Objective To investigate the expression of endometrial developing-related genes,the endometrium-like structure and cells during spontaneous differentiation of hESCs. Methods (1) Embryoid bodies were cultured in suspension for 14 days,fixed in 10% neutral-buffered formalin and embedded in paraffin.Estrogen receptor(ER) was detected by immunocytochemical staining.(2) hESCs were cultured in a 35 mm plastic dish and differentiated spontaneously for 10 days.The expression of ER and Vimentin/Keratin was detected by double immunofluorescence histochemistry.(3) hESCs were cultured in a 35 mm plastic dish and differentiated for 5 days.The expressions of endometrial developing-related genes including Wnt4,Wnt7a,Wnt5a,Hoxa10,Hoxa11 and ER were detected by RT-PCR. Results (1) The endometrium-like structures were detected in 14-day-old EBs and some were positive for ER staining.(2) Some spontaneously differentiated cells from hESCs for 10 days were positive for ER together with Vimentin/Keratin.(3)Wnt7a,Wnt4,Hoxa10,Hoxa11 and ER were detected by RT-PCR in hESCs which were differentiated spontaneously for 5 days.Conclusion During the spontaneous differentiation of hESCs,some cells are liable to differentiate into endometrial stem cells and endometrium-like cells.However,further identifications of the whole development process are needed to be done in the future.

Objective To find a new source to produce hESC lines.Methods D3 embryos with low morphological scores were cultured to blastocyst stage.Trophectoderm cells were separated from the ICMs by immunosurgery and isolated ICMs were cultured for 5-8 days on mitomycin-treated mouse embryonic fibroblasts(MEFs).Colonies derived from the ICMs were passed every 4-7 days and evaluated for cell surface markers including AKP,OCT-4,SSEA-4,SSEA-1,TRA-1-60,TRA-1-81,differentiation potentials and karyotypes.Results A total of 19 blastocysts were obtained from 130 embryos(quality score

Objective To discuss the epidemiological situation of Streptococcus pyogens infection and drug sensitivity results in children in Meilong area in Shanghai,China,and provided scientific pathogen information for clinic infection control and treatment.Methods This was a retrospective study.The group A Streptococcus pyogens strains which were isolated from a total of 1 069 throat swab samples of pediatrics patients between May 2014 and April 2015,the strains were used the method of molecular biology for emm type and MLST and PFGE,part of the strains were used the Kirby-Bauer method for drug sensitivity test.To analyze the infection characteristics,epidemic tendency and drug sensitivity of GAS in different seasons and different age groups.Results A total of 274 S.pyogens strains were detected,the positive rate was 25.63％,the main types of emm were emm1(38.83％)and emm12(52.75％),the others were 8.42％.The main types of MLST were ST-28,ST-36,ST-49.emm types were closely related with MLST and PFGE clust.Among them,emm1/ST28,emm12/ST36,emm75/ST49 were related to each other,the same emm types were mostly the same cluster of PFGE.During this study,the patients were 3-13 years old,and the high infection age were 6-11 years old.The prevalent infection time were May 2014 and June 2014 and between Novemer 2014 to January 2015 and April 2015.The sensitivity rate of beta-lactamase drugs such as penicillin and ampicillin and levofloxacin,vancomycin and linezolid were 100％,the resistance rate of clindamycin and erythromycin and tetracycline were more than 95％.Conclusion The most popular genotype of GAS was emm12,the main age of infected patients were 6-11 years old,and the epidemic season were winter-spring and early summer in Meilong area,Shanghai,China,and beta-lactamase drugs were the first choice for GAS infection.

Objective To investigate the protective effects of Green Tangerine induced hypertension and mild hypothermia on focal cerebral ischemia and reperfusion and local cerebral glucose utilization(LCGU)in the in- farction rim in rats.Methods A total of 64 rats were used and randomly divided into a control group,a Green Tan- gerine induced hypertension group,a mild hypothermia group and a combination therapy group.The neurologic defi- cits,infarct size and LCGU were observed in the rats with focal cerebral ischemia and reperfusion.Results Com- pared with the control group,the neurologic deficits(P

Objective To apply 3C3R model in problem design of PBL for pediatrics teaching.Methods The 3C3R model comprises two classes of components:core components and processing components.Core components of the model are 3C,which are content,context and connection while processing components are 3R,which are researching,reasoning and reflecting.3C3R model was used in the problem design for the PBL case of ‘ Why the mouth of Baobao became purple when he was crying?' Totally 76 eight-year program medical students and 7 tutors were enrolled as teaching object.The anonymous questionnaires from the students were collected for assessment of PBL teaching.Results The percentage of students with scores ≥4 for content in PLB problem design was 90.8％,for context was 80.3％,for connection was 64.5％,for researching was 81.6％,for reasoning was 69.7％ and for reflecting was 40.8％.The percentage of tutors with scores ≥4 for content in PLB problem design was 100％,for context was 71.4％,for connection was 28.6％,for researching was 71.4％,for reasoning was100％,for reflecting was57.1％.Both students and tutors held a positive attitude towards the component of content,context,researching and reasoning in problem design model.But the components of connection and reflection needed to be improved.Conclusion The 3C3R model is helpful for problem design of PBL in pediatric teaching.

OBJECTIVETo investigate the spatial distribution characteristics of pulmonary tuberculosis in Zhejinag province in 2006 by G statistics, so as to find out the hotspot of occurrence of pulmonary tuberculosis in Zhejiang province and provide evidence for control and prevention on pulmonary tuberculosis.

METHODSThe data of pulmonary tuberculosis cases in 90 counties (districts) in Zhejiang province in 2006 were obtained from the tuberculosis surveillance system. Based on the same county field in digital maps and pulmonary tuberculosis incidence database, digital map of Zhejiang province was interrelated with the database of Zhejiang province pulmonary tuberculosis incidence to establish Zhejiang geographic information system database. General and local G statistics were developed to test for spatial heterogeneity by ArcGIS 9.2 software.

RESULTSA total of 43 467 cases of pulmonary tuberculosis were reported in Zhejiang province in 2006, and the reported incidence was 88.74/100,000.G statistics indicated that there were high occurrence of pulmonary tuberculosis (Getis-Ord Gi=0.0764, P<0.05). Local Getis-Ord Gi statistics analysis showed that there were statistically significant hotspots in Yuhuan county, Leqing city, Pingyang county, Dongtou county, Yongjia county, Ruian city and Ouhai, Longwan, Lucheng district in Wenzhou city, other areas were intergradational zone with Z(Gi) value fell in -1.96 to 1.96.

CONCLUSIONPulmonary tuberculosis in Zhejiang province present unrandomly distributed and geographically clustered.

China ; epidemiology ; Cluster Analysis ; Geographic Information Systems ; Geography ; Humans ; Incidence ; Models, Statistical ; Tuberculosis, Pulmonary ; epidemiology

Objective To investigate the effect of small interfering RNA (siRNA) mediated silencing of △Np73 on 5-FU chemotherapy sensitivity in SW620 colocancer cells and provide new treatment approach for the colon cancer.Methods siRNAs were transfected into SW620 colon cancer cells.The expression of △Np73 was observed.Cell viability of colon cancer cells were measured by MTr assay and cell apoptosis was assessed with flow cytometry after treatment of control siRNA or △Np73 siRNA or combined with 5-FU,respectively.The tumorigenesis was assessed by injecting △Np73 siRNA or control siRNA transfeeted SW620 colon cancer cells into nude mice,followed by treatment with 5-FU in the tumors.Results △Np73 siRNA was able to strongly inhibit △Np73 expression,however,it did not inhibit the growth of cells.Combination treatment with △Np73 siRNA and 5-FU produced significant higher apoptotic cell(42.9%) as compared with those with 5-FU treatment(18.9%) alone or those with △Np73 siRNA(8.8%) alone.The treatment with 5-FU in the xenngrafts derived from △Np73 siRNA transfected SW620 cells in nude mice can inhibitor tumor growth significantly (t=15.32,P<0.05).Conclusion △Np73siRNAs can specifically repress the expression of △Np73.Thus it sensitizess the cells to 5-FU chemotherapy in colon cancer.

Objective To investigate the effect of reinfusion of drained bile and pancreatic juice on the outcome of pancreaticoduodenectomy (PD).Methods The clinical data of 51 patients who received PD at the Affiliated Hospital of Binzhou Medical College from June 2005 to March 2009 were retrospectively analyzed.Nineteen patients received external drainage of bile and pancreatic juice ( ED group) and the other 32 patients received external drainage and intraintestinal administration of autologous bile and pancreatic juice (ID group).The daily volume of output of bile and pancreatic juice,intraoperative condition,tolerance of enteral nutrition,liver function and nutritional parameters of the 2 groups were detected.All data were analyzed by using chi-square test,Fisher exact test,independent t test,Mann-Whitney U test and one-way analysis of variance.Results The pulmonary infection rate of ID group was 3％ (1/32) after operation,which was significantly lower than 26％ (5/19) of the ED group (P ＜ 0.05).The output of pancreatic juice in the ID group was significantly lower than that in the ED group since postoperative day 4 ( t =7.143,9.244,8.808,7.915,6.461,14.097,15.038,P ＜ 0.05 ).There was no significant difference in the daily output of bile between the 2 groups.The incidence of diarrhea in the ID group was 9％ (3/32) after nutritional support,which was significantly lower than 37％ (7/19) of ED group (P＜0.05).The duration of achieving targeted enteral feeding in the ID was 3 days,which was significantly shorter than 4 days of the ED group ( U =145.000,P ＜ 0.05 ).The levels of total bilirubin ( TBil),direct bilirubin (DBil) and indirect bilirubin (IBil) were (261 ± 108 ),( 132 + 55 ) and ( 129 + 55 ) μmol/L in the ID group,and (239 ±92),( 12A ±46) and ( 116 ±46) μmol/L in the ED group before operation.The levels of TBil,DBil and IBil were (39 ± 19),(20 ± 10) and ( 19 +9) μmol/L in the ID group,and (55 ±22),(29 ± 12) and (26 ±11 ) μmol/L in the ED group at 12 days after nutritional support.There were significant differences in the decrease of TBil,DBil and IBil between the 2 groups ( t =7.324,8.437,5.827,P ＜ 0.05 ).The levels of serum prealbumin,retinol binding protein and transferrin were (0.261 ± 0.021 ) g/L,(34.3 ± 2.8 ) mg/L,(3.08 + 0.26 ) g/L in the ID group,and (0.263 ±0.021)g/L,(33.8 +3.5)mg/L and (3.10 +0.27)g/L in the ED group before operation.The levels of serum prealbumin,retinol binding protein and transferrin decreased significantly after operation,and then got increased 3 days after nutritional support.The levels of serum albumin,retinol binding protein and transferrin were (0.238 ±0.025)g/L,(30.7 ±2.0)mg/L,(2.78 ±0.19)g/L in the ID group,and (0.222 +0.025)g/L,(29.3 ±2.1)mg/L and (2.63 +0.21)g/L in the ED group at 12 days after nutritional support.The levels of serum albumin,retinol binding protein and transferrin in the ID group were significantly higher than those in the ED group (t=4.615,6.097,4.913,P＜0.05).Conclusion Reinfusion of external drained bile and pancreatic juice after PD could enhance the tolerance of patients in early enteral nutrition,reduce incidence of pneumonia,promote decrease of serum bilirubin and improve the nutritional status.

OBJECTIVETo develop dataset for basic medical information system.

METHODSAll items of basic medical information system dataset were drafted by literature review, brain storm and group discussion. A Delphi study was conducted to evaluate and adjust the candidate items; then items were finalized based on comprehensive scores.

RESULTSThe final dataset developed through two-round Delphi study contained 20 first-level items, 102 second-level items, 40 third-level items and 88 fourth-level items. The response rates were 71.79% and 92.86% in round 1 and 2, respectively. The authority coefficient was both 0.77 and the coordination coefficient of the specialists' opinion was 0.239 and 0.516, respectively.

CONCLUSIONThe dataset for basic medical information system determined from Delphi study is credible and can be used for development of related software.

Delphi Technique ; Information Systems ; Software Design

Objective To determine whether cleavage developmentally retarded embryos have not cleaved during a 24 hour period could develop into blastocysts and produce hESC cell lines.Methods A total of 120 such embryos were cultured to blastocyst stage by sequential culture.Blastocysts formation rate and quality of blastocyst were detected under microscope.The relation between blastocyst formation rate and blastomere number,the fragment of blastomere and blastomere symmetry were analyzed by stepwise Logistical regression analysis.Inner cell masses(ICMs)were isolated by immunosurgery.Colonies derived from the ICMs were passed every 4-7 days and the derivatives were passaged and identified.Results A total of 22 blastocysts were obtained from 120 embryos.The blastulation rate was 18.7%.Early blatocyst, blastocyst,full blastocyst,expanded blastocyst,hatching blastoeyst and hatched blastocyst accounted for 5.9%,23.5%,35.3%,23.5%,5.9%,and 5.9% respectively.The grade of ICM and trophoblast was mostly scored C or B.Blastocyst formation rate was related to cell number and blastomere symmetry but not fragment.Immunosurgery resulted in the formation of 7 ICMs and 3 primary colonies,which produced 2 cell lines.The cell lines satisfied the criteria that characterize pluripotent hESC cells.Undifferentiated cells were positive for AKP,SSEA-4,TRA-1-60,and TRA-1-81.It could continue to proliferate in vitro and form embryoid bodies when cultured in suspension.It had capability to form teratoma in SCID mice.Both cell lines had normal karyotypes after 45 and 34 passages respectively.Conclusions Our results suggest that a subset of developmentally retarded embryos can form blastocysts and give rise to hESC cell lines.