Acta Academiae Medicinae Sinica

ABSTRACT:Tuberculosis (TB), a chronic communicable disease, continues to be a global public health concern. Slow decline of TB incidence and prevalence, human immunodeficiency virus/TB coinfection, growth of multidrug-resistant/extensively-resistant TB have made the control of TB even more challenging. Chemotherapy of TB has developed for decades and now also faces similar challenges.

ABSTRACT:Tuberculosis (TB) is one of the global major public health problems. China has the second highest TB burden worldwide. This article introduces the epidemiology of TB in China and elucidates the progress and challenges in TB prevention and control.

ABSTRACT:Objective To obtain the recombinant rv1837c and rv3803c of Mycobacterium tuberculosis using gene engineering technology and explore their prokaryotic expression, purification, and immunogenicity. Methods The Mycobacterium tuberculosis rv1837c and rv3803c genes were amplified by polymerase chain reaction, and then cloned into the vector pTA2, followed by the subclone into the expression vector pET30a(+). The resulting plasmids, named pET30a(+):rv1837c and pET30a(+):rv3803c, encode recombinant protein containing a hexa-histidine tag on its N-terminus. pET30a(+):rv1837c and pET30a(+):rv3803c were introduced into E. coli BL21 (DE3) by transformation respectively, and the recombinant gene was induced with 0.4 mmol/L isopropyl-D-thiogalactopyranoside. The expressed products were identified by Western blot with hexa-histidine tag antibody and serum from tuberculotic patients. The histidine tagged protein was purified by nickel nitrilotriacetic acid His-Bind resin. Rabbits were immunized with purified recombinant Rv1837c and Rv3803c proteins. Then the purified recombinant Rv1837c and Rv3803c proteins were used to detect antibody in rabbit serum, which had been immunized by Western blot. Results After transformation of the E. coli and induction with 0.4 mmol/L of isopropyl-D-thiogalactopyranoside, recombinant target proteins Rv1837c (relative molecular mass: 92000) and Rv3803c (relative molecular mass: 38000) were expressed in pET30a(+): rv1837c and pET30a(+): rv3803c system. The expressed protein existed in cytoplasm in an unsoluble form and amounted to 30% and 50% of the total proteins of E. coli. The purity of the purified protein reached 90%. The immunogenicity of the recombinant proteins Rv1837c and Rv3803c was strong, as identified by Western blot. Conclusion The prokaryotic expression recombinant plasmids pET30a(+): rv1837c and pET30a(+): rv3803c was successfully constructed and the recombinant proteins Rv1837c and Rv3803c were obtained, which laid a basis for the optimized diagnosis of active tuberculosis.

ABSTRACT:Objective To synthesize two antigens-Ag85b and HspX of Mycobacterium tuberculosis H37Rv with molecular biological methods and to observe their biologic activity after co-administration of adjuvants (aluminum and/or CpG) in mice. Methods Recombinant expression plasmids pET30a-Ag85b and pET30a-HspX were constructed. The objective DNA fragments was characterized with restriction enzyme. Then the recombinant plasmids were transformed into E. coli BL-21, and two proteins were expressed by induction of isopropyl β-D-1-thiogalactopyranoside. After purification with anion exchange column Source30, QHP, and hydrophobic chromatography column, two proteins were identified by amino acid sequencing. After the successful preparation of these two antigens, they were co-administered in mice with adjuvants of aluminum and/or CpG (Ag85b，Ag85b+Al，Ag85b+CpG，Ag85b+Al+CpG；HspX，HspX+Al，HspX+CpG， HspX+Al+CpG); one group received normal saline and served as the control. Splenic lymphocytes were isolated for enzyme-linked immunosorbent spot assay to detect the secreted specific interferon-gamma (IFN-γ); in addition, lymphocytes proliferation test was performed to observe lymphocytes proliferation after in vitro stimulated with two antigens. Results The purity of two proteins reached 95% after purification. The N-terminal amino acid sequence (15 aa) of the purified proteins was same as the target sequence. For Ag85b, the secreted specific IFN-γ from isolated splenic lymphocytes after having been stimulated in vitro with Ag85b (80μg/ml) remarkably increased in Ag85b+CpG group, Ag85b+Al group, and Ag85b+CpG+Al group; the changes were significantly different between these three groups and control group(P<0.05). For HspX, the changes were significantly different between HspX +Al+CpG group and normal sodium group, although remarked increase of IFN-γ was also observed in HspX group，HspX+Al group, and HspX+CpG group. Conclusions Ag85b and HspX were successfully expressed and purified. A cell-mediated immunity may be induced when the antigens are co-administered with adjuvants of aluminum and/or CpG in mice, indicating that the recombinant proteins are bioactive.

ABSTRACT:Objective To study the effect of Mycobacterium smegmatis vaccine on the level of nitric oxide (NO) produced by peritoneal macrophages in immunized mice. Methods Balb/c mice were randomized into low-dose, middle-dose, and high-dose groups (injected with different doses of Mycobacterium smegmatis vaccine) and a control group (injected with normal saline). Then the peritoneal macrophages were cultured with lipopolysaccharide in vitro. The supernatants were collected and the concentrations of NO were analyzed through the reaction with Griess reagents. Results The levels of NO produced by the peritoneal macrophages in the control group, low-dose group, middle-dose group, and high-dose group were (3.50±3.11) ，(16.63±6.47)， (13.97±6.20)，and (7.55±2.26) ng/ml, respectively. The levels of NO in all dosing groups were significantly different from that in control group (P<0.01). Conclusion Mycobacterium smegmatis vaccine can promote the peritoneal macrophages to produce NO in mice.

ABSTRACT:Objective To establish a rapid, inexpensive, and simple drug susceptibility test (DST ) for Mycobacterium tuberculosis (M.tb) and evaluate its feasibility. MethodWe used nitrate reductase combined with mycobacteriophage assay（ PhaB-NRA） to test 49 clinical M.tb isolates of, and the results were compared with those of PhaB-NRA and traditional absolute concentration method. Results The sensitivity, specificity, and accuracy of PhaB-NRA for rifampicin were 89.1%, 91.67%, and 89.8%; on the contrary, those of isonicotinyl hydrazide were 86.21%, 90.0%, and 87.8%, respectively. The coincidence between PhaB-NRA and traditional assay were 0.746 for rifampicin and 0.750 for isonicotinyl hydrazide. Conclusions PhaB-NRA is an inexpensive, rapid, and simple DST method. It is a promising rapid screening technique for DST of M.tb.

ABSTRACT:Objective To explore the influences of Mycobacterium tuberculosis on the levels of human acute monocytic leukemia cell line THP-1 apoptosis and death. Methods Human acute monocytic leukemia cell line THP-1 were infected with Mycobacterium tuberculosis strains H37Ra, H37Rv, or Beijing genotype (BJTB), respectively, to construct the infection models. Cell apoptosis was detected using flow cytometry. The distribution of the apoptotic proteins was detected using immunofluorescent staining assays. The cells late apoptosis was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining assays. The change of cell death was determined by Tyrpan blue staining assays. Results THP-1apoptosis was induced by Mycobacterium tuberculosis strains H37Ra, H37Rv, and BJTB. H37Ra strongly induced THP-1 apoptosis, H37Rv weakly induced THP-1 apoptosis, and BJTB induced THP-1 apoptosis at the lowest level among these three Mycobacterium tuberculosis strains. On the contrary, BJTB strongly induced THP-1 death, H37Rv weakly induced THP-1 death, and H37Ra induced THP-1 death at the lowest level. Conclusions My-cobacterial strains with different virulence induce different levels of apoptosis and death of THP-1 cells. Compared with highly virulent strains, attenuated strains induce more apoptosis and less death.

ABSTRACT:Objective To investigate the relationship between the resuscitation promoting role of resuscitation promoting factor and the initial bacteria amount of dormant Mycobacterium tuberculosis. Methods Mycobacterium tuberculosis (dormant bacteria) was cultured for 100 days, then diluted into 1 mg/ml concentration with 7H9, and further diluted into 0.5, 0.25, 0.125, 0.0625, and 0.03125 mg/ml. Twelve new tubes added with 5 ml 7H9 and divided into two groups: the first group was added with the resuscitation-promoting factor protein, and the second group as control was added with 7H9. In each group the above diluted solutions were added. The tubes were located at 37℃ for culture. Optical density (OD) was detected on day 15, 25, 30, and 35. From each tube 1 μl culture solution was plated on 7H11 medium for colony counting. Results OD detection showed that bacteria proliferation in each group had positive linear correlation (P<0.05, P<0.01), indicating that the resuscitation-promoting factor played a similiar role in solutions with different dilution concentrations. 7H11 results and the OD results show that these two detection methods in each group had linear correlation (P<0.05, P<0.01), indicating that these two methods showed consistent test results. Conclusion The resuscitation-promoting factor has no effect on the resuscitation of dormant Mycobacterium tuberculosis and its initial bacteria amount.

ABSTRACT:Objective To investigate the distribution of the Beijing genotypes of Mycobacterium tuberculosis (M. tuberculosis) and the relationships between Beijing genotype strains and drug-resistant phenotypes in China. Methods Clinical isolates were collected during a 9-month research period from April to December in 2008 in six geographic regions of China. One isolate that had been biochemically confirmed to be a member of the M. tuberculosis complex was collected from each patient. The demographic data of the patients (eg. sex, age, and history of tuberculosis) as well as the drug resistance patterns and sources of the clinical isolates were collected. Drug susceptibility testing was performed using proportion method. Beijing genotypes of M. tuberculosis were identified by spacer oligonucleotide typing or insertion of IS6110 in the genomic dnaA-dnaN locus. Results Among the 410 M. tuberculosis clinical isolates, 67.1% (275/410) isolates were Beijing genotypes of M. tuberculosis. Significantly larger proportions of tuberculosis patients were infected with Beijing genotypes in the northeastern regions of China than that of in the central-western regions (χ2=20.50,P=0.000). No significant associations were found either between Beijing genotype strains and patientsage, sex, or treatment history. Multidrug-resistant isolates and rifampin-resistant isolates were more common among Beijing genotype strains than among non-Beijing strains (P=0.002, P=0.005). Conclusions About two third of the clinical isolates of M. tuberculosis in China are Beijing genotypes. Beijing genotype strains are not correlated with patientsage,sex, treatment history. People living in the northeastern regions of China are more susceptible to Beijing genotypes than those living in the central-western of China.Beijing genotype strains tend to be rifampin-resistant or multidrug-resistant.

ABSTRACT:Objective To explore the influences of intervention on the abilities of detecting pulmonary tuberculosis cases in general hospitals. Methods We selected 6 general hospitals at 3 different levels (A,B, and C). The intervened group included hospitals A1, B1, and C1, and the non-intervened group included hospitals A2, B2, and C2. The results after intervention were compared. Results The report rate of pulmonary tuberculosis, sputum positive rate of reported cases, and sputum check rate of reported cases were significantly higher in hospital A1 than grouping hospital A2 （P=0.000， P=0.045， and P=0.017, respectively). The report rate and sputum examination rate of reported cases were significantly higher in hospital B1 than grouping hospital B2（P=0.000，P=0.024, respectively). The report rate and sputum examination rate of reported cases were significantly lower in hospital C1 than grouping hospital C2 （P=0.000，P=0.001, respectively). In hospital A1, the report rate, sputum positive rate of reported cases, and sputum check rate of reported cases were not significantly different before and after intervention (P=0.182，P=0.116，and P=0.583, respectively). In hospital B1, the report rate were significantly different before and after intervention（P=0.004）,while the sputum positive rate of reported cases and sputum check rate of reported cases were not significantly different (P=0.909， P=0.052, respectively). In hospital C1, the report rate was significantly higher after intervention (P=0.025). In hospital C2, the sputum check rate significantly increased (P=0.000). Conclusions Intervention influences the hospitals abilities to detect pulmonary tuberculosis cases. However, more optimized and long-term intervention mechanism should be established to increase case detection rate of pulmonary tuberculosis.

ABSTRACT:Objective To detect Mycobacterium tuberculosis RD1-encoded antigens-specific, interferon- gamma(INF-γ)-secreting T cells in pleural effusions, ascites, and cerebrospinal fluid. MethodThe early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) peptides-specific T cells in peripheral blood mononuclear cell (MC), ascites MC, pleural effusions MC, and cerebrospinal fluid MC were detected using enzyme-linked immunospot assay (ELISPOT) for INF-γ. Results ESAT-6 or CFP-10 peptides-specific, INF-γ-secreting T cells were detected in peripheral blood, ascites, pleural effusions, and cerebrospinal fluid, which marked the presence of tuberculosis infection. Patients with positive ELISPOT results of INF-γ-release assay were all diagnosed as active tuberculosis. Spot forming cells in ascites and pleural effusions were much higher than those in peripheral blood (up to 6.4 and 31.9 times). Conclusion Detection of RD1-encoded antigens-specific, INF-γ-secreting T cells in pleural effusions, ascites, and cerebrospinal fluid provides a new way to diagnose tuberculosis infection.

ABSTRACT:Objective To compare enzyme-linked immunospot assay (ELISPOT) and tuberculin skin test (TST) and explore their roles in the auxiliary diagnosis of initial pulmonary tuberculosis. Methods Totally 123 patients with initial pulmonary tuberculosis (tuberculosis group) and 102 patients with non-tuberculosis pulmonary disease (control group) were enrolled. The peripheral blood mononuclear cells of all participants were co-cultured with early secretiny antigen target-6/culture filtrate protein-10 fusion protein (ESAT-6/CFP-10 ), and spot forming cells (SFCs) were enumerated by ELISPOT (ESAT-6/CFP-10-ELISPOT). TST was also performed simultaneously. Results ESAT-6/CFP-10-ELISPOT showed significantly higher numbers of SFCs after stimulation in tuberculosis group than in control group (P=0.000). The sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, positive predictive value, and negative predictive value of ESAT-6/CFP-10-ELISPOT were 91.1% （111/123）, 81.4% (82/102),4.60, 0.12, 0.85, and 0.87 respectively, while the above values of TST were 65.6%(59/90), 45.1% (46/102),1.31, 0.76, 0.51, and 0.60, respectively. The sensitivity and specificity of ESAT-6/CFP-10-ELISPOT were significantly higher than those of TST(all P＝0.000). The number of SFCs were not significantly different between smear-positive tuberculosis subgroup and smear-negative tuberculosis subgroup (P=0.166). The sensitivities were 91.8%(67/73) and 88.0%（44/50） in these two subgroups, respectively, (P=0.448). Conclusions ESAT-6/CFP-10-ELISPOT may be a more accurate approach for the auxiliary diagnosis of initial pulmonary tuberculosis; meanwhile, it offers certain diagnostic evidences for smear-negative tuberculosis. However, its specificity may be affected by latent tuberculosis infection. On the contrary, TST has poor value in the auxiliary diagnosis of initial pulmonary tuberculosis.

ABSTRACT:Objective To evaluate the sensitivity of an interferon-gamma release assay T-SPOT.TB in the diagnosis of bacteriologically or histologically confirmed extrapulmonary tuberculosis. MethodTotally 31 patients with bacteriologically or histologically confirmed extrapulmonary tuberculosis in Peking Union Medical College Hospital received T-SPOT.TB assay to detect early secreting antigen target 6 or culture filtrate protein 10 peptides-specific T cells in the peripheral blood mononuclear cells(PBMCs). Results T-SPOT.TB assay showed positive results in 29 patients with extrapulmonary tuberculosis and the sensitivity was 93.5%（95% CI 84.8% - 100%). The median of spot forming cells（SFCs） in response to early secreting antigen target 6 peptides was 196/106PBMCs (interquartile range, 72-532/106 PBMCs), the median of SFCs in response to culture filtrate protein 10 peptides was 276 SFCs/106 PBMCs (interquartile range, 72-568/106 PBMCs), and the median of the incorporate SFCs was 612/106PBMCs (interquartile range, 192-1152/106 PBMCs). Conclusion T-SPOT.TB is highly sensitive in diagnosing extrapulmonary tuberculosis.

ABSTRACT:Objective To highlight the clinical features and diagnosis of chronic disseminated tuberculosis, with emphasizing the usefulness of several recently available diagnostic technologies in this setting. MethodWe presented a case of chronic disseminated tuberculosis diagnosed with the combined application of interferon-gamma release assay T-SPOT.TB,18F-fluorodeoxyglucose (FDG)-positron emission tomography (PET)，and gene chip assay. Results A 53-year-old gentleman who had chronic cough for 7 years and fever for 2 weeks was referred to our hospital for further evaluation.18F-FDG-PET/CT scan showed increased FDG uptake in multiple lesions involving bilateral lungs, supraclavicular, mediastinal and intro-abdominal lymph nodes and bones, mimicking metastatic malignancy. T-SPOT.TB assay revealed significant responses ［early secreting antigen target 6(ESAT-6)：3908spot forming cells (SFCs)/106 peripheral blood mononuclear cells (PBMCs)，culture filtrate protein (CFP-10)：3400SFCs/106 PBMCs］. Subsequent biopsy of supraclavicular lymph node, lung, and ilium revealed granulomas, while culture of the obtained tissue yeilded mycobacteria. Gene chip testing identified M.tuberculosis sensitive to isoniazid and rifampin. After 10 weeks of treatment for tuberculosis, the patients condition was improved and a second T-SPOT.TB assay showed significantly reduced responses（ESAT-6：1528SFCs/106 PBMCs; CFP-10：1460SFCs/106 PBMCs). Conclusions Timely diagnosis of chronic disseminated tuberculosis requires high index of suspicion. T-SPOT.TB assay, PET/CT, and gene chip assay may provide valuable information that facilitates further diagnostic procedures and treatment decision.

ABSTRACT:Objective To explore the osteogenic potential of the nano-hydroxyapatite/collagen/calcium alginate composite implanted in animals. Methods Eighteen 3-month-old New Zealand white rabbits were adopted to prepare 15 mm segmental defect model at the middle part of radius. Rabbit models were randomly divided into experimental group and blank control group. Nano-hydroxyapatite/collagen/calcium alginate was implanted into the defects of experimental group. Four, 8, and 12 weeks after operation, all specimens were examined by X-ray and histological methods. Results All the 18 rabbit models entered the final analysis. X-ray showed that osteotylus was seen in the whole defect area in the experimental group 12 weeks after operation, during which osteogenesis was more obvious than in weeks 4 and 8 and the bridge grafting of defect area was obviously visible. In the blank control group, osteotylus was only observed at the two ends of the defects, and no osteogenesis was found in the central part of the defect area. Histological examination showed that new osteoid formation was seen in internal porous zone in the experimental group in weeks 4 and 8; in week 12, more woven bone-like tissues were visible and trabecular-like structure was formed. Conclusion The nano-hydroxyapatite/collagen/calcium alginate has good osteogenic potential.

ABSTRACT:Objective To investigate the mechanical changes of the degenerated lumbar disc with finite element analysis. Methods A three dimensional finite element model of a human lumbar spine at the L3-L4 disc was established by the software MIMICS and ABAQUS based on computer tomography images. Degeneration was modeled by changes in geometry and material properties. The model was loaded with 0.3 MPa in axial plane. The von Mises stress on the annulus fiber，nucleus pulposus，endplate and facet joints in healthy and degenerated discs was compared. ResultCompared with healthy discs, the von Mises stress of disc distributed in the side of annulus fiber, the stress of nucleus pulposus decreased remarkably, the stress of endplate distributed in the posterior part, and the stress of facet joints increased for the degenerated disc. Conclusion The finite element models can provide a method of understanding the relationship between biomechanical performance of the disc due to disc degeneration.

ABSTRACT:Objective To compare the effects of different types of feeder cells on supporting undifferentiation and high proliferation of human embryonic stem cells (hESC). Methods hESC were seeded on mouse embryonic fibroblasts (MEF), human marrow stromal cells (hMSC), and human foreskin fibroblasts (hFF), respectively. Colony number, cell quantity after digestion, and survival rate were observed by alkaline phosphatase (AP) staining and Trypan blue, and the biological properties of hESC after 5 passages were observed by immunofluorescence staining. Results Although all the three feeder layers could support the formation of hESC colonies and maintain pluripotency, the morphology of colonies on different feeder layers remarkably varied. The stage-specific embryonic antigen-3 and AP staining were positive on three types of feeders. The number of colonies, number of cells produced, and cell survival rates were significantly higher on MEF than on human feeder cells (P<0.01).Furthermore, the number of AP-positive colonies and cell quantity were also significantly higher on hMSC than on hFF (P<0.01). Conclusions All three types of feeder cells are able to support the growth of hMSC, although MEF are more favourable for the proliferation. Two types of human feeder cells lay the foundation for the removal of animal-derived hESC culture system. hMSC is superior to hFF in supporting the proliferation of hESC.

ABSTRACT:Objective To investigate the growth and development of nitric oxide synthase (NOS)-positive neurons in the cerebellum of human fetus in the midanaphase. MethodThe positive expression of the NOS-positive neurons in the cerebellum of midanaphase human fetus was observed by immunohistochemistry. Results By the sixth to seventh month of gestation, NOS-positive neurons were seen in the ependymal layer of the cerebellum. The nucleus was oval-shaped and the neurons had short and small processes. By the eighth to ninth month, NOS-positive neurons were found in the central layer of the cerebellum and the nucleus was round-, oval-, or fusiform-shaped; meanwhile, the neurons grew larger in size with richer cytoplast and heavier staining. The beaded nerve fibers reached the marginal layer and the layer became thickened on the tenth month, which generally was composed of 5 to 6 layers of NOS-positive neurons that were tightly aligned. Some NOS-positive neurons were in smaller size with the cell body and the nerve fibers grew well. Conclusion Nitric oxide generated by NOS of the NOS-positive neurons in the cerebellum plays an important role in the differentiation, proliferation, and migration of neurons and gliacytes.

ABSTRACT:Objective To investigate whether aristolochic acid can be transported into human kidney proximal tubular cell (HKC) and its potential mechanism. Methods Intracellular aristolochic acid was measured by liquid chromatography-tandem mass spectrometry. The release of lactate dehydrogenase (LDH) induced by aristolochic acid in the presence of organic anion transporter inhibitor (probenecid) or organic cation transporter inhibitor (tetraethylammonium) was evaluated. The effects of probenecid on aristolochic acid induced connective tissue growth factor (CTGF) mRNA and protein expression were also examined by real time polymerase chain reaction and Western blot, respectively. Results Aristolochic acid was detected in the suspension of the denatured HKC after incubation with aristolochic acid sodium salt. The release of LDH from HKC, which was induced by 60 mg/L aristolochic acid sodium salt, was significantly inhibited by 1 mmol/L probenecid (P<0.01), but not by 1 mmol/L tetraethylammonium. The increased CTGF mRNA and protein expression in HKC stimulated by 40 mg/L aristolochic acid sodium salt was significantly down-regulated by 1 mmol/L probenecid (P<0.05), with an inhibition rate of 16% and 21%, respectively. Conclusion Aristolochic acid can be transported into HKC by organic anion transport system, and then exerts its biological effects.

ABSTRACT:Objective To investigate the influence of Lewis y antigen on the gene expression of partial drug resistance associated proteins in human ovarian cancer cell line RMG-I-H. Methods RT-PCR was used to determine the gene expressions of partial drug resistance associated proteins in RMG-I-H cell line transfected with α1, 2-fucosyltransferases gene and RMG-I cell line, as well as in RMG-I-H treated with or without anti-Lewis y monoclonal antibody at the concentration of 10 μg/ml. The immunocytochemical method was used to detect the expression of P-glycoprotein (P-gp) in RMG-I and RMG-I-H cell lines. RMG-I and RMG-I-H cells were transplanted into nude mice and the expression of P-gp in the tissues was measured by immunohistochemistry. Results The mRNA expressions of protein kinase C-α(PKC-α), topoismeraseⅠ(Topo Ⅰ), multidrug resistance-associated protein-1(MRP-1), and MRP-2 were significantly higher in RMG-I-H cells than those in RMG-I cells (0.46±0.02 vs. 0.27±0.05, 0.82±0.08 vs. 0.52±0.04, 0.66±0.07 vs. 0.34±0.12, and 0.44±0.08 vs. 0.23±0.05; all P<0.05). However, the mRNA expression of multidrug resistance 1 (MDR-1) was significantly lower in RMG-I-H cells than that in RMG-I cells (0.26±0.05 vs. 0.45±0.08， P<0.05). The P-gp level increased in RMG-I-H cells compared with that in RMG-I cells both in vivo and in vitro (P<0.05). Expressions of MDR-1, MRP-1, MRP-2, PKC-α, and TopoⅠ mRNA decreased by the time in RMG-I-H cells treated with anti-Lewis y monoclonal antibody (all P<0.05), while mRNA expressions of those genes in the control group did not statistically change (P>0.05). In addition, MDR-1, MRP-1, MRP-2, PKC-α, and TopoⅠ mRNA expressions were significantly lower in RMG-I-H cells treated with anti-Lewis y monoclonal antibody than those in the control group at 6 hours (all P<0.05) and the inhibition ratios were 48.55%, 77.50%, 70.18%, 45.86%, and 46.13%, respectively. Conclusion The Lewis y antigen of the human ovarian cancer cell surface is closely correlated with the regulation on the gene expression of partial drug resistance associated proteins.

ABSTRACT:Objective To evaluate the diagnostic specificity of dynamic assessment and monitoring using a portable spirometer in diagnosis and differential diagnosis of asthma. Methods We retrospectively reviewed the results of dynamic monitoring of spirometry in 145 symptomatic patients with physician-diagnosed asthma. Flow-volume curve and simultaneous symptoms and mood were measured in a fixed-time thrice-daily assessment schedule for 2 weeks. Patients were allowed to make additional measurements in case of symptom exacerbations. Results The clinical data of 51 males and 94 females with a mean age of（39.1±13.0） years (ranged from 10 to 65 years) were analyzed. Duration of asthma before study was (6.7±9.9) years. Of 145 patients with physician-diagnosed asthma, 126 (87%) could be conclusively confirmed for a diagnosis of asthma. Asthma was misdiagnosed in 14 patients (9.7%). Overdiagnosis of asthma was observed in 5 patients (3.4%). Conclusion Dynamic assessment and monitoring using a portable spirometer by revealing variability and reversibility of airway obstruction may provide an additional tool for diagnosis and differential diagnosis of asthma.

ABSTRACT:Objective To investigate the changes of insulin resistance and islet β cells function in subjects with euglycemia and high-normal blood pressure. Methods Total 423 subjects were divided into normal blood pressure group and high-normal blood pressure group. Body height, weight, waist and hip circumference, and biochemical data were measured. Homeostasis model assessment of insulin resistance (HOMA-IR), insulin sensitivity index (ISI)-composite, and first-phase (1PH) Stumvoll index were calculated. Results Waist circumference, total cholesterol, triglyceride, low-density lipoprotein cholesterol, HOMA-IR were significantly higher and 1PH Stumvoll index and ISI-composite were significantly lower in high-normal blood pressure group than in normal blood pressure group(P<0.05). Systolic blood pressure (SBP) was positively correlated with HOMA-IR (r=0.122) and negatively correlated with 1PH Stumvoll index (r=-0.159) and ISI-composite (r=-0.131) (P<0.05). SBP and triglyceride were independent factors for 1PH Stumvoll index. Conclusion Insulin resistance and islet dysfunction may exist in subjects with high-normal blood pressure.

ABSTRACT:Objective To assess the diagnostic value of CT enterography in patients with Crohn’s disease. Methods Multi-detector CT enterography and small bowel follow-through were performed in 30 patients with Crohn’s disease. The locations and characteristics of the intestinal and extra-intestinal lesions detected by both two techniques were compared. Results Skip lesions were diagnosed in 16 patients (53.3%) by CT enterography and in 9 patients (30%) by small bowel follow-through (P =0.039). Mucosal changes were detected in 29 patients (96.7%) by CT enterography and in 18 patients (60%) by small bowel follow-through (P =0.001). Among 11 patients whose small bowel follow-through did not show abnormal mucosal changes, 8 patients underwent endoscopy, which showed superficial ulcer with or without mucosal congestion and edema in 5 patients, mucosal congestion and edema in 2 patients, and mucosal erosion in 1 patient. CT enterography and small bowel follow-through consistently depicted fistula in 3 patients and had no significant difference in diagnosing intestinal stenosis. CT enterography also exclusively detected abdominal abscess in one patient. Conclusions CT enterography is superior to small bowel follow-through in diagnosing the disease location and characteristics of Crohn’s disease; furthermore, it can detect more extra-intestinal lesions. CT enterography has potential to replace small bowel follow-through as the imaging examination of choice for patients with Crohn’s disease.

ABSTRACT:T-SPOT.TB is an interferon-gamma release assay to detect T-cell response to early secreting antigen target 6 and culture filtrate protein 10 peptides by enzyme-linked immunospot assay for tuberculosis diagnosis. It is highly sensitive and specific, and will not be affected by the subject’s immune status and Bacillus Calmette-Guerin vaccination. This assay has been licensed for in-vitro diagnosis in Europe and the United States. Its potential roles in distinguishing active tuberculosis from latent tuberculosis infection and predicting active tuberculosis among individuals with latent tuberculosis have been increasingly studies. This article reviews the advances in the clinical application of T-SPOT.TB.

ABSTRACT:Nine proteins encoded by Mycobacterium tuberculosis RD1 region are important protective antigens that become absent in long passaging of Mycobacterium tuberculosis. They only exist in pathogenic Mycobacteria and are absent in Bacille Calmette-Guerin and environmental Mycobacteria. With good immunogenicities, they may play an important role in the diagnosis and prevention of Mycobacterium tuberculosis. This article reviews recent studies on using RD1-encoded proteins as antigens in the diagnosis of active tuberculosis and tuberculous pleurisy.