Abstract: Objective　To investigate the influences of notoginsenoside R1 (NGR1) on cell injury and Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway of alveolar epithelial cells infected by Klebsiella pneumoniae (Kp). Methods　A549 cells were grouped into five groups: control group (C group), infection group (Infect group), infection + low NGR1 group (Infect + L-NGR1 group), infection + high NGR1 group (Infect + H-NGR1 group), and infection+high NGR1+JAK2/STAT3 pathway inhibitor group (Infect+H-NGR1+SD-1029 group). Cell proliferation was measured using CCK8; ELISA kits were applied to detect the contents of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and interferon γ (IFN-γ) in the culture medium; flow cytometry was applied to detect apoptosis; RT-qPCR was applied to detect the expressions of JAK2/STAT3; Western blot was applied to detect JAK2/STAT3 pathway, autophagy protein microtubule-associated protein 1 light chain 3 (LC3), autophagy-relatedgene5 (Atg5), autophagy-related gene (Atg) 6 (Beclin-1), apoptosis protein B-cell lymphoma 2 (Bcl-2), Bcl-2-accociated protein (Bax), cysteinyl aspartate specific proteinase (cleaved-caspase-3) proteins expression. Results　Compared with the C group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the mRNA relative expressions and protein phosphorylation levels of JAK2, STAT3 in the Infect group were obviously decreased (P<0.05); the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-caspase-3 were obviously increased (P<0.05). Compared with Infect group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the mRNA relative expressions and protein phosphorylation levels of JAK2, STAT3 in the Infect+L-NGR1 group and Infect+H-NGR1 group were obviously increased (P<0.05); the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-Caspase-3 were obviously decreased (P<0.05). Compared with Infect+H-NGR1 group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the protein phosphorylation levels of JAK2, STAT3 in the Infect+H-NGR1+SD-1029 group were obviously decreased (P<0.05), and the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-caspase-3 were obviously increased (P<0.05). Conclusions NGR1 can activate the JAK2/STAT3 signaling pathway, promote autophagy of alveolar epithelial cells, and inhibit Kp-induced inflammatory injury and apoptosis of alveolar epithelial cells.

Objective To evaluate the expression and clinical significance of plasma circulating microRNA-23b in patients with myasthenia gravis (MG).Methods Plasma samples from 32 MG patients and 32 healthy people were collected in the Second Affiliated Hospital of Harbin Medical University from October 2015 to August 2016. Expression of microRNA-23b was detected by quantitative reverse transcription polymerase chain reaction .The relationship among microRNA-23b expression level, clinical features and quantitative myasthenia gravis score ( QMG ) was analyzed. Results The expression of circulating microRNA-23b in plasma was significantly higher in MG patients (4.587 (3.654, 5.423)) compared with control group (2.889 (1.968, 4.027), Z=-4.169, P<0.01), meanwhile it was positively related to QMG (r=0.661,P<0.01).But the expression of circulating microRNA-23b had no statistically significant difference in MG subgroups .Conclusion Plasma circulating microRNA-23b is highly expressed in MG and has positive correlation with MG severity .

Objective To investigate the expression level of thymus microRNA-27a-3p in patients with myasthenia gravis (MG) and to explore the pathogenesis of MG.Methods Thymus tissue samples from 36 cases were collected from December 2014 to February 2015 in the Second Affiliated Hospital of Harbin Medical University.Nineteen thymus tissue samples of MG group were collected from department of chest surgery,17 thymus tissue samples of control group were collected from department of chest surgery or congenital heart disease patients from department of cardiac surgery.The expression of microRNA-27a-3p in the thymus from 36 patients was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR),using U6 as housekeeping control.The Wilcoxon Rank-Sum test was used to analyze the relative expression of microRNA-27a-3p of the two groups.Spearman rank correlation was used to determine the correlation coefficient between microRNA-27a-3p and Quantitative Myasthenia Gravis Score (QMGS).Results (1) The expression level of microRNA-27a-3p in thymus was significantly higher in MG group (0.195(0.049,0.714)) compared with control group (0.045(0.004,0.088),Z =-2.646,P =0.008).(2) Nineteen MG patients were included in the study,out of which 7 were ocular myasthenia gravis (OMG) patients and 12 were generalized myasthenia gravis (GMG) patients.The expression of microRNA-27a-3p in GMG patients (0.493 (0.157,1.123)) was significantly higher than that in OMG patients (0.035 (0.008,0.103),Z =-2.620,P =0.009).(3) There was a positive correlation between the expression of microRNA-27a-3p and QMGS (r =0.576,P =0.010).Conclusions The expression of microRNA-27a-3p in thymus is significantly up-regulated in the patients with MG.MicroRNA-27a-3p may be associated with MG severity and significantly elevated in GMG patients compared with OMG patients.

Objective To investigate the association between myasthenia gravis (MG) and single nucleotide polymorphisms (SNPs) of PTPN22 + 1858C/T,CTLA-4 (+ 49A/G;-1772C/T;-1661A/G),KRAS(rs9226),BCL2(rs4987855) and IGF-1R(rs34804698) genes.Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was adopted to detect the gene types of SNPs in 76 MG patients who were enrolled in the Second Affiliated Hospital of Harbin Medical University from July 2011 to June 2015 and 59 healthy blood donors.Results In MG patients,the frequences of CTLA-4 +49A/G(rs231775) (57.9％) and-1772C/T (rs733618) (43.4％) were higher than that in the healthy controls (22.1％) (x2 =35.252,P =0.000; x2 =4.098,P =0.043).The frequence of CTLA-4 +49A/G in MG patients combined with thymoma (25.6％) was higher than other subgroups (thymic hyperplasia group:13.8％; normal thymus group:18.4％)(x2 =7.564,P=0.006; x2 =7.155,P=0.007).Meanwhile,the frequence of the C-1772 allele was higher in thymoma group (19.7％) compared with other two groups (thymic hyperplasia group:9.86％ ; normal thymus group:13.8％) (x2 =5.331,P =0.021 ;x2 =4.411,P =0.036).However,the other SNPs were not associated with the risk of MG.Conclusion There are associations of MG with CTLA-4 + 49A/G and-1772C/T SNPs,but not with PTPN,KRAS,BC12 and IGF-1R SNPs.

Objective To explore the relationship between left ventricular diastolic function and the dynamic changes of myocardial ultrastructure and argyrophilic fiber in diabetic rats on the different periods of lesions(week 4,12 and 24).Methods The diabetes mellitus(DM)in healthy male Sprague-Dawley rats was induced by a single injection of streptozotocin(STZ,Sigma)into intraperitoneal at a dose of 65 mg/kg body weight.The left ventricular diastolic function was measured by Color Doppler Flow Imaging-Pulsed Wave(CDFIPW)and Doppler Tissue Imaging(DTI)echocardiography.Heart tissue at the apex was obtained rapidly for transmission electron microscope study.Argyrophilic staining was used in the study of argyrophilic fiber volume fraction(APFVF)in heart interstitial tissue.Results Diastolic dysfunction of left ventricle was detected in STZinduced diabetic rats by CDFI-PW(E/A ＜ 1)at week 4,and progressed gradually.Pseudonormal filling (E/A ＞ 1)was found in diabetic rats at week 24,which could be identified by DTI(Ea/Aa ＜ 1).Diastolic function of normal rats was not impaired(E/A ＞ 1 and Ea/Aa ＞ 1).Transmission electron microscopy revealed a spectrum of subcellular remodeling in myocardium which was characterized by myofibril content decrease,disorganization,mitochondrial degeneration,sarcoplasmic reticulum,structural disorder.Compared with the control group,APFVF in myocardium was increased significantly in diabetic rats(P ＜ 0.05).Conclusion The diastolic dysfunction in STZ-induced diabetic rats correlates with damage of ultrastructure and increase of myocardial argyrophilic fiber.

Cytoplasmic male sterility is an important way to utilize wheat heterosis. The purpose of thisstudy was to identify cytoplasmic type of three wheat male sterile lines. Amplified fragment length polymorphism (AFLP) marker technique was used to analyze the wheat mitochondrial DNA. We isolated mitochondria by differential centrifugation and density gradient ultracentrifugation. The results show that the extracted mitochondrial DNA was pure. It was suitable for PCR and genetic analysis. We got 4 pairs of specific primers from 64 primers combinations. Primer E1/M7 amplified 3 specific fragments in ms(Kots)-90-110. Primer E4/M2 generated 2 specific fragments in ms(Ven)-90-110. Primer E7/M6 amplified 2 specific fragments in ms(S)-90-110. Primer E6/M4 produced 2 specific fragments in ms(Kots)-90-110. Four specific primers could be used to identify three cytoplasmic types of Aegilops kotschyi, Ae. ventricosa and Triticum spelta. It provided the molecular basis to further study the mechanism of wheat cytoplasmic male sterility.
Amplified Fragment Length Polymorphism Analysis ; methods ; Cytoplasm ; metabolism ; DNA, Mitochondrial ; genetics ; DNA, Plant ; genetics ; Gene Expression Profiling ; Genotype ; Plant Infertility ; genetics ; Triticum ; genetics

Cerebrovascular disease is one of the leading causes of death and disability across the world. Vascular homeostasis and recovery is critical to the body. Klotho is an important endogenous pleiotropic protein involved in a variety of pathophysiological processes,such as aging and calcium and phosphorus metabolism,which is closely associated with vascular disease. Recent clinical studies have demonstrated close links between low Klotho levels and various risk factors for cerebrovascular disease,suggesting that Klotho has the potential of predicting the risk of ischemic and hemorrhagic cerebrovascular diseases and poor prognosis. Basic studies have also shown that Klotho plays a crucial role in the maintenance of vascular homeostasis. Klotho can promote the production of nitric oxide,inhibit the expression of inflammatory factors,and mediate antioxidant and anti-aging effects,which can delay atherosclerosis and vascular calcification. Therefore,Klotho may serve as a new target for the prevention and treatment of cerebrovascular disease. This review provides an overview of the relationship between Klotho and cerebrovascular disease and its potential biological mechanisms.

Objective To improve hand hygiene (HH)compliance of health care workers (HCWs)in neonatal in-tensive care units(NICUs)in China through a series of intervention measures.Methods A multicenter study was conducted,17 tertiary first class hospitals in 9 provinces and cities were selected,HH compliance of HCWs in these hospitals were investigated every month according to HH investigation method of World Health Organization.From October 1 ,2013 to March 31 ,2014 was pre-intervention stage;from April 1 ,2014 to September 30,2014 was post-intervention stage,timely feedback and intensified training were conducted at post-intervention stage.Results HH compliance rates of HCWs before and after intervention were 80.29% and 80.85% respectively,there was no sig-nificant difference (P >0.05).HH compliance rates of HCWs in different sizes of ICUs were significantly different before and after intervention (all P <0.05),in NICUs with<20 beds and 20-30 beds after the intervention were both significantly higher than before intervention (both P <0.05 ),while >30 beds were significantly lower than before intervention(P <0.001 );HH compliance rate of cleaners increased from 58.82% before intervention to 68.09% after intervention (P <0.05 );Of different hand hygiene indications,except before clean/aseptic task, compliance to the other HH indications were significantly different between before and after intervention(all P <0.05).Conclusion HH compliance is high among HCWs in NICUs in China,intervention measures,such as inten-sified training and timely feedback have certain influence in compliance to HH among HCWs at different sizes of ICUs,of different occupations,and at different HH indications.

Objective To investigate the status and risk factors of surgical site infection (SSI)in hospitals in Chi-na,so as to provide theoretical basis for the prevention and treatment of SSI.Methods Four types of surgeries (colorectal surgery,abdominal hysterectomy,femoral neck repair surgery,and vascular surgery)in 29 hospitals were monitored prospectively,risk factors for SSI were analyzed.Results A total of 6 309 surgical procedures were investigated,incidence of SSI was 1 .60%.Incidences of SSI in patients receiving colorectal surgery,abdominal hys-terectomy,femoral neck repair surgery,and vascular surgery were 4.47%(74/1 655 ),1 .03%(22/2 139),0.21 %(5/2 372),and 0.00% (0/143 )respectively.The incidences of SSI were different among different regions (χ2 =114.213,P <0.05).The most common SSI was superficial incisional infection,the next was deep incisional infec-tion.The major pathogens causing SSI were Escherichia coli ,Enterococcus spp .,coagulase negative staphylococ-cus ,Staphylococcus aureus ,and Klebsiella pneumoniae .The independent risk factors for SSI were male patients, long duration of surgery,and high NNIS score.Conclusion The risk of SSI is varied with different types of surger-ies.Male,long duration of surgery,and high NNIS score can increase the risk of postoperative SSI.

Objective To explore the incidence of surgical site infection (SSI)and compliance to bundled interven-tion measures,and evaluate the effect of bundled interventions on controlling SSI.Methods From October 2013 to September 2014,three types of surgeries (colorectal surgery,abdominal hysterectomy,and femoral neck repair sur-gery)in 29 hospitals in China were monitored,October 2013 to March 2014 was baseline investigated stage,April 2014 to September 2014 was intervention stage.Results A total of 6 166 episodes of surgeries were monitored,the incidence of SSI was 1 .64%,incidence of SSI following colorectal surgery,abdominal hysterectomy,and femoral neck repair surgery were 4.47%,1 .03%,and 0.21 % respectively.The P 75 time of three types of surgeries were 3,2,and 2 hours respectively.Compared with the baseline stage,the compliance to most intervention measures im-proved after intervention,the largest increase in the compliance to interventions was disinfection with chlorhexidine-containing disinfectant at surgical sites of colorectal surgery (increased by 29.09%),followed by preoperative shower of femoral neck repair surgery (increased by 26.24%),preoperative shower of colorectal surgery(increased by 22.95%),and skin preparation on the day of operation (increased by 20.75%).Incidences of SSI in three types of surgeries were not significantly different before and after intervention(all P >0.05).Conclusion The incidences of SSI are different among different types of surgeries,the compliance to most bundled intervention measures has im-proved to some extent after intervention,but effectiveness of intervention measures needs to be further observed.