Objective To characterize the relationship between such phenotypes and the patterns of genetic mutations in the corresponding resistance genes in drug-resistant Mycobacterium tuberculosis (MTB)isolates in Shenzhen of China during 2007-2008.Methods According to standard of WHO,International Union Against Tuberculosis and Lung Disease(IUATLD),136 strains of MTB were collected by performing drug sensitivity test(DST)to isoniazid,rifampicin,streptomycin,ofloxacin and kanamycin on Lowenstein-Jensen in 1%proportion method.Genetic mutations in the corresponding resistance genes (rpoB,katG,rpsL,rrs1,gyrAB,rrs2)in these MTB isolates were identified by PCR,followed by DNA sequencing of the purified PCR products.The minimal inhibitory concentration(MIC)values of the aforementioned anti-tuberculosis drugs were determined for these MTB clinical isolates by two-fold dilution method in vitro.Results A total of 123 isolates were collected,73 isolates were drug resistant.50 isolates were drug susceptible.Among the isolates that were resistant to isoniazid,rifampicin,streptomycin.ofloxacin and kanamycin,the proportion of isolates that harboured mutations in the respective genes was 84.6%,93.6%,65.9%,100%,61.1%.For katG gene,the mutation detected were S315T or S315N.For rpoB,the most frequently found changes were S531L(30/44,68.2%)and H526D(9/44,20.5%)or H526R(1/44,2.3%).The reported mutations that K43R and KS8Q were founded in the rpsL locus and 491C→T and 513A→C were founded in the rrs1 gene related with streptomycin-resistant strains.For gyrA,all gyrA mutations were clustered in codons 90,91,and 94 apart from the S95T that was natural polymorphism.accounted for 81.1% of the ofloxacin-resistant isolates,and condon 91 was the most frequently mutated.No mutation were found in gyrB.The most frequent substitution were 1400 A→G(9/11,81.8%)and 1483 G→T(2/11,18.2%)in a specific region of the rrs2 gene related with kanamycin-resistant strains.No mutations except S95T of gyrA detected in the drug-susceptible isolates.The MIC values of clinical drug-resistant strains that the same drug-resistant group contains a different resistance phenotype are basically the same with the relevant resistance genes in the same mutation.Associated resistance mutations in different sites varied significantly with their MIC values.Conclusion The mutation characterization of drug-resistant and drug-suscep-tible isolates of MTB have been shown to vary according to geographic region,phenotypic characteristics exist difference in resistance levels due to different muntants of drug-resistant gene.

Objective To evaluate the performance of DNA microarray for rapid detection resistance to rifampin and isoniazid in Mycobacterium tuberculosis clinical isolates and identify suitable target sites for molecular genetic test.Methods Twenty-four clinical Mycobacterium tuberculosis isolates were collected retrospectively from Shenzhen Center for Chronic Disease Control in 2009 and 127 isolates from project on anti-tuberculosis drug resistance surveillance in Shenzhen during 2007 to 2009.Drug susceptibility to rifampin and isoniazid of the stains were determined by DNA microarray,and results were compared to that obtained with reference proportion method drug susceptibility testing for sensitivity,specificity and accuracy.The consistency of microarray and phenotypic susceptibility testing was evaluated by Kappa test.Genetic mutations in rpoB,katG,inhA,regulatory region of inhA,and regulatory region of ahpC were investigated by DNA sequencing to assess proper loci for rapid molecular diagnosis.Results Compared against results of proportion method,the sensitivity,specificity and accuracy of the DNA microarray assay for rifampin resistance were 94.4％,97.5％ and 96.0％ respectively,and for isoniazid resistance were 79.1％,100％ and 86.8％ respectively.Mutations in resistance-determining region of rpoB were observed in 97.2％ (70/72) of the isolates resistant to rifampin,which contributed in the 531,526,516,511 and 533codon region.Mutations in katG315 codon,inhA-15,and ahpC regulatory region were found in 70.3％ (64/91),11.0％ (10/91) and 9.9％ (9/90) of the isolates resistant to isoniazid,respectively.Mutations of ahpC promoter region consists of ahpC-9 (4 strains),ahpC-10 (2 strains),ahpC-6 (2 strains),ahpC-12 (1 strain),and ahpC-32 (1 strain).Conclusions DNA microarray provided a rapid method for the detection of drug-resistant Mycobacterium tuberculosis isolates,and demonstrated good performance except less sensitive for isoniazid resistance.The mutations in ahpC regulatory region might be good target loci for detection of isoniazid-resistant Mycobacterium tuberculosis,so screening the region may significantly improve the sensitivity for molecular genetic tests.

Objective To explore the feasibility and clinical significance of combined detection of human plasma and feces expression level of microRNA-92a-1 (miRNA-92a-1) as colorectal tumour screening marker.Methods From August to October in 2011,the feces and plasma samples of 60 colorectal cancer (CRC) patients,23 colorectal adenoma (CRA) patients and 30 healthy controls were collected.The expression level of miRNA-92a-1 was determined by quantitative reverse transcription-polymerase chain reaction.Mann-Whitney's U test was applied for the difference test between groups.Then,according to the receiver operating characteristic (ROC) curve analysis the cut-off point was determined and the sensitivity and specificity were analyzed.Results Compared with healthy controls,the expressions of miRNA-92a-1 in plasma of CRC and CRA patients increased (U=288.5 and 151.0,both P＜0.01).Compared with healthy controls,the expression of miRNA-92a-1 in feces of CRC patients increased (U=627.5,P=0.0199).According to ROC curve analysis,when the cut-off point was ＞1.22,the sensitivities of plasma miRNA-92a-1 in CRC patients and CRA patients were 85.0％ (51/60) and 73.9％ (17/23) respectively.The specificity of healthy controls was 76.7％ (23/30).When the cut-off point was ＞1.14,the sensitivities of feces miRNA-92a-1 in CRC patients and CRA patients were 31.7％(19/60) and 26.1％(6/23) respectively.The specificity of healthy controls was 90.0 ％ (27/30).Combined the detection results of plasma and feces,the sensitivities of miRNA-92a-1 in CRC patients and CRA patients was 88.3％ (53/60) and 82.6％ (19/23) respectively.The specificity in healthy controls was 73.3％ (22/30).The sensitivity was higher than that of single sample testing.Conclusions The sensitivity and specificity are high in the combined detection of plasma and feces miRNA-92a-1 expression level in CRC patients and CRA patients,miRNA-92a-1 may be a potential CRC screening marker.

Objective To study the relationship between ABO hemolytic disease of newborn and maternal antibody. Methods The titer of blood group antibody in 122 mothers of O blood group during prenatal diagnosis and blood group serology, bilirubin and hemoglobin level of newborn infants were tested with routine methods. The relationship between ABO hemolytic disease of newborn and the titer of blood group antibody was studied. Results The titer of blood group antibody was remarkably related with ABO hemolytic disease of newborn (P

AIM: To evaluate the relationship between the medial rectus cells counts in concomitant exotropia and surgical results. METHODS: A total of 32 pieces of medial rectus muscle were collected for HE staining in this study, of which 18 pieces were from patients with concomitant exotropia and 14 pieces were from healthy individuals. A method of strabismus score was used to assess the operative effect.RESULTS: The difference of strabismus score before and after the operation in the intermittent exotropia group was significantly higher than that in constant exotropic group (P<0.01). Under light microscope, the loosen muscle fibers and the increased stromal components in the cross sectional area of medial rectus were observed in strabismic group. The muscle cells counts was obviously lower in strabismic group than in control group (P<0.01), which was related to the difference of strabismus score before and after the operation (P<0.05).CONCLUSION: The decreased medial rectus cells counts induce concomitant exotropia directly. It is the crucial causes of the bad surgical results.

AlM: To compare the results of 4 methods for measuring angle of exodeviation in the three types of intermittent exotropia, including when looking at indoor distance target of 6m, looking at indoor distance target of 30m, looking at outdoor far distance target, after 1h diagnostic occlusion test.
METHODS: Prospective case series study. Sixty-five patients with intermittent exotropia between June 2013 and June 2014 were enrolled in the Department of Ophthalmology, Affiliated Hospital to Qingdao University, included 37 males and 28 females with average age ( 12. 5 ± 6. 2 ) years. All the patients were measured when looking at indoor distance target of 6m, looking at indoor distance target of 30m, looking at outdoor far distance target, after 1h diagnostic occlusion test. lntermittent exotropia was divided into basic type, convergence insufficiency type and divergence excess type, which was based on the different result of between the distance and near measurements. The One-way test was applied to analyze the four methods of measuring angle of exodeviation in the three types of intermittent exotropia. LSD - t test was applied to compare the differences between each two methods in each type.
RESULTS: The distance exodeviations tested with looking at indoor distance target of 6m, looking at indoor distance target of 30m , looking at outdoor far distance target, after 1h diagnostic occlusion test were basic type (45. 4 ± 21. 0, 55. 0 ± 15. 0, 64. 68 ± 17. 7, 68. 75 ± 16. 6PD), convergence insufficiency type (33. 3 ± 14. 0, 44. 9 ± 12. 9, 43. 6±11. 8, 54. 6±11. 2PD), divergence excess type (55. 6± 17.4, 66.3±18.8, 76.9±16.4, 78.1±15.6PD). There were obviously differences between each two methods in each type ( basic type F = 9. 649, P = 0. 00; convergence insufficiency type F=6. 886, P=0. 001; divergence excess type F = 7. 989, P = 0. 00 ). Compared with looking at indoor distance target of 30m, looking at outdoor far distance target ( basic type P=0. 044, divergence excess type P = 0. 048 ) and after 1h diagnostic occlusion test (basic type P=0. 04, divergence excess type P=0. 027) had the statistical difference in the basic type and divergence excess type, and there was no obviously difference between looking at outdoor far distance target and after 1h diagnostic occlusion test ( basic type P=0. 353, divergence excess type P=0. 815). Compared with the other three measurements, 1h diagnostic occlusion test can elicit larger angle of deviation in the convergence insufficiency type.
CONCLUSlON: Both measurement with looking at outdoor far distance target and after 1h diagnostic occlusion test can elicit the larger angle of deviation in the basic type and divergence excess type; The measurement with after 1 hour diagnostic occlusion test can elicit the larger angle of deviation in the convergence insufficiency type.

Calcaneus ; diagnostic imaging ; Female ; Humans ; Radiography ; Tuberculosis, Osteoarticular ; diagnosis ; diagnostic imaging ; Young Adult

AIM: To study the protective effect of mese nt eric lymph duct ligation on the functions of liver, kidney and heart, and morpho logy in multiple organ dysfunction syndrome (MODS) rats subjected to two-hit. METHODS: Male Wistar rats were divided into three groups: the me senteric lymph duct ligation group, the non-ligation group and sham group. The M ODS model of two-hit was established by bleeding and LPS administration in both ligation group and non-ligation group. After 24 h, all rats were cannulated to f acilitate blood withdrawal for serum sample, then all rats were killed and organ s including kidney, liver, lung and heart were collected for making microscopic sectio ns. The biochemical indexes of hepatic and renal functions and myocardial enzyme in s erum were determined before and after experiment. RESULTS: After two-hit, the serum contents of AST, ALT, TBA, BUN , Cr and LDH-1 in both non-ligation group and ligation group, and UA content in non-ligation group were obviously increased than those in pre-experiment and sh am group (P

In this study, five rhesus macaques were inoculated intravenously with SIVmac251 to establish a model of simian autoimmune deficiency syndrome (SAIDS). Peripheral blood samples were collected at different time points to monitor changes in the total T cell number and T lymphocyte subset. Plasma viral loads, cytokine expression levels and anti-SIV antibody levels were also assayed to acquire certain basic indexes to evaluate disease progression in the rhesus macaque SAIDS model. During the acute stage of infection, plasma viral loads reached a peak at week 1 post-inoculation and lasted for approximately 3 to 44 weeks. The CD3+ CD4+ T lymphocyte count in peripheral blood also transitorily decreased. During the same period, the level of interferon-gamma show an increasing trend, whereas IL-12 levels decreased; IL-2, IL-4, IL-10 and TNF-alpha were maintained at normal levels or could not be detected. During the asymptomatic and ARC phases, plasma viral loads persisted above 10(4) RNA copies/mL and either increased or declined during the later stages of disease; CD3+ CD4+ counts showed a steadily declining trend and the ratio of CD4 to CD8 decreased during late-stage disease. Moreover, antibodies against viral proteins were detected in the plasma and showed a significant increasing trend, while there were no apparently changes in the levels of IFN-gamma, IL-12, IL-2, IL-4, IL-10 and TNF-alpha. In conclusion, the characteristics of the SIV animal models in our study are similar to those of patients with AIDS. Therefore, the rhesus macaque SIVmac251 infection models can be applied for further studies into AIDS.
Animals ; Antibodies, Viral ; blood ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes ; virology ; Cytokines ; genetics ; immunology ; Disease Models, Animal ; HIV Infections ; genetics ; immunology ; virology ; HIV-1 ; physiology ; Humans ; Macaca mulatta ; Male ; Simian Acquired Immunodeficiency Syndrome ; genetics ; immunology ; virology ; Simian Immunodeficiency Virus ; physiology ; Viral Load

Objective To evaluate the effectiveness of MGIT liquid medium fluorescence instrument manual interpretation method for rapid detection of Mycobacterium. Methods Two hundred sputa with newly diagnosed tuberculosis patients were collected from October 2008 to January 2009 in the district hospitals in Shanghai. Of these 200 sputa, 67 sputa were positive AFB, 133 were negative. All the sputa were isolated by L-J, BacT / Alert 3D system and MGIT liquid medium methods. Results Of the 200 sputa specimens,105(52. 5% ) were isolated as Mycobacterium strains. The positive culture rate of the MGIT, BacT/Alert 3D and L-J method was 49. 5% ( 99/200 ), 48. 0% (96/200) and 45.0% ( 90/200), respectively. The MGIT culture positive rate was significantly higher than that of L-J method (x2 = 5.40, P = 0. 020 1 ). Of the 133 sputa with negative AFB, the positive culture rate was 24. 8% ( 33/133 ), 23. 3% ( 31/133 ) and 18. 8% (25/133) with MGIT, BacT/Alert 3D and L-J method, respectively. The MGIT culture positive rate with the AFB negative sputum was significantly higher than that of L-J method (x2 = 5. 33, P = 0. 020 9 ).The median time of detection with MGIT, BacT/Alert 3D system and L-J method was 11 days, 15 days and 22 days, respectively. Comparing the median time of detection of MGIT with BacT/Alert 3D, the difference was statistically significant ( Z = 3.414 ,P ＜ 0. 01 ). Comparing the median time of detection of MGIT with L-J method, the difference was statistically significant (Z =7.083,P＜0. 01).Conclusions MGIT liquid medium manual method is a rapid detection method of Mycobacterium with a high positive detection rate, and do not need expensive equipment This method may suitable to resource limited medical institutions due to its low cost and short round time.