Table of Content

10 May 2014, Volume 36 Issue 5

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The contribution of Global Fund Tuberculosis Project to China tuberculosis control

JIANG Shi-wen，ZHANG Hui，WANG Dong-mei，WANG Li-xia

Chinese Journal of Antituberculosis. 2014, 36(5):  305-307.  doi:10.3969/j.issn.1000-6621.2014.05.001

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The analysis of 119 cases of multidrug-resistant tuberculosis patients not enrolling for proper treatment in different financing models

RUAN Yun-zhou, LI Ren-zhong, ZHAO Jin, SU Wei, CHENG Jun, ZHANG Can-you, WANG Li-xia, HAO Yang

Chinese Journal of Antituberculosis. 2014, 36(5):  308-312.  doi:10.3969/j.issn.1000-6621.2014.05.002

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Objective  To explore and describe the proportion and the reasons for multidrug-resistant tuberculosis patients nonparticipation of proper treatment. Methods  Five prefectures (Daqing of Heilongjiang, Quzhou of Zhejiang, Puyang of He’nan, Tianjin, Wanzhou of Chongqing) with different financing models were selected as settings by specialists panel of the key national science and technology program. Two hundred and thirty five MDR-TB patients confirmed by traditional DST between Mar 1^st, 2010 and Feb 29^th, 2012 were selected as study objects. All of them received face to face or telephone interview by the trained professionals with half-structured questionnaire including individual information (such as name, gender, age, occupation, income, notification classification, whether enrolled or not) and reasons of nonparticipation (financial risk, health, concerns, transportation, attitude and judgment of doctors, others).  Results  Proportion of patients not enrolled varied by financing models. The non-enrollment proportions were 39.0%（46/118）, 50.0%（25/50）and 71.6%（48/67）respectively（χ²=17.809，P<0.01） in all-free model, only free for drugs model, and all paid by patients model. The main reason of nonparticipation of all-free model is death（37.0%，17/46）, refusing enrollment（21.7%，10/46）, loss of contact（17.4%，8/46）; of only free for drugs model is loss of contact（64.0%，16/25）, transfer-out（16.0%，4/25）, refusing enrollment（12.0%，3/25）; of paid all by patients model is economic difficulties(33.3%，16/48), death(22.9%，11/48), loss of contact(16.7%，8/48)（χ²=62.326，P=0.000）. The reasons for non-enrollment are significantly different by models. The non-enrollment proportion could reduce 39.1%（18/46）, 64.0%（16/25） and 50.0%（24/48） respectively if we could solve refusing and loss of contact issue in all-free model, loss of contact in only free for drug model and economic difficulties and loss of contact for all paid by patients model.  Conclusion  Proportion of nonparticipation for proper treatment of MDR-TB cases will decline with the increase of the scope of free treatment. In addition to the economic difficulties, the study shows that nonparticipation may also relate to other reasons like loss of contact, death during waiting for DST results, refusing enrollment and severe complications.

Establishing indicator system of evaluation on tuberculosis operational research

CHENG Jun, ZHAO Fei, HU Dong-mei, ZHANG Can-you, FAN Hai-ying, LI Meng, CHENG Shi-ming, WANG Li-xia

Chinese Journal of Antituberculosis. 2014, 36(5):  313-317.  doi:10.3969/j.issn.1000-6621.2014.05.003

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Objective  To establish an indicator system to evaluate the whole process of tuberculosis operational research. Methods  Two rounds of Delphi consultation were implemented among 20 selected experts, and the effect of Delphi was evaluated from the aspects of enthusiasm, authority degree and coordination degree of the panel experts. The average score of every indicator was calculated, and the weights of indicators were obtained by percentage weight method. Results  An evaluation indicator system including 4 first-class and 29 second-class indicators with percentage weights was established, covering the whole process of research selection, implementation, output and application. The enthusiasm coefficient of panel experts were 95.0%(19/20) for two rounds, and suggestion were given by 36.8%(7/19) and 21.1%(4/19) of experts respectively. The authority coefficients(Cr) of all second-class indicators ranged from 0.82 to 0.89, and coefficient of variation(CV) from 0.09 to 0.23. The coordination coefficient of se-cond-class indicators was 0.414(χ²=156.541, P<0.01). The highest average score of se-cond-class indicator was 9.60 and the lowest one was 7.37, and average scores for 5 indicators were higher than 9.0, being 9.53, 9.60, 9.30, 9.03 and 9.03. Conclusion  The evaluation indicator system with reliability and operability is suitable for making both the whole process evaluation and stage evaluation on tuberculosis operational research.

Survey on current situation of tuberculosis (TB) infection control in designed medical institutions of tuberculosis in 3 cities

ZHAO Fei, WANG Li-xia, HE Guang-xue, CHENG Shi-ming, ZHANG Hui, QU Yan, HUANG Fei, HU Dong-mei, ZHANG Can-you, FAN Hai-ying, LI Meng, CHENG Jun

Chinese Journal of Antituberculosis. 2014, 36(5):  318-322.  doi:10.3969/j.issn.1000-6621.2014.05.004

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Objective  To strengthen TB infection control and provide some suggestions for reducing the transmission of Mycobacterium tuberculosis by understanding the current situation of TB infection control in designa-ted medical institutions in China. Methods  Field survey on current situation of TB infection control was conducted in 12 designed medical institutions of TB in 3 cites from March to May in 2013. Descriptive analysis was used to analyze the administrative control, environmental control and personal protective equipment. Air change per hour (ACH) and intensity of ultraviolet irradiation were measured in outpatient departments, laboratories and inpatient wards separately. Good ventilation is defined by ACH≥12/h. The requirement of the intensity of ultraviolet irradiation is over 70μW/cm². Results  Of the 12 TB prevention and control institutions, 8 institutions had the regulations of TB infection control. The outpatient departments of 2 institutions were properly designed, of 10 institutions where ACH reached the requirement, and of 6 institutions where the intensity of ultraviolet irradiation reached the requirement. The laboratories of 6 institutions were properly designed, of 6 institutions where ACH reached the requirement, and of 5 institutions where the intensity of ultraviolet irradiation reached the requirement. The inpatient departments of 3 institutions were properly designed, of 11 institutions where ACH reached the requirement. Ultraviolet lights were installed in the wards of 4 institutions and the intensity of ultraviolet irradiation that reached the requirement only in 1 institution. Eleven designed medical institutions could supply the protective face masks for medical use for health care workers and 10 institutions could supply medical surgical masks for TB patients and TB suspects.  Conclusion The TB infection control work is weak and should be strengthened comprehensively in designed medical institutions of TB, especially in the laboratory.

Analysis of information management for 178 immigrants notified as pulmonary tuberculosis by health institutions outside China

LI Jun，LIU Xiao-qiu，LI You，JIANG Shi-wen，ZHANG Hui，WANG Li-xia

Chinese Journal of Antituberculosis. 2014, 36(5):  323-326.  doi:10.3969/j.issn.1000-6621.2014.05.005

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Objective  To understand the situation of information management for patients who entered China after they were notified as pulmonary tuberculosis (PTB) by medical institutions outside China, and to provide evidences for strengthening TB control among immigration in China.  Methods Using a quantitative descriptive analysis method, we analyzed the information and data of 178 PTB patients which were provided by the health institutions outside China from 1^st July 2007 to 31^st December 2013, including the countries where the patients were notified and reported, the provinces that the patients stayed after they entered China, as well as the results of patients tracing, treatment management and information feedback.  Results Among 178 reported PTB cases, 14.6% (26/178) of them were multidrug-resistance PTB and 83.7%(149/178) of the total patients were reported by the health institutions in United States, China Taiwan and Canada, which ranked as the top three counties in terms of the number of reported PTB cases. After the patients entered China, they mainly lived in the eastern part of China, such as Guangdong, Fujian, Shanghai, Jiangsu and Beijing, etc., which accounted for 65.2% (116/178) of total patients. 61.8% (110/178) of reported patients were successfully reached by tracing of the local TB dispensaries, while 14.0% (25/178) of patients could not be contacted due to insufficient or incorrect contact information. The percen-tages of the patients that no any feedback information was received from the local TB institutions were 20.7% (24/116), 36.1% (13/36) and 27.8% (5/18) respectively in the eastern, middle and western areas of China.  Conclusion There is a risk of TB/MDR-TB transmission in China due to the entry of PTB patients, and the eastern part of China should be more concerned. Effective and tailored measures should be adopted to strengthen the information management for notified PTB cases once they enter the country.

The comparative study of health check and passive identification in tuberculosis case detection

LIU Er-yong, ZHOU Lin, CHENG Jun, ZHAO Fei, XIA Yin-yin, FAN Hai-ying, ZHOU Yang, LI Yang-yang, CHENG Shi-ming, WANG Li-xia

Chinese Journal of Antituberculosis. 2014, 36(5):  327-330.  doi:10.3969/j.issn.1000-6621.2014.05.006

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Objective  This study is to compare the TB patients detected by active health check and passive identification, including their geographical and clinical features, laboratory testing and to analyze the significance of active discovery in TB case detection in order to provide a basis for policy formulation.  Methods  We collected the health check information of 221383 people in part of “tuberculosis incidence and intervention pattern” study area (hereinafter referred to as “active discovery methods”)and 7625650 people information among which TB cases were detected by passive identification (hereinafter referred to as “passive discovery methods”). The data of patients detected by active discovery were from the project survey; of passive discovery were from the TB patients management system, out-patient medical records, registration of newly diagnosed patients. Using Excel software for database setting up and double entry, packages SPSS 17.0 for statistical analysis. We compared two groups of patients using a Chi-square test, with P<0.05 as a statistically significant difference. Results  A total of 444 cases of TB patients were detected by active case discovery with the detection rate of 200.56 per 100000. In contrast, 758 cases of TB patients were registered in TB institutions with the registration rates of 9.94 per 100000.In 444 patients with active discovery, the male∶female=2.70∶1; 358 cases elder than 60 years accounting for 80.63%. In 758 cases with passive discovery, male: female=1.86∶1; 300 cases elder than 60 years accounting for 39.58%. The gender and age distribution in active and passive discovery group are statistically different (χ²=5.430 and 203.090 for gender and age respectively, P both less than 0.05). For active discovery, 7.43%（33/444） were sputum smear-positive, lower than passive discovery of 39.31%(298/758). In active discovery patients, the lesions of 251 cases ranged in 1-2 lung fields, accounting for 56.53%;153 cases of 3-4 lung fields accounting for 34.46%;40 cases of 5-6 lung fields accounting for 9.01%.In passive discovery patients, the lesions of 508 cases ranged in one or two lung fields accounting for 67.02%;171 cases of 3-4 lung fields accounting for 22.56%;79 cases of 5-6 lung fields accounting for 10.42%;The difference is statistically significant in two groups(χ²=19.517,P<0.001). 6.53%（29/444） of active discovered patients had lung cavities, significantly lower than 20.84% (158/758) of passive discovered patients (χ²=43.661，P<0.001）. Conclusion  The illness of the patients detected by active discovery may not be light, but symptoms not obvious. The active discovery has an important value for early diagnosis of patients with tuberculosis.

Analysis on process of seeking health care and case-reporting in patients with pulmonary tuberculosis in China in 2010 and its relevant factors

XIA Yin-yin, RUAN Yun-zhou, ZHANG Hui, CHEN Wei, DU Xin, LIU Xiao-qiu, LI Xue, JIANG Shi-wen, CHENG Shi-ming, WANG Li-xia

Chinese Journal of Antituberculosis. 2014, 36(5):  331-336.  doi:10.3969/j.issn.1000-6621.2014.05.007

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Objective  To analyze the process of pulmonary tuberculosis (TB) patients, who were detected during the 5^th National TB Prevalence Survey in 2010, from onset of symptoms and health care seeking to diagnosis with TB, reporting and treatment; to compare the difference of those in urban and rural areas in China. Methods  A face-to-face interview with structured questionnaire was conducted among pulmonary TB patients who were detected during the TB prevalence survey, which was conducted in 77 urban sites and 99 rural sites in China in 2010. Information and data related to socio-economic, onset of symptoms, health care seeking behaviors, process of diagnosis, reporting and treatment were collected; information and data from 1301 patients who completed the interview (373 patients were from urban and 928 patients were from rural) were analyzed andχ² test was used to compare the difference between urban and rural areas (the level of statistical significance was P<0.05); reasons for the difference were explored. Results  49.3% (184/373) of urban patients had TB-related symptoms before the prevalence survey and this proportion was lower than that of rural patients (59.9%, 556/928); the difference had statistical significance (χ²=12.15, P<0.01). Among urban and rural TB patients, the proportions of symptomatic patients who sought health care were 47.3% (87/184) and 46.6% (259/556) respectively, the proportions of patients diagnosed with TB after sought health care were 41.4% (36/87) and 34.0% (88/259) respectively, the proportions of patients who were reported in the National Infectious Diseases Surveillance System after diagnosis with TB were 69.4% (25/36) and 70.5% (62/88) respectively, the proportions of patients who received anti-TB treatment after diagnosis with TB were 91.7% (33/36) and 88.6% (78/88) respectively; those differences between urban and rural patients had no statistical significance (χ²＝0.03, 1.52, 0.01, 0.03; P>0.05). Throughout the whole journey of the TB patients from onset of symptoms to treatment, we could understand the most important reasons to lead patients to remain undetected and untreated, including patients were asymptomatic (42.1%, 548/1301); patients had symptoms but did not seek health care (30.3%, 394/1301); patients had symptoms and sought health care but were diagnosed with other diseases (17.1%, 222/1301). Only 8.5% (111/1301) of TB patients, who sought health care for symptoms and were diagnosed with TB, got anti-TB treatment before the prevalence survey; another 1.0% (13/1301) of TB patients, who did not have symptoms but visited health facilities due to other reasons, were also diagnosed with TB. Conclusion  The following issues should be regarded as the weak points in TB control and need to be addressed, such as the symptomatic patients do not seek health care and TB patients are not timely diagnosed and treated. Compared with TB patients in urban area, the proportion of asymptomatic patients in rural area was significantly lower; however, the proportions of patients had no statistically significant differences in health care seeking, TB diagnosis and anti-TB treatment.

The analysis of TB case detection in College candidates physical examination of Guangzhou, 2009—2013

WU Xiao-ying,TAN Shou-yong, LUO Shao-xia, ZHANG Jin-xin, CHEN Qi-chen, LIU Guo-biao, LIANG Zhi-qiang, SONG Tao, HUANG Dong-sheng, DU Yu-hua，KUANG Hao-bin,ZHOU Hui-xian, GONG Fang, WEN Xing-xuan, HE Xi

Chinese Journal of Antituberculosis. 2014, 36(5):  337-340.  doi:10.3969/j.issn.1000-6621.2014.05.008

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Objective  To investigate the characteristics of tuberculosis detected in College candidates physical examination. Methods This is a retrospective study, we collected data from medical records of College candidates and National Tuberculosis Information Management System between 2009 and 2013, a total of 316386 candidates received the physical examination,the descriptive statistical analysis was used to analyze the 238 cases of active tuberculosis detected, χ² test was used for group comparison, P<0.05 was considered statistically significant. Results  The active tuberculosis detection rate was 75.2/100000 in College candidates between 2009 and 2013,relatively higher in 2010 and 2011, which was 115.0/100000 and 102.0/100000 respectively, lowest in 2012 of 46.5/100000. Among them, males accounted for 55.9% (133/238). In terms of the number of cases detected by original residence, the highest 5 areas were Panyu district of 54 cases, Yuexiu district of 37 cases, Liwan district of 29 cases, Zengcheng district of 25 cases, Tianhe district of 22 cases, Baiyun district of 22 cases. All cases detected were new cases, among which, smear positive pulmonary tuberculosis patients accounted for 18.1% (43/238), smear-negative accounted for 81.9% (195/238). 90.7% (216/238) of them had lesions in 1-2 lung fields. Conclusion The active tuberculosis detection rate is high in College candidates of Guangzhou city, we should strengthen the high school TB control, especially in old town areas and densely populated areas of floating population.

Analysis of delayed health care seeking among patients with pulmonary tuberculosis in Liangping county of Chongqing and its relative factors

JIANG Ling，WANG Li-xia，HUANG Fei，ZHANG Hui

Chinese Journal of Antituberculosis. 2014, 36(5):  341-345.  doi:10.3969/j.issn.1000-6621.2014.05.009

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Objective  To understand the situation of delay in seeking health care among patients with pulmonary tuberculosis (TB) in Liangping county of Chongqing, and to explore the relative factors of patients’ delay. Methods  The individual information of all pulmonary TB patients (3365 patients in total) notified in Liangping county of Chongqing from 2005 to 2012 were exported from the National TB Recording and Reporting (R&R) System. The rate of patients’ delay in seeking health care was analyzed and described. Rank-sum test was used to analyze the differences of patients’ delay by sex, age and occupation of patient, as well as source of patient and type of TB; P<0.05 was considered to have statistical significance. Logistic regression analysis was used to identify if the patients’ delay was related to sex, age, occupation, source of patient and type of TB; the level of statistical significance was P<0.05. Results  The overall rate of pulmonary TB patients’ delay in seeking health care was 73.14% (2461/3365) in Liangping county of Chongqing, and the median of patients’ delay was 32 days; the time of delayed health care seeking was more than one month in 52.84% (1778/3365) of pulmonary TB patients. The delay was related to occupation, source of patient and type of TB: the TB patients with occupation as farmer had the highest delayed health care seeking compared with those in other occupations (OR＝1.513, 95%CI＝1.279-1.778, P＜0.05); compared with the patients who were referred by health facilities at county level, the patients who were referred by township hospital or village doctors as TB suspects were more likely to delay in seeking health care (OR=2.170, 95%CI＝1.798-2.618, P＜0.05); and the relapsed TB patients were more likely to have delayed health care seeking compared with new patients (OR＝1.222, 95%CI＝1.222-0.950, P＜0.05). Conclusion  The rate of delayed health care seeking among patients with pulmonary TB was high in Liangping county of Chongqing, which was associated with patient’s occupation, source of patient and type of TB. Effective measures should be taken to reduce the TB patients’ delay.

Evaluation of new information management model of TB prevention and control in Yichang

FANG Zheng-chao, LIU Xiao-jun, ZHANG Pei, PENG Ping-ping，ZHOU Ping, YUAN Feng-hua

Chinese Journal of Antituberculosis. 2014, 36(5):  346-349.  doi:10.3969/j.issn.1000-6621.2014.04.010

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Objective  To evaluate the implementation effects of integrated information management platform for disease control and prevention institutions, medical institutions and community health service institutions, and provide reference for the new model for TB patients management. Methods  Analysis the overall arrival rate and tracing rate of 1269 （630 cases in 2011，639 cases in 2012）suspected patients, the review on time rate of 451(241 cases in 2011，210 cases in 2012）new smear positive patients and the loss rate of 516 （277 cases in 2011，239 cases in 2012）patients before and after using TB Control Information System (TCIS) in Yichang. Using Delphi to analyze the difference including information delivery methods，reliability, timeliness, accuracy of 8 indicators between the 20 institutions using management platform and other 16 institutions not using it. Results  The overall arrival rate were 85.56%（539/630）and 92.64%（592/639）（χ²=16.45，P<0.01），the tracking in place rate were 63.89%（161/252）and 79.20%（179/226）(χ²=13.61，P<0.01）, the review on time rate after 5 months were 76.76%（185/241） and 94.29%（198/210）(χ²=26.91，P<0.5),the loss rate were 3.97%（11/277） and 0.84%(2/239)(χ²=5.13，P<0.05) respectively before and after using management platform. The differences were all statistically significant. For the 20 institutions used management platform，the 8 indicators including information delivery, reliability, timeliness, accuracy are all better to those not used. Conclusion  Application of TCIS management platform can strengthen the information communication between the institutions, improve work efficiency and patients service capability and management level.

Study on the mechanism of drug resistance in mono ofloxacin-resistant Mycobacterium tuberculosis

WANG Qian，SONG Yuan-yuan，CHI Jun-ying，WANG Yu-feng，ZHAO Yan-lin

Chinese Journal of Antituberculosis. 2014, 36(5):  350-355.  doi:10.3969/j.issn.1000-6621.2014.05.011

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Objective  To investigate the function of the genes related with drug resistance and efflux pump in mono ofloxacin (Ofx)-resistant M. tuberculosis (Mtb) isolates. Methods  Seventeen mono Ofx-resistant Mtb clinical isolates were obtained from National Tuberculosis Reference Laboratory. Direct sequencing was performed to detect the mutations in the gyrA/B genes. The total RNA was extracted from the Ofx-resistant strains without gyrA/B mutations. After RNA reverse transcription, the expression levels of 20 drug efflux genes were detected by Real-time PCR. In comparison with the corresponding MIC results, the Ofx-related efflux pumps were screened from 20 drug efflux genes, and the function of candidates was identified among transgenic E.coli. Results  Of 17 mono Ofx-resistant strains, 4 (4/17) strains were found the mutations of gyrA, in which 1 isolates had mutation at codon 90 and 3 isolate had mutation at codon 94. In addition, the isolates harboring gyrA mutation displayed high-level of drug resistance, their MICs were more than 4 μg/ml. Among the strains without gyrA/B mutations, the expression levels of Rv0933 and Rv2938 were higher in 2 high-level resistant strains, which was 16 and 5 folds higher than that of H37Rv strain, respectively. Furthermore, E.coli BL21 transferred with pEASY-E1-Rv0933 showed higher MIC level, 2μg/ml, compared with the control E.coli. Conclusion  The mutations in gyrA are associated with high-level of Ofx resistance. Rv0933 may be the Ofx-specific efflux pumps in Mycobacterium tuberculosis. The high level of Rv0933 expression may correlated with high-level of Ofx resistance.

Study on diagnostic value for active tuberculosis by detecting Mycobacterium tuberculosis specific antibodies in the peripheral blood

ZHAO Mei-fen, LIU Ying-xia, PENG Zhong-tian, CHEN Chuan-tie, LIU Jing, LIN Yi-min, CHEN Jian-bo

Chinese Journal of Antituberculosis. 2014, 36(5):  356-361.  doi:10.3969/j.issn.1000-6621.2014.05.012

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Objective  To evaluate the diagnostic value for active tuberculosis (TB) by detecting antibodies secreted from Mycobacterium tuberculosis-specific plasma cells in peripheral blood. Methods  The study period was from April 2013 to July 2013, and the study sample included 3 groups of subjects: 104 patients with active TB; 26 cases with latent TB infection (LTBI); and 33 healthy volunteers as a control group. The active TB patients were enrolled from the Outpatient Department of Shenzhen 3^rd People’s Hospital while the LTBI and healthy volunteers were enrolled from Health Examination Clinic of Shenzhen 3^rd People’s Hospital. Peripheral blood mononuclear cells(PBMCs) from all study subjects were isolated and cultured in vitro for 4 days, and then the supernatants of each group were collected. Mycobacterium tuberculosis-specific antibodies in lymphocyte supernatant were tested by ELISA and Western blot and the differences among the three groups were analyzed. GraphPad Prism 5.0 was used for data analysis. The diagnostic value of ELISA was evaluated by receiver operating characteristic (ROC) curve. The Kruskal-Wallis test was used for comparison among multi-groups and the Dunn multiple comparison test was used for comparison between two groups. The difference was considered to be statistically significance if the P-value was <0.05. Results  The A_450nm value of Mycobacterium tuberculosis-specific antibodies measured by ELISA was higher in the group of patients with active TB (0.593±0.206) than those in the group of cases with LTBI (0.342±0.152) and in the group of healthy persons (0.246±0.121) respectively, the differences had statistically significance (H=77.27, P<0.001). Areas under the curve (AUC) of Mycobacterium tuberculosis-specific antibodies which used for distinguish active TB and LTBI, active TB and healthy person were 0.857 and 0.944 respectively; when the value of 0.42 was used as a diagnostic threshold, the sensitivity and specificity for districting active TB and LTBI were 77.9% (81/104) and 80.8%(21/26) respectively, and sensitivity and specificity for distinguishing active TB and health people were 77.9% (81/104) and 93.9% (31/33) respectively. The sensitivity, specificity, and accuracy of Western blot test in detecting active TB patients and healthy person were respectively 79.2% (61/77), 100.0% (11/11) and 81.8% (72/88) (χ²=24.8, P<0.001). Conclusion  The specificity of both ELISA and Western blot tests are high in detecting Mycobacterium tuberculosis-specific antibodies secreted from peripheral blood plasma cells, so they can be used for diagnosis of active TB as new methods in clinical laboratory.

Experimental study on killing fluorescent M. bovis BCG by IFN-γ activated THP-1 derived macrophages

LAI Sai-lin，SUN Yi-fan，CHEN Tao，YANG Xiao， ZHOU Lin，ZHONG Qiu

Chinese Journal of Antituberculosis. 2014, 36(5):  362-368.  doi:10.3969/j.issn.1000-6621.2014.05.013

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Objective  To study fluorescent M.bovis BCG survival in infected IFN-γ activated THP-1 derived macrophage and its killing mechanism whether correlated with autophagy. PMA THP-1 differentiation into macrophages. Methods  THP-1 cells were induced by PMA and differentiated into macrophages, and then were activated by IFN-γ. The macrophages were divided into 4 groups: control, IFN-γ activated, Rapamycin treated and 3-MA+IFN-γ treated, and then subjected to fluorescent BCG infection at MOI 10∶1 for 48 hours. The cells were lysed with hyperpure water. The cell lysate and the culture supernatant were collected and mixed. 10 μl mixtures in 4 groups were diluted with 10-fold serial dilution, then inoculated on MiddleBrook 7H10 agar plates with OADC and cultured at 37℃ biochemical incubator for 3 to 4 weeks. The colonies on plates were counted，and the survival rates were calculated. At different time points of infection process, the macrophages were lysed and extracted the proteins. The expression of an autophagy marker LC3B was detected by Western blot, monitoring the changes of autophagy level. Results  Compared to the control group(inactivated macrophages), BCG survivals both in IFN-γ activated macrophages and Rapamycin treated macrophages decreased remarkably, whereas the levels of autophagy increased remarkably； but BCG survival in 3-MA+IFN-γ treated macrophages did not decrease, and the level of autophagy also did not increase. Conclusion  During the macrophages were infected by M. bovis BCG, or the macrophages phagocytosed BCG strains,the interaction between the macrophages and BCG strains was complex. IFN-γ activated macrophages were able to killed BCG effectively,the killing mechanism was related with the autophagy.

A study on the growth-promoting effect for Mycobacterium tuberculosis from tuberculous pleural effusion with resuscitation promoting factors

XING Ai-ying, LIU Zhong-quan, DU Bo-ping, SUN Qi, JIA Hong-yan, WEI Rong-rong, LIU Yang, CAO Ting-ming, DU Feng-jiao, GU Shu-xiang, MA Yu, ZHANG Zong-de

Chinese Journal of Antituberculosis. 2014, 36(5):  369-374.  doi:10.3969/j.issn.1000-6621.2014.05.014

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Objective  To investigate the growth-promoting effect of resuscitation promoting factors(Rpf) for Mycobacterium tuberculosis(Mtb) from pleural effusion in order to improve culture rate and short culture time of Mtb from clinical samples. Methods  Sixty-two patients with tuberculous pleurisy who admitted in Beijing Chest Hospital from Oct. 2012 to Oct. 2013 were recruited. Pleural effusion drained from every patient was centrifuged and inculated on Lowenstein-Jenson(L-J) media, MiddleBrook 7H11 containing with high- or low-concentration Rpf and 0.9% saline(as negative control). Colony formation unit(CFU) was observed every 3 days. Acid-fast bacilli(AFB), PCR and DNA sequencing for isolation identification were performed when colony formation was positive. Growth-promoting effect of Mtb were compared between media containing high- and low- concentration Rpf. Statistical analysis was performed using the software of SPSS version 17.0. Culture positive rate in different group was analyzed with Chi-square test. The average time of visible colonies and positive colonies were analyzed with paired sample t test. Results  (1) The culture positive rates of Mtb on L-J medium and 7H11 medium containing Rpf were 16.1% (10/62) and 43.5% (27/62), respectively. It was a statistically significant difference （χ²=12.84，P<0.01）. (2) The average colony formation time for isolates was （20.92 ± 0.58）d and （21.69 ± 0.50）d on 7H11 media containing high- and low-concentration of Rpf, respectively. There was no significant difference（t=0.085, P＞0.05）. However, it was more faster than that of negative controls, (38.08 ±0.94)（ t=10.19，t=9.91, P< 0.001）. (3) The number of CFU on the 7H11 media containing high- or low-concentration Rpf were （34.12±4.06）×10² and （37.27±5.63）×10², respectively,There was no significant difference statistically（t=0.45, P＞0.05）. But both of them were much more than the negative controls (3.77±0.88)×10² with statistically significant difference（t=5.88 ，t=7.30, P＜0.001）. Conclusion  MiddleBrook 7H11 medium containing Rpf is an ideal culture medium to improve culture positive rate for Mtb from the tuberculosis pleurisy effusion and short culture time.

Observation on the effect on the retreated tuberculosis patients by levofloxacin combined with rifabutin

LI Shi-lai, CHEN Wei-sheng, HUANG Yao-fan, LIN Shao-yun, WU Shao-yi, CAI Qing-he

Chinese Journal of Antituberculosis. 2014, 36(5):  375-379.  doi:10.3969/j.issn.1000-6621.2014.05.015

Abstract ( 1758 )   PDF (761KB) ( 470 )   Save

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Objective  To observe the effect on retreatment tuberculosis patients by levofloxacin combined with rifabutin. Methods  One hundred and seventy-two retreated TB cases were randomly divided into group A and group B, with 86 cases in each group. The patients in group A were treated with levofloxacin combined with rifabutin (levofloxacin 0.6 g/d, rifabutin 0.3 g/d, qd, PO), and the patients in group B were treated with streptomycin combined with rifampicin (streptomycin 0.75/d, qd, im; rifampicin 0.45 g/d, qd, PO). The sputum negative conversion rates after 8 months’ treatment, the sputum culture positive rates, the absorption rates of lesion and the cavity closure rates of the two groups after treatment were compared. Chi-square test was used to analysis and P<0.05 was set as the criteria of statistical significance. Results  The sputum conversion rate, the sputum culture positive rate, the lesions total absorption rate and the cavity closure rate of group A were 85.7% (72/84), 3.6% (3/84), 95.2% (80/84) and 78.8% (41/52) respectively, while those of group B were 58.5% (48/82), 14.6% (12/82), 61.0% (50/82) and 50.0% (24/48) respectively. The differences were significant between the two groups in all the indexes: the sputum conversion rate (χ²=61.9, P<0.01), the sputum culture positive rate (χ²=25.0, P<0.01), lesions total absorptivity rate (χ²=116.6, P<0.01) and cavity closure rate (χ²=28.8, P<0.01). Conclusion  The effect of levofloxacin combined with rifabutin on retreated TB patients is better than that of streptomycin combined with rifampicin. The sputum negative conversion, lesions absorption and cavity closure is significantly improved with the new treatment regimen.

Quality control and quality assurance of T-SPOT.TB

JIANG Jun

Chinese Journal of Antituberculosis. 2014, 36(5):  380-384.  doi:10.3969/j.issn.1000-6621.2014.05.016

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T-SPOT.TB has been widely used to assay the Mycobacterium tuberculosis infection all around the world. As a new cell biological and immunological detection technology which its operation isn’t complex, T-SPOT.TB also raises a claim strictly on experimental condition, operation technique and quality control. The overall quality control including PBMCs separation, adjusting the number of PBMCs, cell incubation condition and result determination is the key steps to succeed as well as to be ignored. According to the test principle and clinical practice, we described and analyzed the different states of PBMCs separation including PBMCs separation, suspension preparation, incubation and result determination, especially, the error of PBMCs count affected experimental result. In addition, we established “one-step” and “calculation formula” method for PBMCs count according to traditional improved NiuBao leukocyte count plate and solution concentration dilution method, PBMCs count and standard cell suspension final concentration of 2.5×10⁵/100μl.

Research progress of correlation between the polymorphisms of susceptibility genes associated with the function of macrophage and tuberculosis

JIANG Dao-bin,Qimanguli WUSHOUER

Chinese Journal of Antituberculosis. 2014, 36(5):  385-387.  doi:10.3969/j.issn.1000-6621.2014.05.017

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The function of the host macrophage causes different outcomes after Mycobacterium tuberculosis (Mtb) infection. The differences of the function of macrophage among individuals are not only the results of self-regulation in immune system, but also associated with the polymorphisms of susceptibility genes associated with the function of macrophage, including the genes influencing the killing, activation, apoptosis and other abilities of macrophage. This article reviews some achievements about the correlation between the polymorphism of susceptibility genes associated with the function of macrophage and pulmonary tuberculosis. The result shows partial polymorphism of susceptibility genes associated with the function of macrophage is correlated with the susceptibility to tuberculosis, and this correlation has not only ethical difference, but also is related to the clinical types of tuberculosis.