Differential expression of sigma factor of Mycobacterium tuberculosis under in vivo infection and in vitro culture conditions / 公共卫生与预防医学

Objective To investigate the gene expression of sigma factors in vivo, and to explore the sigma factors that may be closely related to the virulence of pathogenic Mycobacterium tuberculosis Methods Tuberculosis (TB) patients diagnosed in the outpatient department of Tianjin Tuberculosis Control Center from January to December 2018 were selected, and 20 sputum-positive specimens were randomly selected from TB patients confirmed with Xpert-positive for the present study. Two immediate sputum specimens were collected from each case of pulmonary tuberculosis before treatment, one for RNA extraction and one for in vitro culture. In vitro cultured strains in the logarithmic phase of growth were harvested for RNA extraction. The specific primers for 13 sigma factors were designed. The differential expression of the 13 sigma factors between sputum isolates and in vitro cultured strains was analyzed by fluorescence quantitative PCR. Taking ribosomal 16s as the reference gene, the transcription level of sigma factors was analyzed by 2ΔCt. Using the stably expressed sigA as the control reference, the expression differences of other sigma factors were analyzed by one-way ANOVA. Results Within 0 days, stress-associated sigma factors have a different expression profile in clinical isolate strains vs H37Rv or in vitro. All the sigma factors induced up regulation in sputum ,while no difference transcription between clinical isolate strains vs H37Rv（P＞0.05）. When compared to in vitro culture ,only sigM transcript highest in sputum(P＜0.05). Conclusion SigM plays an important role in the initial stages of bacterial infection, but its exact role is unclear.We assumed it could have a role in the interplay between the host immune defenses and the bacterial escape mechanisms.

Evaluation of GenoType® MTBDRplus VER2.0 kit for detecting mycobacterium tuberculosis complex in sputum / 公共卫生与预防医学

Objective To evaluate the performance of GenoType®MTBDRplus VER2.0 kit for detecting Mycobacterium tuberculosis complex (MTBC) in sputum. Methods Sputum samples from 177 patients with suspected tuberculosis were collected, and tested by smear, MGIT liquid culture and GenoType® MTBDRplus VER2.0. When the liquid culture was positive, identification of strains was carried out. The results were statistically analyzed using SPSS 22.0, and the consistency of the count data was compared using the Kappa test. Good consistency was defined as K≥0.75. The sensitivity and specificity were applied to evaluate the performance of GenoType® MTBDRplus VER2.0 kit for detecting MTBC in sputum. Results Based on the MGIT liquid culture results,，the sensitivity, specificity, positive predictive value, negative predictive value and total coincidence rate of the GenoType® MTBDRplus VER2.0 kit was 91.67%, 95.58%, 91.67%, 95.58% and 94.22%，respectively．The Kappa test was performed on both methods, K=0.872 (P® MTBDRplus VER2.0 kit had high sensitivity and specificity for detecting MTBC in sputum, and it has good application value for early diagnosis of tuberculosis.

Neuron protective effects of ligustilide on cortex region of cerebral hypoperfusion rats / 第二军医大学学报

Objective To investigate the neuron protective effects of ligustilide (LIG) on cortex regions of cerebral hypoperfusion rats and the possible mechanisms of the protective effect. Methods Bilateral common carotid arteries (two-vessel occlusion, 2VO) of Sprague Dawley rats were occluded to induce the chronic cerebral hypoperfusion models. The sham-operated controls (Sham group) were not occluded. The model rats were treated with LIG (80 mg/kg, by oral) once a day from the 8th day after surgery for 21 d. The rats were sacrificed 7 d after stopping LIG treatment, and the brain tissues of rats were made into frozen sections. Nissl staining, immunohistochemistry and immunofluorescence experiments were performed. Coronal sections of the cortex were stained with cresyl violet or with antibodies for neuronal specific nuclear protein (NeuN) , microtubule associated protcin-2 (MAP-2) and cystcinyl aspartate specific proteinase 3 (Caspasc-3). Results The number of Nissl body and NeuN-positive cells were increased significantly in the cortex region of LIG treatment group compared with those in the model group (P

Current status and urban-rural comparison of clinical agency of detection, management, and health insurance for hypertensive patients in communities of five provinces in China in 2010 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the status of the clinical agency of detection, management, and health insurance for hypertensive patients in urban and rural communities of five provinces in China in 2010, in order to provide fundamental data for implementation and evaluation of community health management of hypertensive patients in basic public health service.</p><p><b>METHODS</b>From Jiangsu, Shandong, Hebei, Sichuan and Gansu provinces, cities and districts (counties) were selected according to economic development level and 10 survey sites were finally determined. In each survey site, 3-4 communities or townships were selected by cluster sampling methods in 2010. A total of 8326 eligible hypertensive patients (4363 in urban and 3963 in rural) were included. The urban-rural difference of clinical agency and health insurance was compared for hypertensive patients.</p><p><b>RESULTS</b>In urban areas, 43.74% (1867/4268) hypertensive patients were first diagnosed at hospitals of district level or above, 25.07% (1070/4268) at community health service centers (CHSC), and 20.20% (862/4268) at community health service stations (CHSS), respectively; 30.72% (1274/4147) and 31.11% (1290/4147) patients chose CHSC and CHSS for their follow-up visiting, respectively; 60.23% (3073/5102) antihypertensive medication was obtained from pharmacies. In rural areas, 54.58% (2133/3908) hypertensive patients were first diagnosed at village clinics, 22.36% (874/3908) at township hospitals, and 18.86% (737/3908) at hospitals of county level or above; 70.49% (2695/3823) patients chose village clinics for their follow-up visiting; 46.23% (2116/4577) antihypertensive medication was obtained from village clinics, and 36.29% (1661/4577) from pharmacies. The main reasons for choosing clinical agency for both urban and rural patients were convenience (45.79%, 6276/13 706) and low cost (11.78%, 1614/13 706). The proportions of reimbursements for hospitalization expenses and total medical expenses for hypertensive patients in urban in the past year were 66.67% and 34.78%, respectively, which were much higher than those in rural (35.71% and 9.50%) (Z value was -12.13 and -17.56, P < 0.01).</p><p><b>CONCLUSION</b>Community-based hypertension detection and routine blood pressure measurement during clinical visiting should be further strengthened to improve early diagnosis of hypertension. The development of community-based clinical agency should be able to provide convenient and low cost health service for hypertensive patients to improve treatment, follow-up and control of hypertension.</p>

The analysis of drug cost and direct medical expense in community health management of hypertensive patients / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the current situation of drug cost, hospitalization cost and direct medical expense in community health management of hypertensive patients, in order to lay foundation for evaluating whether the community health management in basic public health service has cost-effect in Health Economics.</p><p><b>METHODS</b>A total of 8326 hypertensive patients from 10 survey pilots in 5 provinces were selected by cluster sampling methods, including 3967 patients who took part in community health management for over 1 year as management group and 4359 cases who have never taken part in community health management as control group. The essential information of research objects were collected by questionnaire; and the medical cost information in the last year (from November 2009 to November 2010) were collected retrospectively. The different annual medical treatment cost, hospitalization cost and direct medical expense in the two groups were compared and analyzed.</p><p><b>RESULTS</b>The average annual drug cost in hypertension was (621.50 ± 1337.78) yuan per patient; while the cost was (616.13 ± 1248.40) yuan in management group and (626.44 ± 1414.30) yuan in control group respectively. The average annual drug cost of hypertensive patients who took medicine therapy was (702.05 ± 1401.79) yuan per person, while the cost in the management group ((688.50 ± 1300.70) yuan) was much lower than it in control group ((714.64 ± 1489.60) yuan). The annual average drug cost in urban was (731.88 ± 1403.31) yuan per person, which was higher than it in rural as (407.44 ± 1171.44) yuan per person. The average hospitalized rate was 12.2% (1014/8326), and the average annual cost among the hospitalized patients was (9264.47 ± 18 088.49) yuan per person; while the cost was (7583.70 ± 13 267.00) yuan in management group, which was lower than it in control group as (11 028.00 ± 21 919.00) yuan. The average annual hospitalized cost in hypertension was (1064.87 ± 6804.83) yuan per person; while the cost was (936.73 ± 5284.90) yuan in management group, which was lower than it in control group as (1181.50 ± 7937.90) yuan. The average annual direct medical expense in hypertension was (2275.08 ± 8225.66) yuan per person; while the expense was (2165.10 ± 6564.60) yuan in management group and (2375.20 ± 9487.60) yuan in control group. The average annual direct medical expense in urban ((2801.06 ± 9428.54) yuan per person) was higher than it in rural ((1254.70 ± 4990.27) yuan per person).</p><p><b>CONCLUSION</b>The community health or standardized management of hypertensive patients can reduce the average annual drug cost and hospitalization cost (around 26 yuan and 245 yuan separately); and thereby save the annual direct medical expense per capita in hypertension (around 210 yuan). In the reform and development of national medical health system, we should enhance and promote the standardized community health management of hypertensive patients.</p>

Association of drug efflux pump gene expression with Mycobacterium tuberculosis drug resistance / 中华检验医学杂志

Objectives To explore the associations between drug efflux pump gene expression and phenotypic drug resistance as well as gene mutation patterns related to drug resistance of Mycobacterium tuberculosis.Methods Forty-five Mycobacterium tuberculosis isolates resistant to one or more of drugs including isoniazid, rifampicin, streptomycin and ethambutol, and 26 isolates all sensitive to the above four drugs from Tianjin Tuberculosis Control Institute in 2007 were involved in this study. Direct sequencing was applied to detect the mutations in the corresponding resistance genes(isoniazid:katG, inhA, oxyR-ahpC, ndh, rifampicin:rpoB, streptomycin:rpsL, rrs, and ethambutol:embB, embC and embA). After RNA extration and reverse transcription, real-time PCR was conducted to assess the expressions of putative drug efflux pump genes Rv1410c, Rv2136c, Rv0783c and Rv2136c, and Students' t test and ANOVA analysis were used to analyze the expression differences in Mycobacterium tuberculosis with different phenotypic drug resistance and drug resistance related gene mutation patterns.Results Compared to pan-sensitive isolates[(5.67±3.29)×10-5], Rv1410c showed higher expression in streptomycin[(8.48±6.33)×10-5, t'=2.18, P<0.05], isoniazid[(8.43±6.38)×10-5, t'=2.20, P<0.05], rifampicin[(9.59±7.27)×10-5, t'=2.29, P<0.05], multi-drug[(10.37±7.86)×10-5, t'=2.34, P<0.05] resistant isolates, and in isoniazid + streptomycin resistant isolates[(9.39±6.81)×10-5, t'=2.43, P<0.05];Rv2136c showed higher expression in isoniazid resistant[(3.51±2.43)×10-5, t'=2.03, P<0.05], multidrug-resistant isolates[(4.21±2.94)×10-5, t'=2.22, P<0.05] and resistant to isoniazid+streptomycin[(3.81±2.46)×10-5, t'=2.28, P<0.05] isolates . The expression of Rv0783c in rifampicin resistant isolates with rpoB 531 mutations [(5.41±3.03)×10-6] was higher than those with wild type of rpoB 531[(2.29±1.62)×10-6, t=2.81, P<0.05].Conclusions The expression of Rv1410c and Rv2136c are associated with mutiple-drug resistance of Mycobacterium tuberculosis.The expression of Rv0783c in rifampicin resistant isolates is associated with mutation in rpoB 531.

Characteristics of Mycobacterium tuberculosis genotype and the relationship between Beijing genotype and drug-resistant phenotypes in Tianjin / 中华流行病学杂志

Objective To explore the distribution and characteristics on genotype of Mycobacterium tuberculosis and the relationship between Beijing genotype and drug-resistant phenotypes in Tianjin city. Methods 656 clinical strains were collected from Tianjin Center for Tuberculosis Control and ten other Tuberculosis Institute in Tianjin from January 2008 to June 2009.Information regarding administration, clinical as well as laboratory findings of patients were collected.Proportion method was adopted to detect the susceptibility on four anti-tuberculosis drugs, namely streptomycin (SM), isoniazid (INH), rifampicin (RFP) and ehambutol (EMB). Both Beijing and non-Beijing genotypes were differentiated by multiplex PCR. The relationship between Beijing genotype and drug-resistant phenotypes was analyzed. Results In this study, the overall resistance rate of MTB was 26.98%, with multidrug-resistant rate was 6.25%. Among 656 MTB strains, 600isolates (91.46% ) belonged to Beijing genotype. There was significant difference between Beijing and non-Beijing genotype (x2=4.26, P=0.039) among the Tianjin household registered population.Concerning the drug resistance, there was no significant difference between the two groups.Conclusion Beijing genotype strains were the predominant one in Tianjin. The proportion of people infected with the Beijing genotype strains in Tianjin household registration of patients was significantly higher than the proportion of patients in the floating population in the same region.Results from the statistical analysis did not reveal any statistically significant association between Beijing genotype and drug resistance.

Molecular characterization of drug-resistant Beijing family isolates of Mycobacterium tuberculosis from Tianjin, China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Tuberculosis remains a severe public health issue, and the Beijing family of mycobacterium tuberculosis (M. tuberculosis) is widespread in East Asia, especially in some areas in China, like Beijing and Tianjin. This study aimed at determining the mutation patterns of drug-resistant Beijing strains of M. tuberculosis isolated from Tianjin, China.</p><p><b>METHODS</b>A total of 822 M. tuberculosis isolates were screened for drug resistance by an absolute concentration method and the genotype was identified by PCR. 169 drug-resistant isolates of the Beijing family were analyzed for the potential mutations in the rpoB, katG, inhA promoter region and in rpsL, rrs and embB genes, which are associated with resistance to rifampin (RFP), isoniazid (INH), streptomycin (SM) and ethambutol (EMB) respectively by PCR and DNA sequencing.</p><p><b>RESULTS</b>Fifty-eight out of 63 RFP-resistant isolates were found to carry the mutations within the 81-bp RFP resistance determining region (RRDR) of the rpoB gene and the most frequent mutations occurred at codon 531 (44.4%), 526 (28.6%), and 516 (7.9%) respectively. 16 mutation patterns affecting 12 different codons around the RRDR of rpoB were found. Of 116 INH-resistant isolates, 56 (48.3%) had the mutation of katG 315 (AGC-->ACC) (Ser-->Thr), 3 (2.6%) carried S315N (AGC-->AAC) and 27 (16.0%) had the mutation of inhA-15A-->T. 84 out of 122 SM-resistant isolates (68.9%) displayed mutations at the codons 43 or 88 with AAG-->AGG (Lys-->Arg) of the rpsL gene and 22 (18.0%) with the mutations at positions 513A-->C, 516C-->T or 905 A-->G in the rrs gene. Of 34 EMB-resistant isolates, 6 had mutation with M306V (ATG-->GTG), 3 with M306I (ATG-->ATT), 1 with M306I (ATG-->ATA), 1 with D328Y (GAT-->TAT), 1 with V348L (GTC-->CTC), and 1 with G406S (GGC-->AGC) in the embB gene.</p><p><b>CONCLUSION</b>These novel findings extended our understanding of resistance-related mutations in the Beijing strains of M. tuberculosis and may provide a scientific basis for development of new strategies for diagnosis and control of tuberculosis in China and other countries where Beijing strains are prevalent.</p>

Beneficial effect of total flavonoids of Chrysanthemum indicum on adjuvant arthritis by induction of apoptosis of synovial fibroblasts / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect on the extract of total flavonoids of Chrysanthemum indicum (TFC) on adjuvant arthritis synovial cells.</p><p><b>METHOD</b>SD rats were divided randomly into six groups including normal, model, TFC (84, 168, 336 mg x kg(-1)) and control drug Tripterygium glycosides (30 mg x kg(-1)) groups. Adjuvant arthritis rat model was induced by a single intradermal injection of 0.1 mL of the complete Freund's adjuvant into the right hind feet pads of the SD rats. The proliferation of synoviocyte was measured by MT; The apoptosis rates of synovial cells were evaluated using TUNEL and FCM analysis.</p><p><b>RESULT</b>TFC resulted in a dose-dependent way in inhibiting the proliferation of synovial and inducing the apoptosis of synovium and synoviocytes in vivo.</p><p><b>CONCLUSION</b>TFC can inhibit proliferation and induce apoptosis in synovial cells, and exert therapeutical effect on rheumatoid arthritis.</p>

Expression and clinical significance of hepatocellular cancer antigen genes in human hepatocellular carcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of four hepatocellular cancer antigen (HCA) gene mRNA in hepatocellular carcinoma.</p><p><b>METHODS</b>The expression of HCA90, HCA519, HCA520, HCA587 mRNA was detected using RT-PCR in HCC tissues and the corresponding adjacent non-HCC tissues from 46 HCC patients, cirrhosis tissues from 10 samples and normal liver tissues from 10 samples. The relationship between positive expression rate of HCA gene and clinical and lab data was evaluated.</p><p><b>RESULTS</b>Of 46 HCC tissues, HCA90, HCA519, HCA520 and HCA587 mRNA were detectable in 65.2%, 76.1%, 45.7% and 32.6%, respectively. At least one HCA gene mRNA was positive in 82.6% of HCC tissues. Only weak expression of HCA519 could be detectable in 6.5% of the corresponding adjacent non-HCC tissues. None of 10 samples of cirrhosis and normal liver tissues expressed any HCA gene mRNA. No correlation was found between the expression of HCA and clinical date such as age, sex, tumor size, tumor differentiation, serum alpha-fetoprotein level and hepatitis B virus infection or hepatitis C virus infection (P > 0.05). However, in some patients with normal serum alpha-fetoprotein (< 25 ng/L), specific expression of HCA genes was observed.</p><p><b>CONCLUSION</b>HCA gene mRNA is expressed with a high percentage and specificity in hepatocellular carcinomas and their products are new potential promising targets for immunotherapy of HCC.</p>