Background and purpose:It was reported that many microRNAs (miRNAs) have close relation with carcinomas. miR-31 (microRNA-31) shows abnormal change in numerous cancers. China is one of the most high-risk areas of esophageal squamous cell carcinoma (ESCC). The aim of the present study was to investigate the expression of miR-31 in ESCC, and analyze the relationship of its expression with clinicopathological features and prognosis. Methods:The expression of miR-31 in KYSE410, EC1 and EC9706 cell lines, as well as 81 cases of ESCC tissues and adjacent normal esophageal tissues were detected by real-time reverse transcription-polymerase chain reaction (RT-PCR). The result was combined with clinical and follow-up data and statistical analysis was conducted. Results: MiR-31 was up-expression in 3 cell lines and 75.31% of the ESCC tissues. miR-31 up-expression was positively related to severer lymph node metastasis (P=0.043), deeper invasion of tumors (P=0.002) and advanced pathological stage (P=0.027). There was no relationship of miR-31 with other clinicopathological features (P>0.05). Furthermore, high expression of miR-31 was associated with poor progression-free survival (PFS) in 81 ESCC patients by Kaplan-Meier analysis (P=0.014) and by multivariate Cox analysis (P=0.021). Conclusion:Our results identiifed miR-31 may be a new diagnostic criteria and prognostic biomarker for ESCC.

Objective To develop an assay of PCR-produet direct sequencing to detect hepatitis B virus (HBV) YMDD mutation, and compare the results gained by the sequencing and traditional real-time fluorescent PCR assays. Methods Serum samples were collected from 103 patients with chronic hepatitis B. HBV DNA were extracted from sers. YMDD mutation was detected by a commercial real-time PCR assay. Meanwhile, HBV reverse transcriptase-encoding gene was amplified by a nested PCR assay. The PCR products were directly subjected to sequencing at two directions, and the sequencing results were analyzed by NTI program. Using Kappa test, comparison was made between the results of rtM204-site mutations obtained by the direct sequencing and YMDD mutations by the real-time fluorescent PCR. Results The direct sequencing assay proved to be highly effective with bread range of detection in viral load from 500 to 1010copies/ml. And it may simultaneously avoid inhibitory effect caused by high viral load. The coincidence rates between two assays were 100% for YIDD, 97. 1% for YVDD, 76. 2% for YIDD/YVDD coexistence (Kappa = 0. 853, P < 0. 01). Conclusions The direct sequencing assay for HBV drug-resistant mutation detection is highly sensitive with broad dynamic range. It has high coincidence rate with real-time fluorescent PCR assay with advantage of detecting YMDD, YIDD and YVDD mutations simultaneously.

Objective Inflammation is a defensive reaction of body , but excessive inflammatory response can lead to physi-cal injury.The aim of this study was to explore the effects of luteolin on the secretion of inflammatory cytokines from lipopolysaccharide (LPS) and interferon-g(IFN-γ) activated RAW264.7 cells. Methods RAW264.7 cells were divided into 5 groups: control group (without any medicine), M1 group (polarized M1 cells activated by final concentration of 10 ng/mL LPS+20 ng/mL IFN-γ), M1+5L group (simultaneous activation of LPS and IFN-γplus final concentration of 5μmol/L luteolin), M1+10L group(simultaneous activa-tion of LPS and IFN-γplus 10μmol/L luteolin), M1+20L group(simultaneous activation of LPS and IFN-γplus 20μmol/L luteolin). The cell morphological transformation was observed by laser confocal microscope ;the mRNA levels of iNOS , IL-1βand IL-6 were test-ed by real-time quantitative PCR respectively;the secretion levels of TNF-αand IL-6 in culture supernatant were detected by ELISA;the changes of p-STAT3 (ser727) protein pathways were examined by western blot. Results Cellular morphology of activated RAW 264.7 cells changed obviously .Compared with the control group , the mRNA levels of iNOS, IL-1βand IL-6 decreased significantly in the other 4 groups(P<0.05).The iNOS level in M1+20L group significantly de-creased compared with M1 group[(29.52±3.07) vs (98.91±10.65), P<0.01].As to IL-1βlevel, it decreased significantly in M1+10L group(78.38±8.65) and M1+20L group(41.59±6.80) compared with M1 group(110.69±4.12)(P<0.05).While the IL-6 levels decreased significantly in M1+5L group(177.51±19.28), M1+10L group (106.14±5.63), M1+20L group(27.15±1.26), compared with M1 group(394.10±33.47)(P<0.05).LPS+IFN-γcould induce in-creased p-STAT3 (ser727) expression in M1 phenotype of RAW264.7 cells which was proved by its significant increase in M 1 group, M1+5L group and M1+10L group compared with control group (P<0.05).In comparison to M1 group, p-STAT3-ser expression in M1 phenotype downregulated in M1+5L group, M1+10L group, M1+20L group(P<0.05), along with dose-dependent characteristic.Com-pared with control group, the levels of IL-6 and TNF-αincreased significantly in M1 group, M1+5L group and M1+10L group.Com-pared with M1 group, the levels of IL-6 and TNF-αdecreased significantly in M1+5L group, M1+10L group and M1+20L group(P<0.05) , in which IL-6 showed concentration independence and TNF-αshowed no concentration independence . Conclusion Luteolin inhibits the secretion of pro-inflammatory cytokines through the down-regulation of p-STAT3 so as to exert anti-inflammatory effects .

[Abstract ] Objective Chronic renal disease,a common and frequent disease,is the most cause inducing chronic renal failure. There is an important guiding significance for renal biopsy in the diagnosis,treatment and prognosis of renal disease.The aim of this study was to analyze the characteristics of renal biopsy and the relation between pathological types and clinical presentation in Tibet. Methods Between June 2011 and December 2013 in General Hospital of Tibetan Military Command, Lhasa, percutaneous renal biopsy were per-formed in 107 renal patients.In this study, the gender, age, pathologic entities, and clinical manifestation were analyzed retrospectively. The final diagnosis was made by the clinical manifestation, laboratory examination,and pathologic results. Results The mean age at renal biopsy was 29.8 ±12.2(10~66)years in 47 male cases (43.9%) and 69 female cases (56.1%).This includes 75 Tibetan cases (70.1%), 31 Han cases (28.9%), and Bai cases (1.0%).The primary glomerular disease was 95.3%and the secondary glomerular disease was 4.7%in the total 107 cases.Fourty-four point three percent of the primary glomerular disease were male cases and 100%of secondary glomerular disease were female cases.The main pathological type of the primary glomerular disease was podocyte nephropathy (43.9%), followed by membranous nephropathy (18.7%), IgA nephropathy (11.2%), focal segmental glomerulosclerosis (9.3%) and mesangiocapillary glomerulonephritis (5.6%).The clinical manifestations of 107 cases were classified as syndrome of nephrotic syn-dromn(69.1%),urinary abnormalities(14.9%),isolated macrographic haematuria (3.7%),chronic renal failure(2.8%).Among the nephrotic syndrome,podocyte disease was 63.5%,membranous glomerulopathy was 18.9%,focal segmental glomeruolsclerosis was 8.1%,and membranoproliferative glomerulonephritis was 5.4%. Conclusion The primary glomerulonephritis was the most common glomerular diseases at high altitude area in Tibet.The most frequent type of pri-mary glomerular nephritis was podocyte disease, and the most frequent type of secondary glomerular nephritis was Henoch-Schonlein purpura glomerulonephritis.

The Hippo signaling pathway is highly conserved in evolution, and participates in the regulation of cell proliferation, differentiation, and tissular dynamic balance, and plays an important role in regulating tissue, organ size, and cell number. Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary kidney disease and one of the most common causes of end-stage renal disease. Emerging studies have identified the Hippo signaling pathway is closely related to the occurrence and development of ADPKD. The abnormal activity and expression of the main members of the pathway affect the cilia and cell polarity of renal tubular epithelial cells and induce the formation of renal cysts. The review summarizes the potential mechanism of the Hippo pathway in the pathogenesis of ADPKD, the crosstalk with other signaling pathways, and the variances in different species, and discusses the strategies for the treatment of ADPKD based on the Hippo signaling pathway to provide new strategies for the treatment.

Escherichia coli strain DC1515, deficient in glucose phosphotransferase (ptsG), lactate dehydrogenase (ldhA) and pyruvate:formate lyase (pflA), is a promising candidate for the fermentative production of succinate. To further improve the succinate producing capability of DC1515, we constructed plasmid pTrchisA-pyc with heterogenous pyruvate carboxylase (pyc) from Bacillus subtilis 168 under the Trc promoter and introduced it into DC1515. We used lactose as a substitute of IPTG to induce pyc. We optimized the culture conditions such as the lactose addition time, the lactose concentration and the culture temperature after induction for succinate production. We also explored the effect of lactose supplement during the fermentation. The results showed that pyc can be expressed under lactose induction in the fermentative medium with 15 g/L glucose due to the deficient of ptsG in DC1515. Under optimized conditions, the final succinate concentration reached to 15.17 g/L, which was 1.78-fold higher than that of control strain. If complementing lactose twice to the concentration of 1 g/L during the fermentation, the final succinate concentration could further reach to 17.54 g/L. This work might provide valuable information for gene expression in E. coli strains using lactose as inducer for succinate production in a glucose-medium. Due to the reduced cost, E. coli is becoming a more promising strain for succinate production through fermentation.
Bacillus subtilis ; enzymology ; Culture Media ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Lactose ; pharmacology ; Promoter Regions, Genetic ; Pyruvate Carboxylase ; biosynthesis ; genetics ; Succinic Acid ; metabolism

Objective To investigate the influence of early growth response gene-1 (EGR1) on the autophagy of host cells following infection with human T cell leukemia virus type 1 (HTLV-1).MethodsA HTLV-1-positive cell line MT2 was co-cultured with HeLa cells for 24 h to construct the virus early infection model.Immunoblotting assay was used to detect the expression of HTLV-1 core protein p19 and EGR1.Luciferase reporter gene analysis was used to detect the transcriptional activity of 5′-regulatory sequence of EGR1 at different time points after co-culturing.An effective small interfering RNA (siRNA) targeting EGR1 was screened out and transfected into HeLa cells by Lipofectamine 2000.Then the transfected HeLa cells were co-cultured with the HTLV-1-positive cell line MT2 for 24 h.Immunoblotting assay was used to detect HTLV-1 core protein p19, EGR1 and autophagy-related protein LC3.Real-time PCR was performed to detect viral load.Autophagosome was analyzed by immunofluorescence after co-culturing.Results The expression of EGR1 and the transcriptional activity of pEGR1-luc gradually increased after co-culturing HeLa cells with MT2 cells for 8 h (P<0.01).The expression of EGR1 was positively correlated with host cell autophagy following HTLV-1 infection.The effective siRNA for silencing the expression of EGR1 was obtained and named as siE2.The viral load, the expression of HTLV-1 core protein p19 and the proportion of LC3B/LC3A in the co-culture model were markedly down-regulated by RNA interference with siE2, which was concomitant with a persistent decrease of intracellular autophagosome (P<0.01).Conclusion EGR1 is associated with host cell autophagy and viral replication in HTLV-1 infection.

Objective To explore the relationship among depression,anxiety and social support in elderly patients in community outpatient clinic. Methods A total of 551 elderly outpatients from two com-munity health service centers of Hongkou District in Shanghai were evaluated with patient health question-naire-9 (PHQ-9),generalized anxiety disorder-7 (GAD-7),perceived social support scale( PSSS) for de-pression,anxiety,physical health and social support. Results The prevalence rates of depression and anxiety were 26. 1% and 17. 2%,respectively. The scores of PHQ-9 and GAD-7 were 2. 0(4. 0) and 1. 0(2. 0). There were statistically significant differences in the scores of family support,friend support,other support and social support among the elderly patients with different degrees of depression or anxiety (P<0. 01). Fam-ily support(B=-0. 196) and friend support(B=-0. 171) were protective factors of depression in elderly pa-tients in community outpatient clinic. Age,family support and friend support were protective factors of anxiety in elderly patients,while gender and fluctuation of physical diseases were protective factors of anxiety(P<0. 05). Con-clusions The depression and anxiety is intimately related to social support in elderly outpatients. Appropriate measures should be taken to optimize social support,mitigate bad mood negative improve their quality of life.

During the process of dairy farming,various factors such as physical injury and bacterial infection act upon body tissues or organs,leading to the disruption of skin or mucous tissue integ-rity and subsequent tissue injury and trauma.The healing of these injuries is a complex process that necessitates the coordinated efforts of different cells and involvement of diverse cytokines.A-mong them,the interaction between macrophages and myofibroblasts is indispensable for efficient tissue repair.Nod-like receptor protein 3(NLRP3),a pattern recognition receptor in the innate im-mune system,may play a regulatory role in modulating this intricate process.In this study,cow myofibroblasts and M1 type bone marrow-derived macrophages were cultured in vitro,followed by collection of cell culture supernatant for co-culture analysis.Both cytokine secretion levels in M1 type bone marrow-derived macrophages as well as expression patterns levels of myofibroblast growth factor protein and mRNA were detected.The regulatory mechanism underlying NLRP3 in-volvement in mediating interactions between these two cell types was investigated using NLRP3 inhibitor MCC950.The results showed that an effective method for culturing cow muscle fibroblasts in vitro was successfully established and myofibroblast conditioned medium(MFbCM)could regulate M1 macrophage secretion profiles.Moreover,M1 macrophage conditioned medium(M1?CM)was found to influence myofibroblast growth factor expression levels.Our findings sug-gest that NLRP3 plays a significant regulatory role during crosstalk between myofibroblasts and M1-type pro-inflammatory macrophages.

Hyperkalemia is one of the common ion metabolism disorders in clinical practice. Hyperkalemia is defined as serum potassium higher than 5.0 mmol/L according to the guidelines at home and abroad. Acute severe hyperkalemia can cause serious consequences, such as flaccid paralysis, fatal arrhythmia, and even cardiac arrest. The use of renin-angiotensin- aldosterone system inhibitors, β-blockers and diuretics, low-sodium and high-potassium diets, and the presence of related comorbidities increase the occurrence of hyperkalemia. Hyperkalemia risk exist in all clinical departments, but there is a lack of a standardization in the management of multi- department cooperation in hospital. Therefore, a number of domestic nephrology and cardiology department experts have discussed a management model for multi-department cooperation in hyperkalemia, formulating the management standard on hospital evaluation, early warning, diagnosis and treatment, and process. This can promote each department to more effectively participate in nosocomial hyperkalemia diagnosis and treatment, as well as the long-term management of chronic hyperkalemia, improving the quality of hyperkalemia management in hospital.