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.

Diagnosis, Differential ; Ependymoma ; metabolism ; pathology ; Female ; Hemangioblastoma ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Meningeal Neoplasms ; metabolism ; pathology ; surgery ; Meningioma ; metabolism ; pathology ; surgery ; Middle Aged ; Mucin-1 ; metabolism ; Neoplasm Recurrence, Local ; Vimentin ; metabolism

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 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.

To discuss the influence of dexamethasion on lL-1β and TNF - α expression in suture - induced rabbit corneal neovascularization ( CNV ) and analyze the potential mechanism.
●METHODS: For 43 healthy rabbits, 40 were randomly selected for establishing CNV model in corneal stroma. The right eyes (group A) were received no medicine and the left eyes ( group B) were injected dexamethasone after successfully establishing the model. The no modeling 3 rabbits were normal control group. The morphologic change of corneal was observed with slit lamp microscope and the areas of CNV was calculated every day, then 8 rabbits were randomly chosen for sacrificing at 1, 4, 7, 14, 21d respectively. The pathological characteristics of CNV were observed after HE staining, and lL - 1β and TNF - α expression was detected by immunohistochemistry.
●RESULTS: CNV was grown at the 4d after suture, and the 7-14d was vigorous growth period. inflammatory cell infiltration appeared after HE staining, and CNV was located at the superficial stroma of cornea. lmmunohistochemistry results showed that lL - 1β and TNF - α expression was gradually increased with prolonged suture time. Compared with corneal stitch group, the rabbits cured by dexamethasone were found with less inflammatory cells infiltrating and neovescularization, moreover, the expression of lL - 1βand TNF-α decreased. There were statistical significance between the two groups (P<0. 05).
● CONCLUSlON: Dexamethasone can inhibit the CNV growth by controlling the inflammation of corneal and restraining lL-1β and TNF-α expression.

Objective To clarify the role of the virtual reality (VR)in the training of trans-femoral cerebral angiography for novices without any experience of intravascular manipulation. Methods Twenty-four novices without experiences of intravascular manipulation were divided into 2 groups (VR group or control group)with random pair method concerning the age,gender and subjects. The par-ticipants in control group were trained in routine manners while those in VR group practiced on a simulator and subsequently participated in real angiography once as assistants. Real angiography tests were performed for all participants after the training session and additional angiography tests were per-formed for VR group just after the stimulation session on the simulator. Real angiography tests were supervised and scored according to a modified rating scale by neuroradiological experts who were blinded to the novices' training procedures. Difference between the real test performance of the two groups were compared by pared-samples t test,while the correlation between the performance of the real tests and simulator tests of VR group was proved by Pearson correlation analysis. P<0.05 was con-sidered statistically significant. Results The mean scores of VR group were (27.6±3.6)and (26.4± 3.8)in the tests on simulator and real angiography tests respectively,while score of control group was (21.2±3.4)in the real angiography tests. A positive correlation was revealed in VR group between performance on the simulator and that on real patients (r=0.825,P=0.001). Real performance of the VR group was significantly better than that of control group (P=0.010). Conclusions Virtual reality could ensure transfer of acquired endovascular skills from simulators to patients so as to become a valid tool for novices to improve the interventional techniques in tranfemoral cerebral angiography.

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.