Objective To investigate the protective effect of adiponectin on angiotesin Ⅱ (Ang Ⅱ)-mediated cardiomyocyte apoptosis,and explore the related mechanism. Methods Cardiomyocytes isolated from neonatal rats were cultured in vitro. The identity of cardiomyocytes was confirmed by morphological examination and staining with anti-sarcomeric ?-actin antibody,and most (more than 95%) of the cells were identified as neonatal rat ventricular myocytes. After being placed in a serum-free medium for 24 hours,the ventricular myocytes were randomly grouped and received the following treatments respectively:placebo,or Ang Ⅱ (1?mol/L),or Ang Ⅱ (1?mmol/L) plus adiponectin (30mg/L). The protein levels of Bcl-2 and Bax were detected by Western blotting. The apoptotic rate and the levels of intracellular reactive oxygen species (ROS) in the ventricular myocytes were determined by flow cytometry. The survival rate of ventricular myocytes was evaluated by trypan blue staining. Results As compared with placebo group,the apoptosis of neonatal rat ventricular myocytes was significantly promoted in 1?mol/L angiotesin Ⅱ group,the survival rate of cardiomyocytes was significantly reduced,with the protein expression of Bax and the ROS level increased,and the protein expression of Bcl-2 decreased (P

Objective To evaluate the performance of KRAS gene mutation detection in 2014 external quality assessment ( EQA ) program and discuss the problems in clinical laboratories .Methods The sample panel of 2014 EQA program contained 5 artificial formalin-fixed, paraffin-embedded ( FFPE) samples.The participating laboratories were asked to report their results before the deadline .The scores of EQA and the rate of overall coincidence , false positive and false negative were calculated .Results The EQA program for KRAS testing was set twice a year .In 2014, 58 and 57 valid lab results were submitted respectively.About 79.31%(46/58)and 94.73%(54/57) of the laboratories were correct for all samples. The coincidence rate of positive samples were 93.53% ( 217/232 ) and 96.49% ( 165/171 ) . The coincidence rate for negative ones were 100%(58/58) and 98.25% (112/114).The false-negative ratio was 1.29%( 3/232 ) and 0%.The false-positive ratio was 4.14% ( 12/290 ) and 3.15% ( 9/285 ) . Conclusions The results of 2014 EQA for KRAS gene mutation testing suggested that the performance of laboratories had been improved significantly , however , the false-negative and false-positive results had always been the major problems affecting the accuracy of KRAS mutations testing .Laboratories needed to standardize the testing process and manufacturers should optimize the reagents and the way of interpretation , to guarantee the performance of KRAS gene mutation detection .

Objective To investigate the infection of Chlamydia (CT) Untie urea mycoplasma (UU) and person type mycoplasma (MH) in urology outpatients clinic and sexually transmitted diseases clinic and drug resist-ance. Methods A British UNIPATH Clean View Chlamydia rapid immunoassay kits and Tianyang Zhongshan City electronic biosensor Limited production of mycoplasma culture and identification of susceptibility kit were used for the detection of secretion from 3,280 cases of non-gonococcal urethritis (GNU). Results Of 3,280 cases of GNU, only 241 cases were detected with positive CT, accounting for 7.35%, 1163 cases with simply UU positive, ac-counting for 35.46%, only 14 cases with MH positive, accounting for 0.43% ,. Overall detection rate of UU and CT was 42.77% and 11.59% respectively. 122 cases were detected with positive UU + MH, accounting for 3.72%. Female infection rate was higher than that of males (P < 0.01 ) ; UU, MH, MH + UU were more sensitive to azithremycin, Josamycin, doxycycline, roxithromycin, Minocycline. Conclusion The infection rate of urogenital mycoplasma and ehlamydia is higher and drug resistance rate is different. Azithromycin and Josamycin are the best to treat mycoplasma infection, followed by doxycycline, Pyronine Doxycycline, Minocycline.

Circulating tumor DNA (ctDNA) has shown great potential in targeted therapy efficacy prediction, monitoring, high-risk population screening, differential diagnosis, minimal residual disease (MRD) monitoring, and prognosis prediction. The detection of specific gene mutations in ctDNA had been included in clinical practice guidelines for certain tumors to predict the drug efficacy and monitor resistance. A small number of approved companion diagnostic reagents have been used in clinical setting. However, the clinical validity of most ctDNA-related biomarkers it still in the research stage. Besides, the establishment and validation of laboratory-developed tests (LDT) are also problems that need to be solved urgently. Therefore, for the moment, the clinical application of ctDNA analysis is like two sides of a coin, with both opportunities and challenges.

This paper reports 23 cases of the large vestibular aqueduct syndrome.All cases are sensori-neural hearing loss,23.91% is moderately severe hearing loss,54.35% is severe hearing loss,and 21.74% is profound hearing loss.The contour of audiogram in high frequency dctcrioration is 74%,che flat contour of audiogram is 13%,the islanded contour audiogram is 13%.Some cases are progressive hearing loss,and the others are sudden hearing loss,two cases among whole cases were found vestibular dysfunction after examination.After high resolution CT scan in inner ear,27 ears were found pathological changes only in vestibular aqueduct enlargement,in addition to these changes,12 ears were found malformation of vestibule and semicircular canals,and 7 ears were showed cochlea malformation.It is said that the disease is caused by abnormal development on endolymphatic duct in early embryonic stage,and the principal inducement is infectious factor.There is not an effective treatment,but the better results can be reached in deaf children with residual hearing,after they wearing hearing aids and getting the auditory-verbal training in the early period of time.If the hearing aid can not reach effective hearing compensation for these patients,cochlea implantation is considered and suggested.

Objective To evaluate the effect and mechanism of IL‐6 induced Gefitinib resistance in non small cell lung cancer (NSCLC) .Methods The sensitivity of cells to Gefitinib ,the invasion ability of cells and the expression of phosphorylated p‐mTOR was assessed by MTT assay ,Transwell assay and Western blot ,respectively .PC‐9psb388 stable over expressing human recombi‐nant IL‐6(hrIL‐6) cell line was established by transfecting PC‐9 cells with a lentivirus psb388 expressing IL‐6 and stable transfecta‐nts over‐expressing IL‐6 in human lung cancer cell line PC‐9 .The sensitivity of cells to Gefitinib ,the invasion ability ,expression of p‐mTOR were then detected .PC‐9/PC‐9psb388 xenografts were established and the expression of p‐mTOR and IL‐6 in tumor sec‐tions were then detected .Results The sensitivity of PC‐9 cells to Gefitinib was reduced by IL‐6 ,the invasion ability of PC‐9 cells and the expression of p‐mTOR was significantly increased with IL‐6 treatment .The sensitivity of PC‐9 cells to Gefitinib was promi‐nent higher in PC‐9psb388 cells ,while the invasion ability of PC‐9psb388 cells and the expression of p‐mTOR was higher than PC‐9 cells .The sensitivity to Gefitinib was improved and expression of p‐mTOR reduced in rapamycin‐treated PC‐9psb388 cells and IL‐6 stimulated PC‐9 cells .Tumor volume of PC‐9psb388 xenografts was significantly higher than that of PC‐9 cells .The expression of p‐mTOR and IL‐6 in tumor sections of PC‐9psb388 group were higher than that of PC‐9 group .Conclusion IL‐6 could elevate the expression of p‐mTOR to induce Gefitinib resistance in non small cell lung cancer (NSCLC) .

Objective To study the process of foam fractionation of polyphyllin in semi-batch mode. Methods Taking enrichment ratio, recovery rate of polyphyllin, and purity ratio as the performance criteria and using single examining method to examine the operational parameters, i.e. operation mode, air flow rate, initial feed concentration, solution pH value, initial feed height and temperature on separation performance. The optimal conditions of the process were obtained finally. Results The separation performance is good when gas flow rate is 400 mL/min, initial feed concentration (polyphyllins content) is 0.3 mg/mL, pH value is 7, feed height is 30 cm, and feed temperature is 40 ℃. The enrichment ratio is 25.7, recovery ratio is 42.1%, and the foam liquids purity of total polyphyllin is 41.4%, which is 4.5 times higher than that in feed purity. Conclusion Foam fractionation technique could be applied to separate polyphyllin.

Objective To preliminarily investigate the feasibility of MRI-T2* map in evaluating myocardial iron load of myocardial iron overload rabbit models.Methods Eleven rabbits were included in this study and divided into two groups,myocardial iron overload group (n =10) and the control group (n =1).Iron dextrin (dose of 50 mg/kg) was injected in muscles of thigh once a week,totally 12 weeks.Serum iron test and MRI examination were performed before iron injection,and 1 week to 12 weeks after iron injection.MRI scan protocol included short axial T2* map of the left ventricle and cross-section T2* map of the liver.T2* and R2* of the heart and the liver were measured.One rabbit was killed after MRI examination at pre-iron injection,1 week to 8 weeks,11 weeks and 12 weeks after iron injection,respectively.Heart and liver were avulsed to undergo in vitro MRI scan and then paraffin embedded for pathological slices.MRI scan protocol and measurements of the heart and the liver samples were the same to that of in vivo ones.Pearson correlation was used to calculate the relationships between the parameters.Results Myocardial T2* [(32.5 ± 8.3 ms)] and R2* values [(38.4 ± 7.9) Hz] had significant correlation with injecting iron content(1 033.2 ± 673.4 mg),the Pearson coefficients were-0.799 (P =0.001) and 0.770 (P =0.002),respectively.Myocardial T2 had no significant correlation with liver T2* values (r =0.556,P =0.070).T2* values of heart and liver in vivo [(32.5 ± 8.3) ms and (8.8 ± 5.4) ms],respectively had strong correlation with those in vitro [(19.4 ± 6.5) ms and (9.8 ± 5.0) ms],respectively (r =0.757,P =0.007 and r=0.861,P=0.001).T2* and R2* values of the heart and the liver in vivo and in vitro had no significant correlations with serum iron (P ＞ 0.05).On Prussian blue staining slices,blue particles of myocardium,sinus hepaticus and hepatocyte increased with injecting iron content.Conclusions It is feasible for MRI-T2* map to evaluate the myocardial iron load noninvasively.It may provide reliable information for detecting myocardial iron overload in patients with iron overload at an early stage.

BACKGROUND:Dragon’s blood is the main ingredient of traditional medicine prescription for promoting granulation, which has been used in clinical treatment of a variety of refractory wounds and achieved the exact effects. But the Dragon’s blood effect on colagen secretion from normal fibroblasts has not been reported. OBJECTIVE: To investigate the effects of Dragon’s blood extract on the proliferation and secret function of fibroblastsin vitro. METHODS: Dragon’s blood was extracted by extracts chloroform, acetoacetic ester, and alcohol in turn. Normal human fibroblasts were respectively cultured in Dragon’s blood extracts of chloroform, acetoacetic ester, and alcohol, DMEM containing 1% dimethyl sulfoxide, and normal culture medium. Then, the fibroblasts were cultured in vitro in different media containing gradient dilutions of Dragon’s blood extracts (0.002, 0.02, 0.2, 2, 20 g/L), which was folowed by cel proliferation determination assessed with MTT assay. Under the optimal concentration, the cel growth curves were drawn and the flow cytometry was used to determine the changes of cel cycle. The concentration of procolagen type III in the supernatant of the fibroblast culture systems was measured by radioimmunoassay. RESULTS AND CONCLUSION:0.2 g/L-2 g/L dilution of Dragon’s blood extracted by acetoacetic ester enhanced the proliferation of fibroblasts in a dose-dependent manner. The 2 g/L was the optimal dilution of Dragon’s blood extracted by acetoacetic ester, and it increased the ratio of S cels in cel cycle than control group and decreased procolagen type III. These findings indicate that Dragon’s blood acetoacetic ester extract can improve the proliferation of cultured human fibroblastsin vitro, and decrease the secretion of procolagen type III of fibroblasts, and it can be beneficial to improve wound healing and inhibit hypertrophic scar.

OBJECTIVETo investigate the effects of Dragon' s blood extract on proliferation and secret extracellular matrix function of fibroblasts in vitro.

METHODSDragon' s blood was extracted by chloroform, acetoacetic ester, alcohol. Human fibroblast were cultured in vitro in media containing gradient dilutions of Dragon' s blood extracts (0.002, 0.02, 0.2, 2, 20 mg/ml) , which was followed by cell proliferation assessed with MTT assay on 0, 12, 24, 36, 48, 60, 72 h. Under the optimal concentration, the cell growth curves were drawn and the flow cytometry (FCM) was used to determine the changes of cell cycle. On 0, 12, 24, 36, 48, 60, 72 h, the concentration of hyaluronic acid in the supernatant of fibroblast culture was measured by radioimmunoassay.

RESULTS0.2-2 mg/ml Dragon' s blood extracts enhanced the proliferation of fibroblasts in a dose-dependent manner. 2 mg/ml was the optimal dilution of Dragon's blood extract, and it increased the ratio of S cells in cell cycle [(25.80 ± 3.10)%] than control group [(7.50 ± 0.70)%, P < 0.01]. From 12 h to 72 h, in 2 mg/ml Dragon's blood group, concentration of Hyaluronic acid secreted by fibroblasts gradually increased, but were less than control (P < 0.01).

CONCLUSIONSDragon's blood acetoacetic ester extract improved the proliferation of cultured human fibroblasts in vitro, might be beneficial to promote wound healing.

Cell Cycle ; Cell Proliferation ; drug effects ; Culture Media ; chemistry ; Dose-Response Relationship, Drug ; Extracellular Matrix ; Fibroblasts ; cytology ; drug effects ; secretion ; Flow Cytometry ; Humans ; Hyaluronic Acid ; analysis ; secretion ; Plant Extracts ; pharmacology ; Resins, Plant ; Time Factors