Objective To understand the distribution,variation and prevalence of virulence-related gene of the V.cholerae at the molecular level.Methods PCR technique was applied to detect ctxAB,and zot gene of CTX? genetic element,cri gene of TLC cluster of the 176 strains of V.cholerae isolated from 1964 to 2008.Results One hundred and thirty-one strains of V.cholerae carry ctxAB and zot of CTX? genetic element in 176 strains,the positive rates were 72.72%(128/176 strain) and 73.30%(129/176 strain) respectively.69.00%(69/100 strain) of V.cholerae O1 strains isolated from 1964 to 1996 carried cri gene,however the rate was only 1.33%(1/75) in the strains isolated from 1998 to 2008.ctxAB,zot and cri in V.cholerae O139 were all positive.Conclusion Detection of virulence-related genes can reflect the molecular characteristics of strains,and it is important to the molecular epidemiology of V.cholerae research.

Neomangiferin, a natural C-glucosyl xanthone, has recently received a great deal of attention due to its multiple biological activities. In this study, a rapid and sensitive ultra-high performance liquid chroma-tography tandem mass spectrometry (UHPLC–MS/MS) method for the quantification of neomangiferin in rat plasma was developed. Using chloramphenicol as an internal standard (IS), plasma samples were subjected to a direct protein precipitation process using methanol (containing 0.05% formic acid). Quan-tification was performed by multiple reactions monitoring (MRM) method, with the transitions of the parent ions to the product ions of m/z 583.1-330.9 for NG and m/z 321.1-151.9 for IS. The assay was shown to be linear over the range of 0.2–400 ng/mL, with a lower limit of quantification of 0.2 ng/mL. Mean recovery of neomangiferin in plasma was in the range of 97.76%–101.94%. Relative standard deviations (RSDs) of intra-day and inter-day precision were both o 10%. The accuracy of the method ranged from 94.20%to 108.72%. This method was successfully applied to pharmacokinetic study of neomangiferin after intravenous (2 mg/kg) and intragastric (10 mg/kg) administration for the first time. The oral absolute bioavailability of neomangiferin was estimated to be 0.53%7 0.08%with an elimination half-life (t1/2) value of 2.74 7 0.92 h, indicating its poor absorption and/or strong metabolism in vivo.

Objective To prepare the monodisperse,colloidal gold nanoparticles (AuNPs) with controllable sizes (50 nm,65 nm,79 nm and 102 nm) for the qualitative detection of cardiac troponin Ⅰ (cTnⅠ) by immunochromatography assay,and to evaluate the effectiveness of the detection.Methods Four kinds of monodisperse citrate-stabilized AuNPs were prepared using small AuNPs as growth centers (seeds) by a seeded growth thermal aging protocol.As controls,two conventional AuNPs (20 nm,40 nm) were prepared by the traditional citrate-reduction method.The mouse monoclonal antibody against cTnⅠ labeled AuNPs were dropped on polyester mat to make AuNPs conjugate pad.The detection line and quality control line of immunochromatography assay kits for detection of cTnⅠ were coated by mouse anti human cTnⅠ monoclonal antibody paired with antibody in AuNPs and goat anti mouse polyclonal antiboy respectively.The six kinds of AuNPs were employed as color-labels in immunochromatography assay kits for detecting cTnⅠ,and the corresponding detection effects were evaluated in signal intensity,sensitivity,specificity and stability.The assay kit with the best performance was chosen and compared with the commercialized kits for the detection of cTnⅠ in clinical samples.Results Four kinds of monodisperse AuNPs with large sizes of 50 nm,65 nm,79 nm,102 nm respectively were successfully synthesized by the seeded growth thermal aging method.The signal strength of four kinds of kits produced by the four large-sized AuNPs was superior to the kits produced by 20 nm AuNPs in detecting cTnⅠ(all P＜0.01).The signal strength of the kits produced by 65nm AuNPs showed the best performance among the six kinds of AuNPs(all P＜0.01).The lowest detectable limit was 0.50 ng/ml.To compare the agreement of results from chemiluminescent immunoassay versus the results from kits produced by 65nm AuNPs,100 serum samples have been used for detecting cTnⅠ.Their positive coincidence rate was 97.30％ and negative coincidence rate was 100％,the sensitivity and signal strength of the kits produced by 65nm AuNPs was superior to similar products which produced by ABON and Bottests company(all P＜0.01).Conclusions Monodisperse,largesized,citrate-stabilized AuNPs are controllably prepared by a seeded growth-thermal aging method.The development of large-size AuNPs-based immunochromatography assay kits is feasible.65 nm AuNPs can be a suitable candidate for cTnⅠ immunochromatography assay kit.Our findings provides a new idea for the current immunochromatography assay kits which still adopt small-sized AuNPs as color labels.

Objective To explore the relationship between lumbar disc degeneration (LDD) of lumbar spinal stenosis(LSS)and the dural sac cross-sectional area(DSCA)by MRI measurement. Methods 91 patients with central degenerative LSS were randomly selected and 91 age-and sex-matched people without LSS were select-ed as a control group. LDD was classified into five grades by MRI detection according to the method proposed by Pfirrmann and DSCA were measured. Results LDD was not associated with age in LSS. The proportion of severe degenerated disc in lower lumbar levels were higher than that of L2/3 in the two groups;DSCA in severe degenerat-ed disc group was significantly smaller than that in light degenerated group only in L2/3 and L3/4 in LSS. There were no statistical differences in every lumbar level in the control group. Conclusions LDD in L4/5 and L5/S1 of LSS is more severe than that of the normal people. DSCA and LDD are positively correlated in L2/3 and L3/4,but not in L4/5 and L5/S1 for LSS.

AIM:To observe the effects of supraoptic ADH neurons and central diabetes insipidus(CDI)in Wistar rats at different times after hypophysectomy.METHODS:Hypophysectomy was undergone by stereotaxic instrument.Water intake,urine output and urine specific gravity(SG)were observed each day.The survival rate of supraoptic ADH neurons was determined by immunofluorescence after hypophysectomy at different times.RESULTS:The rats manifested triphasic CDI after hypophysectomy.The average water intake in experiment group was 73.9 mL vs 30.9 mL in control group(P0.05),but the cellular body hypertrophy appeared.The survival rate at 10th day was 72%(P0.05),but they were all less than the 10th day(P

Object: To explore the electrocardiographic characteristics of ventricular arrhythmias (VAs) originating from tricuspid annulus region. Methods: Present study included 169 consecutive patients undergoing catheter ablation of VAs from tricuspid annulus origin in our department from August 2007 to September 2016. Based on the origin sites, the patients were divided into two subgroups, the free wall group (81 cases) and septal wall group (88 cases). Based on the location, patients in the free wall group were classified into anterolateral (22 cases), lateral (26 cases) and posterolateral (33 cases) subgroups. Patients in the septal group were classified into anteroseptal (10 cases), midseptal (71 cases) and posteroseptal (7 cases) subgroups. We analyzed the electrocardiographic features of these patients and in 87 patients with PVCs/VT originating from right ventricular outflow tract. Results: (1) A positive R wave inⅠ, aVL, V₅-V₆ leads were found among most of patients, only few cases originating from tricuspid annulus anteroseptum group and tricuspid annulus anterolateral group demonstrated qr or qs pattern in aVL lead. 97.53% (79/81) patients demonstrated rS pattern in V₁-V₃ leads with VAs originating from tricuspid annulus free wall, and 9/10 patients demonstrated rS pattern in V₁ lead with VAs originating from anteroseptum, and 97.44% (76/78) patients demonstrated QS pattern in V₁ lead with VAs originating from midseptum and posteroseptum. Precordial lead transition zone was on or behind V₃ for tricuspid annulus free wall group (96.3%, 78/81), but in front of V₃ for tricuspid annulus septum wall group (47.73%, 42/88) (P<0.01). The S wave's amplitude smaller than-1.81 mV in lead V₂ can be used as a cutoff value to identify if PVC/VT is originating from free wall or septum of TA. R wave in inferior wall leads was found among 98.85% (86/87) patients with PVCs/VT originating from right ventricular outflow tract. Conclusion A positive R wave in Ⅰ, aVL, V₅-V₆ leads was found among most of patients with idiopathic ventricular arrhythmias originating from the tricuspid annulus regions, but VAs originating from different portions of tricuspid annulus area have distinct electrocardiographic characteristics.

Objective To investigate the effect of MMP-8 on cornea. Methods Fifteen C57BL/6J healthy mice were selected. The right eyes corneal stroma was injected by 10μL MMP-8 as the experimental group and the left eyes were injected by same amount of normal saline as the control group. At 0,4,8 h, the two-photon microscope second harmonic generation imaging technology was used to scan mice corneal stroma layer by layer in vivo. The obtained images were performed the 3D reconstruction by Imaris software and the signal intensity of the images were calculated. At 4,8 h, the corneal opacity degree was evaluated under slit lamp. At 8 h,mice were killed and corneas were collected to determine the hydroxyproline concentration. Results The cornea stromal fiber signal strengthes at 0 h in the experimental group and control group were (89.7±11.2) and (85.3±7.0),which at 4 h were (14.5±3.4) and (46.6±14. 0) respectively,which at 8 h were (11.0±4.6) and (34.6±12.5) respectively. The cornea stromal signal strength at 4,8 h in the experiemental group was significantly decreased compared with that in the control group (P<0.05) ;the cornea at 4 ,8 h in the experimental group was significantly turbid than that in the control group (P<0.05);the cornea hydroxyproline concentrations detected at 8h in the experiemental group and control group were (0.433±0. 090) μg/mg and (0. 590±0. 133) μg/mg respectively,the experimental group was significantly lower than the control group (F=7. 193,P=0. 014). Conclusion MMP-8 has obvious degradation and destroy effect on mice corneal stroma collagen,which leads to the decrease of corneal opacity.

Objective:To investigate the level of death anxiety and its related factors in patients with newly diagnosed cancer at different stages.Methods:A total of 266 cancer patients in a tertiary oncology hospital were se-lected and investigated at admission(Tl),discharge(T2),and 1 month after discharge(T3).They were assessed with the self-compiled general information questionnaire,Templer's Death Anxiety Scale(T-DAS),Acceptance and Action Questionnaire Second Edition(AAQ-Ⅱ.),and the Chinese version of the Meaning in Life Questionnaire(C-MLQ).The Kruskal-Wallis H test and generalized estimating equation were used to analyze the level of death anxi-ety and its influencing factors in cancer patients at different time points.Results:The Kruskal-Wallis H test showed that the total T-DAS scores and the difference in the scores of the four dimensions were statistically significant at three time points(Ps＜0.01),with the highest T1 score and the lowest T3 score.The results of generalized estima-ting equation showed that male gender(β=1.25),junior high school education(β=2.59),worker(β=1.46),farmer(β=1.67),respiratory(β=1.74),digestive system tumor(β=2.51),and AAQ-Ⅱ score(β=0.23)posi-tively predicted death anxiety in patients with cancer.Head and neck(β=-1.73),breast tumors(β=-1.84),stage Ⅰ(β=-2.58)and stage Ⅱ tumors(β=-2.11),and C-MLQ score(β=-0.15)negatively predicted death anxiety in patients with cancer.Conclusion:Death anxiety in patients with newly diagnosed cancer is highest on ad-mission and gradually relieves after discharge.

OBJECTIVE：To systematically evaluate the effectiveness and safety of Shenfu qiangxin pills combined with chemical medicine conventional therapy in the treatment of chronic heart failure ，and to provide evidence-based reference for clinical drug use. METHODS ：Retrieved from CNKI ，Wanfang database ，VIP，Google Scholar ，PubMed，the Cochrane Library and Embase database ，RCTs about Shengfu qiangxin pills combined with chemical medicine conventional therapy （trial group ） versus chemical medicine conventional treatment （control group ）were collected during the inception to May 12th，2020. After literature screening and data extraction ，the quality of the literatures was evaluated with risk bias assessment tool recommended by Cochrane 5.1.0 system evaluator manual. Meta-analysis and sensitivity analysis were performed by using Stata 14.0 software. RESULTS：A total of 7 RCTs were included ，involving 596 patients. Meta-analysis results showed that the total response rate of trial group was significantly higher than that of control group [OR ＝4.14，95％CI（2.15，7.97），P＜0.000 01]；the results of sub-group analysis according to the different criteria for determining the efficacy showed that the total response rates of trial group determined by Lee integral method and cardiac function grading method were sig nificantly higher than that of the control group （P＜0.05）. After treatment ，N-terminal pro-B-type natriuretic peptide （NT-proBNP） level of trial group was significantly lower than that of control group [OR ＝－1.33，95％CI（－1.55， qq.com －1.11），P＜0.000 01]. Results of sub-group analysis accor- ding to cardiac failure type showed that NT-proBNP level of patients with chronic heart failure in trial group was lower than control group （P＜0.001）. The level of left ventricular ejection fraction （LVEF）in trial group after treatment [WMD ＝5.76，95％CI （5.05，6.47），P＜0.000 01] was significantly higher than control group ；after treatment ，the level of B-type natriuretic peptide [SMD＝－1.61，95％CI（－2.58，－0.54），P＜0.000 01]，left ventricular end-diastolic diameter （LVEDD）level [WMD ＝ －6.06，95％CI（－6.84，－5.27），P＜0.000 01]，left ventricular end-systolic diameter level [WMD ＝－0.52，95％CI（－5.70，－4.33）， P＜0.000 01] were significantly lower than control group. There was no statistically significant difference in the incidence of ADR between 2 groups（P＞0.05）. Results of sensitivity analysis showed that when NT-proBNP ，LVEF level ，LVEDD level after treatment were used as indicators ，there was no significant difference in the analysis results after eliminating heterogeneity source ， compared with before elimination. CONCLUSIONS ：Shenfu qiangxin pills combined with chemical medicine conventional treatment has good efficacy and safety.

OBJECTIVE：To explo re the clinical characteristics of voriconazole-induced neurological ADR and the occurrence of hypokalemia and hyponatremia before ADR. METHODS ：The medical records of 411 patients receiving voriconazole therapy ， who admitted to our hospital from January 2018 to November 2020，were retrospectively analyzed. The general information of all patients，including sex ，age，body weight ，type of infection ，underlying disease ，type of pathogenic fungal infection and administration route of voriconazole ，maintenance dose ，blood drug concentration ，were collected. The basic information of patients with neurological ADR ，including sex ，age，types of infection ，underlying disease ，drug combination ，occurrence time and clinical manifestations ，were collected . The levels of blood potassium ，blood sodium and liver function indexes （ALT，AST， γ-GT，ALP，total bilirubin ，direct bilirubin ）within 3 days before the neurological ADR were also collected. The relationship of neurological ADR with voriconazole trough concentration ，blood potassium and blood sodium levels was analyzed. RESULTS ： Among 411 patients，31（7.54％）patients suffered from neurological ADR ，which were higher in male （64.52％）than in female （35.48％），mainly in patients aged 50 and over （74.20％）. The major infection type was lung infection （96.77％）. Among 31 patients with neurological ADR ，26 patients suffered from neurological ADR after 1-7 days after voriconazole administration ， accounting for 83.87％. Thirty patients received intravenous drip ，accounting for 96.77％. The incidence of neurological ADR in patients with voriconazole trough concentration ＞5.0 μ g/mL （8.99％）was significantly higher than that in patients with trough concentration ≤5.0 μg/mL（3.42％，χ2＝4.91，P＝0.027）. The clinical manifestations of the patients were mainly 023-68766797。E-mail：cheng7zhu@163.com hallucinations（32.35％），irritability（32.35％）and poor sleep （17.65％），etc. Within 3 days before 30 patients，receiving related indexes test ，suffered from neurological ADR ，16 patients（53.33％）had hypokalemia and 12 patients（40.00％） had hyponatremia ，which w ere significantly higher than the incidence of hypokalemia （24.74％，P＝0.001）and hyponatremia （12.89％，P＜0.001）in those without neurological ADR . There were 8，10，7，13，7 and 10 patients with ALT ，AST，ALP， γ-GT，total bilirubin and direct bilirubin increased. In 31 patients with neurological ADR ，the neurological ADR were relieved or disappeared after reducing the dosage or discontinuing voriconazole. CONCLUSIONS ：The neurological ADR of voriconazole mostly occurs 1-7 days after voriconazole administration ，mainly by intravenous drip ，mostly in male and people aged 50 and over. The occurrence of neurological ADR may be related to trough concentration of voriconazole ，and most patients suffer from hypokalemia or hyponatremia before the occurrence of ADR .