AIM: To investigate the possibility that hepatocyte growth factor (HGF) directly induces differentiation of human embryonic stem cells (hESCs) into neural progenitors (NPs). METHODS: hESCs colonies were induced to form the embryoid body (EB). Four-day-old EBs were randomly divided into 4 groups: control group (EBs were cultured in neural induction medium);G5 supplement group (EBs were cultured in neural induction medium supplied with G5 supplement);HGF group (EBs were cultured in neural induction medium supplied with 10 mg/L HGF), and HGF+G5 group (EBs were cultured in neural induction medium supplied with 10 mg/L HGF and G5 supplement). After induced in suspension system for 7 days, EBs with various treatments were cultured in poly-D-lysine/laminin-coated plates for 7-10 days for selection of NPs. NPs were gathered by 0.3 g/L dispase treatment and characterized by immunofluorescence staining. The percentages of the nestin+ cells in NPs in various groups were detected by fluorescent activated cell sorter (FACS). The multipotency of NPs was determined by immunofluorescence staining after the NPs were cultured without G5 and HGF for 7 days. The expression of region markers of neural progenitors treated with sonic hedgehog (Shh) protein (one of the neural inductive signals), was detected by RT-PCR. RESULTS: HGF+G5 supplement induced hESCs differentiation into neural progenitors. Immunofluorescence staining indicated that NPs differentiated from hESCs expressed NP markers including nestin, Pax6 and musashi-1. FACS data showed that the proportion of nestin positive cells in HGF+G5 supplement group (87.3%±3.9%) was the highest in all treatment groups. The time of HGF and G5 supplement treatment was important to differentiate into NPs, the maximal effect was observed at 7th day. After treated with Shh, the expression of ventral forebrain/hindbrain marker genes (Nkk2.1, and Nkk2.2) and hindbrain progenitor marker gene Gbx2 in NPs were upregulated, while the forebrain progenitor marker genes Otx2 and Bf1 were downregulated. CONCLUSION: The neural induction system containing HGF and G5 supplement effectively induces the differentiation of hESCs into NPs, which might be a potent model for investigating the mechanism of neural development and differentiation.

A typical investigation on third-tier general hospitals, third-tier specialty hospitals, second-tier district or county central hospitals, urban community hospitals, and rural township health centers in Shanghai was conducted so as to examine the responsiveness of outpatients and inpatients to the health system. Most of the patients surveyed said that they could get prompt attention and that they had convenient access to medical information from the community or the mass media. More than half of the inpatients and outpatients thought that they had autonomy with regard to treatment and a greater number of patients made their own choices of doctors and nurses. Most of the patients were willing to talk freely with their doctors about their confidential medical history and could get rational explanations about their conditions from the doctors. The results indicate that reform of the health system in Shanghai has attached importance to the responsiveness of patients and that good effects have been achieved.

Objective To establish a simple ,stable acute liver injury model induced by CCl4 to observe effects of hepatocyte transplantation .Methods CCl4 plant oil with different concentration of 20% and 50% was used in mice by intraperitoneal injection , of which the dose was 2 mL/kg ,and then materials were taken at different time points respectively .Mice survival rate ,alanine amin-otransferase (ALT) ,aspartate aminotransferase (AST) and the pathological changes of the liver were detected .Results Mice sur-vival rate in 20% CCl4 intraperitoneal injection was significantly higher than that of 50% .ALT and AST in experiment group were significantly higher than that of control group ,but there was no significant difference between two experimental groups .Pathologi-cal examination showed that mice liver cells showed typical cytoplasmic ,ballooning ,scattered punctate ,piecemeal necrosis and in-flammatory cell infiltration in 20% CCl4 intraperitoneal injection ;while in 50% CCl4 ,there was obvious fibrosis ,in addition to the mentioned heavier lesions .Conclusion 20% -50% CCl4 intraperitoneal injection in 2 mL/kg dose can induce different degrees of relatively stable liver injury ,and its concentration determines the degree of liver injury .Acute liver injury induced by 20% -50%CCl4 was an ideal model for hepatocyte transplantation experiment .

Objective To evaluate clinical and sexual outcomes in women with severe uterine prolapsed at child-bearing period undergoing laparoscopic high uterosacral ligament suspension combined with cervical amputation.Methods From November 2007 to March 2010,34 patients (≤50 years old) with severe uterine prolapse (prolapse group) who had already given birth and underwent laparoscopic high uterosacral ligament suspension combined with cervical amputation in Peking Union Medical College Hospital were enrolled in this retrospective study.All patients were defined as Pelvic Organ Prolapse Quantification (POP-Q) stage Ⅲ.Follow-up was performed at 1,6,and 12 months,and then annually.Anatomic failure was defined as ≥ 1 pelvic compartment classified as POP-Q ≥ stage Ⅱ.Sexual outcomes were assessed at baseline and 3-year follow-up according to validated Short-Form Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12).Thirty one age-matched women who underwent health examinations during the same time period were selected as a healthy control group.Results Mean age of women at prolapse group were (39 ±5) years and average surgery time were (51 ±8) minutes.No severe intraoperative complications occurred,including urethral twist,bladder and rectum injury,pelvic hemotoma.All patients were followed up for more than 3 years,the mean following up period was 40 months (36-64 months).Based on stage Ⅱ of POP-Q as recurrence criteria,the anatomical success and patient satisfaction rates were both 100％ (34/34).POP-Q point C and D measurements were significantly higher after surgery then those at preoperative values [C:(-5.4±0.6) cm versus (2.2 ±0.7) cm,D:(-6.7 ±0.4) cm versus (-4.0 ±0.7) cm; P ＜0.01 for all].Three years postoperatively,33 patients were sexually active.Among the 94％ (31/33) patients who answered the PISQ-12 questionnaire,there was significant improvement in post-and preoperative total PISQ-12 scores (38 versus 26,P ＜ 0.01) and in all three subscale PISQ-12 domains (P ＜ 0.01 for all).The postoperative PISQ-12 score was similar to that of the control group (38 ± 6 versus 37 ± 3,P ＞ 0.05).Conclusion Laparoscopic high uterosacral ligament suspension combined with cervical amputation conferred satisfying long-term anatomic and sexual function outcomes in women at child-bearing age.

Aim To investigate TY501′s role in bleo-mycin ( BLM )-induced pulmonary fibrosis in rats. Methods Forty rats were randomly divided into 5 groups, including the sham operation, BLM, PFD, TY501(high and low dose) groups. After administra-tion of BLM intratracheally, PFD and TY501 were giv-en in each group daily, according to the dosage de-signed during 21 days. Lung coefficient, PaO2 were tested before killing the rats. The contents of ALB, ALP, LDH, GSH, HYP were detected by regent kit respectively. PCⅢ and COL4 were determined by ELISA. Results ( 1 ) Some indicators of alveolitis in early stage of IPF: the contents of lung coefficient in three treatment groups were lower and PaO2 was higher than those in BLM group ( P<0. 05 ); compared with BLM group, the contents of ALB, ALP, LDH in the treatment groups reduced on 21 st day ( P<0. 05 );the expression of GSH in BLM group was increased for feedback regulation and higher than the treatment groups and the sham operation group (P<0. 05);(2) some indicators of pulmonary interstitial fibrosis in late stage of IPF:the expressions of HYP, PCⅢand COL4 were reduced after the treatment. There were signifi-cant differences compared with BLM group ( P <0. 05 ) . Conclusions TY501 is valuable for the ther-apy of IPF, the same as the positive drug pirfenidone. TY501 attenuates BLM-induced pulmonary fibrosis, which may be related to the affection of TGF-βpathway and inhibition of MMPs.

Objective To investigate the protective effect of hepatocyte growth factor (HGF) on cultured Sprague-Dawley rat cortical neurons injured through hypoxia/reoxygenation.Methods Primary cultured cerebral cortical neurons were isolated from newborn rots.Neurons were pre-incubated with different concentrations (15,30 and 60 μg/L) of HGF.The cell viability was detected by MTT.Apoptosis was measured by Hoechst 33258 staining and flow cytometer.Lactate dehydrogenase (LDH) and caspase-3 activity were determined by colorimetry.Results Compared with normal group,hypoxia/reoxygenation treatment significantly decreased cell viability,increased LDH activity and the percentage of apoptotic cells.Pretreatment of HGF for 12 h could remarkably reverse the decrease of cell viability and the increase of apoptosis rate in neurons induced by hypoxia/reoxygenation treatment.HGF pre-treatment also attenuated the activity of LDH and caspase-3 in a dose-dependent manner.The effects of HGF could be inhibited by a special PI3K/Akt pathway inhibitor,LY294002.Condusion HGF could attenuate rat cortical neuron injury induced by hypoxia/reoxygenation.The neuroprotective effect of HGF may be related to activating PI3K/Akt pathway,and further suppressing the expression of caspase-3.

Objective To investigate the correlation between epicardial fat thickness and non dipper hypertension.Methods A total of 150 subjects was included in the study,of which 50 were in the non dipper hypertension group,the same in the non dipper hypertension group and the healthy control group.History collection and routine laboratory tests,ultrasonic measurement of epicardial fat thickness,and 24 hour ambulatory blood pressure monitoring were carried on all subjects.Epicardial fat thickness between groups was compared to primarily analyze the correlation of epicardial fat thickness and non dipper type hypertension.The optimal screening positive value in epicardial fat thickness of non dipper type primary hypertension was obtained by receiver operating characteristic (ROC) curve and maximum Youden index.Results When non dipper hypertension group and non-dipper hypertension group were compared,epicardial fat thickness was significantly increased [(6.30 ± 0.94) mm vs (5.92 ± 0.75) mm,P ＜ 0.05],as compared dipper hypertension group to healthy group,the epicardial fat thickness was significantly increased [(5.92 ±0.75)mm vs (5.50 ±0.13)mm,P ＜0.05].Epicardial fat thickness and non dipper type primary hypertension were linearly related (r =0.43,P ＜ 0.05),and epicardial fat thickness in diagnosis of non dippers primary hypertension optimal screening positive value was 6.01 mm.Conclusions There is a close relationship of epicardial fat thickness and non dipper hypertension.

Objective To evaluate the relationship between echocardiographic epicardial adipose tissue thickness(EAT) and the presence and severity of coronary artery disease(CAD). Methods One hundredand forty-seven patients (101 patients with CAD and 46 patients with normal coronary arteries by diagnostic coronary angiography) were enrolled. EAT thickness was measured using 2-D echocardiographic parasternal long-and short-axis views. EAT thickness measurements were compared with angiographic findings. Results EAT was significantly higher in CAD group comparison to control group [(7.41 ± 1.63)mm vs (4.41±1.60) mm, P ＜0.01 ]. Furthermore, EAT increased with the severity of CAD [(8.53 ± 1.00)mm vs (6.36 ±1.73)mm, P ＜0.01]. Gensini's score significantly correlated with EAT (r = 0.71, P ＜0.01 ). EAT thickness ≥5.35 mm had 87.13% sensitivity and 80.42% specificity (ROC area 0. 89, P = 0.01,95% CI [0.84 - 0.9;]) for predicting CAD. Conclusions EAT thickness, which is easily and non-invasively evaluated by transthoracic echocardiography, can be an adjunctive marker to classical risk factors for the prediction of CAD, it was significantly correlated with the severity of coronary artery disease.

Objective To analyze the effectiveness evaluation of cardiovascular drugs which have been developed on the CYP2C9 target protein by multi-layer fuzzy evaluation technology .Methods The multi-layer fuzzy evaluation method was used to evaluate the effectiveness of cardiovascular drugs interacting with the CYP 2C9 protein and to construct the index system that affects drug efficacy .Results and Conclusion The index system was used to study such cardiovascular drugs as valsartan and to score the drug effectiveness of individual samples .The results were consistent with actual drug treatment and were well confirmed .The results contribute to evaluation of personalized medication .

BACKGROUND: Whether human embryonic stem cells (hESCs) cultured on different feeder layers can maintain identical or similar characteristics remains unclear.OBJECTIVE: To compare the characteristics of hESCs cultured on human- and mouse- origin feeder layers.DESIGN, TIME AND SETTING: The in vitro cytology observation was performed at the Kunming Institute of Zoology, Chinese Academy of Sciences between September 2007 and February 2009.MATERIALS: Two ICR pregnant mice with 12.5-13.5 embryonic days were provided by Animal Center of Kunming Medical College. Immortalized human adult fibroblast (HAFi) cell line was presented by School of Medicine, Johns Hopkins University (USA). hESCs line was provided by Kunming Institute of Zoology, Chinese Academy of Sciences.METHODS: Mouse embryonic fibroblasts (MEF) were harvested from ICR mice by trypsinization method. HAFi were conventionally cultured. After γ ray treatment, two kinds of cells were incubated on 6-well gelatin-coated plate with density of 2.5×10~4/cm~3. hESCs were cultured on HAFi or MEF feeder cells containing β-mercaptoethanol DMEM/F12 and basic fibroblast growth factor.MAIN OUTCOME MEASURES: Morphology, expressions of specific molecular markers, Oct-4 positive rate, as well as cell doubling time of hESCs cultured by two methods were compared.RESULTS: ①BG02 cells on MEF and HAFi shared the similar morphology and characteristic pluripotency markers, which expressed SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 and Oct-4, but not SSEA-1. However, the proportion of positive Oct-4 cells in hESCs colonies maintained on MEF was lower than that of HAFi (P < 0.05) with shorter doubling time (P < 0.05).CONCLUSION: hESCs cultured on MEF and HAFi represent some differences in the growth and pluripotency characteristics.