Efforts have been directed at the development of efficient protocols for the differentiation of pluripotent stem cells into pancreatic β-cells.This review focuses on the mechanism of RA,BMP,FGFs,IDE1,IDE2,ALK-i 1 and ALKI-2 during the process in which pluripotent stem cells differentiate to pancreatie β-cells and the relationship between the differentiation and the expression of the surface markers including CD14,CD49e,CD238,CD24,EpCAM,SSEA3 and SSEA1 in the different stage of this process.

BACKGROUND:At present, mouse embryonic neural stem cells (NSCs) culture has been skillfully operated by many labs, but there are differences existing about which part are dissociated to get NSCs. Embryonic 14 days (E14) mouse brain tissues are widely used for culturing NSCs, but there are less studies about the detailed percentage and difference of NSCs separated from different brain tissues. OBJECTIVE: To test the proportion and difference of NSCs and neurons percentage from E14 mouse whole brain, cortex and forebrain, providing quantized data for optimizing the isolation of high-purity NSCs. METHODS:E14 C57BL/6 mouse whole brain, cortex and forebrain tissues were separated and dissociated into single cells that were adherently cultured for 3.0-4.0 hours and labeled by DAPI. Then the cells were immunostained with NSCs specific marker, Nestin, and neuron specific marker, Tuj1, to identify NSCs and neurons percentage by calculating Nestin+/DAPI and Tuj1+/DAPI. In addition, real-time PCR assay was used to test Nestin and Tuj1 mRNA expression in the E14 mouse whole brain, cortex and forebrain. RESULTS AND CONCLUSION: (1) Immunocytochemical results showed that there were a large amount of Nestin+ and Tuj1+ cells in the whole brain, cortex and forebrain of E14 mice. NSCs percentage in the forebrain was obviously higher than that in the whole brain (P < 0.01) and cortex (P < 0.05), while the percentage of neurons in the forebrain was significantly lower than that in the whole brain (P < 0.05) and in the cortex (P < 0.001). (2) Real-time PCR results showed that the Nestin mRNA expression in the forebrain was significantly higher than that in the whole brain (P < 0.05) and slightly higher than that in the cortex (P > 0.05); the Tuj1 mRNA expression in the forebrain was significantly lower than that in the whole brain (P < 0.05) and in the cortex (P < 0.05). These findings indicated that the forebrain had the most NSCs and the least neurons compared with the whole brain and the cortex. In summary, E14 mouse forebrain has the highest percentage of NSCs compared with the whole brain and cortex, which is a better source to obtain NSCs for the following cell culture experiments.

Objective To study the availability of escharectomy in shock phase and its role in preventing complications.Methods To make an analysis between the escharectomies and skin grafting of 79 cases in or beyond shock phase in the incidence of sepsis,visceral complications,MODS,mortality,healing time and the expenditure.Result The cases of the early operations revealed much better consequences than those performed beyond shock phase.Conclusion The escharectomy in shock phase proves to be available and significant in reducing post burn complications.

Objective To explore the effect and mechanism of the bone morphologenetic protein 4 (BMP4)/Smad signaling pathway on the apoptosis of mouse primordial follicle oocytes .Methods Three-day-old Kunming mouse ovarine tissues were digested by the two-step enzymatic method to extract and purify oocytes .The cultured oocytes were divided into three groups: the normal culture medium (Con group), the medium with BMP4 (BMP4 group), and the medium with BMP4 and BMP4 inhibitor ( BMP4+inhibitor group ) .TUNEL was used to examine the effects of BMP 4 on the survival of the primordial follicle oocyte;Immunohistochemical staining and Real-time quantitative PCR were performed to investigate the expressions of p-Smad1/5/8, sohlh2, c-kit and foxo3a;siRNA interference, sohlh2 plasmid transfection and LY294002 treatments were performed to explore the mechanism of the BMP 4/Smad signaling pathway on the apoptosis of oocytes . Results TUNEL results demonstrated that the ratio of apoptotic oocytes in BMP 4 group was significantly lower than that in the Con group ( P <0.05 ) and the BMP4 +inhibitor group ( P <0.05 ); BMP4 significantly promoted the nuclear translocation of Smad and inhibited the nuclear translocation of foxo 3a, the mRNA and protein levels of sohlh2 and c-kit remarkably increased in BMP4 group.The effect of BMP4 on the oocyte survival was significantly repressed after sohlh 2 siRNA transfection.Sohlh2 overexpression up-regulated the expression of p-foxo3a, and this activity was abolished by LY 294002.Conclusion BMP4/Smad signaling pathway may inhibit primordial follicle oocyte apoptosis , via up-regulation of the expression of sohlh2 and c-kit, and then down-regulation of the nuclear translocation of foxo 3a.

Objective To observe the features of the changes in the whole blood N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in pregnant patients with complication of hypertensive disorders,its correlation to the severity of the illness,and to investigate the diagnostic value of point-of-care testing of NT-proBNP in patients with hypertensive disorders complicating pregnancy.Methods A prospective observation was conducted.Sixty-nine patients with hypertensive disorders complicating pregnancy admitted to Department of Critical Care Medicine of Liaocheng People's Hospital in Shandong Province from April 2013 to April 2015 were enrolled.All patients were divided into gestational hypertension group (n =16),preeclampsia group (n =30) and eclampsia group (n =23).At the same time,30 age-matched normal pregnant women were enrolled as the control group.The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score of all patients with hypertensive disorders complicating pregnancy were calculated within 24 hours after intensive care unit (ICU) admission.NT-proBNP in venous blood at 1,3,5 days after ICU admission was determined with point-of-care testing,in order to analyze the correlation of changes in NT-proBNP value in each group and the severity of the disorder.Receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic value of NT-proBNP in the whole blood in hypertensive disorder complicating pregnancy.Results The APACHE Ⅱ score of the eclampsia group was significantly higher than that of the preeclampsia group,and it was higher than that of gestational hypertension group (15.91 ± 1.06,13.73 ± 1.09,10.31 ± 1.10,all P ＜ 0.01).The NT-proBNP in normal pregnancy group was lower than 125.00 ng/L,with a mean of 90.00 (79.75,100.00) ng/L.With the aggravation of the disease,NT-proBNP was gradually increased.On the first day in ICU,the NT-proBNP of the eclampsia gronp was significantly higher than that of the preeclampsia group,and it was higher in preeclampsia group than that of gestational hypertension group [ng/L:1960.00 (1 226.00,3 229.00),859.50 (626.75,2439.00),505.00 (171.25,604.05),P ＜ 0.05 or P ＜ 0.01].With the extension of duration of treatment,the levels of NT-proBNP (ng/L) in the eclampsia group,preeclampsia group,and gestational hypertension group was gradually decreased,and had a statistically significant difference on the fifth day as compared with that of the first day [310.00 (210.00,430.00) vs.1 960.00 (1 226.00,3 229.00) in eclampsia group,265.00 (229.50,333.25) vs.859.50 (626.75,2439.00) in preeclampsia group,and 203.00 (115.50,259.25) vs.505.00 (171.25,604.05) in gestational hypertension group,all P ＜ 0.01].APACHE Ⅱ score of the patient with hypertensive disorders complicating pregnancy was positively correlated with the level of NT-proBNP on the first day in ICU (r =0.795,P =0.000).It was shown by ROC curve analysis that the area under the ROC curve (AUC) of NT-proBNP in the whole blood for the diagnosis of the patient with hypertensive disorder complicating pregnancy was 0.986 [95％ confidence interval (95％C/) =0.753-0.924].When the cutoff value was 122.50 ng/L,the sensitivity was 97.1％,and the specificity was 100.0％.No patient died,all the 69 patients recovered and discharged.Conclusions The levels of NT-proBNP in the whole blood in the patients with hypertensive disorder complicating pregnancy,especially those with eclampsia,were significantly higher,and it was correlated with the severity of illness.After treatment,the levels were gradually lowered with the improvement of the disease.Therefore,it is concluded that the point-of-care testing of NT-proBNP in the whole blood has an excellent value for the diagnosis and evaluation of hypertensive disorder complication pregnancy.

Objective To evaluate the applied value of CT in breast lesions.Methods Imaging data of 120 patients with normal breast were reviewed retrospectively in 4 groups of different age(15～25,26～35,36～50 and 51～80),and imaging data of 49 patients with breast lesions biopsy-proved were also analysed.Of 49 breast masses,31 were malignant tumor,18 were benign lesion.All the patients underwent CT scan with supine position whole thoracic region.Results The CT imaging features of normal breast in different age groups were different.65% of normal people had small sporadic axillary lymph nodes.Breast cancer usually showed hillslike and benign tumor showed round oroval-shap.The signs of spiculated lesion,deformed adipose space and thicken of local skin were statistical difference between benign and malignant lesions(P0.05).The metastatic axillary lymph nodes usually showed as the significant enlarged nodes with indefinite margin or multiple small nodes.Conclusion CT scan is valuable in detecting breast disease and differential diagnosis,and also can offer the informations on tumor staging and treatment.

Objective To compare clinical effects of appendicectomy and conservative treatment of uncomplicated acute appendicitis.Methods Searched The Cochrane library,Medline,Pubmed,Embase,CBM,CNKI,VIP and Wan Fang database in any language.RCTs that compared conservative treatment with appendicectomy in patients with uncomplicated acute appendicitis were included from January 1983 to May 2013 and qualities of the trials were evaluated.Statistic analyses were carried out using RevMan 5.1 soft-ware.Results Four randomized trials met our inclusion criteria (821partiCI:pants).Antibiotics compared with appendicectomy resulted in similar outcomes with regards to the incidence of complicated appendicitis (RR:0.89,95％ CI:0.29-2.68) and intra-abdominal infection(RR:0.54,95％ CI:0.13-2.35).Antibiotics carries a lower risk of complications (RR 0.46,95％ CI:0.32-0.67),but requires more length of hospital stay (Mean Difference 0.52,95％ CI:0.16-0.88).Conclusions The effect of conservative treatment is safe and effective.But the recurrent uncomplicated appendicitis recommends operation.

Objective To determine schisandrin,schisandrol B,deoxyschizandrin,and schisandrin B in serum containing drug of Compound Wurenchun Capsula.Methods An HPLC method was set up.Li-chrosphere C18 column(250 mm ?4.6 mm,5 ?m) and Phenomenex Description C18(4.0 mm?3.0 mm)protective column were used.Acetonitrile-water was used as gradient mobile phase.The flow rate was 1.0 mL/min.The column temperature was 30 ℃ and the detection wavelength was 210 nm.Results The linear ranges of schisandrin,schisandrol B,deoxyschizandrin,and schisandrin B were within 0.051 2-0.768 0 ?g(r=0.999 5),0.054 0-0.810 0 ?g(r=0.999 6),0.012 3-0.184 5 ?g(r=0.999 8),and 0.039 8-0.597 0 ?g(r=0.999 6),respectively.The average concentration of these four lignans in serum containing drug were 8.021 1,6.231 0,0.530 8,and 5.851 0 ?g/mL,respectively.Conclusion This method is easy,sensitive,specific,and accurate for the assaying of the four lignans in serum containing drug of Compound Wurenchun Capsula.

Objective To compare the pros and cons of endoscopic papillary balloon dilation (EPBD) with those of endoscopic sphincterotomy (EST) in the treatment of common bile duct stones.Methods We searched The Cochrane library,Medline,Pubmed,Embase,CBM,CNKI,VIP and Wan Fang database in any language.RCTs that compared EPBD with EST for the removal of common bile duct stones were included from January 1983 to September 2012 and qualities of the trials were evaluated.Statistic analyses were carried out using RevMan 5.1 software.Results A total of 18 randomized trials with 2385 participants met our inclusion criteria.EPBD compared with EST resulted in similar outcomes with regards to stone removal on 1 st attempt,overall stone removal,perforation,total short-term complication,long-term cholangitis or mortality.EPBD carries a higher risk of pancreatitis (RR =1.99,95％ CI:1.41-2.81) and severe pancreatitis (RR =4.68,95 ％ CI:1.36-16.11),and requires higher rates of mechanical lithotripsy (RR =1.31,95％ CI:1.14-1.50).Conversely,EPBD not only has statistically significant lower rates of bleeding (RR =0.14,95％ CI:0.06-0.34),but also leads to significantly less long-term cholecystitis (RR =0.38,95％ CI:0.19-0.76),long-term stone recurrence (RR =0.67,95％ CI:0.47-0.96) and total longterm complications (RR =0.52,95 ％ CI:0.40-0.67).Conclusion On the basis of lower rates of bleeding or long-term complications,EPBD should be the preferred strategy over EST for endoscopic management of common bile duct stones,however,the rate of pancreatitis,especially the severe pancreatitis is higher with EPBD.

ObjectiveTo investigate the effect of n-butanol extract from Potentilla anserina (NP) intervention on hypoxia-induced Ca2+ overload and SERCA2 expression of rat cardiomyocytes.MethodsPrimary cultured myocardial cell from SD neonatal rat (1-3 d) was used in the establishment of hypoxia model.After hypoxia for 3 h,the Ca2+ concentration of myocardial cells was measured with fura-2/AM fluorescent probe,and the biochemical indicator intracellular Ca2+-ATPase was examined and the mRNA and its protective protein levels of the sarcoplasmic reticulum (SR) Ca2+-ATPases (SERCA2) were assayed with RT-PCR,Western-blotting,and immune-cytochemical staining in each group.ResultsThe results showed that NP decreased Ca2+ concentration,increased the activity of Ca2+-ATPase,and improved the mRNA and protein expression of SERCA2 in hypoxia-injured myocardial cells as compared with the model group.ConclusionThese results indicate that NP could attenuate the Ca2+ overload.The mechanism might be explained as that NP could elevate the SERCA2 level,increase the activity of myocardium in rats,and further enhance the capacity of SR Ca2+ re-uptake.