Objective To generate a comD gene knock-out mutant of Streptococcus pneumoniae,and determine the correlation of comD gene and the bacterial resistance against β-lactam antibiotics and understand the effect of closantel down-regulating comD, comE and comC mRNA levels. Methods A suicide plasmid pEVP3comD was constructed for comD gene knock-out and a comD gene knock-out mutant (comD-)was generated through homologous recombination and insertion inactivation. PCR and immunofluorescence method were used to identify the comD- mutant and real-time fluorescence quantitative PCR was applied to detect the changes of comD, comE and comC mRNA levels before and after closantel treatment in comD-mutant and wild-type strain. Double agar dilution method was performed to determine the sensitivity of comD- mutant and wild-type strain to penicillin G and cefotaxime. Results The comD gene in genome DNA of the generated comD- mutant was inactivated by sequencing and immunofluorescence detection. 50 μ mol/L or 100 μmol/L closantel had a function to down-regulate the comD, comE and comC mRNA levels ( P ＜ 0. 05) whereas 25 μmol/L closantel did not. Both the MIC values of penicillin G and cefotaxime inhibiting comD- mutant were 32 μg/ml which was significantly higher than that of wild-type strain (0.06 μg/ml and 1 μg/ml). Conclusion In this study a comD gene knock-out mutant of S. pneumoniae was successfully generated. There is a close correlation between comD gene and β-lactam antibiotics resistance of S. pneumoniae. Closantel has a function to inhibit the competence formation of S. pneumoniae through down-regulating the transcription levels of comD, comE and comC genes.

Objective To explore the significance of changes of serum adiponectin and testosterone levels in old male patients with type 2 diabetes mellitus(T2DM) and coronary heart disease(CHD).Methods A total of 122 male patients(more than 60 years old)were enrolled into this study,T2DM patients without CHD(T2DM group,n =40);T2DM patients with CHD (CHD group,n=38);control group(n=44,44 cases of physical healthy men over the age of 60 in Qinghai Provincial People's Hospital from November 2014 to September 2015).The serum of fasting blood-glucose,glycosylated hemoglobin(HbAlc) and blood lipid were detected by Roche cobas 800 full automatic biochemical analyzer,the serum concentration of testosterone(T) was determined by the chemiluminesent immunoassay,serum adiponectin(APN)levels were determined by the enzyme linked immunosorbent assay,the serum insulin(FINS)levels were evaluated by chemiluminescence method,according to the steady-state model evaluation method for calculation of insulin resistance (HOMA-IR),the difference among the three sets of the above indexes indicators were compared,and the correlation between APN and other indicators above were analyzed.Results Compared with control group,the serum T and APN levels in the T2DM group and CHD group were significantly decrease,and the CHD group was the most obviously (P＜0.05).Compared with control group,the FPG,FINS,triglycerides(TG),low density lipoprotein cholesterol(LDL-C)and homeostasis model assessment-insulin resistance (HOMA-IR) levels in T2DM group and CHD group were apparently higher,and the CHD group was the most obviously (P＜ 0.05).The serum adiponectin was negative correlated with TG (r=-0.363),LDL-C(r=-0.417),HOMA-IR(r =-0.602),while positively correlated with HDL-C(r=0.485),T(r=0.624).The serum T was negative correlated with LDL-C(r=-0.457),HOMA-IR(r-0.643),while positively correlated with HDL-C(r=0.478),P＜0.05.Conclusion The level of the serum APN and the serumtin T2DM patients with CHD are significantly lower than the patients without CHD,the serum adiponectin and the serum testosterone may promote the development of T2DM with CHD.

To be on the alert on infants with esophageal foreign body, and to pay more attention to the button battery esophageal foreign body, the clinical data of a 12-month-old infant with button battery esophageal foreign body, which was missed diagnosis for up to 4 months, is analyzed. And the related literature is reviewed. An esophagoscopy was carried out to remove the foreign body. A favorable outcome was achieved. When the infants have unexplained gastrointestinal symptoms, we should consider the possibility of an esophageal foreign body. We should pay attention to the button battery due to its highly corrosive to the esophagus. Timely diagnosis, reasonable operation are the keys to cure.
Electric Power Supplies ; Esophagoscopy ; Esophagus ; Foreign Bodies ; surgery ; Humans ; Infant

Cells, Cultured ; Comet Assay ; DNA Damage ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Lymphocytes ; drug effects ; Styrene ; adverse effects

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Leukemia, Myeloid ; pathology ; physiopathology ; Male

Objective To Identify B-cell epitopes on monkey B virus envelope protein gD.Methods Base on bioinformatics software, secondary structure, hydrophilicity, surface accessibility, antigenic index and flexibility of monkey B virus envelope protein gD was analysed and some potential peptide epitopes were forecasted.Then, the interactions between synthetic peptides and BV positive sera were detected by Enzyme-linked immunosorbent assay ( ELISA) .At last, The sensitivity and specificity of synthetic peptides was evaluated used 20 samples of Standard sera by ELISA.Result Seven epitopes were forecasted by Bioinformatics analysis. Four synthetic peptides, sequence as 46 LPPLEQKTD54 , 106 RGAPEATRSDA116 , 291 PELAPEERGTSRTPGD306 and 361 AVYLVRRRGR370 could be reacted with positive sera pool.The sensitivity of 4 synthetic peptides changed form 40% to 70% and the specificity were 100% for 4 synthetic peptides. Conclusion There are at least four linear epitope on B virus gD protein.

Child, Preschool ; Heart Diseases ; physiopathology ; Humans ; Iliac Vein ; physiopathology ; Leukemia, Lymphoid ; complications ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; complications ; Thrombosis ; etiology ; physiopathology

Background:Screening of colorectal cancer (CRC) and colorectal adenoma (CRA), the precursor of CRC, is crucial for CRC prevention.It is believed that opportunistic screening of CRC in high risk population would narrow down the screening scope and save the health resources, and is suitable for the present status in China.Aims:To analyze the performance of high risk factors in predicting CRC/CRA, for exploring the feasibility of opportunistic screening for CRC in high risk population.Methods:A total of 1 862 outpatients and health examination subjects undergoing colonoscopy from Feb.2015 to Aug.2016 at the PLA 281 Hospital were recruited and asked to complete a questionnaire of high risk factors for CRC and CRA before colonoscopy.The questionnaire was designed based on the updated consensus on the screening and management of early colorectal cancer and precancerous lesion in China.Using the results of colonoscopy as gold standard, the predictive performance of high risk factors for CRC/CRA was analyzed.Results:Four hundred and sixty-eight (25.1%) individuals with high risk factors for CRC/CRA were screened out by the questionnaire.The detection rate of CRC in individuals with high risk factors was significantly higher than those without (17.5% vs.0.9%, P=0.000).The sensitivity, specificity, positive and negative predictive values of high risk factors in predicting CRC were 87.2%, 78.2%, 17.5% and 99.1%, respectively, and the missed diagnosis rate was 12.8%.For CRC/CRA, the corresponding figures were 83.2%, 87.0%, 57.3%, 96.1% and 16.8%, respectively.The risk ratios (RR) of high risk factors for CRC and CRC/CRA were 20.35 and 14.78, respectively.Conclusions:Opportunistic screening of CRC in high risk population is feasible and applicable in China under present condition.

Objective To observe the corneal nerve fibres damage in different stage of diabetic retinopathy (DR) with type 2 diabetes.Methods A cross-sectional study.One hundred and twenty eyes of 120 patients with type 2 diabetes served as diabetes group.According to International Clinical Diabetic Retinopathy Disease Severity Scales (2002),diabetes patients were classified into 4 subgroups:patients without diabetic retinopathy (NDR),patients with mild or moderate non-proliferative diabetic retinopathy (mNPDR),patients with severe non-proliferative diabetic retinopathy (sNPDR) and patients with proliferative diabetic retinopathy (PDR),each subgroup has 30 eyes of 30 patients.Another 30 eyes of 30 healthy participants served as control group.All eyes were scanned with HRT3 in vivo corneal confocal microscopy.Images of sub-basal nerve plexus were quantified including nerve fiber length (NFL),nerve fiber density (NFD),nerve fiber branch density (NFB),and nerve tortuosity (NT).The correlations of corneal nerve fiber with age,duration of diabetes and glycated hemoglobin (HbA1 c) were analyzed using Spearman correlation analysis.Results NFL,NFD and NFB were found to be significantly lower in diabetic patients (F=147.315,142.586,65.898;P=0.000,0.000,0.000),NT was significantly greater in diabetic patients (F=39.431,P=0.000),when compared to control group.In diabetic patients,NFL,NFD and NFB were gradually reduced with DR severity,NT was gradually increased with DR severity.While the difference ofNFL,NFD,NFB,NT was not statistically significant between sNPDR and PDR subgroups (P＞.0.05),but was statistically significant between other subgroups (P＜0.05).Spearman correlation analysis results showed that age (r=-0.071,-0.080,0.001,0.100;P=0.391,0.328,0.991,0.224) and HbAlc (r=-0.109,-0.115,-0.126,0.025;P=0.238,0.211,0.169,0.781) had no correlation with NFL,NFD,NFB,NT.Duration of diabetes was negatively correlated with the NFL,NFD (r=-0.212,-0.264;P=0.020,0.004),positive correlated with NT (r=0.261,P=0.004),and had no correlation with NFB (r=-0.119,P=0.194).Conclusions Corneal nerve fiber loss and nerve tortuosity increased were found in patients with type 2 diabetes,and even without diabetic retinopathy.The progress of corneal neuropathy was correlated with the severity of DR,but it was not change significantly between sNPDR and PDR.

Objective:To investigate the expressions of peripheral lymph node addressin(PNAd)andGlcNAc-6-sulfotransferase(GlcNAc6ST)in endometrium and their impacts on implantation.Methods:PNAd expression in endometrium was examined by immunohistochemistry and Western Blot from 75women(12 from healthy women,in proliferative phase;63 from sterile women,of whom,27 were inearly-secretory and 36 in mid-secretory phase).GlcNAc6ST mRNA was examined by real-time PCR in41 sterile women.The 63 sterile women had underwent ⅣF-ET and were consequently divided into clini-cal pregnant(29 cases)and nonpregnant(34 cases)groups.Results:(1)PNAd localized to the mem-brane and cytoplasm of luminal and glandular epithelia.Staining was patchy and much less intense duringthe proliferative phase than during the secretory phase.In Western Blot of PNAd,four bands appeared,which were Sgp200,CD34,MAdCAM-1,GlyCAM-1 respectively,and each was positively correlatedwith the others significantly.The former three molecular levels were significantly higher during the secre-tory phase as compared with the proliferative phase.Message RNA of GlcNAe6ST was positive in all ca-ses and showed no correlation with any component of PNAd.(2)The expressions of CD34 and GlyCAM-1,but not Sgp200 and MAdCAM-1,were significantly higher in pregnant women than in nonpregnantones.However,the GlcNAc6ST mRNA level did not differ between groups.(3)No significant differ-ence was found in female age,methods of fertilization,thickness of endometrium on day hCG,cumulativeembryo score(CES)and mean score of transferred embryo(MSTE)between the groups.Conclusion:PNAd expression in the human endometrium fluctuates with the menstrual cycle.Elevated CD34 and Gly-CAM-1 during the secretory phase might be stimulative factors for embryo implantation.Defect in PNAdexpression may account for a portion of unexplained infertility.