Objective To construct an shRNA lentiviral vector targeting the MEX3A gene and establish a bladder cancer cell line with stable MEX3A gene knockdown.Methods Real-time PCR was performed to detect the MEX3A gene expression.The recombinant lentiviral vector targeting the MEX3A gene was constructed using the GV115 plasmid.After identification and sequencing,the vectors were co-transfected with the packaging vector into 293T cells to produce lentiviral particles,which were then transduced into bladder cancer cells after viral titer determination.The cell line stably expressing the siRNA was established by antibiotic selection,and real-time PCR was carried out to detect the efficiency of the knockdown.Results Both bladder cancer cell lines,5637 and T24,expressed the MEX3A gene,and its expression was higher in 5637 than in T24.The identification and sequencing results showed that the MEX3A-shRNA lentiviral vector was successfully constructed,and the virus titer was observed to be higher after packaging.The results of the real-time PCR showed that MEX3A gene expression was stably inhibited in 5637 cells after lentiviral transduction.Conclusion Lentivirus-mediated RNAi technology could successfully establish a cell line with stable MEX3A gene knockdown.

Objective To investigate the effect of U50488H on the ultrastructure and organ function in septic shock rats. Methods Forty SD male rats were randomly(random number) divided into 5 groups: sham group, septic shock group, U50488H+septic shock group, nor-BNI+U50488H+septic shock group, and nor-BNI+septic shock group, with 8 rats in each group. Cecal ligation and puncture (CLP) was performed to induce septic shock in the septic shock group. Rats in the U50488H+septic shock group were treated with U50488H injection by intravenous at the shock point, and other procedures were the same as the septic shock group. Rats in the nor-BNI+U50488H+septic shock group were treated with nor-BNI injection by intravenous 3.5 h after abdomen closed, and other procedures were the same as the U50488H+septic shock group. Except for U50488H injection, rats in the nor-BNI+septic shock group received procedures the same as the nor-BNI+U50488H+septic shock group. Albumin, cardiac troponin I (cTnI) and N terminal pro B type natriuretic peptide (NT-proBNP) in serum were measured at abdomen-closed, 3, 6, and 12 h after CLP. The changes of histology and ultramicro structure under electron microscope of lung, heart, liver and kidney of rats were observed at 12 h after CLP. Statistical analysis was performed using SPSS 19.0. Comparison among groups was carried out using ANOVA, and Student's t-test was used for multiple comparisons as post-hoc. Results At 6 and 12 h of CLP, serum albumin of the septic shock group were significantly lower than those of the sham group (P<0.01), while those in the cTnI and NT-pro BNP groups were higher at 3, 6, and 12 h of CLP (P<0.01). Compared with the septic shock group, serum albumin of the U50488H+septic shock group increased significantly (P<0.01), whereas the serum levels of cTnI and NT-pro BNP decreased remarkably at 3, 6 and 12 h of CLP (P < 0.05, P < 0.01, respectively). Compared with the sham group, the alveolar wall was severely damaged, the alveolar septum and blood vessel wall were thickened obviously; the myocardial fiber was swollen, necrotic, and the infiltration of central granulocyte was increased significantly; hepatocyte showed edema, vacuolar-like steatosis, fatty degeneration, spotty and focal necrosis; and slight edema and vacuolar degeneration were found in the glomerulus endothelial in the septic shock group. Compared with the septic shock group, the ultrastructural damage of the lung, heart, liver and kidney of the U50488H+ septic shock group was significantly improved. All the above effects of U50488H could be blocked by nor-BNI (a selective κ-opioid receptor antagonist) (P<0.01). Conclusions U50488H can promote the recovery of serum albumin, and protect organ function in septic shock rats.

Objective To investigate the clinical effect of angioplasty for symptomatic intracranial atherosclerotic stenosis. Methods Eighty-two patients with symptomatic intracranial atherosclerotic stenosis whom underwent angioplasty after the failure of standard medical therapy were enrolled from Nan-jing Stroke Registry Program from September 2010 to June 2013.Nine of them underwent routine balloon angioplasty alone and 73 underwent intracranial stenting.The median time from onset to surgery was 24.5 days.The occurrence of endpoint events (any stroke ≤30 d after procedure,death and ischemic stroke >30 d in guilty vessels or original stenosis had restenosis and needed to be treated again)was assessed. The incidence of restenosis was followed up with imaging (CTA or DSA). Results (1)In the 82 patients, the success rate of operation was 92.7%(n=72 ),and 78 (95.1%)received follow-up,4 were lost to follow-up.The median follow-up time was 22.5 months (range 9 to 29 months ).Ten patients had an endpoint event,7 of them were ischemic stroke,1 was cerebral hemorrhage,and two were severe asymptomatic restenosis who underwent stenting again.The endpoint events of 3 patients occurred at day 30 after procedure (at ≤24 h after procedure).Kaplan-Meier curves showed that the incidences of cumulative endpoint events at 1,6,12,and 24 months were 3.7%,8.6%,11%,and 13%,respectively.(2)60 patients (73.2%)received imaging examination (11 CTA and 49 DSA ).Restenosis occurred in 17 patients (28.3%),among them the incidence of symptomatic restenosis was 5%(n =3 ),and asymptomatic restenosis was 23.3%(n=14). Conclusion After a comprehensive assessment and a rigorous screening, the safety is high and the mid- and long-term efficacy are satisfactory in patients with symptomatic intracranial arterial stenosis who are treated with angioplasty when their medical treatment is invalid.

Objective To study the effects of concentration of glucose on the proliferation of bone marrow mesenchymal stem cells (BMMSCs).Methods BMMSCs passaged to the 4th to 6th were inoculated and cultured.The control group was treated with DMEM/F12 (1∶1) standard medium,and the basal glucose concentration was 5.5 mmol/L.The experimental group was treated with DMEM/F12 (1∶1) high-glucose medium with the concentration of 20.5,25.5,30.5 and 35.5 mmol/L respectively,and continuously cultured for 15 d.The absorbance (A) values of each group once per day were measured by thiazolyl blue (MTT) colorimetry.Results Compared with the control group,the absorbance was increased at the 2nd,3rd,and 5th days in the 20.5 and 30.5 mmol/L groups,and the differences were statistically significant (all P＜0.05).For the 25.5 mmol/L group,the absorbance was increased at the 2nd,3rd,5th and 8th days,and the differences were statistically significant (all P＜0.05).For the 35.5 mmol/L group,the absorbance was increased at the 2nd,3rd,5th and 8th days and decreased at the 8th day,and the differences were statistically significant (all P＜0.05).The results of 20.5,25.5,30.5 and 35.5 mmol/L groups were further compared one another.The results showed that the absorbance at the 4th and 12th days in the 35.5 mmol/L group were lower than that of the 20.5 mmol/L group,and the differences were statistically significant (all P＜0.05).The absorbance at the 11th and 14th days in the 30.5 and 35.5 mmol/L groups were lower than that of the 20.5 mmol/L group,and the differences were statistically significant (all P＜0.05).Conclusions High-glucose environment can promote the proliferation of BMMSCs in a short time.However,with the prolongation of culture time,this effect is gradually weakened because of the inhibiting effect of high-glucose environment to the cell proliferation.

Chemical constituents from the air dried parts of Cichorium glandulosum were studied. The chemical constituents of C. glandulosum were separated and purified by means of silica gel, Sephadex-LH 20, ODS column chromatography and semi-preparative high performance liquid chromatography. The structure was elucidated by physicochemical characteristics and spectral data. One new flavonoid glycoside was isolated from C. glandulosum, and identified as quercetin-3-O-［6″-O-(3-ethoxy-1, 3-dioxopropyl)］-β-D-glucopyranoside(1).

Objective To preliminarily study and establish a method for measurement of the transuranic nuclide ²⁴¹Am in fecal samples, and to provide technical support for internal radiation monitoring of staff. Methods Fecal samples were collected with a self-made stool sampler and treated with a self-made carbonization and ashing furnace. DGA resin was used to separate and purify ²⁴¹Am from fecal samples. With ²⁴³Am as the tracer, the orthogonal method was used for condition optimization. Results The optimum conditions for separation and purification were: the acidity of HNO₃ added into the column, 6 mol/L; column flow rate, 0.6 mL/min; and the volume of analytical solution,12 mL. The method based on inductively coupled plasma mass spectrometry showed a detection limit of 9.79×10⁻⁴ Bq for ²⁴¹Am in fecal samples, which was satisfactory and feasible. Conclusion This method fills the vacancy of ²⁴¹Am measurement in fecal samples to some extent, which is of practical significance for internal radiation monitoring and protection for analysts.

Objective To establish a method for measurement of ²³⁹Pu in fecal samples based on inductively coupled plasma-mass spectrometry (ICP-MS), and to provide a novel method for assessing the internal exposure of workers. Methods Fecal samples were collected from workers and labeled. The samples were pretreated with carbonization ashing and microwave digestion devices, purified on TEVA resin, and measured using ICP-MS. Results The detection limit of ²³⁹Pu in fecal samples based on ICP-MS was 1.91 × 10⁻⁴ Bq. Conclusion In the routine monitoring of class S substances characterized by a 5 μm aerodynamic diameter during 12 months, the committed effective dose corresponding to the detection limit is 0.17 mSv. This value meets the requirements of relevant national standards and ICP-MS can be used as a novel means for accurate evaluation of internal exposure for workers.

Objective To analyze the influence of general anesthesia (GA) on bilateral subthalamic nucleus deep brain stimulation (STN-DBS) in treating Parkinson's disease (PD) through microelectrode recording (MER),and discuss the differences between different modes of anesthesia.Methods A retrospective analysis was performed on clinical data of 31 PD patients accepted bilateral STN-DBS in our hospital from June 2015 to June 2017.Nine patients accepted surgery under GA (A group):4 patients were treated with intravenous anesthesia (A1 group),and 5 patients were treated with inhalation anesthesia (A2 group);22 patients accepted surgery under local anesthesia LA group.MER indexes,including STN discharge frequency,STN recorded length,and maximum target error,and short-term (6 months) efficacy were recorded.A linear regression analysis was performed to find possible influence factors on discharge frequency and improving rate of UPDRS scores.Results The discharge frequencies of B group,A1 group and A2 group were 51.42 Hz±6.28 Hz,35.79 Hz±7.02 Hz and 43.18 Hz±5.87 Hz,respectively,with significant differences (F=12.181,P=0.000);as compared with that in the B group,the discharge frequencies of A1 group and A2 group were significantly lower (P＜0.05).The STN recorded lengths of B group,A1 group and A2 group were 5.48 mm±0.33 mm,5.06 mm±0.15 mm and 5.22 mam±0.16 mm,respectively,with significant differences (F=4.115,P=0.027);as compared with that in the B group,the recorded lengths of A1 group and A2 group were significantly shorter (P＜0.05).A1 group had the maximum target error,but no significant differences were noted among the 3 groups (P＞ 0.05).Six months after the surgery,the UPDRS-Ⅲ scores and Schwab-England scores of A group and B group were decreased and daily levodopa equivalent (LEDD) was decreased.As compared with B group,A group had significantly better improvement in Hoehn & Yahr grading (P＜0.05).Disease durations were positively correlated with discharge frequency (r=0.539,P=0.002);age and improving rate of UPDRS scores were negatively correlated (r=-0.572,P=-0.001);preoperative LEDD and improving rate of UPDRS scores were positively correlated (r=0.725,P=-0.000).Conclusions Bilateral STN-DBS performed under GA in PD enjoys good efficacy,which shows no obvious difference as compared with that under LA.Inhalation anesthesia had less influence on electrophysiology than intravenous anesthesia.

Gout is a metabolic disease closely associated with hyperuricemia and urate deposition. Because of the complex pathogenesis， high morbidity， multiple complications， and increasingly young patients， gout has received worldwide attention. Currently， western medicine mainly treats gout by lowering the uric acid level and reducing inflammation， which， however， causes serious adverse reactions and has contraindications. Phellodendri Chinensis Cortex （PCC） is the dried bark of Phellodendron chinense， with the effects of clearing heat， drying dampness， purging fire， detoxifying， and treating sores. Studies have shown that PCC and its active components have anti-inflammatory， pain-relieving， uric acid-lowering， and anti-gout activities， with extensive sources and high safety. PCC and its active components could prevent and treat gout through multi-targets and multi-pathways， whereas the systematic review remains to be carried out. Therefore， this paper summarized the pharmacological activities and mechanisms of PCC and its active components in the treatment of gout. The available studies have shown that PCC and its active components exert the anti-gout effect by lowering the uric acid level， reducing inflammation， alleviating oxidative stress， and regulationg intestinal flora， and protecting the kidneys. Particularly， the active components represented by alkaloids contribute obviously to the therapeutic effect of of PCC. Herein， we analyzed the problems and future development of the research on PCC， aiming to provide theoretical support and a scientific basis for the research and development of new drugs against gout.