Objective To probe into the optimal concentration of Wnt-11 to induce the differentiation of bone marrow mesenchymal stem cells (BMMSCs)into cardiomyocyte-like cells in vitro.Methods BMMSCs were isolated from the bone marrow of SD rats using whole bone marrow culture method.After cultured for 48 h, BMMSCs of the second generation were utilized for directed induction.Based on the final concentration of Wnt-11 , BMMSCs were divided up into Group A (100 ng/mL),Group B (200 ng/mL),Group C (400 ng/mL)and Group D (blank control).After 72-hour induction,the cells were cultured in complete medium for 4 weeks while cells in Group D were cultured only in the complete medium.The morphological changes were observed under the phase contrast microscope.Surface antigen expression of BMMSCs was identified by flow cytometry.When cells were cultured for 4 weeks,the expressions of Desmin,Connexin43 and cTnI were detected by immunocytochemistry. Meanwhile, the ultrastructural changes were observed using transmission electron microscope. The mRNA expressions of cardiac transcription factors GATA-4,Nkx2.5 andα-MHC in BMMSCs were detected by RT-qPCR at 1,2 and 4 weeks after induction.Results Primary BMMSCs formed cell colonies at 2 weeks;the cells were mainly fusiform or star-shape,and a few irregularly-shaped ones were also visible.The passaged cells were larger than those of primary culture.After induction,the cells exhibited long shuttle-shape and were aligned in parallel. Flow cytometery displayed that the positive rate of the surface antigens of BMMSCs CD29,CD45,and CD90 was 97.9%,0.4% and 99.5%,respectively.When BMMSCs-induced via Wnt-11 were cultured for 4 weeks,Desmin, cTnI and Connexin43 were all positively expressed in induction groups.Whereas in the blank control group they were slightly positive or negative;the positive rate in Group B was the highest (P<0 .05 ).Transmission electron microscopy exhibited that organelles such as rough endoplasmic reticulum,mitochondria,as well as some ribosomes were visible in the cytoplasm of these cells in each induction group.In addition,myofilaments were arranged in parallel in the cytoplasm.The cells in induction groups could express GATA-4 and Nkx2 .5 in the first week,and then the expression of them decreased in the second week,but then increased in the fourth week;gene expression in induction Group B was significantly higher than in the other two induction groups (P<0 .05 ).The expression of GATA-4 and Nkx2 .5 in Group D was 1 ,α-MHC was not expressed in the four groups during the culture period. Conclusion Wnt-11 can induce the differentiation of BMMSCs into cardiomyocyte-like cells in vitro,and the optimal concentration of Wnt-11 is 200 ng/mL.

Objective: To research the effect and mechanism of how chitosan restrain fat-absorbtion, and provide references for developing a kind of nutritional and healthy fastfood. Methods: 1.In vitro digestion: Phosphomolybdic acid colorimetry; 2.In vivo experiment:240 rats were divided into four groups and fed with different diets: basal diet(Group A), high fat diet (Group B), chitosan supplemented diet (Group C), high fat and chitosan supplemented diet (Group D). The fat and chitosan content(%) of diet were A(10,0), B(60,0), C(8,20), D(34.3,14.8) respectively. Determine the blood lipids and fat output in dung, and observe the weight gain and swimming time. Results: 1.In vitro experiment showed chitosan acted on cholic acid at the best proportion 1∶8. 2.In vivo experiment showed chitosan can restrain fat-absorbtion, and increase fat output. Swimming in ice-water showed chitosan improved rats performance. Compared with group B, the blood lipids and weight of group C and D had no obvious change, but the fat-output much increased.

To improve the clinical teaching work, the objective structured clinical examination (OSCE) was first introduced into the periodontal clinical assessment held by Departments of Periodon-titis and Mucosal Disease. With the OSCE exam station and item pool established, standardized pa-tients tralned, the examination form reformed, the knowledge, skill, and attitude of the examinee were assessed comprehensively. Compared with traditional exam,the OSCE evaluated the examinee's skills on clinical admissions, history collection, treatment plan development better. Most examinee have got satisfactory result (pass rate: 86.4 %, 19/22; good rate: 36.4%, 8/22; excellent rate: 9.1%, 2/22). The feedback of the following questionnalre also showed that most of the candidates thought the OSCE helpful to promoting the learning of theoretical knowledge (87.5%, 14/16) and good for converting the learned knowledge and skills into occupation ability (87.5%, 14/16).

Objective To explore the expression of serine/threonine kinase 15 (STK15) gene and its significance for papillary thyroid carcinoma. Methods SP immunohistochemical method was used to detect the expression of STK15 gene in 71 cases of papillary thyroid carcinoma and 45 cases nodular goiter tissue. Results The positive expression rates of STK15 gene in 71 cases of papillary thyroid carcinoma were 100%, and the adjacent of papillary thyroid carcinoma expressions of STK15 gene 8.5%, nodular goiter tissue of STK15 gene expression rates 24.4%. The expression of STK15 gene was positively correlated with that of STK15 gene in papillary thyroid carcinoma (P < 0.01). Conclusion High expression of STK15 gene is confirmed in papillary thyroid carcinoma. The detection of STK15 gene can provide valuable evidence for diagnosis of papillary thyroid carcinoma and evaluation of the malignant potential of nodular goiter.

Objective To investigate the relationship of high sensitivity C reactive protein (hs-CRP), interleukin-6 (IL-6)and soluble IL-6 receptor (sIL-6R)with the severity of clinical symptoms and coronary artery lesions in coronary heart disease (CHD)patients.Methods A total of 522 CHD patients were recruited and divided into three groups:stable angina pectoris (SAP),unstable angina pectoris (UAP)and acute myocardial infarction (AMI)groups.Another 102 healthy individuals served as normal controls (NCs).We calculated Gensini score according to the result of coronary angiography (CAG),collected clinical data and compared the groups. Multiple linear regression analysis was used to investigate the relationship of hs-CRP,IL-6 and sIL-6 R with Gensini score.Results The plasma hs-CRP,IL-6 levels were significantly higher and sIL-6R level was signficantly lower in SAP,UAP and AMI groups than in NC group (P<0 .0 5 ).There was a positive correlation between IL-6 level and Gensini score but a negative correlation between sIL-6 R and Gensini score presented by multiple linear regression analysis (P<0.05 ).Conclusion In CHD patients,plasma hs-CRP,IL-6 and sIL-6R levels are significantly related to the severity of clinical manifestations and coronary artery stenosis.These indicators may help predict the severity of CHD.

Objective To study the diagnostic value of endoscopic ultrasonography (EUS)for benign and malignant bile duct stricture.Methods Data of 51 patients who underwent EUS from January 2007 to March 2015 due to distal biliary stricture were retrospectively studied.And their diagnosis with EUS,IDUS,bile duct cytological brushing and final diagnosis were compared.Results Malignant bile duct stenosis were finally confirmed in 19 cases and benign stenosis were confirmed in 32 cases.The sensitivity (73.7％ VS 57.1％) and positive predictive value (73.7％ VS 57.1％) of EUS for the benign and malignant diagnosis of biliary tract stenosis were significantly higher than that of IDUS (P＜0.05),while their specificity(84.4％ VS 84.2％)and accuracy(80.4％ VS 76.9％)were similar.The sensitivity of EUS was significantly higher than that of bile duct cytological brushing (73.7％ VS 50.0％),while their specificity (84.4％ VS 100.0％)and accuracy(80.4％ VS 81.6％)were similar.EUS findings of hypoechoic mass features had higher diagnostic accuracy for detecting malignant distal bile duct stricture than irregular thickening of the lumen in the distal bile duct (100.0％ VS 64.3％,P＜0.05).Conclusion For patients with lesions of the distal bile duct that CT/MRI could not diagnose,EUS has an important value for the further diagnosis of benign and malignant bile duct stricture.

Cholestatic liver disease refers to a liver disorder caused by cholestasis, which arise from a series of etiologies such as viruses, bacteria, parasites, drugs, poisons, autoimmunity, alcohol, stones, tumors, genetics, and metabolism. This disease has the main manifestations of a change in bile flow and excessive accumulation of bile acid toxicity. In the pathogenesis of cholestatic liver disease, not only does the enterohepatic circulation of endogenous bile acids work, but also the intestinal microbiota plays an important role by regulating metabolism and causing immune responses. In addition, more attention has been paid to the close interaction between intestinal microbiota and the enterohepatic circulation of bile acids. Bile acids can alter the composition of intestinal microbiota, which in turn affects the bile acid pool. In recent years, there has been increasing research on the relationship of the enterohepatic circulation of bile acids and intestinal microbiota with cholestatic liver disease, which may provide new research directions for the pathogenesis and treatment of cholestatic liver disease.

Objects:To investigate the current situation of counselors'implementation and their attitude to informed consent in Beijing.Methods:Eleven counselors who worked in Beijing were interviewed,the average of their working years was (7.4 ± 4.4).A semi-structured interview was used to learn about their practicing processes in and viewpoints on informed consent,and their interview transcripts were analyzed through qualitative method.Results:The results could be categorized into four categories,including the content of informed consent,the influential factors of informed consent,problems and confusions,and the significance of informed consent.The crucial role of informed consent in psychotherapy was generally recognized by counselors,and they could obey the related rules as well.The therapists had some confusion and problems about informed consent,such as the form and length,the consent for special population and online consultation.Conclusion:Basically,the counselors in Beijing could comply with the ethical standards of informed consent.However,they also suggest that certain limitations of these rules should be noticed.

INTRODUCTIONRetinitis pigmentosa (RP) describes a group of inherited disorders characterised by progressive retinal dysfunction, cell loss and atrophy of retinal tissue. RP demonstrates considerable clinical and genetic heterogeneity, with wide variations in disease severity, progression, and gene involvement. We studied a large family with RP to determine the pattern of inheritance and identify the disease-causing mutation, and then to describe the phenotypic presentation of this family.

MATERIALS AND METHODSOphthalmic examination was performed on 46 family members to identify affected individuals and to characterise the disease phenotype. Family pedigree was obtained. Some family members also had fundus photographs, fluorescein angiography, and/or optical coherence tomography (OCT) analysis performed. Genetic linkage was performed using short tandem repeat (STR) polymorphic markers encompassing the known loci for autosomal dominant RP. Finally, DNA sequencing was performed to identify the mutation present in this family.

RESULTSClinical features included nyctalopia, constriction of visual fields and eventual loss of central vision. Sequence analysis revealed a G-to-T nucleotide change in the Rhodopsin gene, predicting a Gly-51-Val substitution.

CONCLUSIONSThis large multi-generation family demonstrates the phenotypic variability of a previously identified autosomal dominant mutation of the Rhodopsin gene.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Genes, Dominant ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree ; Retinitis Pigmentosa ; genetics ; Rhodopsin ; genetics

Objective To evaluate the clinical efficacy of extraperitoneal laparoscopic radical prostatectomy(ELRP)for prostate cancer patients, and to summarize the experience of surgical treatment. Methods The clinical data of 50 prostate cancer patients who underwent ELRP by the same performer from January 2010 to June 2015 were retrospectively reviewed. Results All cases were all successfully completed ,no case was converted to open surgery. The average operation time was 238.8 min,average operative blood loss was 409.1 mL,and intraoperative or postopera?tive blood transfusion was 6(12%). The mean postoperative catheterization time was 23.7(17?38)d. The mean postoperative hospital stay was 15 (10?34)d. The postoperative recovery time of eating was 2?4 d,and the ambulation time was 1?3 d. Totally 3 cases(6%)had lymph node metasta?sis,and 7 cases(14%)had positive surgical margin. Totally 9 cases(18%)had surgery?related complication. Patients were followed up for 6 to 58 months,with an average of 12.5 months. One case(2%)had biochemical recurrence,and the tumor?free survival rate was 84%. At the end of fol?low?up,all of the patients were continent. Conclusion ELRP is safe and effective for the treatment of prostate cancer. With the development of minimally invasive techniques,the applications of RP are increasingly widespread. However,large?scale and long?term follow?up studies are still needed for high?risk prostate cancer patients.