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 detect the antibiotic resistance of four kinds of gram-negative bacilli strains against seven antibiotics and to analyze the differences in antibiotic resistance between the strains isolated in intensive care unit (ICU) and common wards.Methods This study involved 3 238 gram-negative bacilli strains isolated in Beijing Tongren Hospital from January to December 2016.Of all strains, 46.6% were isolated in ICU (severe group) and 53.4% were isolated in common wards (general group).Resistance of these strains to seven kinds of antibiotics was detected and the differences between the two groups were analyzed.Results Antibiotic resistance rates of Pseudomonas aeruginosa strains to ceftriaxone, cefepime and imipenem were 41.7%, 41.2% and 39.5% in severe group and 20.9%, 21.7% and 17.1% in general group.Moreover, the differences between the two groups were all statistically significant (χ2Cefatriaxone=56.72, P<0.01;χ2Cefepime=49.12, P<0.01;χ2Imipenem=69.81, P<0.01).Antibiotic resistance rates of Klebsiella pneumoniae strains to imipenem was 17.2% in severe group and 8.8% in general group, and the difference between the two groups was statistically significant (χ2Imipenem=11.48, P<0.01).Resistance rates of Escherichia coli strains to ceftriaxone and cefepime were 72.9% and 35.8% in severe group and 44.7% and 13.3% in general group, and the differences between the two groups were statistically significant (χ2Ceftriaxone=40.13, P<0.01;χ2Cefepime=41.61, P<0.01).More than 60% of Acinetobacter baumanii strains whether they were isolated in ICU or in common wards were resistant to all the seven antibiotics, and there were no significant differences between the two groups.Conclusion Gram-negative bacilli strains isolated in ICU have higher resistance rates than those isolated in common wards and therefore antibiotics should be used differently.Regular monitoring of drug resistance should be strengthened to provide references for empirical clinical medication.

Objective To investigate the effects of micro alcohol on serum enzymes in vitro and vitro Methods Serum alcohol and ALT?AST?GGT?ALP?CK?LDH?AMY?LIPA activities were measured before and after alcohol consuming (1 ml/kg) in 14 volunteers Meanwhile, the direct inhibitory effects of alcohol on the serum enzymes were studied by comparing the serum enzyme activities with or without alcohol Results Alcohol consuming could depress the serum AST activity from (24 04?3 66) U/L to (22 25?3 27) U/L and LIPA activities from (155 86?93 51) U/L to (128 35?84 85) U/L, whereas increase the other serum enzyme activities, but only serum AMY were found statistic difference [from (48 78?10 66) U/L to (55 50?12 60) U/L] The inhibitory effects of alcohol on all the measured enzymes were found in vitro studies Conclusions Alcohol could obvious influence the serum enzyme activities both in vivo and vitro Avoiding the contamination of alcohol during sample collection and routine laboratory work is necessary

Objective To investigate the clinical significance and biological role of G-protein coupled receptor 49(GPR49) expression in pancreatic carcinoma.Methods GPR49 expression in tumor and adjacent normal tissues of 77 patients with pancreatic cancer was compared by tissue microarray and immunohistochemistry.And then, the GPR49 expression levels in the tumor tissues of patients with different pathological grades and clinical stages were analyzed.GPR49 positive pancreatic cancer cell line CFPAC-1 was taken as cellular model.CFPAC-1 cells were stimulated with roof plate-specific spondin(RSPO)1, the ligand of GPR49, in vitro.The effect of RSPO1 on CFPAC-1 cells proliferation was evaluated with cell counting.The effect of RSPO1 on the expression of membrane molecular CD44 in CFPAC-1 cells was detected by flow cytometry.CFPAC-1 cells incubated with RSPO1 were subcutaneously implanted into nude mice.And then, the time of tumor formation and tumor size were observed.T test and single factor analysis of variance were performed for statistical analysis.Results GPR49 was widely expressed in all 77 pancreatic cancer tissues.By immunohistochemistry, the score of GPR49 expression in pancreatic cancer tissues was 9.0±2.4, which was higher than that of adjacent normal tissues (5.7±2.4), and the difference was statistically significant (t=8.995, P<0.01).There was no correlation between GPR49 expression and tumor sizes, pathological grades, lymph node metastasis, and clinical stages (all P>0.05).The results of experiments in vitro indicated that RSPO1 could promote CFPAC-1 cells proliferation and up-regulate CD44 expression in CFPAC-1 cells.Experiments in vivo demonstrated that after 30 days the tumor volume of mice implanted with RSPO1-pretreated CFPAC-1 cells was (606.0±188.0) mm3, which was larger than that of PBS-pretreated group ((364.2±83.7) mm3), and the difference was statistically significant (t=2.616, P=0.031).Conclusion GPR49 is widely expressed in pancreatic cancer cells and RSPO1/GPR49 pathway has play a role in promoting the proliferation of pancreatic cancer cells, which might be a potential target for interfering pancreatic cancer.

Objective To evaluate the diagnostic value of treponema pallidum particle agglutination (TPPA) and toludine red unheated serum test (TRUST) for the therapeutic effects of early syphilis.Methods Syphilis patients visited the Dermatological Department of Beijing Tongren Hospital, Capital Medical University from January 2012 to October 2016 were recruited, including 189 patients with early syphilis and 39 patients without clinical symptoms but with risky behavior within 3 months.Serum samples were tested by TPPA and TRUST respectively before treatment and at month 3, 6, 12 after treatment.Comparison between groups was done by χ2 test.Results Serum TPPA results of 228 patients with early syphilis prior to the treatment and within 12 months after treatment were all positive.Before treatment, TRUST tests results of 225 patients were positive, and 156 patients turned negative after 12 months of treatment with the negative conversion rate of 69.3%.The TRUST titer ≤1∶4 before treatment was defined as low-titer group, and ≥1∶8 before treatment was defined as high-titer group.There were 93 patients in the low-titer group, including 3 patients with negative results, and 90 patients with titer 1∶1-1∶4.There were 135 patients in the high-titer group.Twenty-seven patients achieved TRUST negative conversion after 3 months of treatment, among which 19.35%(18/93) with the negative conversion rate in the low-titer group and 6.67%(9/135) in the high-titer group.The difference between the two groups was statistically significant (χ2=7.10, P<0.01).Sixty-two patients converted into negative results of TRUST assay after 6 months of treatment, including 36.56%(34/93) in the low-titer group and 11.85%(16/135) in the high-titer group whose titer of 1∶1-1∶4, and 6.45%(6/93) in the low-titer group and 4.44%(6/135) in the high-titer group whose titer ≥1∶8 turned negative.The difference between the two groups was statistically significant (χ2=17.10, P<0.01).There were 67 patients converted negative detected by TRUST assay after 12 months of treatment, including 23.66%(22/93) in the low-titer group with titer of 1∶1-1∶4 and 14.81%(20/135) in the high titer group, and 1.08%(1/93) in the low-titer group and 17.78%(24/135) in the high-titer group whose titer ≥1∶8 were turned negative.The difference between the two groups was not statistically significant (χ2=1.51, P>0.05).Conclusion Serological tests combined with clinical signs can accurately diagnose early syphilis and evaluate the therapeutic effects.

To understand the personality characteristics and temperament type of medical students in Xi'an Jiao tong University,and to provide scientific evidence for their targeted mental intervention by analyzing graphs about Eysenck personality questionnaire revised in China.,310 medical students were investigated by using EPQ(adult version).We found that there were more male students with typical psychoticism personality than female students with the same personality,and there was statistical significant difference in the distribution of the type of temperament between male and female students.Medical students with stable personality probably accounted for two-thirds of the total.Female students were mainly manifested unstable personality with extraversion,while male students were unstable introvents.In future we should take targeted psychological intervention measures.We are supposed to focus on each student's personality characteristic,temperament type and other specific conditions for a variety of education.

Nitric oxide (NO) plays an important pathophysiological role in osteoarthritis and cartilage metabolism. To determine the relationship between NO and the synthesis of type II collagen in cartilage, we measured levels of type II collagen by ELISA and procollagen (II) mRNA by RT-PCR in cultured lapine chondrocytes that were incubated with some kinds of reagents. 0.2mM sodium nitroprusside (SNP, a NO donor) can release high levels of NO, decreasing type II collagen, suppressing the expression of procollagen (II) mRNA (COL2A1).At the same time, chondrocytes showed a large increase in NO synthesis, a decrease in type II collagen and COL2A1 mRNA in response to 100u/ml IL-1. When 1mg/ml N-nitro-L-arginine methyl easter (NAME, an inhibitor of NO synthase) was mixed with IL-1, NO production was inhibited, the amounts of type II collagen recovered partially and COL2A1 mRNA recovered completely.These data indicate NO can inhibit type II collagen synthesis as IL-1 downstream molecule by suppressing procollagen (II) mRNA.

ObjectiveTo assess the effect of comprehensive rehabilitation on facial paralysis.Methods80 patients with facial paralysis were divided into observation group receiving comprehensive rehabilitation(medicine, physical therapy, functional exercise and psychological treatment), and control group with medicine and physical therapy.ResultsThere was a significant difference in the scores for Portmann's Simple Scale between before and after treatment in two groups (P<0.01). There was a significant difference in the clinical effect between observation group and control group (P<0.05).ConclusionComprehensive rehabilitation has a better effect on facial paralysis.

Objective To evaluate the effects of four types of variance reduction techniques ( ring counter grid, high electron cutoff energy, termination of electron tracking in some structures, and emission direction?biased sampling of source) on the efficiency and accuracy of Monte Carlo simulation for the single source channel of the Leksell Gamma Knife. Methods The single source channel of the Leksell Gamma Knife was modeled using Monte Carlo software MCNP . Four types of variance reduction techniques were used to simulate the dose distribution in the water?like phantom. The computation efficiency and simulation result were compared between the four techniques. Results All techniques substantially improved the computation efficiency and had little effect on the accuracy of the simulation ( relative error less than 2. 5%) . However, if the electron cutoff energy was above 50 keV, the simulation became quite inaccurate due to neglect of the scattering of high?energy electrons and their dosimetric contribution to the penumbra. When the scattering of high?energy electrons and their dosimetric contribution to the penumbra were ignored, the dose to the Profile platform was overestimated and the dose to the penumbra was underestimated. Conclusions Rational use of variance reduction techniques can substantially improve the efficiency of Monte Carlo simulation for the single source channel of the Leksell Gamma Knife. However, the impact of variance reduction techniques on the accuracy of the simulation should be carefully evaluated.

BACKGROUND:B-lymphocytes are an important participant in the immunity system. Currently, magnetic beads and complement methods are mainly used to isolate and purify B-lymphocytes. However, these methods are costly or cause large cel damage and low purity, which need further improvement. OBJECTIVE: To explore the isolation and culture methods of B-lymphocytes from mouse spleen and to study suitable conditions for B-lymphocyte isolation and culture in vitro by using interleukin-4, lipopolysaccharide, CD3 monoclonal antibody or their combination. METHODS:B-lymphocytes from mouse spleen were isolated and randomly divided into seven groups, respectively treated with interleukin-4, CD3 monoclonal antibody, lipopolysaccharide, interleukin-4+CD3, interleukin-4+lipopolysaccharide, CD3+lipopolysaccharide, and no stimulation (control group). Flow cytometry was used to detect the changes in the number and proportion of T-lymphocytes, B lymphocytes, and their subpopulations under different culture conditions. RESULTS AND CONCLUSION:The number of lymphocytes peaked at 3-5 days after addition of interleukin-4. In the lipopolysaccharide group, the number of lymphocytes began to increase at 3 days, and then peaked at 5 days. T-lymphocytes disappeared after addition of CD3 monoclonal antibody, so relatively pure B-lymphocytes could be obtained after 2 days and the number of B-lymphocytes reached the peak at 3 days. The number of mature B-lymphocytes (B220+IgD+) increased significantly after addition of CD3 antibody. In al the conditions we tested, transitional B cel subset (B220+CD93+) disappeared completely after 24 hours of culture. Experimental results indicate that after addition of CD3 monoclonal antibody and interleukin-4, T-lymphocytes can be removed in mouse spleen cels cultured, but mature B-lymphocytes remain to survive and proliferate.