Objective： To construct a knockout fliY gene mutant strain of Campylobacter jejuni for determining the role of FliY protein in flagellar movement related to bacterial motility，chemotaxis and colonization.
Methods： The plasmid pBluescript-Ⅱ-SK was used to construct the suicide plasmid; according to homologous exchange principle，the suicide plasmid was utilized to generate fliY gene knockout mutant(fliY) in Campylobacter jejuni strain NCTC11168.The fliY mutant strain was identified by PCR，sequencing and Western blotting.The chemotactic and colonizing abilities of fliY mutant were determined by colony migration test and bacterial chemotactic test in vitro, and colonization test in jejunum of mice.
Results： The fliY^- mutant strain showed a growth curve in medium similar to that of wild-type strain.PCR，sequencing and Western blotting assay confirmed that the fliY gene in fliY^- mutant was deleted.Compared to the wild-type strain，the colonies of fliY^- mutant on semisolid plate were much smaller(P<0.05)，the chemotactic ability of fliY mutant towards sodium deoxycholate and bovine bile was significantly attenuated(P<0.05)，and the number of fliY mutant(CFU) in jejunal tissue specimens of the infected mice was significantly decreased(P<0.05).
Conclusion： The function of C.jejuni fliY gene refers to controlling flagellar movement，which is involved in bacterial chemotaxis and colonization.

Objective： To construct a prokaryotic expression system of groEL gene of Leptospira interrogans serogroup Icterohaemorrhagia serovar Lai strain Lai，and to determine the immunoprotective effect of recombinant GroEL protein(rGroEL) in LVG hamsters.
Methods： The groEL gene was amplified by high fidelity PCR and the amplification products were then sequenced.A prokaryotic expression system of groEL gene was constructed using routine genetic engineering technique.SDS-PAGE plus Bio-Rad Gel Image Analyzer was applied to examine the expression and dissolubility of rGroEL protein while Ni-NTA affinity chromatography was used to extract the expressed rGroEL.The immunoprotective rate in rGroEL-immunized LVG hamsters was determined after challenge with L.interrogans strain Lai.The cross agglutination titers of sera from immunized hamsters with different L.interrogans serogroups were detected using MAT.
Results： The nucleotide and amino acid sequences of the cloned groEL gene were the same as those reported in GenBank.The constructed prokaryotic expression system of groEL gene expressed soluble rGroEL.The immunoprotective rates of 100 and 200 μg rGroEL in LVG hamsters were 50.0% and 75.0%，respectively.The sera from the rGroEL-immunized LVG hamsters agglutinated all the L.interrogans serogroups tested with different levels.
Conclusion： The GroEL protein is a genus-specific immunoprotective antigen of L.interrogans and can be used to develop an universal genetically engineering vaccine of Leptospira.

Objective： To investigate the vasodilating effect of capsaicin(CAP) on rat mesenteric artery and its mechanism.
Methods： The third branch of the superior mesenteric artery in male Sprague-Dawley rat(250-350 g) was excised，the periadventitial fat and connective tissue were removed and the mesenteric artery was dissected into 2 mm rings.Each ring was placed in a 5 ml organ bath of DMT 610M system and the tension was recorded.
Results： CAP(10^-9-10^-5 mol/L) relaxed endothelium-intact and endothelium-denuded mesenteric artery pre-constricted by phenylephrine(10^-5 mol/L)，and the vasodilation in endothelium-intact mesenteric artery was stronger than that in endothelium-denuded one.Pretreatment with either L-NAME(3×10^-4 mol/L)，an inhibitor of nitric oxide synthase(NOS)， or CGRP8-37(2×10^-6 mol/L)，an antagonist of calcitonin gene-related peptide(CGRP)，for 30 min significantly attenuated the relaxation of endothelium-intact mesenteric artery induced by CAP.CGRP(10^-10-3×10^-8 mol/L) relaxed endothelium-intact and endothelium-denuded mesenteric artery pre-constricted by phenylephrine，and the vasodilation in endothelium-intact mesenteric artery was stronger than that in endothelium-denuded one.Substance P did not relax the mesenteric artery pre-constricted by phenylephrine.
Conclusions： CAP has partial endothelium-dependent relaxation effect on rat mesenteric artery，which may be mediated by activating the endothelial NOS-NO pathway.The endothelium-independent relaxation in rat mesenteric artery induced by CAP may be mediated by CGRP.

Objective： To investigate the role of phospholipase C(PLC）in cytoskeleton rearrangement of mouse dendritic cells invaded by Mycobacterium tuberculosis.
Methods： Mouse dendritic DC2.4 cells were co-cultured with Mycobacterium tuberculosis H37Rv.F-actin of DC2.4 cells were strained with palloidin-TRITC，the microtubule was stained with anti-β-tubulin monoclonal antibody and FITC-conjugated affnipure anti-mouse IgG.The changes of cytoskeleton in DC2.4 cells induced by Mycobacterium tuberculosis H37Rv were determined by fluorescence microscopy and the rates of F-actin rearrangements were calculated.The expressions of PLC in cytoplasm and cytomemberane of DC2.4 cells were measured by ELISA.DC2.4 cells were pretreated with PLC inhibitor U73122，then F-actin rearrangements induced by invasion of Mycobacterium tuberculosis were observed.
Results： Bacterial invasion was observed while DC2.4 cells were co-incubated with Mycobacterium tuberculosis H37Rv for 2 h.The rates of invasion were（26.1±4.5）%,（39.9±5.6）%,（51.2±5.9）%,（57.9±6.1）% and（63.9±6.8）% at 4，6，8，10 and 12 h of co-culture，respectively; while those were（13.6±3.1）%,（14.2±3.9）%,（15.1±4.3）%,（16.8±4.0）% and（18.3±5.2）% after blocked by PLC，respectively.The rates of the F-actin rearrangements at 2，4，6，8，10 and 12 h after DC2.4 cells were invaded by H37Rv were（26.9±1.5）%,（59.3±2.8）%,（72.7±4.8）%,（78.2±5.9）%,（63.3±2.9）% and （43.2±2.6）%，respectively; while those were（18.5±1.2）%,（22.3±1.7）%,（23.6±2.5）%,（24.8±2.3）%,（22.3±1.3）% and （23.8±1.8）% after blocked by PLC，respectively.There were no changes of the microtubule observed in DC2.4 cells at the same time points.The rates of the F-actin rearrangements before blocked by PLC were higher than those after PLC blockade at 4，6，8 and 10 h（P＜0.05）.The expressions of PLC in cytomembrane in DC2.4 cells increased after 2 h and reached its highest level at 8 h.The PLC inhibitor U73122 inhibited the expressions of PCL in cytomembrane of DC2.4 cells，but not in cytoplasm.
Conclusions： Mycobacterium tuberculosis can provoke to F-actin rearrangements through PLC molecule，which would further lead to Mycobacterium tuberculosis invasion of DC2.4 cells.

Objective： To compare skin sympathetic response(SSR) between patients with generalized anxiety disorder(GAD) and patients with major depression disorder(MDD).
Methods： The latency and amplitude of SSR wave were measured in 30 GAD patients and 30 MDD patients，before and after 8-week treatment of anti-anxiety or anti-depression drugs.Thirty age and sex-matched healthy subjects served as healthy controls(HC).
Results： Before the treatment，the latency of SSR in GAD patients was significantly shorter than that in HC group，while the amplitude was significantly higher than that in the HC(P<0.05).In MDD group，the latency before the treatment was significantly longer than that in the HC，while the amplitude was significantly lower than that in the HC(P<0.05).After treatment，the latency of SSR in GAD group was extended compared to the baseline level，and close to the level of the HC.The amplitude of SSR in GAD group became lower after treatment，but still higher than that of control group.The latency of SSR in MDD patients was significantly shorter after treatment compared to baseline level(P<0.05).In addition，the latency of SSR in MDD group was still longer than that in GAD group(P<0.05); meanwhile，the amplitude of SSR in MDD group was significantly lower that in GAD group(P<0.001).SSR parameters were positively correlated with HAMA and HAMD scores with a correlation coefficient of 0.57 and 0.73，respectively.
Conclusion： There are significant differences in SSR parameters between patients with GAD and patients with MDD，indicating that SSR can be used as an objective index to distinguish anxiety from depression.

Objective： To determine the effects of ginsenosides Rb1(GSRb1) on learning and memory and expression of somatostatin (SS) in the hippocampus and the frontal cortex in rat model of sleep deprivation (SD).
Methods： Rats were randomized into groups of SD 2 d,SD 4 d,SD 6 d,and SD 0 d,while each group was sub-divided into GSRb1 group and normal saline (NS) sub-groups.Rats were intraperitoneal administered with 30 mg/(kg·d） of GSRb1 or NS for 7 d,then the learning and memory abilities were examined by measuring average swimming speed and mean escape latency using Morris maze.Expression of somatostatin was detected with immunohistochemical method and image analysis in the hippocampus and the frontal cortex.
Results： Compared with SD 0 d rats,SD rats exhibited significant decrease in the average swimming speed and increase in the escape latency (P<0.01).The expression of somatostatin in the hippocampus was decreased significantly in SD 2 d,SD 4 d and SD 6 d rats (P<0.05).However,decrease was only observed in SD 4 d and SD 6 d rats in the frontal cortex (P<0.05).Parallel comparison between NS control and GSRb1 treated rats demonstrated that rats treated with GSRb1 in each subgroup exhibited faster swimming speed and shorter escape latency (P<0.05).Meanwhile,the expression of somatostatin was increased in SD 2 d,SD 4 d and SD 6 d rats in the hippocampus and in SD 4 d and SD 6 d rats in the frontal cortex (P<0.05),respectively.
Conclusion： GSRb1 enhances the expression of somatostatin in sleep deprivation rats and subsequently may improve learning and memory abilities of rats.

Objective： To investigate the metabolic changes on proton magnetic resonance spectroscopy (¹H-MRS) in patients with first left basal ganglia infarction and their relationship with depression.
Methods： Twenty-two patients with first left basal ganglia infarction and 10 matched healthy controls were recruited in the study.Patients and controls underwent ¹H MRS scan and the spectrum of N-acetyl aspartic acid (NAA)，choline (Cho) and creatinine (Cr) was recorded at the first week (D7) and 1 month after onset (M1)，respectively.National Institutes of Health Stroke Scale (NIHSS)，Ability of daily life (ADL)，Hamilton Depression Scale (HAMD) were evaluated at D7，M1 and M6 after onset.The patients were classified into two groups: post-stroke depression (PSD) and non-post-stroke depression (NPSD) according to the result of HAMD.The differences of ¹H-MRS between these two groups were compared and their relationship with NIHSS and ADL was analyzed.
Results： Among 22 patients 10 were diagnosed as PSD and 12 were NPSD.Compared to controls，NAA/Cr was significantly lower and Cho/Cr was significantly higher (P<0.05）in left prefrontal white matter and left basal ganglia in patients with infarct.Patients with PSD showed a lower NAA/Cr and higher Cho/Cr than NPSD patients (P<0.05).At M1 after onset，NAA/Cr in the left prefrontal white matter was significantly correlated with NIHSS (r=-0.551，P =0.032)，while Cho/Cr was correlated with ADL (r=0.682，P=0.005）in PSD patients.
Conclusion： Metabolic changes shown on 1H-MRS in patients with basal ganglia infarction may predict the occurrence of PSD.

Objective： To investigate the efficacy and safety of intravenous ibutilide for conversion of atrial fibrillation(AF) and flutter(AFL) to sinus rhythm.
Methods： Ninety-nine consecutive patients aged 18-75 y with AF/AFL were included.The duration of arrhythmia was <90 d(1 h-90 d) and ventricular rate was >60 beats/min.Patients were assigned randomly into two groups: 49 patients in ibutilide group received ibutilide 1 mg，then repeated if AF/AFL was not converted after 10 min; 50 patients in propafenone group received propafenone 70 mg，then repeated if AF/AFL persisted after 10 min.Two drugs were diluted by 50 ml of 5% glucose and injected intravenously within 10 min.
Results： Ventricular rates were decreased in both groups.AF/AFL were converted in 34 of 49 patients(69.4%) in ibutilide group and in 22 of 50 patients(44.0%) in propafenone group(P<0.05).The converting time of ibutilide was significantly shorter than that of propafenone\[(16.79±12.31)min vs (36.92±11.38)min,P<0.01\].The most serious adverse effect of ibutilide was non-sustained monomorphic ventricular tachycardia(3/49,6.12%).Transient hypotension and heart pause were the main adverse events in patients who received propafenone，acute left heart failure occurred in one patient of propafenone group.
Conclusions： Intravenous ibutilide is a safe and effective agent for cardioversion of recent-onset AF/AFL.Furthermore，strict processing under electrocardio-monitoring is important.

p53 is considered as the “master regulator” in DNA damage-induced cell apoptosis.However，p53 is the most frequently mutated gene in human cancers (more than 50%).Thus the research of p53-independent pathway in cell apoptosis may ultimately provide new therapeutic opportunities for many cancers.It has been shown that Caspase 2，p73，p63，and NF-κB-related signaling pathways are involved in DNA damage-induced，p53-independent cell apoptosis.This article reviews the recent research progress in these signaling pathways.

RF-amide related peptide(RFRP) is the orthologue of gonadtropin-inhibitory hormone(GnIH) in mammals.The bodies of RFRP cell are located in the dorsomedial nucleus of the hypothalamus(DMH) and the fibers project to preoptic area(POA) and median eminence of the hypothalamus.Its receptor mainly distributes in hypothalamus.RFRP fibers project to GnRH cells to regulate mammalian reproduction axis.This paper reviews the progress of current researches on RFRP in regulation of animal behaviors，including reproduction，food intake，anxiety and stress response.

Pectus excavatum is the common congenital chest wall deformity in children，and Nuss procedure is the conventional surgical treatment for this disease.Nuss procedure is superior to Ravitch correction for it is less harmful in terms of the surgical technique.However，Nuss procedure is associated with severe postoperative pain，thus adequate postoperative analgesia is important.In this review，factors that influence the postoperative pain after Nuss procedure，the pain managements，the related complications and the side effects of drugs are discussed.

With the development of assisted reproductive technology(ART)，more and more hepatitis B virus(HBV)-infected couples have their own children successfully; however，vertical transmission of HBV in ART，especially father-to-child transmission，cannot be avoided.The mechanism of attachment and penetration of HBV into human sperm is still not known.Therefore，understanding the state and mechanism of HBV infection of sperm and the impact of HBV on sperm parameters，following up the ART outcome in man with HBV infection are helpful to solve the fertility problem and to control father-to-child vertical HBV infection.