Objective To amplify the 16S RNA fragments of 7 clinically isolated strains of Brucella spp. by PCR-RFLP technique, so as to provide experimental basis for the studies on diagnostics, genetics and epidemiology of Brucella spp. Methods According to the gene sequence of ATCC 25840 standard strain in GenBank, special primers for the 16S RNA conservative area in the Brucella spp. were designed. DNA extraction and PCR amplification of the 16S RNA fragments were performed with the 7 isolated strains. PCR products were then sequenced and RFLP analysis was conducted with appropriate restricted enzymes to study the homology and the mutation sites in those strains. Meanwhile, the clinical data of infected patients were retrospectively analyzed to evaluate the relationship between the clinical features and genotypes of Brucella infection. Results The amplified target fragments were about 1500bp in length and consistent with what was expected. The sequencing and homology analysis showed a 98.88% homology and 11 mutation sites among the 7 isolated strains. Four genotypes were identified by RFLP. Retrospective analysis of the clinical data indicated that no obvious relationship existed between the genotypes and the clinical features. Conclusions Amplifying 16S RNA fragments by PCR technique is a feasible method to make an early diagnosis of Brucella infection. The 7 clinically isolated strains are different in genotypes and 16S RNA fragment is a highly conservative fragment in bacterial genome with some mutations. The research provides evidence for the genetics and epidemiology of brucellosis.

Objective To investigate the potential effects of simvastatin on angiotensin Ⅱ-stimulated secretion and proliferation of adrenocortical carcinoma H295R cells.Methods The H295R cells were divided into control group, Angiotensin Ⅱgroup, simvastatin group and Angiotensin Ⅱ plus simvastatin group.Cortisol in medium was determined by chemiluminescent method , and aldosterone was determined by radioimmunoassay .The mRNA expression of 11 beta-hydroxylase ( CYP11B1 ) and aldosterone synthase ( CYP11B2 ) were examined by RT-qPCR.Cell proliferation was detected by MTS method.Results Compared with control group, angiotensin Ⅱincreased the secretion of cortisol and aldosterone, and the expression of CYP11B1 and CYP11B2.Simvastatin decreased cortisol secretion and CYP11B1 mRNA expression ( P<0.05 ) .Simvastatin also inhibited angiotensinⅡ-induced the secretion of cortisol and aldosterone , and the expression of CYP 11 B1 and CYP11 B2 compared with Angiotensin Ⅱgroup ( P<0.05 ) .Angiotensin Ⅱhad no effect on cell proliferation , while simvastatin significantly inhibited cell proliferation .The inhibitory effect of simvastatin on proliferation was enhanced when simvastatin was prescribed with angiotensin Ⅱ( P<0.05 ) .Conclusions Simvastatin inhibits angiotensin Ⅱ-induced secretion of cortisol and aldosterone in H295R cells.Simvastatin inhibits cell proliferation, which could be enhanced by angio-tensin Ⅱ.

Aged ; Humans ; Lung ; pathology ; Male ; Middle Aged ; Pneumoconiosis ; complications ; diagnosis ; pathology ; Reference Standards

Objective To monitor the constituents and resistant tendency of bacterial pathogens isolated from diarrheal patients in our hospital form 1994 to 2005 to offer the basis for guiding epidemiologic study,vaccination research and clinical treatment. Methods Enteric pathogenic bacteria were cultured and identified to species,group and serotype with biochemical and serologic methods and the susceptibility of bacteria to antimicrobial agents were tested. Results Enteric pathogenic bacteria were isolated predominantly in male patients and mainly in children and youngsters. It reached a peak from July to September every year. Shigella spp.(75.11%) was the most frequendy isolated pathogens and followed by Vibrio spp.(12.7%),Salmonella spp.(6.28%),Aeromonas spp.(4.43%) and Escherichia coli(1.25%).During the period from 1994 to 2005,diarrheal pathogens had a trend of decrease especially Shigella spp.and Salmonella spp.. Of the 6329 isolates of Shigella spp., 75.62% was S. flexneri and S.soanei,S.dysenteriae and S. boydii constituted 23.98%,0.22% and 0.01% respectively.The sensitivity of different species,group or serotype to different antimicrobial agents was not the same.S.flexneri and Aeromonas spp. were highly resistant to most of antibiotics. However, S.sonnei and Vibrio spp.had good susceptibility to antibiotics tested except trimethoprim/sulfamethoxazole and ampicillin. Conclusion There are many species and serotypes of enteric pathogenic bacteria causing infective diarrhea and the distribution changes gradually in Beijing. The resistance rate of enteric pathogenic bacteria to antibiotics is not the same in different species and serotypes.so strict surveillance iS always needed.

Objective To test the level of cell factor interleukin (IL)-21,CXCL13 in the plasma of patients with rheumatoid arthritis (RA),and to analyze the relationship between Follicular helper T cells(Tfh)and clinic features and discuss the possible immunological pathogenesis of RA.Methods The Tfh cells were obtained from patients and healthy controls (NC) and detected by Flow cytometery.While the levels of IL-21,CXCL13 in patients and NC were measured by ELISA tests.Those analysis were performed by student's t-test,one-way ANOVA,SNK-q test,Chi-square test,Spearman's correlation and multiple linear regression.Results The expression of CD4+CXCR5+ICOS+ cells (Tfh) in PBMCs of RA was significantly higher than normal controls (3.0±1.2 vs 1.1±0.4,P＜0.01).Meanwhile,the three RA groups of patients were divided to low,moderate and high disease activity groups,and the results showed that the expression of Tfh were increased accordingly (1.8±0.7,2.5±0.6,4.0±1.2).The expression of Tfh in the three groups were all significantly higher than that of controls (P＜0.01).There was a positive correlation between Tfh and DAS28,ESR,CRP,TJC,and bone erosion,RF and anti-CCP respectively.The expression of Tfh in those patients who had bone destruction was higher than those with no or mild bone destructions (2.7±1.1vs 3.4±1.3).The expression of Tfh in patients with un-treated RA patients,when compared to those RA patients who were treated appropriately and those who were not treated appropriately,was decreased significantly.The expression of Tfh in appropriately treated RA patients was lower than that without appropriately treatment.The level of IL-21,CXCL13 was decreased in patients with RA in the order of high,moderate,low disease activity and NC.Conclusion The expression of Tfh and the levels of IL-21,CXCL13 are increased significantly,and are closely related to disease activity and bone ersions.The expression of Tfh is decreased after relevant treatment.These results indicate that the abnormality of Tfh may play an important role in the pathogenesis of RA.

Objective To investigate the awaken effect of naloxon on dexmedetomidine anesthetized mice and its mechanism. Methods Thirty Kunming mice of clean grade were randomly divided into 3 groups which included NAL group (Naloxon group), ATI group(Atipamezole group)and NS group (Normal Saline group). All groups were given dexme?detomidine 1 mg·kg-1 intraperitoneally. Naloxon 2 mg·kg-1, atipamezole 2 mg·kg-1 and normal saline 10 mL·kg-1 were ran?domly given intraperitoneally to the NAL, ATI and NS group respectively 90 minutes after dexmedetomidine administration. At timepoints prior to dexmedetomidine administration and 5, 15, 30, 60, 90, 95, 105, 120, 180 minutes after it, the sedative and analgesic effects besides recovery time (based on restore of righting reflex loss) were assessed. Results Sedation and analgesia effects became apparent within 5 minutes, and peaked at approximately 60 minutes then spontaneously recovered at 180 minutes after injection of dexmedetomidine. The sedative and analgesic effects were reduced in both ATI and NAL groups. Compared with ATI group, the sedation scores were higher at 95, 105 and 120 minutes after dexmedetomidine admin?istration than those in NAL group (P<0.05) but the scores were not statistically significant at 180 minutes between these two groups. Compared with NS group, the sedation scores were lower at time points of 95, 105, 120 and 180 minutes than those in NAL group (P>0.05). The analgesic scores were not statistically significant at time points of 95, 105, 120 and 180 min?utes between NAL group and ATI group, but they were lower in NAL group compared with NS group at timepoints of 95, 105 and 120 minutes (P>0.05). The recovery time in ATI and NAL group were shorter than that in NS group (F=1 793.368, P<0.05), but it showed no statistical difference between ATI group and NAL group (P>0.05). Conclusion Naloxone had a certain awaken effect on dexmedetomidine anesthetized mice.

Objective:To choose the best vector for the expression of CS3 fimbriae. Methods: The CS3 operon was cloned into different plasmid vectors such as pUC19 and pTrc99A. The expression of CS3 was monitored by whole-cell ELISA and SDS-PAGE analysis. The assembly of CS3 fimbriae was detected with electron microscopy. Results:The expression level of CS3 fimbriae using plasmid pUC19 as carrying vector was the highest, and the insertion orientation of CS3 gene into the plasmid has a little effect on its expression level. The expression of CS3 fimbriae was confirmed by SDS-PAGE analysis and electron microscopy.Conclusions:The promotor of CS3 itself played the key role in the expression of CS3 fimbriae and the copy number of plasmid was the main factor to affect the expression level.

Objective To investigate the expression changes of microRNAs and VEGF-NOTCH in renal ischemic injury in mice, and to explore the potential mechanism associated with renal angiogenesis.Method Male Balb/c mice were subjected to a standard renal ischemia to induce acute kidney injury (AKI) after 45 min of bilateral renal artery clamping. Following 4 h, 24 h of reperfusion or sham operation, kindey tissues were collected and subjected to detect the expression changes of microRNAs which relatived with angiogenesis and VEGF, Flk-1, Notch1 mRNA by Quantitative Real-time RT-PCR. Flk-1 protein was detected by Western blotting analysis at 24 h and 72 h following Ischemia/Reperfusion(I/R) injury. The expression of CD31 was examined in tissue sections by immunohistochemistry staining, and the microvessels in ischemic region of each group were counted. Results miRNA-210 and miRNA-92a expression increased significantly, with prominent changes at 4 h and 24 h after reperfusion( P ＜ 0.05 ). VEGF and Flk-1 mRNA expression and Flk-1 protein were increased in renal I/R compared with control group respectively (P＜0.05 ).Immunohistochemistry staining results of CD31 showed a significant increase of microvessels in renal ischemic region. Conclusion This study first reported the changes in miRNAs expression in response to kidney I/R in mouse. our results implied that miRNAs may be involved in targeting VEGF-Notch pathway signaling to regulate angiogenesis after renal I/R injury. It provided novel insights into the angiogenesis mechanism of renal ischemic injury.

Aged ; CD5 Antigens ; metabolism ; Chromosomes, Human, Pair 11 ; Chromosomes, Human, Pair 14 ; Cyclin D1 ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Lymphoma, B-Cell ; metabolism ; pathology ; Lymphoma, Follicular ; metabolism ; pathology ; Lymphoma, Mantle-Cell ; genetics ; metabolism ; pathology ; surgery ; Pseudolymphoma ; metabolism ; pathology ; Translocation, Genetic

Objective:To analyze KCNJ5 mutation of adenomas in patients with aldosterone-producing adenoma (APA) companying with hypokalemia, and to compare the clinical characteristics of patients with and without KCNJ 5 mutations.Methods:Clinical data of 144 APA patients were retrospectively analyzed. DNA were extracted from adenoma tissues, and amplified and sequenced for KCNJ5 gene. The serum potassium level and cardiac complications in patients with and without KCNJ5 gene mutation were compared.Results:Among 144 tumors, 131 tumors (91%) had KCNJ5 mutation, including 68 tumors with G151R, 56 tumors with L168R, 5 tumors with E145Q, and two tumors with novel mutations, V156_K160delITE and G151delinsVR. Compared with patients without KCNJ5 mutation, patients with KCNJ5 mutation had lower preoperative serum potassium levels, more cardiac complications, lower postoperative systolic blood pressure, and better postoperative hypertension relief. There were no statistical differences in age, gender, blood pressure, serum potassium level, plasma renin activity or plasma aldosterone concertration.Conclusion:91% adenomas in patients with APA and hypokalemia had KCNJ5 mutation, suggesting that KCNJ5 mutation is the main cause in these patients.