Objective: Trying to find the ways to enhance the expression of cyp71av1 gene encoding cytochrome P450 mono-oxygenase which is a key enzyme in artemisinin biosynthesis pathway accelerating the artemisinin synthesis, the promoter of cyp71av1 was isolated and characterized. Methods: 5′ untranslated regions of cyp71av1 were isolated from Artemisia annua with thermal asymmetric interlaced PCR. For functional characterization, the isolated fragments were fused with β-glucuronidase GUS reporter gene and introduced into Nicotiana tabacum by Agrobacterium-mediated transformation. The GUS expression regulated by 5′ untranslated regions of cyp71av1 in transgenic N. tabacum under the normal or stressed conditions were detected by histochemical staining and quantitative spectrophotometry assay. Results: Two DNA fragments upstream of cyp71av1 coding sequence, a long fragment and a truncated fragment, were isolated from A. annua and introduced into N. tabacum respectively. Histochemical staining showed that two isolated fragments confered stable GUS expression in transgenic plants, and no significant difference was found between the two fragments on the GUS activity. The quantitative results also showed that the GUS activity in transgenic tobacco plants treated by dehydration, low-temperature (4 °C), and ultraviolet irradiation were 1.4 to 2.7 folds higher than that in the controls. Conclusion: It suggests that the isolated fragments has promoter activity and may be responsive to adverse environmental stresses.

Objective To observe the effects of Shenqi Mixture on sLOX‐1 ,SOD ,MDA ,NO ,ET and lipid level among the patients with different types of hyperlipidemia and to investigate the mechanism of blood vessel protection .Methods Sixty‐four patients with hyperlipidemia were divided into high TC group (20 cases) ,high TG group(12 cases) ,mixed type hyperlipidemia group(23 cases) and low HDL group(9 cases) .Every patients were treated with Shenqi Mixture 10 g/d ,treatment course was 6 weeks .Other 25 healthy people of normal blood lipid were selected as the control group .The levels of sLOX‐1 ,SOD ,MDA ,NO ,ET and blood lipid were detected before and after treatment .Results The levels of sLOX‐1 was increased by high TC and mixed type of hyperlipidemia(P<0 .01) .There was a positive correlation between sLOX‐1 with TC and LDL‐C(r=0 .616 ,P<0 .05 ,r=0 .537 , P<0 .05);each type of hyperlipidemia could decrease the level of SOD and NO (P<0 .01) and increased the level of MDA and ET (P<0 .01) .But the effect of SOD ,NO ,ET and MDA on types of hyperlipidemia had difference .Shenqi Mixture decreases the ex‐pression of sLOX‐1 ,regulates the balance of lipid oxidation and peroxidation level TC ,LDL‐C ,MDA and ET (P< 0 .01) and in‐creased the levels of SOD ,NO and HDL‐C(P<0 .01) ,but had no effect on TG(P>0 .05) .Conclusion Shenqi Mixture has good effects on blood vessel protection and protects the vascular endothelial cells ,thus plays the protective role on blood vessel .

Heterotrimeric G proteins are classes of signal-transducing proteins that bind to guanine nucleotides and possess GTP hydrolase activity. G proteins are composed of three subunits α, β, and γ, and are considered as the "molecular switch" in the GPCR signaling pathway. The abnormal activation of G protein is strongly related to diseases such as uveal melanoma, asthma, et al., making directly targeting G protein as an promising strategy for combating diseases. In this review, the classification and physiological functions of G protein are briefly described, and the research progress of G proteins in diseases, G protein modulators are reviewed.

Objective To investigate the effect and mechanism of soluble epoxide hydrolase inhibitor (sEHI) for NF-κB pathway and cell circle arrest of tubular epithelial cell in unilateral ureteral obstruction (UUO) mice model.Methods Thirty-two healthy C57BL/6 male mice performed UUO surgery to induce renal interstitial fibrosis.Animals were randomly divided into 4 groups:sham group (n=8),sEHI (1 mg· kg-1·d-1) group (n=8),UUO group (n=8) and UUO+sEHI (1 mg· kg-1· d-1) group (n=8).Daily sEHI [1-(1-methylsulfonyl-piperidin-4-yl)-3-(4-trifluoromethoxy-phenyl)-urea,TUPS] or 2％ DMSO was applied to mice by oral gavage from day 1 to day 14 after surgery.All mice were sacrificed at day 14 and kidneys were harvested for further analysis.The changes of renal tissue morphology and pathology were observed by Hematoxylin and eosin (HE) and sirius red staining.The expressions of sEH,nuclear factor κB p65 (NF-κB p65) and IκB were measured by Western blotting.The expressions of TNF-α,IL-1β,MCP-1,IL-6,TGF-β,CTGF,collagen-Ⅳ and α-SMA were analyzed by real-time PCR.Immunofluorescence staining of phospho-histone H3 (p-HH3) and Ki67 was performed to determine the stage of cell cycle G2/M arrest.Results The expression and activity of sEH increased in UUO group (P ＜ 0.05).Administration of sEHI inhibited activity of sEH and infiltration of inflammatory cell in tubular interstitial,as well as attenuated tubular damage and tubular interstitial fibrosis.Western blotting analysis revealed administration of sEHI inhibited up-regulated NF-κB p65 and down-regulated IκB in UUO group (P ＜ 0.05).Real-time PCR demonstrated that administration of sEHI obviously decreased the mRNA expression of cytokines and fibrosis markers,including of TNF-α,IL-1 β,MCP-1,IL-6,TGF-β,CTGF,Collagen-Ⅳ,α-SMA (P ＜ 0.05).Immunofluorescence staining showed that there were much more p-HH3 and Ki67 double positive nuclear tubular epithelial cells and interstitial cells in UUO group,compared with Sham group (P ＜ 0.05).Administration of sEHI reduced the number of double positive nuclear cell only in tubular epithelial cells (P ＜ 0.05),but not in interstitial cells.Conclusions In UUO tubular interstitial fibrosis model,sEHI inhibits the activation of NF-κB pathway by down-regulating p65 and up-regulating IκB and ameliorates the infiltration of inflammatory cells.In addition,sEHI plays anti-fibrosis effect by moderating cell cycle G2/M arrest and reducing the excrete of pro-fibrosis factors of tubular epithelial cells.

Objective To explore prognosis and remission-related factors in lupus nephritis (LN) patients.Methods Patients diagnosed as LN by renal biopsy in Tongji Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology between Jan 1,2011 and July 31,2016 were enrolled.All related baseline clinical data was recorded and regular follow-up was performed.Kaplan-Meier curves was used to analyze partial remission and complete remission rates.Log-rank test was performed to compare remission rates of patients with nephrotic-range proteinuria (24-hour proteinuria≥3.5 g) and without nephrotic-range proteinuria (24-hour proteinuria＜3.5 g).Univariate and muhivariate Cox regression analyses were performed to evaluate the remission-related factors in different periods.Results A total of 115 patients,with 88.7％ female and (31.5±9.5)years mean age,were followed up for up to 5 years.During follow-up period 2 patients died and 1 dialyzed.The 6-,12-,24-and 36-and 48-month renal partial remission and complete remission rates were 33.3％,58.2％,71.5％,84.0％,89.6％,and 18.9％,40.5％,67.3％,79.4％,87.0％,respectively.Patients without nephrotic-range proteinuria had higher complete remission than patients with nephrotic -range proteinuria (HR=2.01,95％CI 1.15-3.34,P=0.014),but there was no difference in their partial remission (HR=1.33,95％ CI 0.74-2.43,P=0.341).Multivariate Cox regression model indicated that every 1 g/L increase in baseline level of serum albumin was associated with increased 8％ and 9％ risk,respectively,in partial remission (HR=1.08,95％CI 1.01-1.15,P=0.024) and complete remission (HR=1.09,95％CI 1.01-1.07,P=0.038).Conclusions Around half of LN patients reach remission during 1 year.Patients without nephrotic-range proteinuria have higher complete remission,and serum albumin is a remission-related factors.

Objective To investigate the significances of monitoring urine neutrophil gelatinase -associated apolipoprotein (NGAL)and kidney injury molecule -1 (KIM-1)levels before and after coronary intervention in early predication of contrast -induced nephropathy.Methods The clinical data of 249 patients with coronary heart disease undergoing percutaneous coronary intervention were collected.All patients were divided into contrast -induced nephropathy group(n =21 )and non -contrast -induced nephropathy group(n =228)according to whether had contrast -induced nephropathy.Before surgery and 4h,12h,24h,48h,72h after surgery,the levels of serum creatinine were tested.Before surgery and 4h,12h,24h,48h after surgery,the levels of urinary NGAL and KIM-1 were detected by using enzyme -linked immunosorbent assay(ELISA).Results Compared with before surgery,the serum creati-nine level of contrast -induced nephropathy patients after surgery 48h[(101.7 ±20.3)μmol/L]was elevated,the difference was statistically significant(t =15.972,P <0.05).Compared with before surgery,the urinary NGAL levels of contrast -induced nephropathy patients after surgery 4h ~48h were (12.3 ±1.6)μg/L,(14.5 ±1.5 )μg/L, (14.1 ±1.2)μg/L and (14.3 ±1.4)μg/L,which were significantly elevated(t =8.672,11.817,15.942 and 17.641,all P <0.05),and the urinary KIM-1 levels after surgery 24h and 48h were (5.1 ±0.9)μg/L and (5.5 ± 1.3)μg/L,which were elevated,the differences were statistically significant(t =9.672,14.381,all P <0.05).The urinary NGAL levels of contrast -induced nephropathy patients after surgery 4h ~48h were higher than non -contrast-induced nephropathy patients,and the urinary KIM-1 levels after surgery 24h and 48h were higher than non -contrast -induced nephropathy patients,the differences were statistically significant(t =17.838,19.370,13.996, 18.172,2.792,3.307,all P <0.05).Pearson correlation analysis showed that the urinary NGAL levels after 4h and urinary KIM-1 levels after 24h were positively correlated with serum creatinine levels after surgery 48h(r =0.698, 0.576,all P <0.05).ROC curve analysis showed that urinary NGAL levels in predicting contrast -induced nephrop-athy,the area under the curve was 0.963 (95% CI:0.931 ~0.995 ),sensitivity was 85.7%,and specificity was 94.3%,for urinary KIM-1 levels,those were 0.839 (95%CI:0.768 ~0.909),81.0% and 72.8%.Conclusion The urinary NGAL levels of contrast -induced nephropathy patients after interventional treatment 4h were increased, and the urinary KIM-1 levels appeared increased after surgery 24h,which were earlier than serum creatinine.They were expected to be early indicators for determining acute kidney injury and predicting contrast -induced nephropathy after intervention treatment.

OBJECTIVETo investigate the role of endogenous hydrogen sulfide (H2S) in erectile dysfunction (ED) induced by androgen deficiency.

METHODSWe randomly divided 30 eight-week-old healthy male SD rats into six groups: 2-week control (A), 4-week control (B), 2-week castration (C), 4-week castration (D), 2-week castration + androgen replacement (E), and 4-week castration + androgen replacement (F), those in groups E and F subcutaneously injected with testosterone propionate (TP) at the physiological dose of 3 mg/kg per day after castration, while those in the other groups with isodose oil instead. At 2 and 4 weeks after operation, we determined the level of serum testosterone (T) , intracavernous pressure (ICP) , mean carotid arterial pressure (MAP) of the rats, measured the concentration of H2S in the plasma and corpus cavernosum tissue, and detected the expressions of cystathionine-P3-synthase (CBS) and cystathionine-gamma-lyase (CSE) by immunohistochemistry and Western blot.

RESULTSThe serum T level was significantly lower in group C ([0.63 +/- 0.15] nmol/L) than in A ( [ 16.55 +/- 4.17] nmol/L) and E ( [ 18.99 +/- 4.62] nmol/L) (P <0.05), as well as in group D ([0.70 +/-0.22] nmol/L) than in B ([15.44 +/-5.18] nmol/L) and F ([20.99 +/-6.41] nmol/L) (P <0. 05) , and so were ICP/MAP after 5 and 7 V electrical stimulation of the pelvic ganglia (P <0. 05) , H2 S concentration (P <0.05), and the expressions of CBS and CSE (P <0.05). The expressions of CBS and CSE proteins were also significantly decreased in group C as compared with D (P <0.05).

CONCLUSIONThe reduced expressions of CBS and CSE may inhibit the H2 S signaling pathway, which might be one of the mechanisms underlying androgen deficiency-induced ED in rats.

Androgens ; deficiency ; Animals ; Cystathionine beta-Synthase ; metabolism ; Cystathionine gamma-Lyase ; metabolism ; Erectile Dysfunction ; metabolism ; Hydrogen Sulfide ; metabolism ; Male ; Orchiectomy ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To investigate the pharmaeokinetics of levofloxacin in rat's pancreatic tissue. Methods Pancreatic tissue and blood were sampled in vivo by microdialysis simultaneously. The concentrations of levofloxacin in beth blood and tissues were measured by high performance liquid chromatography. All date were analyzed by WinNonlin software. Results The maximum concentration of free levofloxacin in blood and pancreatic tissue were (65.23 ± 12.9) μg/ml at 10min and (30.56±3.22) μg/ml at 20 min, respectively, then beth continuously decreased. Concentration of free levofloxacin in pancreatic tissue was higher than that in blood from 20min to 100min, then returned to similar level. The area under the concentration curve(AUC)of unbound levofloxacin was(2465.11±258.56)min·μg~(-1)·ml~(-1) in pancreas,and (2914.38±205.73)min·μg~(-1)·ml~(-1) in blood.Conclusions Microdialysis with reversed phase high performance liquid chromatography established in this essay could be used to determine the pharmacokinetics of levofloxacin objectively. High concentration of levofloxacin in pancreatic tissue and blood was observed.

Objective To investigate the feasibility of buccal mucosa swab method to isolate genomic DNA for au-tism spectrum disorders (ASD)-related genetic screening. Methods Buccal mucosa swabs and blood were collected from 41 children with ASD. Genomic DNA was extracted from either blood by using a commercial genomic DNA kit or buccal mucosa swab by using phenol-chloroform-isoamyl alcohol method. The concentration, total quality and purity of genomic DNA were compared between these two methods. Genotyping of the ASD-related methylenetetra-hydrofolate reductase (MTHFR) gene C677T locus was analyzed using PCR-restriction enzymatic digestion and sanger sequencing was per-formed for validation. Results The total quality [(5.87±2.58)μg vs. (2.00±0.92)μg] and concentration [(143.25±72.78) mg/L vs. (66.68±24.43) mg/L] of genomic DNA extracted from buccal mucosa swab were higher than that form blood (P<0.05), while the purity was not significantly different between these two methods (P>0.05). Genotyping analysis of MTHFR was also consistent between these two methods. Conclusion Buccal mucosa swab is a simple, non-invasive and reliable meth-od to obtain genomic DNA, which can partially replace blood for analysis of ASD-related gene polymorphisms.

[Summary] Drug naive, newly diagnosed type 2 diabetic subjects were randomized to Saxagliptin/Metformin / Rosiglitazone(Triple Therapy, n=23) or insulin 70 30 mix group(Intensive Insulin Therapy) (n=21) for 24 weeks. How did the 2 therapies influence fasting blood glucose, fasting insulin, C-peptide, and glucagon levels and the change of body weight were compared. This study was aimed to explore the comparative glycemic efficacy and impact on α/ β-cell function of two different antidiabetic therapies, triple combination therapy using saxagliptin/metformin/ rosiglitazone and intensive insulin therapy, for newly diagnosed type 2 diabetes mellitus. The results indicated that fasting blood glucose, HbA1C , insulin resistance index 2(HOMA 2-IR), glucagon and body mass index level were significantly decreased, and insulin secretion index 2 ( HOMA 2-% β) was increased significantly( P <0. 05) in triple therapy group, and the decreasing extent of HOMA 2-IR, glucagon, and body mass index were significantly greater than that in the intensive insulin group(P<0. 05). Triple therapy group has a stronger effect of reducing insulin resistance, as well as on inhibiting glucagon and promoting weight loss.