Objective To establish a rapid genotyping method of for methicillin-resistant Staphylococcus aureus(MRSA) based on polymerase chain reaction(PCR)-high resolution melting (HRM ) curve analysis and staphylococcal protein A (SPA ) classifica-tion .Methods 71 strains of MRSA clinically isolated were collected as test strains .Gene sequencing and HRM curve analysis were employed to conduct SPA gene typing .Results According to gene sequencing method ,SPA gene of 71 strains of MRSA was divided into four types ,namely t570 ,t030 ,t002 and t588 .The most predominant type was t570 (74 .65% ) ,followed by t030 and t002(both 7 cases) .The result of SPA gene typing by HRM analysis were basically consistent with that by gene sequencing .Con-clusion PCR-HRM analysis is expected to become a fast ,efficient genotyping for MRSA SPA gene ,providing the basis for hospital infection control .

Objective To construct the recombinant plasmid pET32a-AKT1 and express human AKT1 protein using prokaryotic expression system .Methods Reverse transcriptase-polymerase chain reaction(RT-PCR) was employed to amplify the gene AKT1 in coding region and integrated it with pET 32a plasmid ,following by transforming it into Escherichia coli DH5α and prokaryotic strains BL21(DE3) .Isopropyl-beta-D-thiogalactopyranoside(IPTG) was adopted to induce its expression .Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE ) and Western-blot were used for protein identification .Results Complete fusion of target gene and plasmid was observed .The recombinant plasmid pET32a-AKT1 was successfully transferred into the strain DE3 . After IPTG induction ,protein with relative molecular mass 70 000 was expressed by DE3 .Conclusion The recombinant plasmid pET32a-AKT1 is constructed successfully and AKT 1 protein is completely and efficiently expressed by prokaryotic strain DE 3 .

Objective To detect serum leucine aminopeptidase (LAP) level in the patients with hyperthyroidism for investiga‐ting the changes of LAP in the patients with thyroid dysfunction and its clinical significance .Methods 117 patients with hyperthy‐roidism were taken as the test group and 109 healthy people as the control group .The differences of various laboratory indexes were comparatively analyzed and the correlation between LAP with FT 3 ,FT4 and TSH was analyzed .117 patients with hyperthyroidism were divided into the liver damage group(52 cases) and non‐liver damage group(65 cases) and performed the LAP determination . Results The serum LAP ,FT3 ,FT4 and TSH levels in the hyperthyroidism whole group ,hyperthyroidism liver damage group and hyperthyroidism non‐liver damage group had statistically significant differences compared with those in the control group (P<0 .01) ,serum LAP was positively correlated with FT 3 and FT4 ,and negatively correlated with TSH ;LAP ,FT3 ,FT4 and TSH had no statistical differences between the hyperthyroidism live damage group and the hyperthyroidism non‐liver damage group (P>0 .05) .Conclusion The serum LAP content is closely related with the thyroid hormones and has certain clinical reference value for the diagnosis ,disease condition evaluation and medication .Lap can serve as an independent diagnostic indicator .

Objective To evaluate the clinical application significance of the direct drug susceptibility test and the conventional drug susceptibility test in blood bacterial infection .Methods 280 specimens of positive blood culture in our hospital from March 2012 to April 2014 were randomly selected and performed the the direct drug susceptibility test and the conventional drug suscepti‐bility test .The correlation of the bacterial identification results and the drug susceptibility test results between these two kinds of method was investigated .Results The consistence of the direct bacterial identification method and bacterial identification method was 92 .5% ,the consistence rate of sensitivity ,drug resistance and moderate sensitivity had no statistical differences between the two kinds of method (P>0 .05) .Conclusion The direct drug susceptibility test can shorten the report time of blood culture posi‐tive specimen and effectively guide the rational use of antibiotics in clinic .

Objective To investigate the human papillomavirus (HPV) infection situation among female patients and its subtypes distribution rule for providing the measures of diagnosis and prevention .Methods The nucleic acid hybridization was adopted to de‐tect HPV subtypes of cervical exfoliated cells in 12 422 female outpatients and inpatients in a hospital .The HPV infection situation among different patients were analyzed .Results The HPV infection rate was 17 .0584% ,the HPV infection rate had statistical difference among different age groups (P<0 .01) .Totally 20 subtypes of HPV were identified .HPV16 ,HPV52 and HPV58 were the most prevalent high‐risk subtypes .Conclusion The HPV subtype detection can be used in the screening and condition judgment of cervical cancer .The key for the HPV positive population is early diagnosis and early treatment for blocking the HPV persistent infection and preventing the cervical cancer occurrence .

Objective To investigate the correlation between the serum levels of homocysteine (Hcy) ,folate acid (FA) and vita‐min B12 (VitB12 ) with cerebral infarction .Methods The enzymatic cycling method and chemiluminescence immunoassay were used to detect the serum Hcy ,FA and Vit B12 levels in 60 patients with cerebral infarction ,and then the detection results were compared the results of 40 people undergoing physical examination in the same period .Results The serum Hcy level in the cerebral infarction group was (23 .16 ± 8 .17)μmol/L ,which was significantly higher than (8 .09 ± 2 .45)μmol/L in the control group(P<0 .01) ,and the serum FA and Vit B12 levels in the cerebral infarction group were (6 .12 ± 2 .19) ng/mL and (323 .92 ± 133 .79)μmol/L ,which were in the control group lower than (10 .57 ± 5 .70) ng/mL and (530 .85 ± 168 .56)μmol/L in the control group (P<0 .01) .The serum Hcy level in the cerebral infarction group showed the negative correlation with FA and VitB 12 levels ( r1 = - 0 .444 , r2 = -0 .535 ,P<0 .05) .Conclusion High hyperhomocysteinemia is an independent risk factor of cerebral infarction and is related to the decrease of FA and VitB12 .

Objective To investigate the distribution situation of human papillomavirus (HPV) genotypes profile in cervical cells among women in Xuzhou area and its clinical significance .Methods 23 kinds of HPV DNA were extracted in cervical cell samples from 8 010 women in Xiuzhou area .The gene‐chips technique of PCR combined with reverse dot blot was adopted to detect the HPV genotypes .Results Among 8010 cervical cell samples ,there were 1 852 HPV infected cases ,the total HPV infection rate was 23 .12% ,the HPV infection rates of single type accounted for 17 .17% and its predominant types were 16 type (4 .35% ) ,followed by 58 type (2 .12% ) and 52 type (1 .82% ) ,The detection rate of multiple HPV infection was 5 .96% ,in which the predominant types were HPV16+58(4 .40% ) ,16+52(2 .94% ) ,11+16(2 .52% ) .Conclusion The single HPV infection of HPV16 ,58 ,52 and the multiple HPV infection of HPV16+58 ,16+52 ,11+16 are the main genotypes of cervical cells among women in Xuzhou area , this gene chip technique is suitable for the cervical cell sample ,its once detection can detect 23 kinds of HPV genotypes with high specificity and high sensitivity ,which has an important significance for the molecular epidemiologic survey study of HPV genotypes distribution among women in our country .

Objective To evaluate the significance of the level change of urinary conductivity (Cond) on the disease progress in the patients with type 2 diabetic nephropathy(DN) .Methods 138 patients with type 2 diabetes mellitus (T2DM) in our hospital were selected and divided into the normoalbuminuria (NUA) ,microalbuminuria (LUA) and macroalbuminuria (MUA) group;then among them 107 cases were re‐divided into the DN group and the diabetes mild renal injury (DC) group .The levels of urinary Cond were measured by using the Sysmex UF‐1000i urine flow cytometer .The urine specific gravity (SG) was detected by the ARKRAY AUTION MAX AX‐4280 analyzer ,and the urine albumin (U‐Alb) was tested by the Siemens BNⅡ automatic protein analyzer .Re‐sults The Cond level in the MUA group was (14 .1 ± 4 .5)ms/cm ,which was lower than (15 .7 ± 4 .3)ms/cm in LUA group(P<0 .05) ,while the Cond level in the LUA group was significantly lower than (17 .6 ± 5 .7) ms/cm in the NUA group(P<0 .05);the SG levels in the NUA group and the LUA group were 1 .014(1 .010-1 .019) and 1 .015(1 .010 -1 .020) respectively ,both were higher than 1 .011(1 .009-1 .012) in the MUA group SG (P<0 .05) .Cond was positively correlated with SG (r=0 .63 ,P<0 .05) and negatively correlated with 24 h‐UAE (r= -0 .183 ,P<0 .05) .The Cond level in the DN group was(13 .2 ± 4 .3)ms/cm ,which was significantly lower than (15 .0 ± 4 .4) ms/cm in the DC group (P<0 .05) ,there was no statistically significant differences in the SG level between the DN group and DC group (P>0 .05) .The area under curve (AUC) of ROC for Cond was 0 .612 (0 .502-0 .723) .When setting the cut‐off vales of Cond as 11 .85 ms/cm ,then the sensitivity was 43 .8% ,and the specificity was 78 .0% . Conclusion The urine Cond level change can reflect the disease progress of DN in T 2DM ,but can not be used as its early screening indicato r .

Objective To observe the effectiveness and safety of basal insulin plus oral antidiabetic drugs (OADs) in treating the patients with type 2 diabetic mellitus (T2DM ) ,who were poor blood sugar control by premixed insulin with or without OADs . Methods 32 cases of T2DM and poor blood sugar control by premixed insulin combinated with or without OADs stopped the premixed insulin therapy and changed to insulin glargine plus OADs for 16 weeks .Glycosylated haemoglobin (HbA1c) ,fasting blood glucose(FBG) ,postprandial blood glucose(PBG) ,body mass index (BMI) and mean daily insulin dose were compared among patients .And the episodes of hypoglycemia events were recorded .Results After 16‐week treatment ,HbA1c ,FBG ,PBG and BMI were all significantly decreased comapared with before treatment (P<0 .01) .The mean insulin glargine daily dose was significantly decreased compared with the premixed insulin dose at admission .2 cases (6% )appeared twice hypoglycemia episodes during the treatment period ,all were general hypoglycemia .Conclusion Basal insulin once daily can effectively improve the sugar metabolism in T2DM patients failed by premixed insulin with or without OADs ,moreover the body mass is not increased .

Objective To evaluate the performance of self‐established angiotensin converting enzyme (ACE) detection system . Methods The performance evaluation of ACE reagent kit produced by the Jiuqiang Company including precision ,linearity ,correla‐tion with the reference reagent and reportable range were conducted by using the automatic biochemical analyzer according to the re‐quirements of the EP5‐A2 and EP9‐A documentation in the Clinical and Laboratory Standards Institute (CLSI) .Results Intra‐as‐say CV of the system were 6 .87% ,2 .39% and inter‐assay CV were 6 .09% ,1 .81% ,respectively .During the day CV were 8 .00 %and 2 .8% respectively ,which were less than those provided by the manufacturer (<10% );the lenearity result was R2 =0 .99 .The correlation coefficient (r) of the system comparing with the reference reagent was 0 .990 56 ,moreover the average bias was 8 .55% , showing good correlation ;the repotable range was 9 .0U/L‐600U/L .Conclusion Self‐established ACE detection system can meet the requirements for clinical application .