BACKGROUND: The long-term cryopreservation of embryonic stem cells is an important link in its researches and application. Traditional protectants of cryopreservation often contain dimethyl sulfoxide, glycerine, animal serum, etc., but they cannot effectively ensure the survival rate of some cells, and even damage some functions of the cells.OBJECTIVE: To develop the suitable protectant for the cropreservation of mouse embryonic stem cells. DESIGN: An observational study.SETTINGS: Department of Physiology, Conghua College of Guangzhou Medical College; Sinocells Bio Technologies Co.,Ltd.MATERIALS: The experiment was carried out in the laboratory of Sinocells Bio Technologies Co.,Ltd. from October 2004 to September 2005. Mouse cell line mES-1 was given as a present by the Research Room of Cell Biology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences of Chinese PLA.METHODS: The cells collected from cultured mES-1, a mouse embryonic stem cell line. Dimethyl sulfoxide was used as the main protectant for cryopreservation. There were four different groups according to different protectants. In the control group, the protectant for cryopreservation contained DMEM (high sugar), 10% dimethyl sulfoxide, 10% FBS,10 mg/mL ascorbicacid, 0.18 mg/mL inositol and 0.44 mg/mL folic acid; Besides, 7.56 mg/mL mannose in the mannose group, 34.23 mg/mL mycose in the mycose and 41.94 mg/mL saccharu in the saccharu group were supplemented respectively. The effects of different polyols on the survival rate and ability of multiple differentiation of mouse embryonic stem cells after cryopreservation were observed.MATN OUTCOME MEASURES: Formation rate of mouse embryonic stem cells colony; Formation rate of embryoid bodies.were all significantly decreased as compared with that before cryopreservation [control group: (24.0±8.8)%; mannose group: (42.0±10.1)%; mycose group: (84.0±8.2)%; saccharu group (70.0±14.2)%; before cryopreservation: (95.0±4.7)%,embryoid bodies in the mycose group was higher than those in the control group and mannose group [(90.0±5.2)%,(80.0±6.9)%, (82.0±9.6)%, P＜0.05], but had no significant difference as compared with that in the saccharu group.CONCLUSION: The protectant for cryopreservation by taking mycose as the main component can effectively maintain the abilities of self-renewing and multiple differentiation of mouse embryonic stem cells.

Objective To investigate the diagnosis of Crohn's disease(CD)in Ruijin hospital of School of Medicine of Shanghai Jiaotong University according to the criteria made by World Health Organization(WHO)and Chinese Medical Association.Methods The consecutive patients with CD diagnosed in Ruijin hospital between 1998 and 2007 were retrospectively analyzed.The clinical manifestation.endoscopic,radiologic and pathologic findings of all patients were re-evaluated accordingto the diagnostic criteria of CD proposed by WHO and Chinese Medical Association in 2007.The diagnostic level of CD was assessed.Results Only 5 and 10 patients were confirmed to be well consistent with the criteria of WHO and Chinese Medical Association,respectively.Two hundred and thirty-six patients were confirmed as suspected diagnosis to the criteria of Chinese Medical Association and 20 patients to WHO.The level of endoscopic diagnosis was raised,wherease the level ofpathologic diagnosis was decreased in recent years compared with that before 2004.Conclusion The pathologic diagnosis rate of CD is still at a low level.The current diagnostic level of CD is lagging behind the demand of the guidelines.

Recently,to assess the reproductive effects of bisphenol A (BPA),the environmental estrogen,has become a focus of stusy. Investigations,to date,have focused on the effects on the reproductive system including the morphogenesis of tissues and organs,endocrine and the toxic mechanisms of action,particularly in males. Based on the study results concerning BPA’s impacts on male reproduction,the further study directions in this field were presented in this paper.

Objective To investigate folic acid and vitamin B12 on the plasma homocysteine (Hcy) levels and neurological function in young and middle-aged acute ischemic stroke patients with hyperhomocysteinemia (HHcy).Methods Young and middle-aged acute ischemic stroke patients with HHcy were enrolled.They were randomly divided into either an intervention group or a control group.The patients with cerebral infarction in both groups were treated with conventional treatment.At the same time,the patients in the intervention group were treated with folic acid 5 mg and vitamin B12 25 μg,3 times a day for 4 weeks.Those in the control group were given placebo.The next day after admission and thereafter,the plasma Hcy levels were determined once every week.At the time of admission and after 4-week treatment,the National Institutes of Health Stroke Scale (NIHSS) was used to evaluate neurological deficits.Results A total of 78young and middle-aged acute lschemic stroke patients with HHcy were enrolled (n =39 in each group).The baseline plasma Hcy levels and the NIHSS scores in all patients had significant linear correlation (r =0.717;P＜0.01).Four weeks after treatment,the plasma Hcy levels of the intervention group were significantly lower than those of the control group (15.07 ± 2.01μmol/L vs.21.29 ± 2.48 μmol/L; t =4.539; P ＜0.05),but there was no significant difference in the NIHSS scores (4.87 ± 2.17 vs.5.13 ± 1.67; t =0.585,P =0.560).Conclusions The baseline plasma Hcy level was positively correlated with the severity of stroke.Folic acid in combination with vitamin B12 could effectively decrease the plasma Hey level in young and middle-aged acute ischemic stroke patients with HHcy,but it had no significant effect on the improvement of neurological function.

Objective To explore the expression of glucose-regulated protein (Grp) 78 in ovarian cancer tissues and its clinical significance.Methods The expression of Grp78 in 60 cases of ovarian cancer tissue,15 cases of ovarian borderline tumor tissue,10 cases of normal ovarian tissue,10 cases of ovarian cyst tissue,was detected by immunohistochemistry,and analyzed the relationship between its expression and clinicopathological features of ovarian cancer.Results The expression of Grp78 in ovarian borderline tumor and ovarian cancer tissue was significantly higher than that in normal ovarian and ovarian cyst tissue[7/15 and 68.3％ (41/60) vs.1/10 and 1/10] (P ＜0.05).The expression of Grp78 was positively correlated with clinicopathological staging and lymphatic metastasis of ovarian cancer (P ＜ 0.05),negatively correlated with histological differentiation (P ＜ 0.05).No correlation with age and ascites (P ＞ 0.05).Conclusions The levels of Grp78 are elevated in ovarian cancer specimens; high expression of Grp78 maybe participate in the occurrence,development,and prognosis of ovarian cancer.Increased expression of Grp78 might be a useful marker for predicting the carcinogenesis and progression of ovarian cancer.

Objective To investigate the effect of atorvastatin calcium combined with sitagliptin phosphate on β2-microglobulin (β2-MG ) in peripheral blood and serum uric acid ( UA) levels in patients with type 2 diabetes.Methods A total of 78 patients with type 2 diabetes in endocrinology department from Hangzhou Xiasha Hospital were collected and randomly divided into the control group and the experimental group, with 39 cases in each group.The two groups of patients were treated by conventional treatment, life intervention, the control group were treated by metformin hydrochloride sustained-release tablet and sitagliptin phosphate, the experimental group were treated on the basis of control group with atorvastatin calcium.Both groups were treated for 3 cycles, one cycle for 7 days.The clinical curative effect,β2-MG in peripheral blood and urine, serum UA levels and adverse reactions were compared between two groups after treatment.Results After treatment, compared with control group, the clinical total effective rate in experimental group was higher (P<0.05).The serum IgG,β2-MG, urineβ2-MG and UAlb, serum UA levels were lower in two groups post-treatment compared with pre-treatment(P<0.05).Compared with control group, serum IgG, β2-MG, urine β2-MG and UAlb, serum UA levels were lower (P<0.05).There was no significant difference between two groups.Conclusion The atorvastatin calcium combined with sitagliptin phosphate has a significant efficacy in the treatment of patients with type 2 diabetes, it could down-regulate β2-MG in peripheral blood and urine and serum UA levels, improve immunity and prevent patients from cardiovascular complications.

This paper discussed the roots of American ethical review system, the development of twists and turns, and puts forward the enlightenment to our country, the ethical review system mainly include:perfecting the ethical review at the top of specification design, improve specification status;improve the system of education and training, to strengthen the construction of information communication channels, establish standard operating proce-dures, improve the regulatory system;improve the system of continuous review, formulate specific implementation specification.

The clinical data of a case of Weave syndrome admitted in the Department of Endocrinology, Children′s Hospital of Nanjing Medical University in October 2018 were retrospectively analyzed.The patient was a 9 years and 2 months old girl, who was hospitalized because of " growing too fast for 9 years" . After birth, the child is found to grow fast and have mental retardation, slurred speech, a blurred vision, a long face, a protruding forehead, ocular hypertelorism, epicanthus, nasal bridge pit, finger pads on both hands and feet, uncoordinated gaits, and intoeingpigeon toes.A novel heterozygous c. 1720A>G (p.K574E) mutation was detected in the exon 15 of the EZH2 gene of the patient.This mutation has not been reported at home and overseas.Sanger sequencing revealed that the patient′s parents did not carry the mutation.The disease is an autosomal dominant genetic disorder, and the parents and sibling of the patient have no corresponding symptoms, so it is inferred that the mutation is spontaneous.Based on the peculiarity of the face, clinical manifestations and the results of molecular genetics, the child was diagnosed as Weaver syndrome.

Objective The purpose of this study was to evaluate the protective potential of the Aspergillus fumigatus thiore -doxin reductase GliT ( TR) antigen by establishing and optimizing ELISPOT assay for TR antigen-specific T cells ( TR/AST) secreting IFN-γand IL-4 in peripheral blood mononuclear cells ( PBMCs ) and explore the role of TR/AST in invasive aspergillosis ( IA ) . Methods We optimized the reaction conditions of ELISPOT by preliminary checkerboard titration and determined the frequencies of positive spot-forming cells ( SFCs) specifically secreting IFN-γand IL-4 in the PBMCs of 20 healthy individuals with TR as specific stimulant and with PHA and PMA as positive controls ,. Results Checkerboard titration demonstrated the best result of ELISPOT with the TR antigen at the final concentration of 10μg/well and PBMCs at 3 ×105/well.The median frequency of IFN-γSFCs was sig-nificantly higher (15 [3.5, 59.5]) than that of IL-4 SFCs (0 [0, 0]) (P<0.001).TR induced IFN-γresponses in all the 20 healthy donors, including 9 cases of strong IFN-γresponse (SFCs>20/3 ×105 PBMCs), accounting for 45%, but failed to induce IL-4 response in 19 of the healthy individuals . Conclusion The Aspergillus fumigatus TR antigen could induce an immunodominant Th1 response , and therefore might be a potential protective antigen .

Purpose The mean pulmonary artery (MPAP) has been widely used as an important parameter to diagnose and evaluate pulmonary hypertension (PH).The purpose of this paper is to compare the efficacy of two methods in evaluating PH,including estimating pulmonary artery systolic pressure (PASP) using Doppler ultrasonography to measure tricuspid regurgitation (TR) velocity,and directly using the peak velocity of TR.Materiasl and Methods From January 2012 to June 2013,eighty patients with left-to-right shunt congenital heart diseases (CHD) planned for closure procedure in Tangdu Hospital of the Fourth Military Medical University and the General Hospital of Shenyang Military region were included in this prospective study,who underwent right heart catheterization to measure pulmonary artery pressure,and underwent Doppler ultrasonography to measure the peak velocity ofTR.Results Using catheter-measured MPAP of≥ 25 mmHg as diagnostic reference,the false positive rate was 62.96％,and the false negative rate 0％ when the estimated PASP of ＞30 mmHg determined by TR method was used to diagnose PH.There was high diagnostic agreement when peak velocity of TR was used to diagnose PH.When 320 cm/s and 340 crn/s were used as diagnostic cutoff values,false positive rates were 14.81％ and 7.41％,and false negative rates were 15.91％ and 20.45％,respectively.Conclusion In patients with left-to-right shunt CHD,peak velocity of TR measured on echocardiography can be used to diagnose PH which overcomes the high false positive rate in estimation method.It is more suitable to diagnose PH when the MPAP is used as the diagnostic criterion.