Objective To explore the role of miRNA-148a in bladder tumorous development and progression.Methods Ex-pression of miRNA-148a was assessed in 35 bladder carcinoma tissues and 16 non-carcinoma tissues by fluorescence quanti-tative real time PCR,and correlation with clinical features was evaluated.Target genes and transcription factors of miRNA-148a were predicted using bioinformatic analysis,then TF-miRNA-148a-target genes network diagram was built and the tar-get genes was analyzed of gene ontology enrichment and KEGG pathway.Results Expression of miRNA-148a was lower in bladder carcinoma tissues than in non-carcinoma tissues(0.000 8±0.000 2 vs 0.002 1±0.000 5)(t=2.46,P <0.05),but its expression was no statistical significance in different groups of gender,age,pathological classification,clinical stage, lymph node metastasis (P >0.05).268 target genes of miRNA-148a were predicted by three softwares at the same time,60 transcription factors were predicted and the binding sites with combination scroes above 80 was 657.The target genes of miRNA-148a was enriched in many biological processes,such as neuron differentiation,generation of neurons,neuron projec-tion development,cytoplasmic mRNA processing body,cytoplasm(P <0.001).They also participated in p53 signaling path-way,proteoglycans in cancer-homo sapiens,pathways in cancer,prostate cancer,protein processing in endoplasmic reticulum, focal adhesion and so on(P <0.05).According to TF-miRNA-148a-target genes network diagram,miRNA-148a was regula-ted by SP1,ESR1,AP1,MYC and BRCA1,genes of IGF1,P27kip1 ,NCOA1,PTEN,SERPINE1 might be regulated by miR-NA-148a.Conclusion miRNA-148a which was significantly down-regulation in bladder carcinoma tissues may be participate in bladder tumorous development and progression,bioinformatics analysis provides some ideas for further research.

Objective:To establish the bacterial endotoxin test method and the abnormal toxicity test method for reduced glutathi -one for injection.Methods:According to the requirements and methods in Chinese Pharmacopoeia (2015 edition, part IV), the bacte-rial endotoxin test and the abnormal toxicity test for reduced glutathione were studied .Results:The limit of bacterial endotoxin for re-duced glutathione was 0.125 EU· mg-1 , and the limit of abnormal toxicity was 1.0 g· kg-1 .Conclusion: The bacterial endotoxin test method and the abnormal toxicity test method are feasible .The abnormal toxicity should be supplemented in the quality standard for reduced glutathione , and the bacterial endotoxin test can replace the pyrogen test .

BACKGROUND: Studies have demonstrated that the freeze-dried and irradiation-sterilized allogeneic bone is an ideal material for bone transplantation, they are present with good biocompatibility and biomechanical property, also maintains some necessary enzymes for bone morphogenetic protein and morphogenesis protein in bone matrix with some osteninductivable potentials. OBJECTIVE: To observe the effect of the compound of placenta tissue injection and allogeneic lyophilized bone on the reconstruction of jaw bone defects of dogs, and to compare with single allogeneic lyophilized bone. DESIGN, TIME AND SETTING: A comparative observational trial was performed in the Animal Experimental Center of Harbin Medical University between December 2007 and September 2008. MATERIALS: Eight healthy hybred adult dogs; allogeneic lyophUized bone was offered by Hubei Osteolink Biomatedals Co.,Ltd; placenta tissue injection was purchased from Livzon Pharmaceutical Factory Zhuhai (2 mL per injection); allogeneic lyophilized bone: placenta tissue injection=(4-5):1.METHODS: A total of 96 experiment areas from hemisphere jaw defect models at 1.0 cm diameter were established in dog jaw bone site corresponding with central incisor, canine teeth and root apex of the first molar. In the experiment group, the allogeneic lyophilized bone and bone particles were soaked in placenta tissue injection and under saturation state, then the compound of placenta tissue injection and allogeneic lyophilized bone were implanted to jaw bone defect. In the positive control group, the allogeneic lyophilized bone and bone particles were soaked in sodium chloride injection and under saturation state, then implanted to jaw bone defect. In the negative control group, nothing was implanted to jaw bone defect. Each experiment area comprised four materials in each group.MAIN OUTCOME MEASURES: The radiological and histological observations were performed at 2, 4, 8 and 12 weeks after operation.RESULTS: In the experiment group, there was obvious cartilaginous osteogenesis in the earlier period and intramombranous osteogenesis in the late period. The new bone was well integrated with the surrounding tissues. In the positive control group, new recovered bone existed but the combination between the new bone and the original bone was not well. In the negative control group, jaw bone defects were not filled with bone trabecula. Histological examination results showed that there were more new bones in the experiment group than the control groups at 2, 4, 8 and 12 weeks after operation. Statistical difference could be observed among them (P < 0.05, P < 0.01 ).CONCLUSION: The compound of placenta tissue injection and allogenalc lyophilized bone can promote recovery of jaw bone defect actively and shorten recovering time effectively.

Objective:To investigate the role of melatonin in calcium overload-induced heart injury.Methods:Thirty-two rats were divided into 4 groups:a control group (Control),a melatonin control group (Mel),a calcium overload group (CaP),and a calcium overload plus melatonin group (Mel+CaP).Isolated Sprague Dawley male rat hearts underwent Langendorffperfusion.Left ventricular developed pressure (LVDP) was calculated to evaluate the myocardial performance.Triphenyltetrazolium chloride staining was used to measure the infarct size of myocardium.Lactate dehydrogenase (LDH) activity in the coronary flow was determined.The expressions of caspase-3 and cytochrome c were determined by Western blot.The pathological morphological changes in myocardial fiber were analyzed by HE staining.Results:Compared with the control group,calcium overload significantly induced an enlarged infarct size (P＜0.01),accompanied by the disordered arrangement of myocardial fiber,up-regulation of cytochrome c and caspase-3 (P＜0.01),and the increased activity of LDH (P＜0.01).T hese effects were significantly attenuated by 10 μmol/L melatonin (P＜0.01).Conclusion:Melatonin can alleviate calcium overload-induced heart injury.

Objective To investigate the effects of lovastatin on MAPK activity in MCF 7 breast cancer cells. Methods After MCF 7 cells were treated with 4, 8 and 16 ?mol/L lovastatin for 48～72 h, the expressions of ERK1 and p38 MAPK proteins were detected by Western blotting. Their phosphorylation levels were observed by kinase activity assay. Results MCF 7 cells treated with different doses of lovastatin for 48～72 h had no significant effect on the protein level of ERK1, but the activities of ERK1 and the P38 MAPK proteins were down regulated at 72 h after treatment. Conclusion Lovastatin may down regulate the activity of MAPK in MCF 7 cells by means of inhibiting the mevalonate metabolism pathway and/or affecting phosphorylation levels.

A total of 121 subjects comprising 40 normal subjects,58 patients with overweight or obesity and 23 patients with Cushing's syndrome were recruited in the study.The modified radioimmunoassay (RIA) for salivary cortisol test was established'and its normal range was determined.Then the diagnostic value of the salivary cortisol for the initial diagnosis of Cushing's syndrome was evaluated and single midnight salivary cortisol test demonstrated a sensitivity of 100.0% and specificity of 91.4 %.Salivary cortisol test can be recommended as a first-line diagnostic parameter for Cushing's syndrome.

Abstract? AlM: To evaluate macular thickness and macular volume changes in people with diabetes mellitus but no significant decrease of visual acuity.?METHODS:A total of 87 eyes were collected in diabetic group. According to the international stage of diabetic retinopathy, these cases were divided into two subgroups:DR0 stage 54 eyes and DR1 stage 33 eyes. All the cases were received optical coherence tomography (OCT) scan in macular area;the scanning model is 512× 128; recording macular average thickness and macular volume, and compared with healthy subjects.? RESULTS: Macular average thickness and macular volume were higher in DR1 group than those in DR0 stage and control group, and differences were having statistical significance. But DR0 group and control group differences of the two indexes were not statistically significant.? CONCLUSlON: With the aggravation of diabetic retinopathy, the macular thickness tends to be thicken. Although without obvious visual loss, there have been slight morphological changes. Using OCT scan can find fundus changes earlier in patients with diabetes mellitus, and provide clinical basis for both early diagnosis and treatment.

Objective To investigate the feasibility of macrophages as cell carriers to deliver floate modified oxygen loaded ultrasound contrast agent . Methods The phagocytic activity of macrophages was analyzed by ink phagocytose test , and the expression of folate recepters ( FRs ) on macrophages cell membrane surface was tested by immunofluofluorescence assay . Oxygen/paclitaxel loaded lipid microbubbles( OPLMB) and folate‐targeted OPLMB ( TOPLMB) were synthesized by mechanical shock method and incubated with macrophages in vitro . According to different treatment conditions ,the cells were divided into three groups:group A ( OPLMB) ,group B ( free folic acid + TOPLMB) and group C ( TOPLMB) , the fluorescence intensity of the cells were observed under fluorescence microscope ,and the phagocytic percentage and the phagocytic index of macrophages uptake OPLMB and TOPLMB were observed by bright field microscope . Results The phagocytic percentage of macrophages phagocytose ink was (99 .3 ± 1 .0)% ,FRs was highly expressed on macrophages cultured in vitro . After incubation for 30 minutes ,the fluorescence intensity of group C was significantly higher than those of A and B ,the phagocytic percentage in three groups were (19 .5 ± 0 .2)% ,(21 .0 ± 0 .2)% and (81 .2 ± 10 .0)% respetively . The phagocytic percentage of group C were significantly higher than those in group A and group B ( P <0 .05) . Conclusions Macrophages cultured in vitro possess highly phagocytic activity and these cells highly express FRs ,and can be used as cell carriers to deliver floate modified oxygen loaded multimodality ultrasound contrast agent .

The ?glutamyltranspeptidase encoding gene(ggt) from Bacillus subtilis SYU 20016 was amplified by PCR. The ggt gene was inserted in pBV220 to yield the recombinant expression vector pBV220ggt. Overexpression of ggt in E.coli JM109 was achieved with pBV220ggt. SDSPAGE analysis showed an overexpressed recombinant product at about 65kDa,consistent with the molecular weight predicted from gene sequence. The ferment conditions of r-glutamyltranspeptidase were also discussed. The optimum temperature and pH for the enzyme were determined as 30℃ and 7.2 respectively.The cultures were incubated at 42℃ for 4h with broth volume 20ml/250ml flask and the yield of 6U/ml was obtained, enzyme activity of B. subtilis NX2 was only 3.2 U/ml.

Objective To investigate the possibility of using well-differentiated human airway epithelial cells (HAE) to isolate and identify human influenza A virus from a stale respiratory tract specimen.Methods The stale specimen used in this study was a nasopharyngeal swab specimen collected from a patient with unexplained pneumonia in Qinghai in 2010.It was positive for influenza A virus (H3N2) RNA, but negative for hemagglutination.Equal amount of the specimen was inoculated on HAE and on Madin-Darby canine kidney (MDCK) cells for virus isolation and passage.Cytopathic effects were observed daily after inoculation.Hemagglutination inhibition test was performed at every passage.Electron microscope was used to observe viral morphology.Viral genome was sequenced, followed by molecular evolutionary analysis.Results No progeny virus was isolated in MDCK cells, while a influenza A virus subtype H3N2 strain [A/Qinghai/178/2010(H3N2)] was isolated in HAE with a typical morphology and cytopathic effect of influenza A infection.The hemagglutination inhibition activity was 1∶16.Results of the molecular evolutionary analysis of viral genome showed that the influenza A virus (H3N2) strain was highly homologous to the A/Nanjing/1655/2010(H3N2) strain, which was isolated during the 2010 influenza pandemic in Nanjing.Conclusion HAE can be used for isolation and identification of virus from stale respiratory tract specimens.It is more sensitive than MDCK cells with regard to human influenza virus isolation.