Objective To study ischemic tolerance induced by acanthopannx senticosus saponins(ASS) on ischemia in PC12 cells and involve expression of hypoxia inducible factor-1?(HIF-1?).Methods An ischemic model was developed in PC12 cell line with treatment of oxygen glucose deprivation(OGD).The protective effects of ASS pretreatment on ischemic tolerance of PC12 cells and whether protective effects of ASS could be inhibited by LY294002(PI3K inhibitor) were analyzed through MTT assay.The induction of phospho-glycogen synthesis kinase-3?(p-GSK-3?) and HIF-1? after pretreatment of ASS and possible blocking effects of LY294002 were detected by Western-blot.Results MTT results showed that after 9 h ischemia,the viability of PC12 cells decreased dramatically(P

Overlapping PCR technology was employed to splice IFN?and HSA genes in vitro. The spliced gene was inserted into Pichia pastoris secretory vector pPIC9K. The IFN?-HSA gene was designed for secretory expression under the control of promoter AOX1 and Mat a signal peptide in pPIC9K. The recombinant plasmid pPIC9K/IFN?-HSA was linearized by restriction enzyme SalI and transformed into Pichia pastoris KM71 by electroporation. The recombinant strains identified by G418 selection and confirmed by PCR analysis were induced by methanol to express fusion protein IFNp-HSA. SDS-PAGE and Western blot analysis of the fusion protein showed that the expressed fusion protein IFNp-HSA with an apparent 90kDa molecular weight had the antigenicity of HSA. The specific activity of culture supernatant was about 640IU/ml assayed by the standard amiviral activity test on WISH cells challenged with VSV virus.

Objective To summarize and discuss the diagnostic and treating experiences of chronic pancreatitis with mass in the head. Methods Eight patients of chronic pancreatitis with mass in the head who were misdiagnosised as carcinoma of head of pancreas were analyzed retrospectively in the past 10 years. Results All the patients exhibited abdominal pain,5 of whom were with jaundice and 3 with anorexia. All the patients were misdiagnosised as carcinoma of head of pancreas before the operation,but the pathology after operation indicated chronic pancreatitis. The pancreaticoduodenectomy was performed in 5 patients,the choledochojejunostomy in 2 patients,while the exploratory laparotomy in 1 patient. After the operations,the abdominal pain was relieved in 7 patients, while 2 patients who accepted pancreatoduodenectomy suffered from pancreatic fistula,1 of whom died in the end. Conclusion It’s hard to differentiate the chronic pancreatitis with mass in the head from the carcinoma of head of pancreas before operation. If the carcinoma of head of pancreas can’t be excluded during the operation,the pancreatoduodenectomy should be performed,while the duodenum-preserving total resection of the head of the pancreas or any intra-drainage operations should be done if chronic inflammation is found in the whole pancreas with a negative result of the biopsy of the pancreas through the needle aspiration.

Objective To evaluate the feasibility and safety of embedding the totally implantable venous access port (TIVAP) via the access of right brachiocephalic vein (BCV).Methods The clinical data of 493 patients,who underwent the placement of TIVAP by using right BCV route during the period from March 2013 to December 2015,were retrospectively analyzed.The patients included 137 males and 356 females,with a mean age of (47.3±13.2) years old (ranging from 29 to 78 years old).The puncture success rate and TIVAP indwelling procedure-related complications were analyzed.Results The technical success rate was 100％,the success rate of initial puncturing was 99％ (488/493).The mean operation time was (22.5± 8.3) minutes (range of 18-35 minutes).Mis-puncturing of artery happened in 3 patients (0.61％,3/493);and no severe complications such as hemothorax or pneumothorax occurred.After implantation,the patients carried TIVAP for 124-986 days,with a mean of (271.1±53.8) days.The incidence of complications was 2.25％ (11/488),including hemorrhage at port site (n=2),catheter-related infection (n=l),partial thrombosis (n=2),and formation of fibrous protein sheath (n=6).No serious complications such as displacement or rupture of catheter,or catheter pinch-off syndrome (POS),etc.were observed.Conclusion The implantation of TIVAP by using right BCV route has high puncturing success rate,the technique is safe and reliable,and it can provide another option of catheter access for the clinical performance of TIVAP implantation.

Objective To explore the clinical value of guide-wire shaping in subclavian vein catheter-ization.Methods Totally 400 patients requiring right subclavian vein catheterization were equally divided into two groups according to the clinic date: intervention group ( with guide-wire shaping , n =200 ) and control group (without guide-wire shaping, n=200).The catheterization was carried out by the same doctor .The rates of ectopic wire were compared between the two groups .Results The overall success rate of catheteriza-tion was 98.25%(393/400) [98.5% (197/200) in intervention group and 98.0% (196/200) in control group, P=0.500].The incidence of catheter displacement was 1.02%(2/197) in intervention group, which was significantly lower than that [7.14% (14/196)] in control group (P=0.002).Conclusion As a sim-ple procedure , guide-wire shaping can effectively prevent catheter displacement during catheterization .

OBJECTIVETo investigate the clinical effectiveness of sertraline hydrochloride combined with four-spot caress in the treatment of primary premature ejaculation (PE).

METHODSWe randomly assigned 90 primary PE patients to three groups of equal number. The patients in group A (aged [28.1 ± 5.2] yr and with a disease course of [3.1 ± 1.9] yr) were treated with oral sertraline hydrochloride at 50 mg qd, those in B (aged [27.8 ± 4.1] yr and with a disease course of [3.2 ± 2.0] yr) by four-spot caressing (caressing the tongue, breasts, and vulva prior to intercourse), and those in C (aged [27.1 ± 4.7] yr and with a disease course of [3.1 ± 2.0] yr) by the combination of oral sertraline hydrochloride and four-spot caressing, all for 12 weeks. Before and after 4, 8, and 12 weeks of treatment, we obtained the intravaginal ejaculatory latency time (IELT) and Chinese Index of Sexual Function for Premature Ejaculation-5 (CIPE-5) scores and compared them among the three groups of patients.

RESULTSThe IELT was dramatically prolonged in groups A, B, and C after 4 weeks ([1.08 ± 0.29], [0.93 ± 0.28] and [1.21 ± 0.27] min), 8 weeks ([1.43 ± 0.30], [1.20 ± 0.33] and [1.72 ± 0.42] min) and 12 weeks of treatment ([2.12 ± 0.63], [1.90 ± 0.65] and [2.67 ± 0.82] min) as compared with the baseline ([0.63 ?0.14] , [0.60 ?0.14] and [0.62 ?0.11] min) (P < 0.05), even longer in group C than in A and B (P < 0.05). The CIPE-5 scores were markedly improved in groups A, B and C after 4 weeks ([15.17 ± 1.74], [14.57 ± 1.94] and [15.60 ± 1.63] min), 8 weeks ([17.13 ± 1.63], [16.37 ± 1.97] and [18.00 ± 1.05] min) and 12 weeks of intervention ([18.93 ± 1.57], [18.53 ± 1.67] and [20.00 ± 1.46] min ) as compared with the baseline ([12.57 ± 2.05], [13.20 ± 2.51] and [13.07 ± 2.01] min) (P < 0.05), even higher in group C than in A and B (P < 0.05).

CONCLUSIONSertraline hydrochloride combined with four-spot caressing, with its definite efficacy and rare adverse reactions, deserves wide clinical application in the treatment of primary PE.

Adult ; Coitus ; Ejaculation ; Female ; Humans ; Male ; Premature Ejaculation ; drug therapy ; Sertraline ; therapeutic use ; Young Adult

Objective By the method of multiple polymerase chain reaction (PCR),we intend to amplify different regions (DFR) of Yersinia pestis DNA,and to establish a multiple DFR genotyping technique for detection of Yersinia pestis.Methods According to the product size of 23 DFRs and pMT plasmid,24 primers were optimized and combined,then multiple primers in one PCR reaction system were added,and positive template DNA was amplified.Meanwhile,200 wild strain DNAs were amplified by multiple PCR and normal PCR,to verify the coincidence rate of the two methods.Results Totally 24 target segments were amplified through the positive DNA template.Through different permutation and combination,24 primers were optimized and combined into 9 groups.Totally 200 wild strain DNAs were used for verification,the coincidence rate of multiple PCR and normal PCR was 100％.Conclusions Multiple PCR is applicable and feasible for DFR genotyping of Yersinia pestis.It is an efficient,economic and high accuracy experimental method for large quantities of Yersinia pestis DFR genotyping.

It is now well established that inflammation plays an important role in the development of numerous chronic metabolic diseases including insulin resistance (IR) and type 2 diabetes (T2DM). Skeletal muscle is responsible for 75% of total insulin-dependent glucose uptake; consequently, skeletal muscle IR is considered to be the primary defect of systemic IR development. Our pre- vious study has shown that rutaecarpine (Rut) can benefit blood lipid profile, mitigate inflammation, and improve kidney, liver, pan- creas pathology status of T2DM rats. However, the effects of Rut on inflammatory cytokines in the development of IR-skeletal muscle cells have not been studied. Thus, our objective was to investigate effects of Rut on inflammatory cytokines interleukiri (IL)-1, IL-6 and tumor necrosis factor (TNF)-α in insulin resistant primary skeletal muscle cells (IR-PSMC). Primary cultures of skeletal muscle cells were prepared from 5 neonate SD rats, and the primary rat skeletal muscle cells were identified by cell morphology, effect of ru- taecarpine on cell proliferation by MTT assay. IR-PSMC cells were induced by palmitic acid (PA), the glucose concentration was measured by glucose oxidase and peroxidase (GOD-POD) method. The effects of Rut on inflammatory cytokines IL-1, IL-6 and TNF-α in IR-PSMC cells were tested by enzyme-linked immunosorbent assay (ELISA) kit. The results show that the primary skeletal muscle cells from neonatal rat cultured for 2-4 days, parallel alignment regularly, and cultured for 7 days, cells fused and myotube formed. It was shown that Rut in concentration 0-180. 0 μmol x L(-1) possessed no cytotoxic effect towards cultured primary skeletal muscle cells. However, after 24 h exposure to 0.6 mmol x L(-1) PA, primary skeletal muscle cells were able to induce a state of insulin resistance. The results obtained indicated significant decrease (P < 0.05 to P < 0.001) IL-1, IL-6 and TNF-α production by cultured IR-PSMC cells when incubating 24 hours with Rut, beginning from 20 to 180.0 μmol x L(-1). IL-1, IL-6 and TNF-α in the Rut treated groups were dose-dependently decreased compared with that in the IR-PSMC control group. Our results demonstrated that the Rut promoted glucose consumption and improved insulin resistance possibly through suppression of inflammatory cytokines in the IR-PSMC cells.
Animals ; Cell Proliferation ; drug effects ; Cytokines ; metabolism ; Female ; Glucose ; metabolism ; Indole Alkaloids ; pharmacology ; Inflammation ; metabolism ; Insulin Resistance ; Male ; Muscle, Skeletal ; cytology ; drug effects ; metabolism ; Quinazolines ; pharmacology ; Rats

Objective To explore whether paraquat (PQ) can induce the formation of neutrophil extracellular traps (NETs) in human peripheral blood.Methods Neutrophils were isolated from healthy human peripheral blood,and the cells were identified by hematoxylin-eosin (HE) strain.The cells were treated with different concentrations of PQ [0 (as control),200,400,600,800,1 000 and 1 200 μmol/L],and the cell viability was measured by cell proliferation and CCK-8 cytotoxicity detection kit,and the median lethal concentration of PQ was selected.The cells were treated with the median lethal concentration of PQ (PQ poisoning group),and the untreated cells were served as the control.Immunofluorescence staining was adopted to evaluate NETs formation.PicoGreen dye was used to determine the quantitative content of circulating free DNA.Western Blot was used to determine the expressions of citrullinated histone 3(H3Cit) and myeloperoxidase (MPO) in the supernatant.Results The purity of neutrophils was about 95％ by HE staining.The cells were treated with different concentrations of PQ,and the result showed that the viability of cells was (58 ± 2)％ with 800 μmol/L PQ for treatment.The immunofluorescence showed that there were few expressions of H3Cit and MPO in neutrophils in the control group,and there was no NETs formation,which was composed of DNA,H3Cit and MPO.Compared with the control group,a large amount of NETs was generated from neutrophils stimulated by 800 μmol/L of PQ.Meanwhile,quantitative result showed that the content of cell free DNA in the supernatant was significantly increased in PQ poisoning group as compared with that of control group (μg/L:2 235 ± 462 vs.561 ± 87,P ＜ 0.01).The protein expressions of H3Cit and MPO in the supernatant were also significantly increased as compared with those of control group [H3Cit protein expression (gray value):0.23 ± 0.03 vs.0.11 ± 0.01,MPO protein expression (gray value):0.47 ± 0.05 vs.0.21 ± 0.04,both P ＜ 0.05].Conclusion 800 μmol/L of PQ can induce the formation of NETs in human peripheral blood.

Brain Neoplasms ; diagnosis ; diagnostic imaging ; radiotherapy ; Glioma ; diagnosis ; diagnostic imaging ; radiotherapy ; Humans ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; Positron-Emission Tomography ; Radiotherapy, Conformal ; Tomography, Emission-Computed, Single-Photon