Objective To study the effect of shenfu injection on the neuron apoptosis in hippocampal CA1 region of newborn with hypoxic-ischemic brain damage(HIBD).Methods The experiment included 2 parts.One was to measure the apoptosis rate by the flow cytometry,and the other was to investigate the expression of Bcl-2 and Bax of neurons in left hippocampal CA1 region.Models of postnatal 7-day Sprague-Dawley(SD)rats with HIBD were established,and were equally divided into 4 groups:sham operation(group S),control group(group C),shenfu injection pretreatment(group P),and shenfu injection treatment(group SF).The neuron apoptosis rate in hippocampal CA1 region in every group was measured at 2 h before and 2,12,24 h,3,7,14 and 28 d after hypoxic ischemic(HI) insult.The expressions of Bcl-2 and Bax were performed by immunohisochemistry.Results The apoptosis of neuron in hippocampal CA1 region in group P and group SF after HI insult was significantly less than that of group C(P

Objective To clone and express the encoding sequence of glyceraldehydes-3-phosphate dehydrogenase(GAPDH)from periodic Brugia molayi(Bm).Methods Total RNA was extraeted from periodic Brugic malayi.The BmGAPDH gene was amplified by RT-PCR.The PCR product was cloned and then subeloned into pcDNA3.1(+)vector.The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification,and were transformed into COS-7 cell subsequently.The expressed protein was identified by SDS-PAGE.Results BmGAPDH mRNA was highiy expressed in transfected COS-7 cell.The deduced amino acid sequence was identical with that of BmGAPDH.The recombinant pnotein wag about Nr 43 000.Conclusion The recombinant plasmid peDNA3.1(+)-BmGAPDH has been constructed and the protein has been expressed correctly.

Objective To understand clinical distribution and antimicrobial resistance of clinically isolated Serratia marcescens(S .marcescens ),and provide basis for rational use of antimicrobial agents,as well as prevention and control of infection.Methods 427 S .marcescens strains isolated between January 1 ,2012 and December 31 ,2015 were analyzed,antimicrobial susceptibility testing were performed by disk diffusion method.Results 427 S . marcescens strains were mainly from respiratory tract (70.26%),among which the majority were from sputum (64.87%).S .marcescens were primarily from intensive care unit(ICU,19.44%),department of integrated tradi-tional Chinese and Western medicine(15.46%)as well as rehabilitation department (13.58%).The resistance rates of S .marcescens to cefoperazone/sulbactam,ertapenem,cefepime,ceftazidime,amikacin,imipenem,levofloxacin, and piperacillin/tazobactam were all<10%;resistance rates to ciprofloxacin,gentamicin,tobramycin,ceftriaxone, sulfamethoxazole/trimethoprim (SMZ/TMP),and aztreonam were 10%-30%.Difference in the resistance rates of S .marcescens to cefoperazone/sulbactam,ciprofloxacin,ceftriaxone,amikacin,aztreonam,and SMZ/TMP dur-ing 4 years were statistically significant (P <0.05).In 2012-2013,resistance rates of S .marcescens to cefopera-zone/sulbactam,ciprofloxacin,ceftriaxone,aztreonam,and SMZ/TMP increased obviously,then resistance rates tend to be stable,while resistance rates to cefoperazone/sulbactam decreased.Conclusion Susceptibility of S.marcescens to most antimicrobial agents are high,but resistance had increasing tendency;susceptible rates of S .marcescens to ertapenem,ceftazidime,levofloxacin,and piperacillin/tazobactam are all high,and can be used as the empirical medication for the treatment of related infection.

Objectives Selecting and constructing the biofilm -model of Pseudomonas aeruginosa in vitro.Observing the antibacterial activity of using Micafungin alone , or combined with Meropenem against Pseudomonas aeruginosa ( plankton-grown and biofilm-grown ) . Methods Ten clinical isolates of Pseudomonas aeruginosa were collected in July 2012, constructing the biofilm-model by microwell plate from Xiangya Hospital, Central South University.The ability of biofilm-formation of these strains was estimated by crystal violet colorimetric method, and optical microscope was used to observe the shape of the biofilm .MICs of Micafungin and Meropenem against plankton -grown and biofilm-grown Pseudomonas aeruginosa were tested by broth microdilution method, and the changes of MICs were compared.Using broth microdilution method, and connecting with the crystal violet colorimetric method , to observe the antibacterial effect of using Micafungin alone, or combined with antibiotics in the inhibition of the biofilm formation and destruction of mature biofilm of Pseudomonas aeruginosa.SPSS18.0 and t-test were used in comparing the differences between both treatment group and control group.P <0.05 showed the difference was statistically significant . Results Ten strains of Pseudomonas aeruginosa were successful in forming biofilms.Comparing with their planktonic counterparts, biofilms became more resistant to Meropenem , with the MIC raised 4-128 times. However, MIC of Micafungin could not be measured.Micafungin can inhibit the formation of biofilm in 9 experimental strains (PA1-PA9), where the minimum effective concentration of Micafungin were 156.25, 625, 10 000, 2 500, 1 250, 2 500, 1 250, 625 and 10 000 mg/L respectively.The absorbance values of the minimum effective concentration group and its positive growth control group were 0.342 ±0.020 vs 0.491 ±0.027, 0.512 ±0.018 vs 0.627 ±0.043, 0.862 ±0.021 vs 1.155 ±0.027, 0.731 ±0.028 vs 0.863 ± 0.017, 0.311 ±0.003 vs 0.447 ±0.021, 0.435 ±0.021 vs 0.597 ±0.011, 0.520 ±0.012 vs 0.605 ± 0.027, 0.611 ±0.059 vs 0.734 ±0.017, 0.223 ±0.011 vs 0.343 ±0.037 respectively, where the P values were 0.02, 0.03, 0.00, 0.01, 0.01, 0.00, 0.03, 0.01 and 0.03 respectively.The differences are statistically significant.Micafungin can damage the mature biofilm of 7 strains (PA1, PA2, PA4 -PA8), where the minimum effective concentration of Micafungin were 2 500, 2 500, 5 000, 2 500, 5 000, 2 500, 5 000 mg/L respectively.The absorbance values of the minimum effective concentration group and its positive growth control group were 1.459 ±0.014 vs 1.534 ±0.020, 1.279 ±0.020 vs 1.431 ±0.007, 1.365 ±0.024 vs 1.467 ±0.065, 1.322 ±0.028 vs 1.530 ±0.090, 0.920 ±0.004 vs 1.047 ±0.013, 1.860 ±0.005 vs 1.953 ±0.055, 1.407 ±0.005 vs 1.553 ±0.045 respectively, where the P values were 0.01, 0.01, 0.02, 0.01, 0.00, 0.03, 0.02.The difference is statistically significant.Micafungin combined with Meropenem applied in multiple drug resistant strains , which can inhibit the formation of biofilm better.Conclusions Micafungin can inhibit the formation Pseudomonas aeruginosa biofilm and damage the mature biofilms.Micafungin combined with Meropenem can act on multiple drug resistant strain , which may get a higher inhibition rate of the biofilm.

Field studies were conducted for a season to determine potential for alterations in the rhizosphere fungi quantity dynamics and dominant populations of 4 wild sugarcane clones(S.spontaneum L.) with different UV-B sensitivity under an enhanced ultraviolet-B(UV-B,280 nm～310 nm) radiation.The quantity of rhizosphere fungi was most in tillering stage,second in seedling stage and lest in elongating stage and maturing stage,the sequence wasn’t alternated by UV-B radiation.The rhizosphere fungi quantity of tolerant clone was obviously increased and greater than the sensitive clone under the enhanced UV-B radiation,however,the number of dominant populations decreased and Penicillium was the dominant population during the periods of 4 wild sugarcane clones.

Objective To study the influence of nursing intervention on compliance behavior and living quality of the patients with CHF. Methods The CHF patients who were treated in our hospital during January 2006 to January 2008 were selected as the study objects. All of the patients were divided into control group and intervening group. The patients in control group were instructed with the conventional guidance when leaving hospital, while the patients in intervening group were given with overall nursing intervention after leaving hospital. Results The rates of complying with doctor of the five aspects in the intervening group were higher than those in the control group(P<0.05). The scores of the four aspects of QOL of intervening group were lower than those of the control group(P<0.05). Conclusions Nursing intervention can play a very important role in promoting the behavior of complying with doctor and hying quality of the CHF patients.

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.

Objective To investigate the relationship between cervical intraepithelial neoplasia (CIN) and high-risk (HR)HPV infection among late pregnant women.Methods From Aug.2007 to Feb.2010,168 women at 13 to 32 gestational weeks undergoing prenatal examination in Beijing Obstetrics and Gynecology Hospital went through three stage cervical disease screening,including 21 women with cervicitis and 147 women with C1N (42 women with CIN Ⅲ,37 women with CIN Ⅱ and 68 women with CIN Ⅰ ).Hybrid capture assay version Ⅱ ( HC- Ⅱ ) test was used to measure HR-HPV DNA load,and the logarithmic transtormation (log10) was performed.All 168 women were followed up to postpartum 3 -6 months.HR-HPV infections rates of cervicitis and different CIN,the rate of HR-HPV infection turned naturally negative at postpartum of 3 to 6 months,and HR-HPV load at pregnancy and 3 -6 months postpartum were observed.Results ( 1 ) HR-HPV infection rate:CIN Ⅲ,Ⅱ,Ⅰ and cervicitis pregnant women's HR-HPV positive infection rates were 98％ (41/42),86％ ( 32/37 ),76％ ( 52/68 ) and 62％( 13/21 ) respectively,which reached statistical difference (P =0.002).(2) HR-HPV naturally negative:the rate of pregnant women with different levels of CIN who turned HR-HPV naturally negative within 3 -6 months of postpartum were CIN Ⅲ 5％ (2/41),CIN Ⅱ 47％ (15/32),CIN Ⅰ 52 ％ (27/52) and cervicitis 10/13,which also reached statistical difference among those four groups (P =0.000).(3) HR-HPV load:pregnant women with different grade of CIN and cervicitis HR-HPV DNA load were CIN Ⅲ 2.02 ng/L(1.53,2.67 ng/L),CIN Ⅱ 1.94 ng/L ( 0.75,2.75 ng/L),CIN Ⅰ 2.04 ng/L (0.08,2.95 ng/L) and cervicitis 1.98 ng/L( -0.07,2.47 ng/L).There was no significantly different HPV load in women with cervicitis and different CIN (P =0.719).At 3 -6 months postpartum,HR-HPV load was CIN Ⅲ1.55 ng/L(0.90,2.10 ng/L),which was significantly higher than the amount of CIN Ⅱ 0.09 ng/L(-0.69,1.74 ng/L),CIN Ⅰ 0.48 ng/L( -0.56,2.2 ng/L) and cervicitis -0.46 ng/L ( -0.78,1.40 ng/L,P =0.036).Conclusions With the increasing of CIN grade,the rate of HR-HPV infection in pregnant women was increased,however,the rate of HR-HPV turning negative naturally at 3 -6 months postpartum decreased.With different CIN grade during pregnancy,HR-HPV DNA load did not change significantly,but HR-HPV DNA load increased at 3 -6 months of postpartum.HR-HPV DNA loads with the same grade of CIN and cervicitis during pregnancy higher than that of postpartum among pregnant women.

OBJECTIVETo investigate the changes of serum soluble CD 163 (sCD 163) level, to assess the severity of critical illness and to evaluate the immune status of sepsis or severe sepsis in children.

METHODA prospective study was conducted. The sCD 163 was determined in 50 cases with sepsis or severe sepsis in pediatric intensive care unit (PICU) and 23 cases of age- and gender-matched healthy children were enrolled as control during the period from April 2010 to March 2011. Double-antibody sandwich ELISA was used for sCD 163 measurement. The relationship with sCD 163 level and disease severity score (pediatric critical illness score, PCIS; and pediatric risk of mortality III, PRISM III), lymphocyte subsets, C-reactive protein (CRP), tumor necrosis factor α (TNFα) were analyzed.

RESULTThe sCD 163 in sepsis/severe sepsis groups (171.04 ± 177.85) mg/L was significantly higher than that in control group (44.19 ± 86.48) mg/L (P < 0.01).sCD 163 in sepsis group [(105.32 ± 145.87) mg/L] was significantly lower than that of severe sepsis group [(233.32 ± 171.78) mg/L] (P < 0.05). sCD 163 level was significantly higher in lower PCIS score patients. (P < 0.01). The sCD 163 levels was higher in PRISM III ≥ 10 than the PRISM III < 10 group. The sCD 163 levels were higher in death group than the survival group. The sCD 163 was negatively correlated with CD4 +, CD4 +/CD8 + (R = -0.820, P < 0.05; R = -0.839, P < 0.01).

CONCLUSIONDetection of sCD 163 was helpful in predicting the severity of sepsis and severe sepsis, and sCD 163 may reflect the immune status of critically ill children with sepsis.

Adolescent ; Antigens, CD ; blood ; Antigens, Differentiation, Myelomonocytic ; blood ; Biomarkers ; blood ; C-Reactive Protein ; analysis ; Case-Control Studies ; Child ; Child, Preschool ; Critical Illness ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Infant ; Intensive Care Units, Pediatric ; Lymphocyte Subsets ; immunology ; Male ; Prognosis ; Prospective Studies ; Receptors, Cell Surface ; blood ; Sepsis ; blood ; immunology ; mortality ; Severity of Illness Index ; Tumor Necrosis Factor-alpha ; blood

The importance and feasibility was analyzed of the teaching BBS for aiding classroom teaching based on campus network. The design, technique, content, advantages and deficiencies were presented of agricultural microbiology teaching BBS. The prospect also was discussed of teaching BBS based on campus network in this paper.