In order to study the analgesic effect of Shaoyao Gancao Decoction, this paper discussed material basis and mechanism from the perspective of macromolecules in traditional Chinese medicine. Inspired by the phenomenon of turbidity after boiling Chinese medicine, this experiment took Shaoyao Gancao Decoction as the research object to study the formation process of precipitation during boiling. The results showed that aggregates with a certain shape were formed in the solvent during the boiling process, and the precipitate was obtained by standing and centrifuging. Analysis found that the precipitation was mainly composed of small molecules such as paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid, isoliquiritin and gallic acid, and macromolecules such as protein and polysaccharide. The composition of precipitate was consistent with that of Shaoyao Gancao Decoction, but the analgesic effect of Shaoyao Gancao Decoction after removing the precipitate was significantly reduced. Based on these results, we isolated small molecular compounds, polysaccharides and protein from Shaoyao Gancao Decoction and their contents are 60.4, 700.7 and 207.2 mg·g^-1 respectively. We get the ratio, polysaccharide: small molecule = 11.6∶1, protein: small molecule = 3.4∶1, the precipitate is prepared in the state of boiling. The characterization results showed that the particle size of the precipitate will change significantly after co-heating, and the content determination results showed that the content of the six small molecular compounds which was free in solvent was significantly reduced after the formation of the precipitate. The acetic acid writhing experiment proved that the precipitate has a good analgesic effect, and effectively reduced the levels of inflammatory factors prostaglandin E2 and nitric oxide, and increased the level of anti-inflammatory factor interleukin-10. These results proved that the precipitate in Shaoyao Gancao Decoction is an important material basis for analgesic effect, and macromolecules such as protein and polysaccharide are the main components of the precipitate. The study of macromolecules in the precipitate of Shaoyao Gancao Decoction not only provides new ideas and methods for elucidating the pharmacodynamic material basis of Shaoyao Gancao Decoction, but also provides a reference for analyzing the scientificity of traditional decoction.

Objective To understand health seeking behavior and its influential factors to reproductive tract infections (RTIs) on women at reproductive age in the rural areas. Methods 54 540 fertile women aged 15-49 were surveyed by a stratified-cluster-random sampling method and gynecological examination were conducted in two steps: converging at the clinics, and then visiting their households, later, 31 624 women who had at least one RTI symptom were chosen. Results Among all the women at reproductive age, the rate of having at least one RTI symptom was 59.8% with the means of RTI symptom as 1.66±0.89. 15 989 women went to see doctors out of the 31 624 women who had RTI symptoms, with a proportion of 50.6 %. The results of logistic regressy showed that those women whose husbands having higher education level, higher income, more RTI symptoms and better knowledge on RTI were more easily to go to the hospitals. However, those women whose husbands working out of the county, having older first bearing age and more numbers of pregnancy were less likely to go to the hospitals. Reasons that refrained them from going to see a doctor would include: 2137(13.7%) did not know that RTI was a disease; 7443(47.6%) of them thought that every woman were bound to have at least one symptom and it did not matter; 1629 (10.4%) of them felt shameful; 349 (2.2%) learned that the diseases were incurable; 975 (6.2 % ) felt the cost of treatment was too expensive; 2101 (13.4 %) had no time; 1001 (6.4 %) would treat themselves through buying medicines over the counter. Conclusion RTI symptoms were quite prevalent among women at reproductive age but the rate of seeing a doctor was low and caused by multi-factors. Health education and gynecological census in increasing the curable rate of RTIs should to be strengthened.

Objective To assess the efficacy and safety of mifepristone combined with oral or vaginal misoprostol for termination of pregnancy between 8 and 16 weeks of gestation. Methods This was a randomized, multi-center, open clinical trial. A total of 625 women at 8-16 weeks of gestation were randomized to receive 200 mg oral mifepristone followed by either oral misoprostol 400 μg every 3 hours or vaginal misoprostol 400μg every 6 hours for a maximum of 4 doses 36-48 hours later. There were 417 women in oral group with 198 at 8-9 weeks and 219 at 10-16 weeks, while 208 women in vaginal group with 99 at 8-9 weeks and 109 at 10-16 weeks. The outcome measures were the success abortion rate, induction to abortion interval, the amount of bleeding, reoccurrence of menstruation and adverse events. Results Abortion rate was significantly higher in vaginal group [98.1% (202/206)] than that in oral group [94.0%(390/415), P=0.023]; concerning termination of pregnancy at 8-9 weeks and 10-16 weeks respectively, there were no significant differences between oral and vaginal groups (P=0.156, P=0.073). The induction to abortion interval was no significant difference in oral and vaginal group in different gestational weeks ( P=0.238, P=0.273). The average induction to abortion interval was (4.1 ± 6.6) hours and (6.0 ± 4.5) hours respectively in terminating 8-9 weeks and 10-16 weeks of gestation. Concerning the amount of bleeding within 2 hours of placenta expulsion, there was significant difference between oral group [(63±46) ml] and vaginal group [(55 ± 45) ml] in terminating 8-9 weeks of gestation (P=0.047), while there was no significant difference between groups in terminating 10-16 weeks of gestation [oral group (76 ± 52) ml versus vaginal group (76 ± 61) ml, P=0.507]. The reoccurrence of menstruation was about 37 days in both oral and vaginal groups. Two cases of incomplete abortion were serious adverse events (SAE) relating to treatment. The common adverse events (AE) of nausea and vomiting were significantly higher in oral group [57.2% (239/417), 36.3% (151/417)] than those in vaginal group [45.4% (94/208), 26.1% (54/208); P=0.005, 0.011]. Conclusion Oral or vaginal misoprostol combined with mifepristone, is effective and safe for termination of pregnancy between 8 and 16 weeks of gestation.

BACKGROUNDMono-(2-ethylhexyl) phthalate (MEHP), the metabolite of di-(2-ethylhexyl) phthalate (DEHP), was suspected to be toxic to human embryos. This study contributes to investigating its toxic effects by an embryonic stem cell test (EST) based on two human embryonic stem cell (hESCs) lines.

METHODSCH1 established in our own lab and H1, a federally registered cell line were two human embryonic stem cell lines used in this test. Four endpoint measurements were performed consisting of cell viability, proliferation ability, apoptosis as well as changes of gene expression patterns after spontaneous differentiation were determined. For measuring effects on the first three endpoints, the cells were treated with various concentrations of MEHP dissolved in dimethyl sulfoxide (DMSO) and only with DMSO which served as control and harvested after 5 days. For measuring effects during spontaneous differentiation, the RNA of embryoid bodies (EBs) formed after 8 days' MEHP exposure was collected and changes in differentiation specific gene expression patterns were analyzed by quantitative real time RT-PCR.

RESULTSAs a result the viability and proliferation ability of both cell lines decreased significantly at 1000 µmol/L MEHP, while there was no effect on apoptosis or cell morphology. In addition MEHP also changed the gene expression pattern in the EBs of both cell lines.

CONCLUSIONMEHP in a high dose was cytotoxic and affected the development of hESCs, which indicates its embryo toxicity in human embryos.

Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Diethylhexyl Phthalate ; analogs & derivatives ; toxicity ; Dose-Response Relationship, Drug ; Embryonic Stem Cells ; drug effects ; pathology ; Humans

Objective: To elucidate the possible mechanism responsible for the improved protection of terminal warm blood cardioplegia (TWBC) after hypothermic cardiopulmonary bypass (CPB) through analysis of tubulin (TB) components changes in myocardial cells exposed to TWBC. Methods: Stable animal models of CPB were established in cats, which were then randomly divided into 2 groups. Group Ⅰ was subjected to intermittent cold blood cardioplegia (ICBC) whereas group Ⅱ to ICBC followed by TWBC before uncross-clamping. Left ventricular performance was then monitored and evaluated by LVSP, LVEDP, ±dp/dtmax and t-dp/dtmax in both groups and semi-quantitive analysis was conducted with Western blot method as to the content and constitution of TB in myocardial cells at 15 min, 120 min after aortic crossclamping (ACC) and 5 min,15 min, 60 min,120 min after reperfusion. Results: Within 120 min after reperfusion, systolic and diastolic functions decreased significantly in group Ⅰ as compared with group Ⅱ(P<0.05). At 115 min after ACC and 15 min after reperfusion, the content of free and polymerized TB in both groups had no difference (P>0.05). At 120 min after ACC and 5 minutes after reperfusion, there was a significant difference between groupⅠ andⅡ (P<0.01). Conclusion: TWBC accelerates the repolymerization of myocardial TB during hypothermic CPB, which may mediate the improved cardiac performance in the early stage of myocardial reperfusion.

Objective To study the effect of bone marrow mesenchymal stem cells (BMMSCs) combined with normothermic mechanical perfusion (NMP) on biliary epithelial cells (BEC) after DCD donor liver transplantation in rats.Methods The third generation of BMMSCs and the BMMSCs modified by Ad/HO-1 (Ad/HO-1/BMMSCs) were cultured,identified and expanded in vitro.To establish a stable NMP system device in vitro.The DCD liver transplantation models were constructed in rats after cardiac ischemia for 30 minutes,220 SD recipient rats were randomly divided into sham operation group (S group,n=44) static cold storage (SCS group,n =44) group,and simple NMP group (P group,n =44),BMMSCs combined with NMP group (BP group,n =44) and BMMSCs modified by Ad/HO-1 combine with NMP group (HBP group,n =44),NMP group,BP group and HBP group were subjected to vitro perfusion for 4h.The group were taken at 0,1,7 and 14 days after transplantation and the relevant indicators were detected,n =6 in each group.The survival rate of the recipient rats,liver function and pathological changes of the bile duct were observed.The expression of cytokeratin 19 (CK19) protein in BEC was detected by immunohistochemistry and Western blot.Apoptotic biliary epithelial cells were detected by TUNEL staining and the expression of apoptosis-related protein caspase-3 was detected by immunohistochemistry.Results The survival time of HBP group was significantly prolonged for (5.6 ±0.8) d in SCS group vs.(18.4 ±2.0) d in NMP group,(20.5 ± 1.5) d in BP group,(82.5 ±3.2) d in HBP group,the differences were statistically significant (all P ＜ O.05).Compared with other groups,the HBP group and the BP group were significantly improved in liver function and biliary pathology,and the expression of CK19 protein in BEC was significantly increased [(0.81 ±0.02) in S group vs.(0.35 ±0.03) in SCS group,(0.47 ±0.02) in NMP group,(0.63 ± 0.02) in BP group,(0.77 ± 0.01) in HBP group on postoperative day (POD) 14],the differences were statistically significant (all P ＜ 0.05).The number of apoptosis and the expression of apoptosis-related protein caspase-3 in HBP group were significantly decreased [(10.0 ± 1.2) in S group vs.(57.3 ±5.5) in SCS group,(40.1 ±4.6) in NMP group,(32.0 ± 2.2) in BP group,(13.7 ± 3.1) in HBP group on POD 14],the difference was statistically significant (all P ＜ 0.05).Compared with the BP group,the protective effect of the HBP group was more obvious,and the difference was statistically significant (P ＜ 0.05).Conclusion By the method of the BMMSCs modified by Ad/HO-1 combined with NMP in vitro preservation of rat,DCD donor liver can significantly improve the effect of BEC on rats and the survival rate after liver transplantation.

Objective:To explore the role of heme oxygenase-1 (HO-1) modified bone marrow mesenchymal stem cells (BMMSCs) in improving hepatic microcirculation after reduced-size liver transplantation (RLT) in rats.Methods:BMMSCs were isolated and cultured in vitro by adherence method. Then HO-1/adenovirus (Adv) was transfected for constructing HO-1/BMMSCs. The "dual-sleeve" method was employed for establishing an acute rejection model after 50% RLT. Immediately post-operation, 1 ml normal saline (NS) and BMMSCs or HO-1/BMMSCs single cell suspension were injected. The changes of surviving rats were observed by parameters at Day 3/7/14 post-operation. Five rats were observed at each timepoint. The serum level of mitochondrial aspartate aminotransferase (mAST) was detected; Na+ -K+ -ATPase in transplanted liver was measured by chemical colorimetry; mitochondrial ultrastructural changes were observed under a transmission electron microscope. Portal vein pressure was detected by Power Lab at Day 7 post-operation; the expressions of endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS) and von Willebrand factor (vWF) in liver tissues were detected by Western blot. Liver histopathological changes were observed by hematoxylin & eosin stain. The expression of vWF was detected by immunohistochemistry and serum level of hyaluronic acid (HA) detected by enzyme-linked immunosorbent assay (ELISA).Results:HO-1/BMMSCs could significantly lessen the pathological injury and rejection of 50% reduced-size transplanted liver, improve mitochondrial damage and energy metabolism, promote the expression of eNOS, suppress the expression of iNOS, reduce portal pressure, up-regulate the expression of hepatic sinus vWF and HA degradation, protect hepatic sinusoidal endothelial cells (SECs) and ultimately improve hepatic microcirculation. And the differences were statistically significant as compared with NS/BMMSCs group ( P<0.05). Conclusions:HO-1/BMMSCs may play an important role in protecting rat liver by improving hepatic microcirculation during RLT.

Objective To explore the effect and mechanism of heme oxygenase-1 (HO-1) gene modified bone marrow mesenchymal stem cells (BMMSCs) on rat reduced-size liver transplantation.Methods 50 male Brown Norway (BN) rats were used to prepare BMMSCs.Male Lewis and BN rats were 75,respectively.BN rats were randomly divided into model group (n =25),stem cell group (n =25) and combined group (n =25).Acute rejection models following 50％ reduced-size transplantaton were established in rats using two-cuff technique,1 ml of normal saline,BMMSCs suspension,or HO-1/BMMSCs suspension were injected immediately after surgery.Rats were executed at an instant,3rd,7th and 14th day after surgery to identify BMMSCs and HO-1/adenovirus infection efficiency.Evaluated hepatic pathology by HE staining.Liver function indexes were detected.Portal vein pressure on 7th day after surgery was detected.The levels of endothelin-1 (ET-1) and nitric oxide (NO) in serum were detected using ELISA.The expressions of ET-1 in liver were detected by immunohistochemistry staining and Western blotting.Results High purity BMMSCs were obtained and HO-1/BMMSCs were successfully infected.Compared with model group,liver tissue injury and rejection were alleviated in stem cell group and combined group,liver function was improved,and the combined group was superior to stem cell group.The portal vein pressure in model group,stem cell group,and combined group were 21.3±0.2 mmHg,11.2±0.2 mmHg,and 10.1±0.1 mmHg,respectively.The portal vein pressure in three groups showed a decreasing trend,difference was statistically significant (P＜0.05).On the 3rd,7th and 14th day after surgery,compared with model group,the expression levels of ET-1 and NO in the stem cell group and the combined group were decreased,and the combined group was significantly lower than stem cell group (P＜ 0.05).Conclusion HO-1/BMMSCs improved liver function and portal vein pressure after reduced-size liver transplantation in rats,and may play a protective role by regulating ET-1/NO expression.

ence of precancerous pathological changes.

BACKGROUNDFew reports have evaluated the efficacy of re-operation for relapse after initial surgery for hepatocellular carcinoma (HCC) with bile duct thrombosis (BDT). The aim of this study was to investigate the efficacy of initial surgery and subsequent re-operation for HCC with BDT, and their effects on prognosis.

METHODSThe clinical data of 880 patients with HCC, including 28 patients with BDT, who underwent radical hepatectomy between 1998 and 2008 in our hospital, were reviewed. The effects of BDT and re-operation on prognosis were retrospectively analyzed.

RESULTSThe 1-, 3- and 5-year survival rates were 89.3%, 46.4% and 21.4%, respectively, in 28 patients with BDT versus 91.4%, 52.9% and 20.9% in 852 patients without BDT (P>0.05). Six patients with BDT underwent re-operation after disease relapse, and their survival time was significantly longer than those who did not undergo re-operation (P<0.05). Multivariate analysis indicated that portal vein invasion and tumor size were independently associated with tumor relapse and prognosis (P<0.05). Univariate analysis and multivariate analyses showed that obstructive jaundice was not significantly correlated with tumor relapse or prognosis (P>0.05).

CONCLUSIONSHepatectomy plus BDT removal is an effective treatment option for HCC with BDT. Obstructive jaundice is not a contraindication for surgery. Re-operation after relapse can provide good outcomes if the cases are appropriately selected.

Adult ; Bile Ducts ; pathology ; Carcinoma, Hepatocellular ; diagnostic imaging ; surgery ; Female ; Hepatectomy ; Humans ; Liver Neoplasms ; diagnostic imaging ; surgery ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography ; Thrombosis ; surgery ; Treatment Outcome ; Ultrasonography