Objective To investigate the expression of nicotinic acetylcholine receptor (nAchR) subunits, and the effects of smoking on its expression in peripheral blood mononuclear cells (PBMCs). Methods Blood was collected from 92 healthy male smokers and 36 healthy male non-smokers. Peripheral blood monocytic cells were obtained, and reverse transcription-polymerase chain reaction (RT-PCR) was employed to study the gene expression of 11 nAchR subunits (?2-?7, ?9, ?10, ?2-?4). Results The results showed that the PBMCs of smokers and non-smokers appeared to express ?2, ?5, ?6 and ?7 only out of 11 nAchR subunits. However, the expression of ?5 and ?7 subunits was lower in smokers′ PBMCs than in non-smokers′ (P

Multilocus sequence typing (MLST) is a molecular genotyping method based on nucleotide sequencing. The procedure of this method characterizes isolates of bacterial species using the DNA sequencing of multiple housekeeping genes(usually seven). For each housekeeping gene, the different sequences present within a bacterial species are assigned as distinct alleles.For each isolate, the alleles at each of the loci define the allelic profile or sequence type (ST). MLST has the advantages of being robust (based on genetic data) and electronically portable to generate data that allow rapid and global comparisons between different laboratories. In this paper, the principle, method, data analysis, application, advantages and flaws of MLST are introduced.

Objective To investigate the effect of resveratrol (Res) on paraquat (PQ)-induced acute lung injury (ALI) and mortality in mice and the mechanism of nuclear factor-κB (NF-κB) inflammatory pathway. Methods Sixty-eight healthy male ICR mice with grade SPF were enrolled, among them 20 mice were used for mortality observation (n = 10), and other 48 were used for determination of related parameters (n = 6). The mice were randomly divided into four group s: normal saline (NS) control group, Res control group, PQ group and PQ + Res group. The mice in the latter two groups were subdivided into 6, 24, 72 hours subgroups. The PQ poisoning model of mice was reproduced by one injection of 30 mg/kg PQ intraperitoneally. The mice in PQ + Res group were given 60 mg/kg Res intraperitoneally on the contralateral side after PQ injection. The mice were sacrificed at 6, 24, 72 hours after PQ poisoning, and lung tissue was harvested. The serum levels of tumor necrosis factor-α (TNF-α), interleukins (IL-6 and IL-1β) were determined by enzyme linked immunosorbent assay (ELISA). The pathological changes in lung tissue were observed with electron microscopy. Apoptosis cells in the lung were identified by terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) for the estimation of apoptosis rate. The protein expression of NF-κB p65 was determined by Western Blot. Results Compared with PQ group, the death number of mice at 48, 72, 96 hours in PQ + Res group was slightly decreased (0 vs. 2, 2 vs. 5, 4 vs. 6) but without statistically significant difference (all P > 0.05). Under electron microscope, the lung injury in PQ group was severer than that in NS control group, and Res was found to be able to alleviate the lung injury. Compared with NS control group [(2.45±0.61)%], the apoptosis rate at 6 hours in PQ group was significantly increased [(8.42±1.48)%], and peaked at 72 hours [(21.23±3.47)%]. Res could decrease the apoptosis rate after PQ poisoning [6 hours: (5.56±1.31)% vs. (8.42±1.48)%, 24 hours: (11.14±2.07)% vs. (16.88±2.96)%, 72 hours: (13.28±2.32)% vs. (21.23±3.47)%, all P < 0.05]. The serum levels of TNF-α, IL-6, and IL-1β, and NF-κB p65 in lung tissue were all markedly increased after PQ poisoning, and they were significantly decreased after Res intervention as compared with those of PQ group [TNF-α (ng/L): 2.62±0.29 vs. 4.06±0.74 at 6 hours, 3.98±0.41 vs. 6.79±0.80 at 24 hours, 5.06±0.75 vs. 11.00±0.75 at 72 hours; IL-6 (ng/L): 14.19±1.54 vs. 16.55±1.24 at 6 hours, 13.21±1.37 vs. 19.73±0.85 at 24 hours, 13.72±0.56 vs. 22.45±0.72 at 72 hours; IL-1β (ng/L): 8.54±1.64 vs. 12.59±0.66 at 6 hours, 10.15±0.29 vs. 16.24±1.03 at 24 hours, 16.14±0.70 vs. 19.55±0.56 at 72 hours; 6-hour NF-κB p65: (1.34±0.07) folds vs. (1.86±0.11) folds when the expression in NS control group was represented as 1, all P < 0.05]. Conclusions Res cannot lower the mortality in mice with PQ poisoning, but it seems to be able to attenuate PQ-induced ALI and cell apoptosis. The mechanism responsible for the latter maybe the inhibitive effect of Res on NF-κB p65 translocation and cytokines production.

[Abstract ] Objective The purpose of this study was to observe the effects of dl-3n-butylphthalide (NBP) sodium chloride injection post-processing on the expressions of X-inhibitor of apoptosis (XIAP) and Bcl-2/adenovirus E1B19kDa interacting protein 3 (BNIP3) in the hippocampus CA1 neurons of focal cerebral ischemia reperfusion (IR) rats, and to investigate the brain-protection mechanisms of NBP. Methods A total of65 adult male Sprague-Dawley rats were divided into five groups of equal number, sham op-eration, IR, and low-,medium -and high-dose NBP, according to the random number table. The IR models were established by modified ligation of the middle cerebral artery.The animals in the NBP groups received intra-abdominal injection of NBP at 2, 4, and 6 mg/kg, re-spectively.All the rats were sacrificed at 24 hours after modeling,neurological scores obtained by Zea Longa, the volume of infarction measured by TTC staining, the number of apoptotic cells counted by TUNEL, and the expressions of XIAP and BNIP3 detected by immunohistochemistry and real-time PCR. Results The neural function defect scores were markedly lower in low-, medium-and high-dose NBP groups than in IR model rats (P<0.05), with statis-tically significant differences among the three dose groups (P<0.05).The volume of infarction was remarkably higher in the low-dose than in the medium-and high-dose NBP groups (P<0.05).The number of apoptotic cells in the hippocampus CA1 neurons was de-creased in the NBP groups as compared with the IR models (P<0.05).The XIAP-and BNIP3-positive cells were significantly in-creased in the IR model rats as compared with the sham operation group ([22.31 ±0.94] and [60.13 ±2.59]/HP vs [3.07 ±1.43] and [5.78 ±0.44]/HP, P<0.05).In comparison with the IR models, the NBP-treated rats showed a progressively increased number of XIAP-positive cells in low-, medium-, and high-dose groups ([28.70 ±1.18], [32.79 ±0.88], and [37.01 ±1.24]/HP) (P<0.05) but a decreased number of BNIP3-positive cells in the three dose groups ([52.07 ±1.02], [40.30 ±2.00], and [31.04 ± 0.43]/HP) (P<0.05).Similarly, the expression of XIAP mRNA was up-regulated while that of BNIP3 mRNA down-regulated in the NBP treatment groups as compared with the IR model rats, both in a dose-dependent manner (P<0.05). Conclusion NBP post-processing has a neuroprotective effect on IR rats, which is associated with its impact on the expressions of XIAP and BNIP3.

Objective To investigate clinical features,treatments and prognostic factors of the patients with necrotizing fasciitis caused by vibrio infections and thus provide reference for the early treatment and prognostic assessment.Methods A retrospective analysis was conducted on clinical data of 56 patients with vibrio necrotizing fasciitis admitted to the emergency center of our hospital from May 1995 to June 2011.The clinical characteristics and treatments of the patients were summarized,and differences of clinical factors between the survival group and death group were compared.The possible influencing factors for prognosis were also analyzed.Results The main clinical manifestations included fever (61％),shock (84％) and organ dysfunction,of which renal insufficiency (88％) was the most common,with case fatality of 43％.Early pathological changes of limbs were only local swelling and pain,while skin ecchymosis,tension blood blisters,necrosis and subcutaneous crepitation were the signs of advanced stage.Comprehensive treatment regime including early administration of sensitive antibiotics plus surgical incision and drainage and medicine support was given.A series of factors were significantly different between the survival and death groups including the duration from the presentation of symptoms to hospital admission (P ＜ 0.05 ),limb lesions involving the trunk (P ＜ 0.01 ),creatine kinase level (P ＜ 0.05 ),and emergency incision and drainage ( P ＜ 0.01 ).Conclusions The most prominent clinical manifestations of vibrio necrotizing fasciitis are rapidly progressive local symptoms and signs,and sharp deterioration of systemic conditions.Delayed visiting,severe local lesions,and failure to emergency surgery may be the factors for poor prognosis.

Highly Active Anti-Retroviral Therapy (HAART) has effectively inhibited the prevalence of HIV-1 and reduced the death rate caused by AIDS. In recent years,the emergence of resistance-conferring RT gene mutations in HIV-1 strains has become the major reason for HAART failure. The detection of drug resistance is important for the HAART regimen choice and novel drug development. A novel assay for the detection of HIV-1 RT drug resistance mutations was developed. HIV-1 drug resistance and wild strains in B subtypes were investigated using Two-Step Mutagenically-Separated PCR (MS-PCR),and point mutations including M41L,K70R,K103N,Y181C,T215F were detected. A longer mutant type primer was designed,using microplates hybridization and ELISA technique to detect several point mutations within a mixed mutant-wild type population. The results indicate that the Two-Step MS-PCR is as sensitive and specific as that in the traditional MS-PCR and MS-PCR combined with ELISA can give a good P/N quotient with better sensitivity,low cost,relatively less time consumption and high-throughput screening. It will be used in clinic usage for the detection of HIV-1 drug resistance mutations as well as other point mutations.

HIV-1 envelope glycoprotein gp41,which is a hopeful target for HIV-1 fusion inhibitors,plays a critical role in the fusion of viral and cellular membranes.In order to build up the screening assay of HIV-1 fusion inhibitors targeting gp41,HIV-1 gp41 5-helix and 6-helix were expressed in prokaryotic cells.Gp41 5-helix and 6-helix recombined plasmids were constructed by using PCR,enzyme digestion and ligation taking the clade B HIV-1 genome as a template.The plasmid was transferred into E.coli BL21(DE3)and then induced by IPTG.The expressed protein was purified by affinity chromatography after denaturation and renaturation.The SDS-PAGE analysis was used during expression and purification.Native-PAGE was used to identify the interaction between gp41 5-helix and T-20.The result will be helpful to build up the screening assay of HIV-1 fusion inhibitors targeting gp41.

Objective To discuss the clinical management and significance of the prenatal diagnosis of Fetal Choroid Plexus Cysts(CPC).Methods From May 2004 to March 2007,55 cases of fetal CPC diagnosed by B-ultrasound during second trimester were prospectively studied.Each case was studied regarding fetal chromosome karyotype,disappearance weeks of the cyst,the clinical outcome and follow-up results respectively.Result The cases were diagnosed during 16-25 gestational weeks.The diameters of the cysts varied from 0.2 cm to 2.4 cm.There were 25 cases of bilateral cysts and 30 cases of unilateral or 50 cases of isolated CPC and 5 cases of complicated CPC.The cysts of all cases who continued pregnancy disappeared before 28 weeks.Fetal chromosome karyotypes were obtained in 50 cases.Among them,two cases were 18-trisomy,and one case was 21-trisomy.Five cases were terminated pregnancy because of abnormal chromosome karyotype or malformation during second trimester.One neonate was diagnosed as ventricular septal defect among 50 cases of follow up.Among these six cases,three were from advanced-age pregnant women,five cases were with abnormal fetal structure and five cases were with the diameter of bilateral or unilateral cysts more than 1.0 cm.Conclusion(1)Fetal CPC can be diagnosed during second trimester,and the majority disappear before 28 gestational weeks.(2)High risk factors for fetal abnormal chromosome karyotype may be:advanced-age pregnant women,abnormal structure of fetus,and the diameter of bilateral or unilateral cyst more than 1.0 cm.It is suggested that fetal CPC with the high risks should receive fetal chromosome karyotype test during pregnancy.

OBJECTIVETo explore the expressions of HO-2 and CO in the corpus cavernosum of castrated rats in order to further study the pathogenesis of erectile dysfunction (ED).

METHODSWe randomly divided 72 male SD rats into four groups: normal control, sham operation, castration, and castration + ZnPP. We detected intracavernous pressure (ICP) and penile erection in the basic condition and after apomorphine (APO) induction, determined the expression of the HO-2 protein in the corpus cavernosum by laser scanning confocal microscopy, and measured the level of CO by spectrophotometry during different periods of penile erection.

RESULTSThe ICP in the basic condition and that after APO induction and the rate of penile erection were decreased significantly in the castration group ([11.68 ± 0.69] mmHg, [54.81 ± 3.86] mmHg, and 33.3%) and the castration + ZnPP group ([11.20 ± 0.71] mmHg, [41.17 ± 5.41] mmHg, and 22.2%) as compared with the normal control ([22.83 ± 2.66] mmHg, [66.92 ± 7.77] mm-Hg, and 100%) and the sham operation group ([23.35 ±2.22] mmHg, [70.43 ?7. 22] mmHg, and 100%) (all P <0. 01). After APO induction, ICP in the castration + ZnPP group was remarkably reduced in comparison with that in the castration group (P < 0.01), and so was the expression of the HO-2 protein before and during penile erection in the castration (445.4 ± 23.7 and 847.4 ± 35.0) and the castration + ZnPP group (390.1 ± 29.7 and 526.0 ± 52.5) compared with the normal control (512.7 ±57.4 and 1145.2 ± 89.8) and the sham operation group (583.7 ± 8.0 and 1016.3 ± 79.8), the expression of the HO-2 protein significantly decreased in the castration group (445.4 ± 23.7 and 847.4 ± 35.0) (P < 0.05 or 0.01), markedly lower in the castration + ZnPP than in the castration group during penile erection (P < 0.01) but with no significant differences among the four groups after it. Before, during and after penile erection, the levels of CO were remarkably decreased in the castration ([20.59 ± 1.01], [32.53 ± 1.26], and [18.71 ± 1.22] x 10(-7) nmol/L) and the castration +ZnPP group ([12.52 ± 1.05], [21.90 ± 1.02], and [16.56 ± 0.55] x 10(-7) nmol/L) as compared with the normal control ([26.76 ± 1.41], [48.25 ± 1.01], and [27.10 ± 1.58 ] x 10(-7) nmol/L) and the sham operation group ([25.41 ± 2.09], [ 47.90 ± 1.22], and [25.67 ± 1.20] x 10(-7) nmol/L) (P < 0.05 or 0.01), significantly lower in the castration + ZnPP than in the castration group during penile erection (P < 0.01).

CONCLUSIONDecreased expressions of HO-2 and CO may correlate with erectile dysfunction in castrated rats.

Animals ; Apomorphine ; pharmacology ; Carbon Monoxide ; metabolism ; Dopamine Agonists ; pharmacology ; Erectile Dysfunction ; etiology ; Humans ; Male ; Molecular Chaperones ; metabolism ; Orchiectomy ; Penile Erection ; drug effects ; Penis ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To study the first-time killing efficacy and the chain-killing efficacy of four gel baits against Blattella germanica: 1% chlorpyrifos, 0.05% fipronil, 2.15% imidacloprid, and 0.5% dinotefuran and provide a basis for drug selection in controlling Blattella germanica. Methods Laboratory killing efficacy test was conducted according to the national standard GB/T 13917.7-2009 and the chain-killing efficacy test was conducted for three rounds.The first round of chain efficacy test was conducted by feeding the cockroaches killed in the laboratory efficacy test, and each next round by feeding the cockroaches killed in the last round.Median lethal time (LT₅₀), 95% confidence limit, and toxicological regression equation of each test were calculated by software DPS V9.01. Results The LT₅₀ of the efficacy test with 1% chlorpyrifos gel bait was 0.745 5 (0.603 4-0.890 3) d.The LT₅₀ of the first, second and third chain experiments increased by 3.30, 2.18 and 2.76 times, respectively.The LT₅₀ of the efficacy test with 0.05% fipronil gel bait was 0.846 5(0.464 7-1.228 0)d, and increased by 5.42, 2.09 and 1.48 times, respectively, in the first, second and third chain experiments.The LT₅₀ of the efficacy test with 2.15% imidacloprid gel bait was 3.192 1(2.865 0-3.506 0)d, and increased by 1.13, 1.65 and 1.15 times, respectively in the first, second and third chain experiments.The LT₅₀ of the efficacy test with 0.5% dinotefuran gel bait was 0.997 1(0.805 8-1.191 6) d, and increased by 3.85, 1.37 and 1.78 times, respectively in the first, second and third chain experiments. Conclusion In the laboratory killing efficacy test, 1% chlorpyrifos, 0.05% fipronil, and 0.5% dinotefuran gel baits are better than 2.15% imidacloprid gel bait.In the chain-killing efficacy test, 2.15% imidacloprid and 0.5% dinotefuran gel baits are better than 1% chlorpyrifos and 0.05% fipronil gel baits.Based on our results, we recommend the use of 0.5% dinotefuran gel bait for comprehensive and sustained killing effect.