Immunohistochemical Studies on Anti-keratin Autoantibody

On the basis of the detection of IgG anti-keratin autoantibody (AK auto Ab)in human body fluids,it was shown that IgG AK auto Ab could hardly pass the blood-brain barrier,but it could easily penetrate the capillary walls into epithelial tissues.Un-der electron microscope,the interaction between AK auto Ab and keratinocyte is largelylocated in tonofilaments and desmosomes and is irrelevant to other organellae.Dynamicobservations of AK auto Ab suggest that the accumulation of AK auto Ab in keratinocy-te cytoplasm and transitory decrease of IgG AK auto Ab in serum of rabbits with sodiumsulfide dermatitis are related predominantly to keratinocytic injury.     

An Immunohistochemical study of Antikeratin Autoantibodies in Human Sera

With frozen sections of normal human skin as antigens,the antikeratinautoantibodies (AK auto Abs) in the sera of 161 normal persons and 184 patients of variousdiseases were studied immunohistochemically by means of indirect immunofluorescence(IIF) staining technique.The positive rates of IgG,IgM,and IgA AK auto Abs in normalsera examined were 66.67%,96.43% and 100% respectively.In psoriasis and other skindisorders,as well as in various types of carcinomas,the positive rates were much lower,while in internal diseases and nonepithelial tumors they were very similar to those ofnormal persons.There is no definite correlation either between different AK auto ABs orbetween AK auto Abs and Ig levels in the sera.These phenomena indicate that the AKauto Abs react to different antigens,and that their increases and decreases are notinfluenced by Ig levels in the sera,but are probably related to keratin polypeptides.     

Influence of anti-keratin autoantibodies on telomerase activity of squamous cell carcinoma

Objective: To investigate the influence of anti-keratin autoantibodies (AK auto Abs) on telom-erase activity of squamous cell carcinoma cultured in vitro and the mechanisms of the inhibitory effects of AK auto Abs on squamous cell carcinoma. Methods: Influence of AK auto Abs on the proliferation of Tca cells was observed by MTT colorimetry. Telomerase activity of cultured Tca cells and human keratinocytes was determined by telomeric repeat amplication protocol-ELISA (TRAP-ELISA) and polyacrylamide gel elec-trophoresis (PAGE). After being treated with AK auto Abs for 36 h at a concentration of 4, 8, 16 mg/L respectively, the changes of telomerase activity of Tea cells were also detected by TRAP-ELISA and PAGE. Results: MTT colorimetric determination showed that the capacity of proliferation of Tca cells correlated negatively with the concentration of AK auto Abs (r= -0. 74, P<0. 01). TRAP-ELISA and PAGE showed that telomerase activity of Tca cells increased significantly compared to that of cultured human     

Double cortical line in the acetabular roof of paraplegic patients.

The pelvic radiographs of 60 patients with paraplegia or quadriplegia were reviewed and double cortical line in the acetabular roofs on both sides was found in 35 patients. It was concluded that the double cortical line was a radiologic sign of disuse osteoporosis after spinal cord injury. It was noticed that the acetabular double cortical line in incomplete paralysis was not so prominent as that in complete paralysis (P<0.05), suggesting that the degree of acetabular osteoporosis may be related to the severity of spinal cord injuries. The etiology of double cortical line and related factors were discussed.     

Purification and identification of natural antikeratin autoantibody from normal human sera and its effects on the cultured keratinocytes

Objective: To purify the natural antikemtin autoantibody (AK auto-Ab) and observe its effects on the proliferation of the cultured keratinocytes. Methods: Natural AK auto-Ab was purified by using keratin affinity column, and then the titre and specificity of the Abs were studied by ELISA, immunoperoxidase staining and immuno-electronicmi-croscope. The effect of the purified Abs on the cultured keratino-cytes was assayed by ^3H-TdR incorporation. Results:Natural AK auto-Ab was obtained. The binding activity oflgG AK auto-Ab in purified Ab remained similar to that in pooled human sera, and the specificity of the obtained antibody is strong. The purified antibody could decrease the ^3H-TdR incorporation of the cultured keratinocytes in a dose-dependent manner. Conclusion: The method of purifying AK auto-Ab is simple, practicable and reliable. Natural AK auto-Ab, existing in normal human individuals, has inhibitory effect on the proliferation of the cultured keratinocytes.     

Expression of Angiopoietin-1/-2 in the Process of Mouse Embryo Implantation

This study examined the expression and distribution of angiopoietin-1/-2 (Ang-1/-2) in the endometrium of early pregnant mice. The expression of Ang-1/-2 was detected by immunohisto- chemical staining and in situ hybridization respectively. Computerized image analysis system was used to measure the average optical intensity of Ang-1/-2 in endometria at different time points after gestation. Mice were randomly divided into 5 groups: control group, D2 group (2 days after preg- nancy), D4 group (4 days after pregnancy), D6 group (6 days after pregnancy) and D8 group (8 days after pregnancy), each containing 15 mice. The results showed that the expression of Ang-1 and Ang-2 was very different among 4 groups (P<0.01). Immunohistochemical staining revealed that Ang-1 was localized in the cytoplasma of stromal cells 2 days after pregnancy (day 2), and in luminal epithelial cells on day 4. The protein of Ang-2 was mainly expressed in the cytoplasma of glandular epithelia and stromal cells. With gestation time, the positive reactions of Ang-1/-2 were stronger in the endometria of the pregnant mice (P<0.01). In situ hybridization showed Ang-1 mRNA in stromal cells on day 2. Hybridization signal was localized in both stromal cells and vessel epithelial cells on day 4; Ang-2 mRNA was expressed in stromal cells and glandular epithelia on day 2; high mRNA levels appeared in stromal cells, glandular epithelia and vascular endothelia on day 4; an increasing in mRNA expression of Ang-1/-2 was observed on day 6 and day 8 (P<0.01). It is suggested that Ang-1/-2 may play an important role in the cross-talk between blastocyst and maternal endometrium during the process of embryo implantation.     

Clinical Observation on the Treatment of Patients After Abdominal Operation with Exogenous Energy of Qi Gong

84 patients after abdominal operation were evenly divided into two groups,theexperimental group and the control group to observe the effect of the exogenous energy of QiGong (EEQG) on gastroenteric peristalsis.Beginning at the 8th h after operation,EEQGwas delivered to points Dantian,Zhongwan (Ren 12) and Zusanli (St 36) of the patients inthe experimental group.The results showed that bowel sounds were recoveredearlier and maintained for a longer time,and the occurrence of the first aerofluxus fromanus (OFAA) after surgery was on an average of 29.4 heartier than that in the controlgroup.It was found that EEQG might promote significantly the recovery of entericperistalsis.     

Relationship between the Changes of VEGF Level and Dendritic Cells in Peripheral Blood of Patients with Hepatocellular Carcinoma after Transcatheter Arterial Chemoembolization

In order to investigate the relationship between the VEGF level and the counts of den-dritic cells (DCs) in peripheral blood of patients with hepatocellular carcinoma (HCC) before and af-ter transcatheter arterial chemoembolization (TACE), the peripheral blood was obtained from 37 pa-tients with HCC who treated by TACE. The blood was obtained on the day before TACE, the first day, the 7th day and the 15th day after TACE respectively. The counts of DCs were quantified by flow cytometry. The plasma VEGF level was measured by ELESA kit. It was shown after TACE, the counts of DCs in peripheral blood were decreased significantly (P<0.05), and the VEGF level in pe-ripheral blood was increased significantly (P<0.05). The counts of DCs in peripheral blood had an inverse correlation with the plasma VEGF level (r=-0.57, P<0.05) after TACE. It was concluded that in patients with HCC after TACE, the increased plasma VEGF level appeared to have the effect to suppress the maturation of DCs, which may contribute to reduction of the body's anti-tumor im-munity effect, with a consequence of recur and metastasis of tumor.     

Experimental Study on the Inhibitory Effects of Verapamil on the Proliferation of Meningiomas Cells

In order to investigate the effects of verapamil on the proliferation of meningiomas cells in vitro and in vivo, the cultured meningiomas cells were cultured with verapamil at different concen-trations for 24 h and the inhibitory effects of verapamil on cell proliferation were observed by MTT method. The meningiomas model was established by implanting the newly removed tumor fragments into the nude mice subcutaneously. The nude mice with tumors were divided into two groups: vera-pamil-treated group and control group. Tumor volumes were measured and after 12 weeks the tumors were taken out and examined histologically. The expression of proliferating cell nuclear antigen (PCNA) in the tumors was detected by using immunohistochemistry. It was found that verapamil could inhibit the growth of cultured meningiomas cells in a concentration-dependant manner. The in-hibitory effect could be observed in the concentration of 1 μmol/L verapamil and the most obvious effects appeared in the concentration of 100 μmol/L. Tumor volume in the verapamiltreated group was obviously smaller than that in the control group (211.40±5.50 vs 163.94±3.62, P<0.01) and the expression of PCNA was also lower (1.52±0.24 vs 2.86±0.53, P<0.05). Tumor inhibition rate was about 22.45%. It was suggested that verapamil could inhibit the proliferation and growth of men-ingiomas cells in vitro and in vivo.     

A new method for intraoral tissue defect repair:diepithelialized tissue flap with vascular anastomosis

Diepithelialized tissue flap(DTF)with vascular anastomosis was de-signed in August 1987,and was used for the repair of 12 cases of large intraoraltissue defect.All the operations were successful.The clinical data were reportedand the procedure to incise and transplant a DTF described.When the healingprocess and changes of DTF after transplantation were observed,it was foundthat a layer of new smooth epidermis was formed on the surface of the DTF.Theorigin of this new epidermis was discussed.Ten out of the 12 cases have been fol-lowed up for from half a year to 2 and a half years,and the results weresatisfactory.The new epidermis showed no hair growth and the patients did nothave any rough feeling,which usually occurred after ordinary skin grafting.It isbelieved that DTF transplantation is likely an ideal method for the repair andreconstruction of intraoral soft tissue defect.Its disadvantage was that the DTFcontracted severely after healing.Finally the authors put forward the problemsconcerning the transplantation of DTF,which should be investigated further.     

Proliferation and Differentiation of Duct Epithelial Cells after Partial Pancreatectomy in Rats

The proliferation and differentiation of pancreatic duct epithelial cells in remnant pancreas during regeneration after partial pancreatectomy in rats were studied, and the source of pancreatic stem cells was characterized. Partial (90 %) pancreatectomy was performed on 4- to 5-week-old Sprague-Dawley rats, and different duct epithelial cells and acinar cells were detected by immunohis-trochemical stain method and scored using 5-bromo-2'-deoxyuridine (BrdU) labeling index (LI) at various time points after partial pancreatectomy. It was found that at 24 h after partial pancreatectomy proliferation started in the main, large and small duct cells, and persisted in small duct cells to day 5. There was significant difference between the experimental group and the control group (P<0.001). Acinar cells positive for BrdU were greatly increased and reached the peak LI on day 5. The destroyed lobular architecture almost totally recovered on day 7, and the newly islet cells appeared around the pancreatic ducts. These results suggest that regeneration after partial pancreatectomy is involved in proliferation and differentiation of pancreatic stem cells, and pancreatic stem cells may locate in the pancreatic ductules.     

Experimental study on vasoactive intestinal peptide—like immunoreactivity in plasma and brain in high—velocity missile injuries of extremity

Experimental high-velocity missile injuries were produced on both hind legs of 15 dogs.It was found that the concentration of vasoactive intestinal peptide-like immunoreactivity(VIP-LI)significantly elevated in both plasma and brain at the early stage after injuries.However,thechanges of VIP-LI concentration in the plasma and brain did not coincide with each other.The re-sults suggest that VIP-LI may participate in the post-traumatic general reaction.The significanceand machanism of the elevation of cerebral VIP-LI level after a severe non-cerebral trauma remainto be clarified.     

Changes of length of elastic fibers in rabbit lungs after smoke inhalation injury.

Pulmonary morphometry was determined in rabbit lungs after smoke inhalation. Twenty-four hours after injury, changes of the total lung volume (TLV) and the total length of all parenchymal elastic fibers (LT) were measured. Concomitant changes in arterial blood gas levels and pathomorphology of lung tissues were also studied. It was found that alveolar walls were broken and became thinner, and that alveoli were dilated. TLV was markedly increased and LT was markedly decreased as compared to normal lungs (P less than 0.05). These findings suggest that proteinases, especially elastase, released from phagocytes may play an important role in acute lung injury after smoke inhalation.

     

Effects of Activated Complement on Lung Fluid Exchange in Conscious Goats:Role of Superoxide Anion

The effects of zymosan-activated plasma(ZAP)on lung fluid exchangeand on superoxide anion production by granulocytes were studied in 8 consciousgoats with chronic lung lymph fistula,It was found that after ZAP infusion, thecirculating granulocytes were activated and the release of superoxide anions wasmarkedly enhanced. There was an increase in lung lymph flow and proteinconcentration.It suggests that the superoxide anion released by the stimulatedgranulocytes can play a role in inducing lung injury by the activated complement.     

The effects of tumor necrosis factor on cultured hepatocytes and non-parenchymal liver ceils in the mouse

The effects of tumor necrosis factor(TNF)on the cultured mouse hepa-tocytes and non-parenchymal liver cells were observed.It was found that therewere no significant changes of the morphological integrity and viability of thehepatocytes and the aspartate transferase level in the culture supernate after theaddition of TNF into the culture medium as compared with those of the normalcontrol,which indicates that TNF exerts no obvious cytotoxocity on the culturedmouse hepatocytes. In addition,there were also no significant changes of theabove mentioned parameters after TNF was added to the cocultures of hepato-cytes and non-parenchymal liver cells,which implies that the unactivated non-parenchymal liver cells are not involved in the TNF-related hepatocyte injury.     

Antisperm Antibodies on the Surface of Spermatozoa before＇Ejaculation from Vasectomized Men

The antisperm antibodies (AsAbs) coated on spermatozoa of the proximal vas deferens (sperm before ejaculation, SBE) from 48 fertile men who were volunteers of vasectomy and 24 vasectomized men who asked for vasovasotomy,were determined by immunobead test (IBT) and sperm cervical mucus contact test (SCMC). The results showed that in fertile men there were no positive sam ples of SBE in IBT and SCMC. In vasectomized men positive samples of SBE were found in 79.4% for IgG, 38.2% for IgA and 35.5% for SCMC. The AsAbs on SBE could be found at the time of less than one year to more than 3 years after vasectomy. The AsAbs were still found on the semen samples at 1-3 months after vasovasotomy. Our results also indicated that the incidence of AsAbs on SBE from vasectomized men could not predict the levels of AsAbs on their ejaculated sperm after vasovasotomy. There was no significant correlation between the levels of AsAbs in serium before vasovasotomy and those on SBE from vasectomized men.     

Effects of tetrandrine on the changes of free calcium level in neutrophils after smoke inhalation injury in the rabbit

The changes of the cytoplasmic free calcium level in the neutrophils after smoke in-halation injury were observed in rabbits and then the effects of tetrandrine,a calcium antago-nist,on the changes of free calcium level were studied.It was found that the number of neu-trophils increased significantly preceded by a transient decrease in the blood and also increasedin the bronehoalveolar lavage fluid after smoke inhalation.and the level of cytoplasmic free calci-um in the blood neutrophil increased likewise.Administration of tetrandrine resulted in a reduc-tion of the neutrophils number in the lungs and the free calcium level in the blood neutrophils toalleviate the pulmonary injury due to smoke inhalation.It is believed that there is a close rela-tionship between the activation of neutrophils and the pathophysiological changes of the lungs,and tetrandrine can exert its therapeutic effects on the injury by decreasing the free calcium levelin the neutrophils to modulate their functions.     

Changes of Tumor Necrosis Factor-α and the Effects of Ulinastatin Injection during Cardiopulmonary Cerebral Resuscitation

The changes of tumor necrosis factor-a (TNF-a) and brain ultrastructure during cardiopulmonary resuscitation and the effects of ulinastation injection were observed, and the mechanism was investigated. Twenty-four adult healthy Sprague-Dawley rats were randomly divided into.control group (8 rats), resuscitation group (8 rats) and ulinastatin (UTI) group (8 rats). Rats in control group underwent tracheotomy without clipping the trachea to induce circulatory and respiratory standstill. Rats in resuscitation and ulinastatin group were subjected to the procedure of establishing the model of cardiopulmonary cerebral resuscitation (CPCR). Rats in ulinastatin group were given with UTI 104 U/kg once after CPCR. In the control group, the plasma was collected immediate,30 min, 2 h, 4 h, and 6 h after tracheotomy. In resuscitation group and UTI group, plasma was collected immediate after tracheotomy, 30 min, 2 h, 4 h and 6 h after successful resuscitation. The plasma levels of TNF-a were determined by radioimmunoassay (RIA). At the end of the experiment, 2 rats were randomly selected from each group and were decapitated, The cortex of the brain was taken out immediately to observe the ultrastructure changes. In control group, there were no significant differences in the level of TNF-a among different time points (P＞0.05). In resuscitation group, the level of TNF-a was increased obviously after resuscitation (P＜0.01) and reached its peak 2 h later after resuscitation. An increasing trend of TNF-a showed in UTI group. There were no differences in TNF-a among each sample taken after successful resuscitation and that after tracheotomy. The utrastructure of brains showed the injury in UTI group was ameliorated as compared with that in resuscitation group. In early period of CPCR, TNF-a was expressed rapidly and kept increasing. It indicated that TNF-a might take part in the tissue injury after CPCR. The administration of UTI during CACR could depress TNF-a and ameliorate brain injury. By regulating the expression of damaging mediator, UTI might provide a protective effect on the tissue injury after CPCR.     

Effect of Amygdalin on the Proliferation of Hyperoxia-exposed Type Ⅱ Alveolar Epithelial Cells Isolated from Premature Rat

The pathogenesis of hyperoxia lung injury and the mechanism of amygdalin on type 2 alveolar epithelial cells (AEC2) isolated from premature rat lungs in vitro were investigated. AEC2 were obtained by primary culture from 20-days fetal rat lung and hyperoxia-exposed cell model was established. Cell proliferating viability was examined by MTT assay after treatment of amygdalin at various concentrations. DNA content and the proliferating cell nuclear antigen (PCNA) protein expression of AEC2 were measured by using flow cytometry and immunocytochemistry respectively after 24 h of hyperoxia exposure or amygdalin treatment. The results showed that hyperoxia inhibited the proliferation and decreased PCNA protein expression in AEC2 of premature rat in vitro. Amygdalin at the concentration range of 50--200 μmol/L stimulated the proliferation of AEC2 in a dose-dependent manner, however, 400 μmol/L amygdalin inhibited the proliferation of AEC2. Amygdalin at the concentration of 200 μmol/L played its best role in facilitating proliferation of AEC2s in vitro and could partially ameliorated the changes of proliferation in hyperoxia exposed AEC2 of premature rat. It has been suggested that hyperoxia inhibited the proliferation of AEC2s of premature rat, which may contribute to hyperoxia lung injury. Amygdalin may play partial protective role in hyperoxia-induced lung injury.     

Effects of Shenqi Fuzheng Injection(参芪扶正注射液)on Changes of Matrix Metalloproteinases in Patients with Acute Myocardial Infarction

Objective: To investigate the effects of Shenqi Fuzheng injection (参芪扶正注射液, SQFZI)on ventricular remodeling (VR) after acute myocardial infarction (AMI). Methods: Sixty patients with AMI were randomly divided into two groups and received conventional therapy and conventional therapy plus SQFZI separately. Matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-9 (TIMP-9) in plasma was measured on the 1st and 14th days after AMI. Results: MMP-9 and MMP-9/TIMP-9 were increased after AMI, after 2-week treatment, the level of MMP-9 and MMP-9/TIMP-9 decreased more significantly in SQFZI group than that in the conventional group (P＜0.05). Conclusion: Early treatment with SQFZI after AMI could reduce the level of MMPs and the production of collagen. It may be useful for the prevention of VR and heart failure after AMI.