Contribution of Water and Lipid Soluble Substances in the Relaxant Effects of Tymus Vulgaris Extract on Guinea Pig Tracheal Smooth Muscle in Vitro

Objective:To examine the relaxant effects of hydro-ethanolic,macerated aqueous(MA) and lipidfree macerated aqueous(LFMA) extract of Tymus vulgaris on tracheal chains of guinea pigs.Methods:The relaxant effects of five cumulative concentrations of each extract(0.4,0.8,1.2,1.6 and 2.0 g/100 mL) were compared with saline as negative control and five cumulative concentrations of theophylline(0.1,0.2,0.4,0.6 and 0.8 mmol/L) on precontracted tracheal smooth muscle of guinea pig with 60 mmol/L KCI(group 1) and 10 μmol/L methacholine(group 2,n=6 for each group).Results:In group 1 all concentrations of theophylline,three higher concentrations of hydro-ethanolic,two concentrations of LFMA and last concentration of MA extracts showed significant relaxant effects compared with that of saline(P0.05 or P0.01).Two lower concentrations of LFMA and all concentrations of MA except higher one caused contraction compared with saline(P0.05 or 0.01).In group 2 experiments,all concentrations of theophylline,hydro-ethanolic,MA and LFMA extracts showed significant relaxant effects compared to that of saline(P0.05 or P0.01).In both groups,the relaxant effect of all concentrations of hydro-ethanolic extract were significantly higher than most concentrations of others(P0.05 or P0.01).The relaxant effect of different concentrations of three extracts were significantly greater in group 2 compared with group 1 experiments(all P0.01).There were significantly positive correlations between the relaxant effects and concentrations for theophylline and ail extracts in both groups(P0.05 or P0.01).Conclusion:Hydro-ethanolic extract has a potent weaker relaxant effect for other extracts from Tymus vulgaris on tracheal chains of guinea pigs.     

Effects of Senegenin against Hypoxia/Reoxygenation-Induced Injury in PC12 Cells

Objective:To investigate the effect and the potential mechanism of Senegenin(Sen) against injury induced by hypoxia/reoxygenation(H/R) in highly differentiated PC12 cells.Methods:The cultured PC12 cells were treated with H/R in the presence or absence of Sen(60 μmol/L).Four groups were included in the experiment:control group,H/R group,H/R+Sen group and Sen group.Cell viability of each group and the level of lactate dehydrogenase(LDH) in culture medium were detected for the pharmacological effect of Sen.Hoechst 33258 staining and annexin V/propidium iodide double staining were used to analyze the apoptosis rate.Moreover,mitochondrial membrane potential(△Ψm),reactive oxygen species(ROS) and intracellular free calcium([Ca~(2+)]i) were measured by fluorescent staining and flow cytometry.Cleaved caspase-3and activity of NADPH oxidase(NOX) were determined by colorimetric protease assay and enzyme linked immunosorbent assay,respectively.Results:Sen significantly elevated cell viability(P0.05),decreased the leakage of LDH(P0.05) and apoptosis rate(P0.05) in H/R-injured PC12 cells.Sen maintained the value of△Ψm(P0.05) and suppressed the activity of caspase-3(P0.05).Moreover,Sen reduced ROS accumulation(P0.05) and[Ca~(2+)]i increment(P0.05) by inhibiting the activity of NOX(P0.05).Conclusion:Sen may exert cytoprotection against H/R injury by decreasing the levels of intracellular ROS and[Ca~(2+)]_i,thereby suppressing the mitochondrial pathway of cellular apoptosis.     

Potent Therapeutic Effects of Shouwu Jiangqi Decoction(首乌僵芪汤)on Polycystic Ovary Syndrome with Insulin Resistance in Rats

Objective:To investigate the effect of Shouwu Jiangqi Decoction(首乌僵芪汤,SJD) on polycystic ovary syndrome(PCOS) with insulin resistance(IR) in rats and to explore the underlining molecular mechanisms.Methods:A total of 51 female Sprague-Dawley rats were randomly divided into 6 groups:control group(n=7),model group(n=8),SJD high-dose group(n=9),SJD medium-dose group(n=9),SJD low-dose group(n=9) and DMBG group(n=9).Radioimmunoassay was used to measure serum follicle-stimulating hormone(FSH),luteinizing hormone(LH) and testosterone concentrations and qRT-PCR and western blot were used to examine the expression levels of mRNA and protein respectively of insulin receptor substrate 1(IRS-1)and phosphatidylinositide 3-kinases(PI3K) p85α in different groups.Results:FSH level significantly decreased in the model group compared with the normal control(P0.01),and high-dose SJD and DMBG can significantly increase FSH level(P0.01).LH level showed a mild increase without statistic significance in the model group compared with the control and different dosages of SJD had no significance effect on LH level,while DMBG can significantly decrease LH level(P0.01).Testosterone level significantly increased in the model group compared with the control group(P0.01),and high-dose SJD and DMBG can significantly decrease testosterone level(P0.01).The expression of IRS-1 as well as PI3Kp85α were significantly decreased in the model group compared with the normal control group at both mRNA(P0.001) and protein(P0.01) level,and both high-dose SJD and DMBG can enhance IRS-1 and PI3 K expression(P0.05).Conclusions:SJD has potent therapeutic effects on PCOS with IR in rats.The therapeutic effects of SJD on IR and ovulatory dysfunction are probably achieved through correcting the defective insulin signaling transduction.     

Synchronous Dynamic Research on Respiratory and Intestinal Microflora of Chronic Bronchitis Rat Model

Objectives

To investigate the mechanism of the Chinese medicine theory that Fei (Lung) and Dachang (Large Intestine) are exteriorly and interiorly related via synchronous observation on the dynamic changes of the respiratory and intestinal microflora.

Methods

Forty specific pathogen free Sprague-Dawley rats were selected and randomly divided into blank (10 rats) and chronic bronchitis model groups (30 rats). The blank group rats were put into the smoke-free environment and the model group rats were put into the smoke environment in order to establish pulmonary disease (chronic bronchitis) model. Then the corresponding changes of the respiratory and intestinal microflflora of the model on 20th, 50th and 70th days were synchronously observed.

Results

The respiratory tract microflflora showed an increase in the total aerobic and Staphylococcus aureus and reduced anaerobic amount signifificantly on 20th day in the respiratory tract microflflora (P<0.05 or 0.01). On 50th day, total aerobic, total anaerobic amount and bififidobacterium signifificantly increased (P<0.05). On 70th day, Staphylococcus aureus reduced and lactobacillus increased signifificantly (P<0.01). The intestinal microflflora showed an increase in the total aerobic, Clostridium perfringens, enterobacter and enterococcus significantly increased on 20th day (P<0.05 or 0.01). Staphylococcus aureus on 50th day increased significantly (P<0.05). Total aerobic and enterococcus increased, total anaerobic and Clostridium perfringens reduced signifificantly on 70th day (P<0.05 or 0.01).

Conclusion

The microecosystem of respiratory tract and intestine of rat model during the pathological process showed a dynamic disorder, indicating an interaction between the lung and large intestine which may be one of the connotations as they exteriorly and interiorly related.

     

Effect of Health Education Based on Integrative Therapy of Chinese and Western Medicine for Adult Patients with Type 2 Diabetes Mellitus: A Randomized Controlled Study

Objective: To investigate the effects of health education based on integrative therapy of Chinese and Western medicine for type 2 diabetes mellitus(T2DM) from the aspects of knowledge, attitude and practice(KAP), health-related quality of life(HRQo L), body mass index(BMI) and glucose control. Methods: Patients were individually randomized into intervention group(receiving integrative education, n=120) and control group(receiving usual education, n=120). The primary outcome was the changes in glycosylated hemoglobin A1c(HbA1c) levels after 3, 6, 9 and 12 months from baseline. Hierarchical linear models(HLMs) were used to assess within-group changes in outcomes over time and between-group differences in patterns of change. Secondary outcomes were KAP scores, HRQo L scores and BMI after 6 and 12 months, paired-sample t test was used to assess within-group changes in outcomes in 6 and 12 months, independent-sample t test was used to assess between-group differences in patterns of change. Results: HbA1c decreased statistically from baseline to 3 months, from 3 to 6 months, from 6 to 9 months and from 9 to 12 months in the intervention group(all P0.01); and decreased significantly from baseline to 3 months, and from 3 to 6 months in the control group(P0.01). There were significant between-group differences from baseline to 3 months(P=0.044), from 6 to 9 months(P0.01) and from 9 to 12 months(P0.01). Significant improvements in the intervention group along with significant between-group differences were found in KAP and HRQo L scores respectively(all P0.05). The number in the intervention group of normal weight increased from 56 at baseline to 81(6 months), 94(12 months), the number in the control group were 63(baseline), 69(6 months), 70(12 months), the χ~2 of hierarchical analysis of BMI were 6.93(P=0.075), 10.31(P=0.016), 15.53(P0.01), respectively. Conclusion: Health education based on integrative therapy of Chinese and Western medicine is beneficial to the control of T2DM and should be recommended for T2DM.     

Effects of Shenfu Injection (参附注射液) on Cerebral Metabolism in A Porcine Model of Cardiac Arrest

Objective:To investigate the effects of Shenfu Injection(参附注射液,SFI) on cerebral metabolism in a porcine model of cardiac arrest(CA).Methods:Thirty Wuzhishan minipigs were randomly assigned to the control group(n=6),epinephrine group(EP group,n=12) and SFI group(n=12).After8 min of untreated ventricular fibrillation(VF),pigs in the EP group or SFI group were administered with either EP(0.02 mg/kg) or SFI(1.0 mL/kg),respectively.After successful resuscitation,cerebrospinal fluid(CSF) levels of glucose,pyruvate,lactate,glutamate and glycerol were measured at 1,6,12 and 24 h after recover from spontaneous circulation(ROSC).In addition,neurologic deficit score(NDS) was calculated at 24 h after ROSC.Surviving pigs were killed at 24 h after ROSC,and the brain tissue was obtained for ultra-microstructure examination.Results:Compared with the EP group,CSF glucose and pyruvate levels were higher(all P0.01),and lactate levels were lower in the SFI group(P0.01).Meanwhile,CSF glutamate and glycerol levels in the SFI group were lower in comparison to the EP group(all P0.05).In addition,SFI decreased NDS at 24 h after ROSC(P0.01),and alleviated the histopathological damage of the brain.Conclusions:SFI could alleviate brain injury after CA,which may be associated with improving cerebral metabolism.     

Protective Effects of Xiongshao Capsule (芎芍胶囊) on Anti-inflammatory Function of High-Density Lipoprotein in An Atherosclerosis Rabbit Model

Objective: To observe the effects of Xiongshao Capsule(芎芍胶囊, XSC) on anti-inflammatory properties of high-density lipoprotein(HDL), myeloperoxidase(MPO) and paraoxonase 1(PON1) in serum of atherosclerosis(AS) rabbit model and explore the anti-inflammatory protective effects of XSC on HDL. Methods: Sixty rabbits were randomized into the control, the model, XSC low-, medium-and high-dose(Rhizoma Chuanxiong + Radix Paeoniae rubra : 0.6+0.3, 1.2+0.6, 2.4+1.2 g·kg-1·day-1, respectively), and simvastatin(1 g·kg-1·day-1) groups. The model rabbits were fed with high-fat diet and respective drugs for 15 weeks. The blood and thoracic aortas samples were collected at the end of 15 weeks. The levels of serum MPO and PON1 as well as total cholesterol(TC) and free cholesterol(FC) in aorta wall cells were tested by enzyme linked immunosorbent assay. Results: TC and FC in the model group were significantly higher than those in the control group(P0.01). Compared with the model group, TC and FC in the XSC groups were significantly lower(P0.05 or P0.01), so was simvastatin group(P0.01). There was no significant difference in PON1 level between groups(P0.05), even between model and control groups(P0.05). The serum MPO level in the model group was significantly higher than that in the control group(P0.05), which was significantly lower in XSC groups as well as simvastatin group(P0.05 or P0.01), and no difference was found between XSC groups and simvastatin group(P0.05). Conclusions: XSC can reduce the serum MPO level in AS rabbits to protect the anti-inflammatory function of HDL, maintaining the normal lipid transport function. TC and FC levels in aorta cells decline, and this process initiated by XSC plays an anti-AS role.     

Clinical Research on Intelligence Seven Needle Therapy Treated Infants with Brain Damage Syndrome

Objective

To assess whether the intelligence seven needle therapy administered in infants with perinatal brain damage syndrome (BDS) as early intervention would improve patients' neural development.

Methods

A randomized controlled trial was conducted. Sixty-four infants with BDS were randomly assigned to two groups: the comprehensive group and the control group. Both groups received routine early intervention; in addition, the comprehensive group received intelligence seven needle therapy. Before and after treatment, the Bayley Scale of Infant Development (BSID), Gesell Developmental Schedules, Gross Motor Function Measure (GMFM), transcranial doppler ultrasound (TCD), and cranial imaging examination were tested for contrast.

Results

After treatment, the comprehensive group showed significant difference in the Mental Development Index (MDI) scores of BSID compared with the control group (P<0.05), however, no significant discrepancy in psychomotor development index (PDI,P>0.05) was observed. The children's development quotients (DQ) of the comprehensive group exhibited a significant superiority in improving the social adaptation DQ of Gesell Developmental Schedules compared with the control group (P<0.01), as well as GMFM and linguistic and social intercourse (P<0.05). Again, no discrepancy in the fine movement DQ was found (P>0.05). The total scores of GMFM in the comprehensive group were higher than those in the control group (P<0.05). Comparing the two groups, the comprehensive group showed a significantly greater recovery rate than the control group on TCD after treatment (P<0.05). After 6-month follow-up, some recovery in both groups, specifically on broadening of brain outside space by cranial imaging examination were observed. The comprehensive group demonstrated a significantly greater recovery rate than the control group (P<0.05).

Conclusion

The developmental level of intelligence, motion function, linguistic competence and social intercourse can be promoted for infants with perinatal BDS by treating with the intelligence seven needle therapy. This approach can improve the brain blood supply and promote the growth of frontal and parietal lobes.

     

Effect of Kuanxiong Aerosol(宽胸气雾剂)on Patients with Angina Pectoris:A Non-inferiority Multi-center Randomized Controlled Trial

Objective: To evaluate the effect and safety of Kuanxiong Aerosol(宽胸气雾剂, KA) on patients with angina pectoris. Methods: Block randomization was performed to randomly allocate 750 patients into KA(376 cases) and control groups(374 cases). During an angina attack, the KA group received 3 consecutive sublingual sprays of KA(0.6 m L per spray). The control group received 1 sublingual nitroglycerin tablet(NT, 0.5 mg/tablet). Log-rank tests and Kaplan-Meier estimations were used to estimate the angina remission rates at 6 time-points after treatment(1, 2, 3, 4, 5, and 5 min). Logistic regression analysis was performed to observe the factors influencing the rate of effective angina remission, and the remission rates and incidences of adverse reactions were compared for different Canadian Cardiovascular Society(CCS) classes of angina. Results: The 5-min remission rates in the KA and control groups were not significantly different(94.41% vs. 90.64%, P0.05). The angina CCS class significantly influenced the rate of remission (95% confidence interval = 0.483–0.740, P0.01). In the CCS subgroup analysis, the 3-and 5-min remission rates for KA and NT were similar in the CCSⅠ and Ⅳ subgroups(P0.05), while they were significantly better for KA in the CCSⅡ and Ⅲ subgroups(P0.05 or P0.01). Furthermore, the incidence of adverse reactions was significantly lower in the KA group than in the control group for the CCSⅡ and Ⅲ subgroups(9.29% vs. 26.22%, 10.13% vs. 20.88%, P0.05 or P0.01). Conclusions: KA is not inferior to NT in the remission of angina. Furthermore, in CCSⅡ and Ⅲ patients, KA is superior to NT, with a lower incidence of adverse reactions.     

Effects of Modified Qing'e Pill (加味青娥丸) on Expression of Adiponectin, Bone Morphogenetic Protein 2 and Coagulation-Related Factors in Patients with Nontraumatic Osteonecrosis of Femoral Head

Objective: To observe the regulation of Chinese herbal medicine, Modified Qing'e Pill(加味青娥丸, MQEP), on the expression of adiponectin, bone morphogenetic protein 2(BMP2), osteoprotegerin(OPG) and other potentially relevant risk factors in patients with nontraumatic osteonecrosis of the femoral head(ONFH). Methods: A total of 96 patients with nontraumatic ONFH were unequal randomly divided into treatment group(60 cases) and control group(36 cases). The treatment group were treated with MQEP while the control group were treated with simulated pills. Both groups were given caltrate D. Six months were taken as a treatment course. Patients were followed up every 2 months. The levels of plasma adiponectin, BMP2, OPG, von Willebrand factor(vWF), von Willebrand factor cleaving protease(vWF-cp), plasminogen activator inhibitor 1(PAI-1), tissue plasminogen activator(tPA), C-reactive protein(CRP), blood rheology, bone mineral density(BMD) of the femoral head and Harris Hip Score were measured before and after treatment. Results: After 6 months of treatment, compared with the control group, patients in the treatment group had significantly higher adiponectin and BMP2 levels(P0.01 and P=0.013, respectively), lower vWF, PAI-1 and CRP levels(P=0.019, P0.01 and P0.01, respectively), and lower blood rheology parameters. BMD of the femoral neck, triangle area and Harris Hip Score in the treatment group were significantly higher than those in the control group. Moreover, plasma adiponectin showed a positive association with BMP2(r=0.231, P=0.003) and a negative association with PAI-1(r=–0.159, P0.05). Conclusions: MQEP may play a protective role against nontraumatic ONFH by increasing the expression of adiponectin, regulating bone metabolism and improving the hypercoagulation state, which may provide an experimental base for its clinical effects.     

Efficacy and Safety of Qiaoshao Formula(翘芍方) on Patients with Lifelong Premature Ejaculation of Gan(Liver) Depression and Shen(Kidney) Deficiency Syndrome:A Randomized Controlled Trial

Objective: To observe the efficacy and safety of Qiaoshao Formula(翘芍方, QSF) on patients with lifelong premature ejaculation(LPE) of Gan(Liver) depression and Shen(Kidney) deficiency syndrome. Methods: A total of 60 LPE patients were randomly divided into treatment(QSF) and control(dapoxetine) groups. The treatment group received QSF twice a day and the control group received dapoxetine 1 to 2 h prior to planned sexual intercourse for 4 weeks. The outcomes included intra-vaginal ejaculation latency time(IELT), premature ejaculation diagnostic tool(PEDT), clinical global impression of change(CGIC), scores of Chinese medicine symptoms(CMSS), sex life satisfaction(SLS) and adverse events(AEs). Results: In the treatment group, the median IELT was 3 min vs. 1.5 min before and after treatment(P0.05). PEDT in the treatment group was reduced to 11.76±1.68 from 15.83±2.30 after treatment(P0.05). Besides, patient's SLS was improved from 1.30±0.05 to 6.30±0.04(P0.05), and spouse's SLS was increased from 1.30±0.08 to 6.10±0.06(P0.05); CMSS was decrease from 14.86±3.02 to 9.62±2.87(P0.05). In addition, no significant AE was observed in both groups. Conclusion: QSF may be effective and safe on LPE patients with Gan depression and Shen deficiency syndrome.     

Contribution of Borneolum syntheticum to the Intervention Effect of Liuwei Dihuang Pill (六味地黄丸) on Experimental Retinal Degeneration

Objective: To observe the contribution of Borneolum syntheticum to the intervention effect of Liuwei Dihuang Pill (六味地黄丸, LDP) on experimental retinal degeneration, and initially investigate the mechanism of Borneolum syntheticum as meridian-lead-in drug. Methods: A total of 180 sodium iodate-induced retinital degeneration rats were randomly divided into three groups, including distilled water group, LDP group, and LDP+Borneolum syntheticum (LDP+BS) group. Twenty normal rats were fed regularly without any treatment as normal control. On day 7 and 14 after treatment, histopathological study and transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) test were performed to evaluate the retinopathy. Claudin-5 expression at blood-retina barrier (BRB) was detected by Western blot at different time points from 0.5 to 8 h after gavage. Results: On day 7 and 14 after treatment, the retinal lesion grades were significantly different among the three groups (P<0.05). The grade in the LDP+BS group was significantly less than the LDP and distilled water groups (both P<0.05), no significant difference was observed between the LDP and distilled water groups (P>0.05). The apoptosis rates in the LDP+BS group was significantly less than the distilled water and LDP groups (both P<0.05), while there was no significant difference between LDP and distilled water groups (P>0.05). Expression of claudin-5 in LDP+BS group was significantly less than the other two groups at 0.5, 1 and 2 h after gavage (P<0.05). There was no apparent difference among the three groups at 4 and 8 h after gavage (P>0.05). Conclusion: Borneolum syntheticum could strengthen the effect of LDP on experimental retinal degeneration, indicated that Borneolum syntheticum might play the role of meridian-lead-in drug in the formula. The mechanism may be due to Borneolum syntheticum could promote the physiologically openness of blood-retina barrier through transiently affecting the expression of claudin-5.     

A Multi-center,Randomized,Controlled and Open Clinical Trial of Heyan Kuntai Capsule (和颜坤泰胶囊) and Hormone Therapy in Perimenopausal Women

Objective: To evaluate the efficacy and safety of Heyan Kuntai Capsule(和颜坤泰胶囊, HYKT) and hormone therapy(HT) on perimenopausal syndromes(PMSs). Methods: From 2005 to 2008, 390 women with PMSs were recruited from 4 clinic centers. The inclusion criteria included ages 40 to 60 years, estradiol(E2) below 30 ng/L, and follicle stimulating hormone(FSH) above 40 IU/L, etc. The patients were randomly assigned to HYKT group or HT group by random number table method, administrated HYKT or conjugated estrogen with/without medroxyprogesterone acetate tablets for 12 months. During treatment, the patients were interviewed quarterly, Kupperman Menopausal Index(KMI) scores, hot flush scores, insomnia scores, Menopause-Specific Quality of Life(MENQOL) scores and adverse effects were used for evaluating drug efficacy and safety respectively. The last interview was made at the end of 12-month treatment. Results: After treatment, KMI scores of HYKT group and HT group were both significantly decreased compared with baseline(P0.01) and there was no significant difference between groups(P0.05), except that KMI of HYKT group was higher after 3-month treatment(P0.05). After treatment, hot flush and insomnia scores were both improved significantly in two groups(P0.01); and HT had a better performance than HYKT in improving hot flush(P0.05). MENQOL were significantly improved in both groups after treatment(P0.01); but there was no significant difference between two groups(P0.05). The incidence of adverse event in the HYKT group was much lower than that in the HT group(P0.01). Conclusions: HYKT could effectively relieve PMSs and improve patient quality of life without severe adverse reactions. Although HYKT exerted curative effects more slowly than hormone, it possessed better safety profile than hormone.     

Jinmaitong(筋脉通)Alleviates the Diabetic Peripheral Neuropathy by Inducing Autophagy

Objective:To observe the deregulation of autophagy in diabetic peripheral neuropathy(DPN) and investigate whether Jinmaitong(筋脉通i,JMT) alleviates DPN by inducing autophagy.Methods:DPN models were established by streptozotocin-induced diabetic rats and Schwann cells(SCs) cultured in high glucose medium.The pathological morphology was observed by the improved Bielschowsky's nerve fiber axonal staining and the Luxol fast blue-neutral red myelin staining.The ultrastructure was observed by the transmission electron microscopy.Beclinl level was detected by immunohistochemistry and Western blot.The proliferation of cultured SCs was detected by methylthiazolyldiphenyl-tetrazolium bromide.Results:Diabetic peripheral nerve tissues demonstrated pathological morphology and reduced autophagic structure,accompanied with down-regulation of Beclinl.JMT apparently alleviated the pathological morphology change and increased the autophagy[in vivo,Beclinl integral optical density(IOD) value of the control group 86.6±17.7,DM 43.9±8.8,JMT 73.3 ±17.8,P0.01 or P0.05,in vitro Beclinl IOD value of the glucose group 0.4710.25 vs the control group 0.88 ±0.29,P0.05].Consequently,inhibition of autophagy by 3-methyladenine resulted in a time- and concentrationdependent decrease of the proliferation of SCs(P0.05,P0.01).Conclusions:Down-regulation of autophagy in SCs might contribute to the pathogenesis of DPN.JMT alleviates diabetic peripheral nerve injury at least in part by inducing autophagy.     

Puerarin Up-regulates Methyl-CpG Binding Protein 2 Phosphorylation in Hippocampus of Vascular Dementia Rats

Objective: To observe the effect of puerarin on methyl-CpG binding protein 2(MeCP2) phosphorylation(pMeCP2) in the hippocampus of a rat model of vascular dementia(VD). Methods: Thirty-six healthy Sprague-Dawley rats were randomly assigned to the sham-operated group, dementia group and puerarintreated group using a random number table(n=12 per group). The modified permanent bilateral common carotid artery occlusion method was used to establish the VD model. The sham-operated and dementia groups were given 2 m L/d of saline, while the puerarin-treated group was given 100 mg/(kg·d) of puerarin for 17 days. The learning and memory abilities were evaluated by the Morris water maze test. Hematoxylin-eosin staining, immunohistochemical(IHC) staining and Western blot analysis were carried out to observe changes in neuron morphology and in level of pMeCP2 in the hippocampus, respectively. Results: The morphologies of rat hippocampal neurons in the puerarintreated group were markedly improved compared with the dementia group. The escape latency of the dementia group was significantly longer than the sham-operated group(P0.05), while the puerarin-treated group was obviously shorter than the dementia group(P0.05). Cross-platform times of the dementia group were significantly decreased compared with the sham-operated group(P0.05), while the puerarin-treated group was obviously increased compared with the dementia group(P0.05). IHC staining showed no significant difference in the number of MeCP2 positive cells among 3 groups(P0.05). The number of pMeCP2 positive cells in the CA1 region of hippocampus in the dementia group was significantly increased compared with the sham-operated group, and the puerarin-treated group was significantly increased compared with the dementia group(both P0.05). Western blot analysis showed no significant difference of MeCP2 expression among 3 groups(P0.05). The expression of pMeCP2 in the dementia group was significantly increased compared with the sham-operated group, while it in the puerarin-treated group was significantly increased compared with the dementia group(P0.05). Conclusion: Puerarin could play a role in the protection of nerve cells through up-regulating pMeCP2 in the hippocampus, improving neuron morphologies, and enhancing learning and memory ablities in a rat model of VD.     

Effect of Artemisia annua Extract on Treating Active Rheumatoid Arthritis: A Randomized Controlled Trial

Objective

To investigate the effect and safety of the complementary use of the extract of Artemisia annua L. (EAA) on treating active rheumatoid arthritis (RA).

Methods

A randomized controlled clinical trial was performed. All the 159 participates with active RA were randomly assigned to the control group (80 cases) and EAA group (79 cases) using concealed random allocation method. In the control group, patients were medicated with leflflunomide and methotrexate for 48 weeks; and patients in the EAA group were administrated with leflflunomide, methotrexate plus EAA (30 g/d). At the time points of 0, 12, 24 and 48 weeks, the clinical outcome measures, including objective pain score, tenderness score, number of painful joints, number of swollen joints, health assessment questionnaire (HAQ) score for quality of life, levels of serum rheumatoid factor (RF), anti-cyclic citrullinated protein antibodies (CCP-Ab), erythrocyte sedimentation rate (ESR), C reactive protein (CRP), visual analogue score for pain (VAS), and the overall effificacy were detected and recorded.

Results

The objective pain score, number of painful joints and ESR at 12 weeks, tenderness score and HAQ at 24 weeks, and the tenderness score, number of painfull joints, number of swollen joints, HAQ, CRP, RF and CCP-Ab at 48 weeks were signifificantly improved in the EAA group compared with the control group (P<0.01 or P<0.05). At 24 and 48 weeks, the overall effificacy of the EAA group was signifificantly higher than the control group (P<0.01). There were signifificantly higher withdrawal rate of corticosteroids within 12 weeks post-treatment and lower incidence rate of adverse effects in the EAA group compared with the control group (P<0.01 or P<0.05).

Conclusion

EAA plus methotrexate and leflflunomide were more effective and safer than the routine use of methotrexate and leflflunomide in the treatment of active RA.

     

Antipyretic Effect of Herba Ephedrae-Ramulus Cinnamomi Herb Pair on Yeast-Induced Pyrexia Rats: A Metabolomics Study

Objective

To investigate the antipyretic mechanism of Herba Ephedrae (Eph)-Ramulus Cinnamomi (RC) herb pair on yeast-induced pyrexia in rats.

Methods

Totally 30 qualified male SD rats were randomly assigned to the normal control (NC) group, the pyrexia model (model) group, the Eph, RC and Eph-RC treatment groups by a random digital table, 6 rats in each group. Each rat received a 20% aqueous suspension of yeast (10 mL/kg) except the NC group. The 3 treatment groups were administered 8.1, 5.4 and 13.5 g/kg Eph, RC and Eph-RC respectively at 5 and 12 h after yeast injection, the NC group and the model groups were administered equal volume of distilled water. Rectal temperatures were measured at 0, 6, 8, 10, 12, 15, 18, 24 and 30 h and urine was collected prior to yeast injection and at 6, 10, 18, 24, 30, and 36 h after yeast injection. Then urine metabolomic profiling by gas chromatography tandem mass spectrometry, coupled with multivariate statistical analysis and pattern recognition techniques were used to explore the antipyretic effects of Eph-RC. Partial least squares discriminate analysis was used to analyze the metabolomics dataset including classification and regression in metabolomics plot profiling.

Results

Compared with the NC group, rectal temperatures were significantly higher in the model group (P<0.01), while 3 treatment groups decreased significantly compared with the model group (P<0.05 or P<0.01). Rectal temperatures of Eph-RC-treated rats started to go down at 6 h, and markedly decreased at 8, 12, 15, 18 and 24 h (P<0.05 or P<0.01), while those of the Eph and RC groups had decreased firstly at 8 h and were markedly lower at 12 h (P<0.05 or P<0.01). Seventeen potential biomarkers related to pyrexia were confirmed and identified, including pyruvic acid, L-phenylalanine, L-tyrosine, phenylacetic acid, hippuric acid, succinic acid, citrate and so on. Eight potential alterations of metabolic pathways including phenylalanine metabolism, citrate cycle, tryptophan metabolism, biosynthesis of valine, leucine and isoleucine, were identified in relation to the antipyretic effects of Eph-RC using MetPA software.

Conclusion

The antipyretic effect of Eph-RC herb pair on yeast-induced pyrexia in rats involved correction of perturbed amino acid, fatty acid, and carbohydrate metabolism according to the metabolic pathway analysis with MetPA.

     

Gefitinib plus Fuzheng Kang'ai Formula(扶正抗癌方) in Patients with Advanced Non-Small Cell Lung Cancer with Epidermal Growth Factor Receptor Mutation:A Randomized Controlled Trial

Objective: To evaluate the effect of Fuzheng Kang'ai Formula(扶正抗癌方, FZKA) plus gefitinib in patients with advanced non-small cell lung cancer with epidermal growth factor receptor(EGFR) mutations. Methods: A randomized controlled trial was conducted from 2009 to 2012 in South China. Seventy chemotherapynaive patients diagnosed with stage ⅢB/Ⅳ non-small cell lung cancer with EGFR mutations were randomly assigned to GF group [gefitinib(250 mg/day orally) plus FZKA(250 m L, twice per day, orally); 35 cases] or G group(gefitinib 250 mg/day orally; 35 cases) according to the random number table and received treatment until progression of the disease, or development of unacceptable toxicities. The primary endpoint [progression-free survival(PFS)] and secondary endpoints [median survival time(MST), objective response rate(ORR), disease control rate(DCR) and safety] were observed. Results: No patient was excluded after randomization. GF group had significantly longer PFS and MST compared with the G group, with median PFS of 12.5 months(95% CI 3.30–21.69) vs. 8.4 months(95% CI 6.30–10.50; log-rank P0.01), MST of 21.5 months(95% CI 17.28–25.73) vs. 18.3 months(95% CI 17.97–18.63; log-rank P0.01). ORR and DCR in GF group and G group were 65.7% vs. 57.1%, 94.3% vs. 80.0%, respectively(P0.05). The most common toxic effects in the GF group and G group were rash or acne(42.8% vs. 57.1%, P0.05), diarrhea(11.5% vs. 31.4%, P0.05), and stomatitis(2.9% vs. 8.7%, P0.05). Conclusion: Patients with advanced non-small cell lung cancer selected by EGFR mutations have longer PFS, MST with less toxicity treated with gefitinib plus FZKA than gefitinib alone.