Objective To express and purify the human renin(REN)and mature renin(REN-340)in prokaryotic cells,and detect the concentration. Methods According to the human REN gene sequence(5972)registered in NCBI,REN and REN-340 gene fragments were artificially synthesized and cloned into vector p ET-28a(+)after E.coli codon optimization,and the recombinant expression plasmids 6 × His-REN-p ET and 6 × His-REN-340-p ET were constructed. The recombinant plasmids were transformed into competent E.coli BL21(DE3)and induced at different induction temperatures(20,25,30and 37 ℃)and different final concentrations(0. 1,0. 2,0. 4 and 0. 5 mmol/L)of IPTG. Recombinant REN and REN-340were induced under the optimal induction conditions,and the concentrations were detected by ELISA after nickel ion affinity chromatography with Ni-NTA. Results The recombinant expression plasmids 6 × His-REN-p ET and 6 × His-REN-340-p ET were constructed correctly as identified by restriction analysis and sequencing. The optimum induction temperature of recombinant REN and REN-340 was 37 ℃ and 20 ℃,and the best final concentration of IPTG was 0. 1 and 0. 2 mmol/L,respectively. The concentrations of purified recombinant REN and REN-340 were 9 148. 21 and 15 115. 31 m IU/m L,respectively. Conclusion The prepared recombinant REN and REN-340 proteins have high concentration with short production cycle,which are expected to be used as candidate quality control products and standard substances for external quality assessment(EQA).

Objective To explore the expression and significance of vascular endothelial growth factor (VEGF),tissue polypeptide specificity antigen (TPS) and tumor type M2 pyruvate kinase (TuM2-PK) before and after interventional therapy in lung cancer (LC).Methods Sixty cases of LC treated by interventional treatment in our hospital from January 2013 to January 2015 were analyzed.The levels of VEGF,TPS and TuM2-PK on preoperative 1 d,postoperative 1,3,7,14,28 d were detected by enzyme-linked immunosorbent assay(ELISA).The change of VEGF,TPS and TuM2-PK levels in the patients with different treatment outcomes was observed.Contemporaneous 40 individuals undergoing healthy physical examination served as the healthy control group.Results The levels of VEGF,TPS,TuM2-PK on preoperative 1d,postoperative 3,7,14,28 d had statistical differences between the observation group and control group (P<0.05,P<0.01).Compared with pre-treatment,the levels of VEGF,TPS and TuM2-PK in partial remission patients were significantly improved,which in the patients with stable and progressive condition,the levels of VEGF,TPS and TuM2-PK were significantly increased (P<0.05).Conclusion The levels of VEGF,TPS and TuM2-PK are significantly elevated before and after interventional treatment in LC patients,moreover the worse the therapeutic efficacy,the higher the levels of VEGF,TPS and TuM2-PK,which can be used as the detection indexes of LC interventional treatment.

Stem cells are characterized by the ability to renew themselves and differentiate into a di-verse range of specialized cell types.Bmi1 gene has been determined to play critical roles in the self-renewal of stem cells.Many of the tumor originated in stem cells.Enhanced self-renewal capability in stem/progenitor cells changes its biological characteristic and drives tumor initiation

Gene associated with retinoid-IFN-induced mortality-19(GRIM-19)is originally isolated as a growth suppressive gene using a genetic screen.GRIM-19, initially found in nucleus and mitochondria, is essential for the assembly and functioning of mitochondrial complex I.GRIM-19 involves the regulation of cell proliferation and apoptosis, and low expression or mutation of GRIM-19 contributes to abnormal proliferation.During viral oncogenesis, GRIM-19 may be a general target protein similar to other cellular tumor suppressors.GRIM-19 plays an important role in tumor formation and apoptosis inhibition, and may be as a new tumor marker to early cancer screening.

Aim To observe the expressions of KLF4 in the myocardium of diabetic mice and their changes under Tongxinluo capsule intervention. Methods For-ty KK/Upj-Ay mice were randomly divided into diabet-ic model group(n=10)and diabetic model with Tongx-inluo(TXL low,middle,high) groups(n =10,respec-tively). C57BL/6 mice were selected as control group ( n=10 ) . At the end of the 3 th month the mice were sacrificed and were weighed. The fasting blood-glucose (FBG),glycosylated hemoglobin(HbA1c),triglyceride ( TG) ,total cholesterol ( TC ) and insulin ( FINS ) were measured to calculate HOMA-IR. Radioimmunoassay was used to measure serum TNF-αand IL-6 . The path-ological changes in the myocardium of mice were ob-served by HE staining. KLF4 mRNA was examined by Real-time PCR, while KLF4 and NF-κB protein were measured by Western blot. Results Compared to the control group,FBG,HbA1c,TG,TC,FINS,HOMA-IR,
TNF-α and IL-6 in model group were markedly in-creased;the expressions of myocardial KLF4 were markedly decreased and the expression of nuclear NF-κB protein were markedly increased ( P < 0. 01 ) . Tongxinluo capsule could significantly reduce myocar-dial pathological damage, FINS, TG, TC, TNF-α, IL-6 level and the expression of nuclear NF-κB protein and up-regulate the expression of KLF4 ( P <0. 05 ) , but had no effect on FBG and HbA1c(P>0. 05). Conclu-sions KLF4 may be involved in the development of myocardial injury during diabetes. Tongxinluo capsule can ameliorate the myocardial damage and improve the function of diabetic myocardium by up-regulating the expression of KLF4 and inhibiting the activation of NF-κB inflammatory signal pathway.

Objective To compare the detection rate of Xpert MTB/RIF test and sputum smear and culture method, and to investigate the value of the detection of the smear negative pulmonary tuberculosis.Methods 69 patients with tuberculosis diagnosed in Tianjin Haihe Hospital from July 2014 to July 2015 were examined by fiberoptic bronchoscopy ( and check of tubercle bacillus in sputum smear negative three times ) .With final diagnosis of pulmonary tuberculosis patients the number of statistical smear, culture method and Xpert MTB/RIF test their tuberculosis detection rate.Results A total of 69 patients with sputum smear negative patients with final diagnosis of 54 cases of pulmonary tuberculosis, confirmed 54 cases of tubercle bacillus in sputum smear negative pulmonary tuberculosis patients, Xpert MTB/RIF test was positive in 33 cases accounted for 61.11%(33/54).Among the 35 cases, 17 patients with Xpert MTB/RIF positive, can improve the detection rate of bacterial negative tuberculosis 31.48%(17/54), 19 cases of patients with negative smear positive, 16 cases of Xpert MTB/RIF positive, and the culture method was 84.21%(16/19).BALFXpert MTB/RIF test of tuberculosis found in 61.11%(33/54), the culture method of tuberculosis found 35.19%(19/54).The report is more than 50%, also has reached to 80%, and the selection has a great relationship.Xpert method and culture method, the difference was statistically significant (P<0.05).The positive rate of Xpert test was positive in 2 cases of positive MTB/RIF in the patients with positive smear positive 100%.Conclusion It is a simple, rapid and accurate method to do Xpert MTB/RIF test for BALF, and it can improve the smear negative tuberculosis patients found rate of 48.75%.The positive rate of the sputum smear negative is 59.62%(14+17/35+17), which is better than that of sputum smear and culture.

Pulmonary arterial hypertension is characterized by elevated pulmonary arterial pressure with high disabled rate and lethality.Since many causes could lead to pulmonary arterial hypertension,the mechanism studies were developed rapidly,and some new therapeutic targets have been explored according to the multiple pathogenic mechanisms and pathways.We summarized the potential and novel biomarkers of the mechanisms associated with pulmonary arterial hypertension,and categorized based on their relationship to endothelial cell dysfunction,inflammation,epigenetics,cardiac function,oxidative stress,extracellular matrix,etc.The biomarkers can help for diagnosis,sorting,disease severity assessment of PAH,in order to provide the basis for precision treatment and new therapeutic target development.

[ ABSTRACT] AIM:To explore the effects of transcription factor HOXB7 silencing on the viability , invasion and migration of SGC-7901 cells.METHODS:The siRNA was designed to specifically interfere the expression of HOXB7.To assess the silence efficiency of the siRNA , the expression of HOXB7 at mRNA and protein levels was detected by real-time PCR and Western blot in the SGC-7901 cells transfected with siRNA .The cell viability was measured by MTT assay .Cell cycle-related proteins such as cyclin D 1 and CDK4 were determined by Western blot .Transwell assay was performed to an-alyze the migration ability of the SGC-7901 cells.The mRNA and protein levels of the tumor invasion-and metastasis-relat-ed molecules, such as VEGF, PTEN and p-AKT, were also detected by real-time PCR and Western blot .RESULTS:The expression of HOXB7 at mRNA and protein levels was higher in the gastric carcinoma tissues than that in the normal gastric tissues.The expression of HOXB7 at mRNA and protein levels was knockdown by siRNA .After silencing of HOXB7 gene expression, the viability of the SGC-7901 cells decreased, the expression of cyclin D1, CDK4 and VEGF was down-regula-ted, and the phosphorylation level of AKT was also obviously decreased .CONCLUSION:HOXB7 effectively promotes the viability, invasion and migration of gastric carcinoma cells by up-regulating the expression of cyclinD 1, CDK4 and VEGF, and increasing the phosphorylation level of AKT to inhibit PTEN function .

Urotensin-Ⅱ is a vasoactive 'somatestatin-like' cyclic peptide. Recently, human urotensin-Ⅱ has been cloned and demonstrated to be the most potent vasoconstrictor identified so far. The receptor of urotensin-Ⅱ has now been identified as the orphan receptor GPR 14 . This peptide may influence cardiovarscular homeostasis, pathology and also influence respiratory system, central nervous system and endocrine function.

At the end of last century,the snlall-molecule selective kinase inhibitor,imatinib mesylate (IM),Was successfully exploited.This resulted in a revolutionary step in the treatment of chronic myeloid leukemia(CML).However,along with the expansion of its clinical application and the process,drug resistance Was commonly observed in patients, especially during accelerated or blast phases, and has become a significant therapeutic problem in clinical application.The problem has prompted the appearance of novel small molecule tyrosine kinase inhibitors,and its development is reviewed.