Ethanol Extract of Phellinus merrillii Protects against Diethylnitrosamine- and 2-Acetylaminofluorene-Induced Hepatocarcinogenesis in Rats

Objective: To study whether the ethanol extract of Phellinus merrillii (EPM) has chemopreventive potential against liver carcinogenesis. Methods: Thirty male Spraque-Dawley rats were randomly divided into control group, EPM control group, hepatocarcinoma control group, low-dose EPM group and high-dose EPM group, 6 in each group. Using the Solt and Farber protocol in a rat model of hepatocarcinogenesis, the chemopreventive effect of EPM on diethylnitrosamine (DEN)-initiated, 2-acetylaminofluorene (2-AAF) and partial hepatectomy (PH)-promoted liver carcinogenesis in rats was evaluated. Basic pathophysiological and histological examinations, together with the serum levels of glutamic oxaloacetic transaminase (sGOT), glutamic pyruvic transaminase (sGPT) and gamma-glutamyl transpeptidase (γ-GT) were measured. Results: Treatment of EPM at the concentration of 2 g/kg body weight in the diet for 8 weeks clearly prevented the development of carcinogenesis and reduced the levels of sGOT, sGPT, and serum γ-GT of rats as compared with the hepatocarcinoma control group (P<0.05 or P<0.01). These phenotypes were accompanied by a significant increase in natural killer cell activity. Conclusion: EPM showed a strong liver preventive effect against DEN+2-AAF+PH-induced hepatocarcinogenesis in a rat model.     

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.     

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.     

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.     

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.     

Chinese Medicine for Outcomes in Colorectal Cancer Patients: A Retrospective Clinical Study

Objective: To investigate the effect of Chinese medicine(CM) on survival of patients with stage Ⅱ and Ⅲ colorectal cancer(CRC). Methods: A total of 295 patients who received chemotherapy were assigned to Group 1. The other 171 patients received the same chemotherapy treatment combined with the usage of CM Jianpi Jiedu Formula(健脾解毒方, JPJD) for more than 3 months(Group 2). Patients' survival time, relapse and metastasis, and cause of death were observed. Cox proportional hazard regression models were established for the analysis of the effect of independent factors on the survival prognosis of patients with CRC. Results: The survival rate of patients in Group 2 was higher than that of Group 1(P0.05). Compared with Group 1, the mean survival time was prolonged by 5.594 months and the median survival time was prolonged by 6 months in Group 2(P=0.004). Cox regression analysis indicated that CM combined with chemotherapy provided significant protective effect, as observed with the improvements in the survival rates of CRC patients(P0.01). Conclusion: CM can improve the survival rate in patients with stage Ⅱ and Ⅲ CRC.     

Shen-Fu Injection(参附注射液) Alleviates Post-resuscitation Myocardial Dysfunction by Up-regulating Expression of Sarcoplasmic Reticulum Ca2~+ -ATPase

Objective: To compare the effect of Shen-Fu Injection(SFI) and epinephrine on the expression of sarcoplasmic reticulum Ca2~+ ATPase 2a(SERCA2a) in a pig model with post-resuscitation myocardial dysfunction. Methods: Ventricular fibrillation(VF) was electrically induced in Wu-zhi-shan miniature pigs. After 8 min of untreated VF and 2 min of cardiopulmonary resuscitation(CPR), all animals were randomly administered a bolus injection of saline placebo(SA group, n=10), SFI(0.8 mg/kg, SFI group, n=10) or epinephrine(20 μg/kg, EPI group, n=10). After 4 min of CPR, a 100-J shock was delivered. If the defibrillation attempt failed to attain restoration of spontaneous circulation(ROSC), manual chest compressions were rapidly resumed for a further 2 min followed by a second defibrillation attempt. Hemodynamic variables were recorded, and plasma concentrations of catecholamines were measured. Adenylate cyclase(AC), cyclic adenosine monophosphate(c AMP) and the expressions of β1-adrenoceptor(AR) and SERCA 2a were determined. Results: Cardiac output, left ventricular dp/dt_(max) and negative dp/dt_(max) were significantly higher in the SFI group than in the SA and EPI groups at 4 and 6 h after ROSC. The expression of β1-AR and SERCA2 a at 24 h after ROSC were significantly higher in the SFI group than in the SA and EPI groups(P0.05 or P0.01). Conclusions: The administration of epinephrine during CPR decreased the expression of SERCA2 a and aggravated postresuscitation myocardial function(P0.01). SFI attenuated post-resuscitation myocardial dysfunction, and the mechanism might be related to the up-regulation of SERCA2 a expression.     

Effect of Bushen Yisui Capsule(补肾益髓胶囊) on Oligodendrocyte Lineage Genes 1 and 2 in Mice with Experimental Autoimmune Encephalomyelitis

Objective: To study the effects of Bushen Yisui Capsule(补肾益髓胶囊, BSYSC) on the oligodendrocyte lineage genes(Olig) 1 and Olig2 in C57BL/6 mice with experimental autoimmune encephalomyelitis(EAE) in order to explore the remyelination effect of BSYSC. Methods: The mice were randomly divided into normal control(NC), EAE model(EAE-M), prednisone acetate(PA, 6 mg/kg), BSYSC high-dose(3.02 g/kg) and BSYSC low-dose(1.51 g/kg) groups. The mice were induced by immunization with myelin oligodendrocyte glycoprotein(MOG) 35-55. The neurological function scores were assessed once daily. The pathological changes in mice brains were observed with hematoxylin-eosin(HE) staining and transmission electron microscope(TEM). The protein expressions of myelin basic protein(MBP), Olig1 and Olig2 in brains were measured by immunohistochemistry. The m RNA expressions of Olig1 and Olig 2 was also determined by quantitative real-time polymerase chain reaction. Results: Compared with the EAE-M mice,(1) the neurological function scores were significantly decreased in BSYSC-treated mice on days 22 to 40(P0.01);(2) the inflammatory cells and demyelination in brains were reduced in BSYSC-treated EAE mice;(3) the protein expression of MBP was markedly increased in BSYSC-treated groups on day 18 and 40 respectively(P0.05 or P0.01);(4) the protein expression of Olig1 was increased in BSYSC(3.02 g/kg)-treated EAE mice on day 40(P0.01). Protein and m RNA expression of Olig2 was increased in BSYSC-treated EAE mice on day 18 and 40(P0.01). Conclusion: The effects of BSYSC on reducing demyelination and promoting remyelination might be associated with the increase of Olig1 and Olig2.     

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.

     

Suppression of Oxidative Stress of Modified Gongjin-Dan (WSY-1075) in Detrusor Underactivity Rat Model Bladder Outlet Induced by Obstruction

Objective

To investigate the anti-oxidative stress and preventive effect of modified Gongjin-dan (WSY-1075) in a detrusor underactivity rat model.

Methods

Rats were randomly allocated to three groups: shamoperated (control), bladder outlet obstruction-induced detrusor underactivity (BOO-DU), and BOO-DU with WSY-1075 (WSY) groups. WSY-1075 was orally administrated to rats 200 mg daily for 2 weeks prior to the operation and 4 weeks after the operation. Bladder outlet obstruction was surgically induced in rats by ligation around the urethra avoiding total obstruction. Cystometrography was conducted on rats in each group for examination of bladders.

Results

Compared with the control group, bladder outlet obstruction led to a significant increase in oxidative stress with consequent changes to molecular composition, and decrease in maximal detrusor pressure (P<0.05). WSY-1075 treatment significantly suppressed oxidative stress and prevented degenerative and dysfunctional changes in bladder, as compared with BOO-DU group (P<0.05).

Conclusion

WSY-1075 had beneficial effect on prevention of BOO-DU.

     

Chinese Medicine Yishen Jiangu Granules(益肾健骨颗粒)on Aromatase Inhibitor-Associated Musculoskeletal Symptoms

Objective: To assess the effectiveness of Yishen Jiangu Granules(益肾健骨颗粒, YSJGG) on aromatase inhibitor-associated musculoskeletal symptoms(AIMSS). Methods: A single-arm, open-label study was conducted in 34 postmenopausal women with breast cancer who experienced AIMSS. Patients were treated with YSJGG for 12 weeks(12.4 g orally twice daily). The primary outcome was a change in the mean worst pain score of Brief Pain Inventory-Short Form(BPI-SF) over 12 weeks, and the second outcomes included changes in pain severity and pain-related interference of BPI-SF and Western Ontario and McMaster Universities Osteoarthritis Index(WOMAC), Modified Score for the Assessment of Chronic Rheumatoid Affections of the Hands(M-SACRAH), the Functional Assessment of Cancer Therapy-Breast(FACT-B), bone mineral density(BMD) and blood indices such as calcium(Ca), phosphate(P), and alkaline phosphatase(ALP). Results: Of 37 women recruited, 30 initiated the therapy and 24 were evaluable at 12 weeks. The primary outcome(BPI-SF worst pain scores) achieved a 2.17-point reduction compared with baseline(5.75±1.87 vs 3.58±2.15, P0.01). There were reductions in pain severity(decreased 1.65, P0.01) and pain-related interference(decreased 2.55, P0.01). The changes in WOMAC and M-SACRAH scores were similar to BPI-SF(P0.05). In the FACT-B, only physical wel-being and functional wel-being were improved compared with baseline(P0.05). No clinical differences were found in BMD, Ca, P and ALP. Conclusion: YSJGG is an effective and wel-tolerated agent to reduce AIMSS.     

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.     

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.