after ischemia in successful rat models. ②After reperfusion, the brain tissue was quickly removed at 24 or 72 hours and the slices were obtained from optic chiasma to funnel manubrium. The changes of the number of apoptotic cells were observed using the terminal deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling method. The expressions of Bcl-2 protein were determined with immunohistochemical staining. The activity of calcineurin was determined by the inorganic phosphorus method. The content of excitatory amino acid was detected by high performance liquid chromatography. MAIN OUTCOME MEASURES: ① Glutanate content in brain tissue; ② Conditions of apoptosis; ③ Calcineurin activity in brain tissue; ④ Bcl-2 expression in brain tissue. RESULTS: Totally 30 SD rats were used, 5 died and the other 25 were involved in the analysis of results. ① Changes of apoptosis: There were 0–3 apoptotic cells in the sham-operated group. In the model group, the numbers of apoptotic cells were obviously increased at 24 and 72 hours of reperfusion (P < 0.01), and it was markedly reduced at 72 hours as compared with 24 hours (P < 0.01). ② Changes of glutanate content: The glutamate contents at 24 and 72 hours of reperfusion in the model group were obviously higher than those in the sham-operated group (P < 0.01); In the model group, it was obviously increased at 24 hours as compared with 72 hours (P < 0.01). ③ Changes of Bcl-2 protein: In the model group, the Bcl-2 protein expression had no obvious changes at 24 hours of reperfusion, and it was obviously enhanced at 72 hours, which was obviously different from that in the sham-operated group and that at 24 hours (P < 0.01). ④ Changes of calcinerin activity: In the model group, the activity of calcineurin in brain tissue had no obvious changes at 24 hours of reperfusion; The activity of calcineurin at 72 hours was obviously higher than that in the sham-operated group and that at 24 hours (P < 0.01). CONCLUSION: The brain cyto-apoptosis action at different time points following reperfusion incompletely depends on the glutamate levels, while it depends on the interaction of some apoptosis related factors, such as amino acid, calcineurin, and Bcl-2, etc.     

Clinical characteristics, neurophysiological grade and outcome of patients with carpal tunnel syndrome——A retrospective study

BACKGROUND: Carpal tunnel syndrome (CTS) is diagnosed mainly according to clinical symptoms, physical sign and neurodiagnostic laboratory examination. The therapeutic effect of conservative management and surgical operation in treating CTS need to be further observed and evaluated. OBJECTIVE: To analyze the clinical characteristics, neurophysiological grade and outcome in patients with CTS. DESIGN: Retrospective case-analysis. SETTING: Department of Neurology, First Affiliated Hospital, Medical College, Xi'an Jiaotong University. PARTICIPANTS: Totally 161 patients with suspected CTS from National Neuroscience Institute of Singapore referred to the Neurodiagnostic Laboratory for the confirmatory testing between January and September 2002. The involved patients, 137 male and 24 female, were aged 21–85 years. METHODS: ①The condition of diabetes mellitus complicated by abnormal thyroid function was observed. ② The effect on predominant hand, and paraesthesia were observed. ③Neuroelectrophysiological studies were performed and the results were graded into mild, moderate and severe CTS according to the American Association of Electrodiagnostic Medicine (AAEM) criteria. ④ Conservative management and surgical intervention were followed up 3 months later, and symptoms and physical sign basically disappeared, and function was basically recovered, which indicated that disease condition improved. MAIN OUTCOME MEASURES: ①Condition of CTS complicated by metabolic disease; ②Effects on predominant hand and paraesthesia; ③Electrophysiological grading; ④Prognosis. RESULTS: Totally 161 patients participated in the final analysis. ①Condition of CTS complicated by metabolic disease: Among 161 patients, 17.4% (28/161) were documented to have diabetes mellitus and 7(4.3%) had hypothyroidism. ②Effects on predominant hand and paraesthesia: Dominant hand involvement was present in 134 patients (83.2%) and more than 75% had onset of symptoms in the dominant hand. Sensory symptoms like numbness and paresthesias were the predominant symptoms, accounting for 89.1% (134/161), this discomfort was felt in all 5 digits of the hand in 47.6%, and lateral three and half digits in 21.4%. The noctural symptoms were present in 30.4% (49/161) patients. ③Electrophysiological typing: The most frequent abnormality was that of the prolonged mid-palm median and ulnar latency difference in 146(54.7%) hands; 103(38.6%) hands had prolonged median motor distal latency. Absent response from thenar muscle was present in 35 (13.1%) hands. Nerve conduction study showed bilateral CTS in 105 (65.2%) patients and unilateral CTS in 56 (34.8%) patients. Sixteen patients with bilateral CTS had symptoms in one hand only. Overall, 36.8% had mild, 49.2 % had moderate and 13.9 % had severe CTS, with median duration of symptoms of 6, 9 and 14 months, respectively. ④Delay in diagnosis: 37(22.9%) patients delayed in diagnosis from 1–4 months, 16(43.2%) were misdiagnosed as cervical spondylosis; 6(16.2%) were ignored due to their condition by busy work; 15(40.5%) were unware of their symptoms. ⑤Prognosis: Follow up data was available for only 72.7% (117/161) patients. Conservative management was conducted in 73.5% (86/117). Clinical symptoms were resolved or improved in 65.1% (56/86) patients with 17 mild CTS, 29 moderate CTS, and 10 severe CTS. 26.5% (31/117) patients underwent surgery for CTS release, and clinical symptoms were improved in 12(38.7%) with moderate CTS and 2 (6.5%) with severe CTS at 3 months of follow up. CONCLUSION: ①Sensory symptoms in CTS are more in severe and common in dominant hand. ②Conservative management showed resolution or improvement for mild and moderate CTS. Surgical intervention shows either resolution or improvement in clinical symptoms in moderate CTS. ③ The common reasons for delay in diagnosis were due to misdiagnosis as cervical spondylosis and lack of awareness of the condition. ④Assessment on severity of CTS by electrophysiological grade is of important significance for determining therapeutic mean.     

Neurovascular compression and decompression of the intracranial vagus for blood pressure

BACKGROUND: Previous researches demonstrated that neurovascular decompression could cure hypertension; however, whether it could effectively control refractory hypertension after hypertensive cerebral hemorrhage should be further studied. OBJECTIVE: To observe the effect of neruovascular compression on intracranial vagus for blood pressure of dogs and investigate the effect of neurovascular decompression on blood pressure of patients with hypertensive cerebral hemorrhage. DESIGN: Randomized controlled animal study, clinical effects and retrospective analysis. SETTING: Department of Neurosurgery, Changzheng Hospital Affiliated to the Second Military Medical University of Chinese PLA. MATERIALS: The experiment was carried out in the Department of Neurosurgery, Changzheng Hospital Affiliated to the Second Military Medical University of Chinese PLA from May to October 2006. A total of 15 healthy adult dogs of both genders were randomly divided into experimental group (n =10) and control group (n =5). Clinical observation: A total of 41 patients with hypertensive cerebral hemorrhage were selected from the Department of Neurosurgery, General Hospital of Nanjing Military Area Command of Chinese PLA and the Department of Neurosurgery, Changzheng Hospital Affiliated to the Second Military Medical University of Chinese PLA from October 1999 to October 2006. Among them, one patient had brain stem hemorrhage. There were 27 males and 14 females aged from 41 to 66 years. Inclusion criteria: All patients were diagnosed with CT examination once or several times. Volume of hematoma ranged from 50 to 120 mL and had obviously operative indication. All patients provided consents. In addition, another 281 patients with hypertensive cerebral hemorrhage who received traditionally internal and surgical therapies in our departments of neurosurgery, neurology and emergency room were selected in the control group. METHODS: ① Animal experiments: 20 cm autochthonous great saphenous vein was taken from dogs in the experimental group and coincided with tip of facial artery to form arterial loop so as to oppress left vagus and lateral bulb abdomen. In addition, 20 cm autochthonous great saphenous vein was taken from dogs in the control group and coincided with tip of facial artery to establish arterial loop so as to oppress left cerebellum to observe changes of blood pressure before and at 1, 2, 3 and 4 weeks after operation. ② Clinical observation: Among 41 patients with hypertensive cerebral hemorrhage including one with brain stem hemorrhage, they received microvascular decompression of vagus immediately after getting rid of intracerebral hematoma and stopping bleeding to observe its effect of depressurization. All patients and their relatives provided consents. ③ A total of 281 patients with hypertensive cerebral hemorrhage who discharged after the treatment of traditionally internal and surgical therapies were studied retrospectively to observe changes of blood pressure after routine treatment and compare the results with neurovascular decompression. MAIN OUTCOME MEASURES: ① Changes of blood pressure of experimental dogs; ② effect of vascular decompression of vagus for blood pressure of patients with hypertensive cerebral hemorrhage after clearing intracerebral hematoma; ③ different effects of neurovascular decompression and routinely internal and surgical therapies on hypertension. RESULTS: ① Results of animal experiments: Nine dogs in the experimental group survived. At 1, 2, 3 and 4 weeks after operation, blood pressure of dogs in the experimental group was (139.77±4.06), (149.11±4.90), (148.10±4.16), (147.76±4.15) mm Hg (1 mm Hg=0.133 kPa), which was higher than that of dogs in the control group [(117.20±2.74), (116.65±3.74), (116.26±1.8), (115.81±3.76) mm Hg, P < 0.01]. ② Results of clinical observation: Among 41 patients, 8 (20%) cases died during the operation. In addition, among other 33 (80%) survival patients, 11 (33%) cases had normal blood pressure; blood pressure of 14 (43%) cases was improved or closed to normal value; blood pressure of 8 (24%) cases was not changed obviously as compared with that before operation. ③ The results demonstrated that, by using traditionally internal and surgical therapies, among 281 patients with hypertensive cerebral hemorrhage, blood pressure of about 15% cases was recovered or closed to normal value. Those mentioned above did not have history of hypertension before hemorrhage. However, patients who had history of hypertension before hemorrhage received the traditionally internal or surgical therapies, and the blood pressure was not improved to the normal value after the treatment. CONCLUSION: ① Neurovascular compression in left intracranial vagus can cause obvious increase of blood pressure of dogs, and the increasing volume was 30 mm Hg. ② Vascular decompression of vagus has a great effect on refractory hypertension, and the improvement of blood pressure is superior to traditionally internal and surgical therapies in clinic.     

Correspondence of CT perfusion imaging to pathological manifestations in rabbit models of hyperacute cerebral infarction

BackgroundCould the infarction be diagnosed quickly and accurately at the acute stage by CT perfusion imaging (CTPI) technology? Whether the images of CTPI will correspond with the pathological changes or not? All the questions need to be solved by experimental and clinical studies.ObjectiveTo reveal the rules of perfusion map changes and guide the early diagnosis of hyperacute cerebral infarction by analyzing the correlation of CTPI with pathological manifestations for hyperacute cerebral infarction.DesignA randomized controlled animal experiment.SettingExperimental Center of Medical Radiology, Longgang Central Hospital of Shenzhen City.MaterialsForty-two adult New Zealand rabbits of (2.6±0.5) kg, either male or female, were randomly divided into experimental group (n =36) and control group (n =6). Six rabbits in the experimental group were observed after ischemia for 0.5, 1, 2, 3, 4 and 6 hours respectively, and 1 rabbit in the control group was observed at each corresponding time point.MethodsThe experiments were carried out in the Experimental Center of Medical Radiology, Longgang Central Hospital of Shenzhen City from March 2003 to July 2004. Rabbit models of cerebral infarction were established by modified O'Brein method.

The rabbits in the experimental group were scanned at 0.5, 1, 2, 3, 4 and 6 hours after ischemia respectively. The dynamic CT scan slice was 13 mm from the anterior edge of the frontal cortex, and six fake color functional images were obtained, including cerebral blood flow map (CBF map), cerebral blood volume map (CBV map), peak to enhancement map (PE map), flow without vessels map, time to peak map (TP map), time to start map (TS map). The manifestations and changes of the functional maps in different interval were observed.

Bilateral symmetric ranges of interest (ROI) were drawn separately on the CBF map, CBV map, TP map and TS map. The blood flow parameters of focal and contralateral cerebral tissues could be obtained to calculate relative cerebral blood flow (rCBF, rCBF=focal CBF/contralateral CBF), relative cerebral blood volume (rCBV, rCBV= focal CBV/contralateral CBV), a relative time to peak (rTP, rTP= focal TP–contralateral TP), a relative time to start (rTS, rTS= focal TP–contralateral TP).

The perfusion maps were input into AutoCAD software. The percents of ischemic cores and peri-ischemic areas accounting for contralateral cerebral hemisphere were calculated.

The animals were anesthetized and killed, then the cerebellum and low brain stem were taken out. The brain tissues were cut on coronal plane at 14 mm from the anterior edge of the frontal cortex, a 2-mm piece anterior to the incision, and a 3-mm piece posterior to the incision. The anterior piece was fixed, stained and observed. A 1-mm slice was cut from the front of the posterior piece tissues as electron microscope sample, the remnant was fixed and then taken out, and the location and size of stained “white” areas were observed as the reference for electron microscope sample.

The correlation between CTPI and pathological manifestations was observed.Main outcome measures

Laws of time and spatial changes of ischemic areas;

Pathological changes of the ischemic tissues;

Correspondency between CTPI and pathological manifestations.Results

Laws of time and spatial changes of ischemic areas: Relative ischemic-core areas were consistent in each perfusion map, increased incessantly along with the ischemic times. Relative peri-ischemic areas were inconsistent in each perfusion map, on CBF map from 1 to 6 hours after ischemia, the area of ischemic core increased from (1.503±0.523)% to (7.125±1.054)%, the ascending trend occurred. But the peri-ischemic areas showed a descending trend on CBF map, the areas decreased from (8.960±0.719)% to (5.445±0.884)% from 0.5 to 6 hours; The relative areas were the largest one on TP maps, the average value was (32.796±3.029)% at 0.5 hour after ischemia happening (60.540±1.683)% at 6 hours. The trend of ischemic areas was increased. No obvious change was observed on TS maps.

Pathological changes of the ischemic tissues: Under light microscope, there was no obvious change at 0.5–2 hours after ischemia, edema at 3 hours, karyopycnosis at 4 hours and eosinophilous changes at 6 hours; Under electron microscope, there was edema in ischemic cores within 4 hours after ischemia, whereas karyopycnosis or structure vanished after 4 hours; Edema was observed in peri-ischemic areas.

Correlation between CTPI and pathological manifestations: On CTPI maps, the ischemic core was blue on CBF and CBV maps, black on TP and TS maps. Along with the ischemic times, the rCBF and rCBV decreased, whereas the rTP and rTS prolonged. Hemodynamic parameters were not significantly different within 2 hours of ischemia and 2 hours after ischemia. The rTP and rTS became 0 after 1 and 2 hours respectively. On CTPI maps the peri-ischemic area was red on CBF and CBV maps, red and yellow on TS maps, red on TP maps. Along with the ischemic times, the rCBF decreased, and the lowest level was always at about 20%, whereas the rTP and rTS prolonged.Conclusion

CTPI manifestations corresponded well with pathological findings, and it is a sensitive, stable and reliable technique to diagnose hyperacute cerebral infarction.

TP map was more sensitive than CBF map and TS map in exhibiting the peri-ischemic areas, thus TP maps could be a good choice for observing peri-ischemic areas.     

Huayu capsule enhances limb-catching capability of rats with experimental open traumatic brain injury

BACKGROUND: It is hard to cure the open traumatic brain injury (TBI), especially for the brain functional recovery after brain injury. In this regard, traditional Chinese medicine (TCM) has a wide prospect. OBJECTIVE: To observe the effect of Huayu capsule on limb-catching capability of rat models of open TBI, and investigate its possible mechanism. DESIGN: Randomized and controlled study. SETTING: Grade 3 Pharmacological Laboratory of TCM, State Administration of TCM, Chengdu University of TCM. MATERIALS: This study was performed from October 2005 to January 2006. Fifty Sprague-Dawley rats of either gender, aged 3 months old, weighing from 190 to 220 g, were involved in this study. Huayu capsule was made and supplied by the Department of TCM Processing of Chengdu University of TCM, Lot No. 050121; Xuefuzhuyu oral liquid was manufactured by Jilin Aodong Yanbian Pharmaceutical Industry Co.,Ltd., Lot No. 050406. METHODS: Open right parietal lobe TBI rat models were made as described in references. The involved rat models were randomized into 5 groups according to gender and body mass: model group, high-, middle-, low-dose Huayu capsule groups and Xuefuzhuyu oral liquid group, with 10 rats in each. Rats in the model group were administrated with distilled water of 5 mL/kg; Rats in the high-, middle- and low-dose Huayu capsule groups were administrated with 1.030, 0.515, 0.258 g/kg raw herbs; Rats in the Xuefuzhuyu oral liquid group were administrated with Xuefuzhuyu oral liquid of 5 mL/kg, intragastrically once a day for 7 days successively for all after recovering consciousness from anesthetization. ① One hour after administration on the 6th day, rats in each group were placed on a 100 cm fine straight iron wire paralleling to the ground and 20 cm above the operational table. The time of the rats keeping on the wire was counted and it indicated the nerve-muscle catching capability. The longer the remained time, the better the nerve-muscle catching capability.② Twenty-four hours after the administration on the 7th day, the samples of the whole brain were carefully taken out and stained by toluidine blue for observing the morphology of cells in the injured brain tissue. ③ The nerve cells in 4 visual fields from 4 directions (upper, lower, left, right) of injured area of brain tissue were counted. The amount was the total number of the four visual fields. The nerve cells in the injured brain tissue were measured by the same way. ④ On the basis of nerve cell counting, the content of Nissl's body in the corresponding nerve cells was measured. MAIN OUTCOME MEASURES: ①Nerve-muscle catching capability. ② Histopathomorphological examination of the injured areas in brain. ③Measurement of nerve cells and macrophages in the injured areas of brain. ④Measurement of content of Nissl's body in the injured areas of brain. RESULTS: All the 50 rats were involved in the final analysis. ①The catching time of rats in the high- and middle-dose Huayu capsule groups as well as Xuefuzhuyu oral liquid group was extended to （23.6±10.12）,（18.6±8.17） and (22.6±9.43) s, respectively, which was significantly higher than that in the model group [ (12.1±4.15) s, P < 0.05–0.01]. ② The injured areas of brain tissue of rats in the Huayu capsule-treated groups and Xuefuzhuyu oral liquid group were decreased to different extents. The nerve cells adjacent to brain injured area were increased. ③ The number of macrophages around the brain injured area of rats in the high- and middle-dose Huayu capsule groups as well as Xuefuzhuyu oral liquid group was 63.9±7.99, 59.7±7.41 and 62.9±7.37, respectively, which was significantly larger than that in the model group（49.2±8.00,P < 0.01）. The number of nerve cells adjacent to brain injured area of rats in the high-, middle-, and low-dose Huayu capsule groups as well as Xuefuzhuyu oral liquid group was 86.2±25.93, 93.5±31.79, 92.1±14.54 and 125.2±34.25, respectively, which was significantly larger than that in the model group（62.5±16.98,P < 0.05–0.01）. ④ The total area, the total and integral absorbance , the average gray degree of Nissl's body in the cytoplasm of nerve cells of rats in the Huayu capsule-treated groups and Xuefuzhuyu oral liquid group were all significantly increased (P < 0.05–0.01). CONCLUSION: Huayu capsule at different doses can promote the limb-catching capability of rat models of open TBI to different extent. This promoting effect may be related to increasing macrophages in the injured area, lessening the apoptosis of nerve cells and increasing the content of Nissl's body in the nerve cells.     

Changes of serum high sensitive C-reactive protein in patients with acute cerebral infarction

BACKGROUND: Serum high sensitive C-reactive protein (hs-CRP), which regards as a high sensitive mark of systemic inflammatory response syndrome, can provide a lot of valuable information for the treatment and prognosis of cerebrovascular disease. OBJECTIVE: To observe the differences of blood glucose, lipid, homocysteine and previous disease history among patients with acute cerebral infarction at various levels of hs-CRP and compare changes of hs-CRP of patients with various degrees of neurologic impairment. DESIGN: Contrast observation. SETTING: Department of Neurology, Shenzhou Hospital, Shenyang Medical College. PARTICIPANTS: A total of 102 patients with acute cerebral infarction were selected from Department of Neurology, Shenzhou Hospital of Shenyang Medical College from February 2005 to September 2006, including 55 males and 47 females aged from 55 to 86 years. All accepted patients met the diagnostic criteria of cerebral infarction established by the Fourth National Cerebrovascular Disease Academic Meeting and were diagnosed with CT or MRI examination. All patients provided the confirmed consent. Based on clinical criteria of neurologic impairment established by the Fourth National Cerebrovascular Disease Academic Meeting, patients were randomly divided into mild group (0–15 points, n =46), moderate group (16–30 points, n =38) and severe group (31–45 points, n =18). In addition, based on hs-CRP level within 72 hours, patients were divided into normal group (hs-CRP ≤ 3 mg/L, n =53) and increasing group (hs-CRP > 3 mg/L, n =49). METHODS: ① 2 mL venous blood was selected from hospitalized patients in the next morning to separate serum. Quantitative measurement of hs-CRP was dealt with Latex Enhnced Turbidimetric Immunoassay (LETIA). ② Fasting venous blood was colleted from hospitalized patients in the next morning to measure numeration of white blood cells, fibrinogen, blood glucose, total cholesterol (TC), triacylglycerol (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and homocysteine. ③ Measurement data were compared with t test or analysis of variance. MAIN OUTCOME MEASURES: ① Comparisons of serum biochemical indexes among patients with various levels of hs-CRP; ② comparisons of risk factors among patients with various levels of hs-CRP; ③ comparisons of levels of hs-CRP among patients with various degrees of clinical neurologic impairment. RESULTS: A total of 102 patients were involved in the final analysis. ① Plasma fibrinogen and numeration of leucocytes were more in the increasing group than those in the normal group (t =4.39, 3.54, P < 0.01); while, there were no significant differences of blood glucose, TC, TG, HDL-C, LDL-C and homocysteine between the two groups (P > 0.05). ② Percentage of patients with hypertension and diabetes mellitus (DM) was higher in the increasing group than the normal group (χ2=3.98, 4.23, P < 0.05); while, percentage of patients with smoking in the increasing group was not significantly different from that of patients in the normal group (P > 0.05). ③ Level of hs-CRP of patients with severe neurologic impairment was higher than that of patients with moderate neurologic impairment (t =2.273, P < 0.05); that of patients with moderate neurologic impairment was higher than that of patients with mild neurologic impairment (t =2.586, P < 0.05); that of patients with severe neurologic impairment was obviously higher than that of patients with mild neurologic impairment (t = 4.913, P < 0.01). CONCLUSION: ① With the increase of hs-CRP, plasma fibrinogen and numeration of leucocytes of patients with acute cerebral infarction is increased, especially, they are increased remarkably among patients who have history of diabetes mellitus and hypertension. ② Increase of level of hs-CRP can be regarded as one of marks to evaluate severity of acute stroke.