Objective To explore a rapid method for classification of microorganisms.Methods The electrophorese fingerprinting, direct sequence of 16S rDNA and 16S-23S rDNA ISR after PCR, multiplex PCR for 16S rDNA and antibiotic resistance genes, were utilized to explore fast approaches of extracting total DNA from different clinical specimens.Results The specific 16S-23S rDNA ISR fingerprinting fragments were shown on the genus or species level in bacteria and fungi.So fingerprinting can be used to identify pathogenic microorganisms, to differentiate the evolution relations or to set the phylogenetic tree by comparing their DNA banding patterns with those of standard strains (NCCLS). Multiplex PCR was able to examine the special genes of genus or species, mecA gene, TEM, SHV and CTX gene in staphylococcus and ESBLs(E.coli or K.pneumoniae) at the same time.Conclusion The part of 16S rDNA sequencing and 16S-23S rDNA ISR genotypes by gel electrophoreses were useful for bacterial species identification in addition, it was clearly more rapid and accurate than culture technique, and the large numbers of strains can easily be examined.Multiplex PCR could provide a good method for identification of microorganisms and analysis of antibiotic resistance at the same time.

Objective To study the mechanism of the resistance in Acinetobacter baumannii to common antibiotics.Methods Bacterial susceptibility test to ?-lactamatic antibiotics,quinolones,aminogiycosids were done by the Kirby-Bauer method and agar dilution method for 35 isolates.Penicillin-beta-lactamase,AmpCs,Metallo-beta-lactamase,ESBLs were detected by iodine-starch test,3-dimension test,microbiology sensitivity synergic test,disc agar diffusion method respectively.Outer membrane protein was analyzed by SDS-PAGE.Accumulation of ciprofloxacin was determined by direct Fluorescence method.The gene of Tem-1,aac-4 and gyrA were amplified by PCR while the gyrA was sequenced.Results 28 isolates were multi-resistant to common antibiotics in 35 isolates.16 isolates produced Penicillinase,10 isolates produced Cephalosporinase,2 isolates produced metal-beta-lactamase,3 isolates produced ESBLs.The analysis of outer membrane proteins showed that a protein of 29kD disappeared and 26kD protein enhanced in resistant isolates.The accumulation of ciprofloxacin in resistant isolates decreased.After treatment with NaN_3,the drug uptake increased to the normal level.Most of [QX(Y8]Tem-1 gene were positive except 2 drug resistant and 3 sensitive isolates.All of [QX(Y8]aac-4 were negative while gyrA were positive.DNA sequencing analysis revealed there had point mutation in the gyrA gene.Conclusion Beta-lactamase,active drug efflux,outer membrane protein permeability decreasement and gene mutation were the factors contributing to the antibiotics resistance of Abaumannii.

Objective To study the influence of general anesthesia and epidural anesthesia on postoperative short-term cognitive function in the elderly patients undergoing orthopedics surgery.Methods 120 patients with hip joint displacement or internal fixation after femur fracture and American Society of Anesthesiology Ⅰ and Ⅱ were randomly divided into general anesthesia and epidural anesthesia groups(n=60 for each group).Artery blood pressure(ABP)and heart rate were recorded before operation,pre-operation after anesthesia,during 30 min operation,during main operation and at operation end.Cognitive function was detected by mini-mental state(MMS)before induction of anesthesia and 6 h,12 h,24 h and 72 h after anesthesia.Results There were no differences in ABP and heart rate during operation between the two groups(P＞0.05).The scores of MMS in general anesthesia group at 6 h(26.5±0.5),12 h(25.4±0.7)and 24 h(27.4±0.3)were decreased as compared with pre-induction of anesthesia(29.5 ± 0.3)(P＜ 0.05),while no difference was found at 72 h(29.3±0.3).The scores of MMS in epidural anesthesia group at 6 h(26.6±0.4)and 12 h(25.6±0.8)were lower(P＜0.05),while had no difference at 24 h(29.1±0.4)and 72 h (29.5±0.4)(P＞0.05)as compared with pre-induction of anesthesia(29.4±0.4).At 24 h after anesthesia,the MMS scores were higher in epidural anesthesia group(29.1±0.4)than in general anesthesia group(27.4±0.3)(P＜0.01).Conclusions General anesthesia may contribute to more obvious influences on cognitive function than epidural anaesthesias within 12h after operation in the elderly patients undergoing orthopedics surgery.

Objective To investigate the expression of 4-hydroxynonenal (4-HNE) in the kidney of diabetic rats and the effect of probucol.Methods The rats were being intraperitoneal injected with STZ (60 mg/kg) to establish diabetic models.Then diabetic rats were randomly divided into diabetic group (group D,n =24),probucol treated group (group P,n =24).Normal rats were taken as control group (group C,n =24).Rats in group P were treated by probucol (110 mg·kg-1·d-1); rats in group D and group C were given equal volume water instead.Scr,BUN,triglyceride (TG),total cholesterol (TC) and 24-hour urinary proteinin were measured at the 4th,8th and 12th week.PAS staining and HE staining were used to evaluate the pathological changes of the kidney.The immunohistochemistry and Western blotting were used to detect the expression of 4-HNE in renal tissue.Results Levels of Scr,BUN,TG,TC and 24-hour urinary protein in group D were higher than those in group C at the 4th,8th and 12th week(all P ＜ 0.05); Levels of Scr,BUN,TG,TC and 24-huor urinary protein in group P were lower than those in group D at 4th,8th and 12th week (all P ＜ 0.05).The pathological changes of the kidney in group D were more serious than that in group P.The expression of 4-HNE in group D were higher than group C at the 4th,8th and 12th week (all P ＜ 0.05);The expression of 4-HNE in the kidneys of group P decreased significantly compared to that of group D at the same time (P ＜ 0.05).Conclusions As an indicator of lipid peroxidation,the expression of 4-HNE significantly increases in the kidney of diabetic rat.Probucol may protect the diabetic kidney through decreasing the expression of 4-HNE and the level of lipid peroxidation.

Objective To observe the levels of serum 25‐hydroxy(25OH) vitamin D and bone mineral density(BMD) in appar‐ently healthy middle‐aged and elderly people in Zhongshan area and investigate the correlation between each other .Methods The BMD of 200 participants was measured by ultrasound BMD detector ,the participants were divided into 3 groups according BMD re‐sults .Simultaneously ,their serum 25OH vitamin D levels were measured by ECLIA .Results Among 200 participants ,the preva‐lence of vitamin D severe deficiency ,deficiency ,insufficiency and sufficiency w ere 7 cases (3 .5% ) ,35 cases (17 .5% ) ,102 cases (51 .0% )and 56 cases(28 .0% ) ,respectively .There was no significant difference in the BMD values among subgroups of different vitamin D levels(P>0 .05) .The prevalence of low BMD and osteoporosis were 60 cases(30% ) and 10 cases(5% ) ,the levels of ser‐um 25OH vitamin D in normal BMD ,low BMD and osteoporosis group were (67 .31 ± 18 .28) ,(65 .62 ± 15 .41) and (64 .95 ± 19 .86)nmol/L ,respectively .There was no significant difference in the levels of serum 25OH vitamin D among BMD subgroups (P>0 .05) .Serum 25OH vitamin D levels were not directly correlated with BMD(P>0 .05) .Conclusion Vitamin D deficiency and insufficiency are quite serious problems in apparently healthy adults in Zhongshan area .There is no evidence that the status of ser‐um 25OH vitamin D is correlate with BMD ,for laboratory diagnosis of osteoporosis ,more sensitive laboratory markers are needed .

Objective To study the genotypic and biological characteristics of catheter-related MRSE isolates and to further provide information for the diagnosis and prevention of catheter-related bloodstream infection. Methods Thirty strains of catheter-related MRSE isolates were collected from venosus blood and whole blood of 30 inpatients including 20 males and 10 females from Beijing Tongren Hospital, Capital Medical University from January 2006 to December 2009. The genetic features of these strains were determined by MLST. PCR was used to detect the icoA gene (encoding the polysaccharide intercellular adhesion associated with pathogenicity), and the antimicrobial susceptibility test was detected by disc diffusion test. Results A total of 15 STs were obtained from 30 strains ST259, ST20, ST2 and ST235 were common clones obtained from 17 strains. Four novel STs were found and uploaded to the MLST database (http://www. mlst. net), including ST259 (6 strains), ST260 (1 strain), ST261 (1 strain) and ST262 (1 strain). The ST259 was the dominant clone of catheter-related MRSE isolates in this hospital, and 3 strains carrying icaA gene were detected in this study. Conclusion Some ST259 isolates express high pathogenesis among the four novel STs, which may make them as the pandemic strains in nosocomial infection, and this would increase the difficulty of the prevention and control of nosocomial infection.

Objective To study the clinical application of the ITS and β-tubulin gene regions in identification of Aspergillus spp. Methods One hundred and twenty-four Aspergillus strains that isolated from fungal rhino-sinusitis specimens were collected in Beijing Tongren Hospital, Capital Medical University from July 2007 to January 2010. They were identified by morphological and molecular methods. The first one included traditional culture, slide culture, and microscopic examination after lactophenol cotton blue stain and KOH digestion. The second one was amplifying and sequencing the part of ITS and β-tubulin gene and aligned all the sequences in the GenBank, European Molecular Biology Laboratory nucleotide sequence database, and DNA Data Bank of Japan. Results Of the 56 Aspergillus flavus identified by morphological features, fifty-five isolates were identified as Aspergillus flavus and 1 isolates was Aspergillus parasiticus by the ITS and β-tubulin gene region sequence analysis. In the 37 Aspergillus fumigatus identified by morphological method, and all the 37 isolates were identified as species complex of Aspergillus fumigatus by the ITS region sequence analysis, but through the sequence analysis of β-tubulin gene region, thirty-five isolates were identified as Aspergillus. fumigatus and 2 were Aspergillus lentulus. Twenty-one isolates were identified as Aspergillus versicolor by morphological method, but 16 of them were identified as Aspergillus. versicolor and 5 can not be identified to species level by the ITS region sequence. And by comparative-sequence analysis of β-tubulin gene region, the 5 isolates were identified as Aspergillus sydowii,the other 16 isolates were Aspergillus. versilcolor. Ten isolates were identified as Aspergillus nidulans by morphological features, the ITS and β-tubulin gene region sequence analysis. Conclusions β-tubulin gene sequencing is more suitable for identifying Aspergillus, and could identify Aspergillus spp. to species level Sequences of ITS region could only identify Aspergillus spp. to species complex.

Objective To study the effects of using the polysiloxane impression material in the repair of the precise attachment.Method Using the Rapid polysiloxane impression material to make 37 work impressions,29 un-work impressions were made by alginate impression material.Results All the work impressions were eligible while there were 4 un-work impressions not eligible at the first time. Conclusion The effects of using the polysiloxane impression material in the repair of the precise attachment is satisfactory.

Objective To analysis the human T lymphotropic virus (HTLV) infection status in Wenzhou among voluntaty blood donors.Methods Selected 72 417 voluntary blood donors of Wenzhou from from March,1,2016,to November,30,2016,to screen HTLV-Ⅰ / Ⅱ antibody by ELISA method.The positive samples were reexamined two times,two test results of samples were determined positive by ELISA.HTLV positive samples was confiemed by Western Blotting (WB).Results Screened 23 cases of anti-HTLV positive by ELISA method,then confirmed 9 cases of HTLV positive by Western Blotting (WB).HTLV infection rate of Wenzhou blood donors was 0.01％ (9/72 417).Conclusions HTLV infection was found among volunteer blood donors in Wenzhou,but the HTLV infection rate of volunteer blood donors in Wenzhou is still at a relatively low level.

Objective To study the action of puerarin on Caspase-3 and Bcl-2 expression of hippocampal CA1 neurons in ovariectomized rats, and explore its mechanism. Methods SD female rats were randomly assigned into sham operation group, model group, premarin group and puerarin groups (120, 60, 30 mg/kg). The model group and sham operation group were injected NS intraperitoneally, other groups were treated with corresping drugs for 30 d. Immunohistochemistry and RT-PCR were used to determine Caspase-3 and Bcl-2 expression of hippocampal CA1 neurons. Results Caspase-3 expression of hippocampal CA1 neurons was significantly decreased in high-dose puerarin group (P <0.05). Bcl-2 expression of hippocampal CA1 neurons was significantly increased in high- and medium-dose puerarin groups (P<0.05, P<0.01). Bcl-2 mRNA level of hippocampal CA1 neurons in high-dose puerarin group was significantly increased (P<0.05). Conclusion Puerarin can decrease Caspase-3expression and increase Bcl-2 expression of hippocampal CA1 neurons in ovariectomized rats, and has protective effect on neuronal structure.