[Diagnosis and treatment of bronchial tuberculosis].
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Bronchi↗
Biomedical subjects
Y Zhuang
Publications and source records attributed to Y Zhuang.
[The immunogenic characteristics of the recombinant 38,000 antigen from Mycobacterium tuberculosis].
OBJECTIVE: To study the ability of recombinant 38,000 protein of Mycobacterium tuberculosis (rMT38) to induce antibodies and its value as a serological diagnostic reagent. METHODS: Rabbits were divided into six groups, namely: PPD + physiological saline(group A), rMT38 + physiological saline(group B), rMT38 + adjuvent(group C), 1/2 rMT38 + 1/2 rMT16 + physiological saline(group D), 1/2 rMT38 + 1/2 rMT16 + adjuvent(group E), rMT16 + adjuvent(group F). The sera from the immunized rabbits and the sera immunized by mycobacteria were evaluated by enzyme-linked immunosorbent assay(ELISA) and bilateral agar diffusion assay (BADA). 389 human sera, including 145 patients with pulmonary tuberculosis, 105 healthy controls, 31 non-tuberculosis respiratory patients, 78 BCG-vaccinated healthy controls and 30 patients with M. abscessus infection, were detected by ELISA. RESULTS: Titers of specific antibodies in sera from C and E groups were 1:16 and 1:4 in BADA, and 1:6,400 and 1:3,200 in ELISA, respectively. Precipitate lines of sera from A, B, D, F groups reacting with rMT38 were not found by BADA. Sera titer of A, B, D, F groups were negative, 1:400, 1:400, and negative by ELISA, respectively. Sera from E group had no reaction with PPD from thirteen mycobacteria, however, sera from E group reacted with PPD from Mycobacterium tuberculosis and BCG. Sera titer of all rabbits decreased with time. After two months, serum titers in B and D groups became negative. However, antibodies were still found in C and E groups after five months. The sensitivity of rMT38 and PPD in detecting pulmonary tuberculosis were 69.0% and 75.2%, respectively. The specificity of rMT38 and PPD in detecting healthy controls, non-tuberculosis respiratory patients, BCG-vaccinated healthy controls and M. abscessus-infected patients were 97.1%, 93.5%, 86.0%, 53.6% and 94.3%, 87.1%, 67.9%, 39.3% respectively. CONCLUSION: rMT38 has a strong immunogenicity, which may be used as an immunodiagnostic reagent for tuberculosis.
Animals↗
[The studies on the molecular mechanism of rifampin-resistant Mycobacterium tuberculosis].
OBJECTIVE: To evaluate the molecular mechanism of rifampin (RFP)-resistance of M. tuberculosis, and to provide evidence of molecular genetics for basic and clinical research of tuberculosis. METHODS: rpoB gene mutation of M. tuberculosis was detected with PCR-SSCP from 262 clinical isolates, including 74 drug sensitive strains, and 188 RFP-resistant only or multiple drug-resistant strains of M. tuberculosis, collecting from Henan, Hebei and Anhui province, Beijing and Shanghai cities. Plasmids were extracted from 76 clinical isolates of M. tuberculosis with alkaline bacteriolytic methods. RESULTS: Mean mutation rate of rpoB gene was 92% in the RFP-resistant clinical isolates of M. tuberculosis. Respectively, the mutation rate was 90.0% in Henan province, 94% in Hebei province, 91% in Anhui province, 91% in Beijing city and 92% in Shanghai city. There was no significant difference between results from the provinces and the cities (chi 2 = 0.42, P > 0.05). However, mutation of rpoB gene was not detected in 74 sensitive clinical isolates of M. tuberculosis. Moreover, in 76 clinical isolates of M. tuberculosis the plasmids were not found, including 29 sensitive strains, 33 RFP-resistant only or multiple drug-resistant strains and 14 RFP, isoniazid, streptomycin and ethambutol-resistant clinical isolates. CONCLUSIONS: This study confirmed the strong correlation between the RFP-resistance and the mutation of the rpoB gene, which might be a major molecular mechanism of RFP-resistance in M. tuberculosis. No evidence shows that M. tuberculosis has the plasmid, which mediates RFP resistance to M. tuberculosis.
Antibiotics, Antitubercular↗
[Delayed type hypersensitivity responses to recombinant mycobacterial antigen 38,000 in guinea pigs sensitized to Mycobacterium tuberculosis].
OBJECTIVE: To evaluate the ability and influencing factors that recombinant 38,000 antigen induces delayed-type hypersensitivity(DTH) reaction in guinea pigs. METHODS: In one skin test, physiological saline as negative control and PPD as positive control, guinea pigs were shaved on the back and given intracutaneous injection with 0.1 microgram, 0.2 microgram, 0.4 microgram of recombinant 38,000 antigen. Skin reactions were measured 48 h after antigen injection, each measuring two transverse diameters of the induration. In the other skin test, each guinea pig was intracutaneously injected in four sites, respectively with 5 IU PPD and recombinant 38,000 antigen (0.1 microgram, 0.2 microgram and 0.4 microgram) or 5 IU PPD and biantigen(combination 38,000 with 16,000 antigen, 0.1 microgram, 0.2 microgram and 0.4 microgram). Single antigen and biantigen were tested in three separate experiments. Each experiment needs six sensitized guinea pigs. Skin reactions were measured 24 h, 48 h and 72 h after antigen injection. RESULTS: DTH responses induced by 0.4 microgram were weaker than 0.1 microgram and 0.2 microgram in 24 h. 0.1 microgram and 0.2 microgram elicited DTH responses similar in strength to that of 5 IU PPD. From 0.1 to 0.4 microgram, similar dose of recombinant 38,000 antigen and combination antigen elicited similar DTH responses. DTH responses induced by recombinant 38,000 antigen had better be measured at 48 h, however that induced by combination antigen at 24 h. CONCLUSIONS: Recombinant 38,000 antigen can induce DTH responses in guinea pigs. Dose of antigen is important factor that affect DTH responses in guinea pigs.
Animals↗
[Studies on application for classification and identification in Mycobacterium by analysis of PCR amplification of 16S-23S ribosomal DNA spacer sequences].
The 16S-23S ribosomal DNA spacer sequence of mycobacterium were amplified by PCR. The products were visualized by PAGE, and evaluate the possibility for classification and identification of mycobacterium at gene level. The sensitivity of PCR in annealing temperature 45 degrees C was 500 fg/microL, whereas 50 degrees C was 5 pg/microL. The results showed that: the amplified bands ranging from 300 to 600 bp, most of rapid-growing Mycobacterium and Nonmycobacterium tested have more bands and the bands molecular weights were larger than slow-growing Mycobacterium. The relativity of mycobacterium < 70%, most of them < 50%. This experimental method might be rapid and effective for differentiation of Mycobacterium at species level.
Cluster Analysis↗
[Studies on detection methods of two DNA probes in Mycobacterium tuberculosis].
Two different biotinylated DNA probes which are highly specific to M. tuberculosis(MT) were made and studied. One probe is a 20 bp oligonucleotide labeled with biotin at 5' end, the other is a long DNA probe produced by PCR amplification procedure allowed for the incorporation of biotin labeled UTP. The two probes were hybridized with MT genome DNA and a 317 bp PCR product amplified from IS6110 sequence of MT, and then detected by alkaline phosphatase conjugates through colorimetric reaction. The detection sensitivity and specificity of the two probes were comparatively studied. The hybridization condition including concentration of probe, temperature of hybridization and washing filter thereafter were also investigated preliminary. The detection limit of the oligonucleotide probe and the 188 bp PCR probe were 100 ng and 6 ng of DNA respectively in detection of M. T genome, and 400 pg and 50 pg of DNA respectively in detection of PCR products of MT. The two probes can be only hybridized to MT and BCG, but not with other 24 mycobacterium or non-mycobacterium tested. The optimal hybridization temperature and washing filter temperature of oligonucleotide were 42 degrees C and 60 degrees C respectively; and that of 188 bp probe, 68 degrees C and 60 degrees C-68 degrees C. Generally the specificity of two probes were all high, but the sensitivity of 188 bp DNA probe was 7-16 times that of the oligonucleotide probe. The higher sensitivity, lower hybridization background and faster revelation of the 188 bp DNA probe made it a better choice in detection of MT.
Biotin↗
Induction of nitric oxide-dependent apoptosis in motor neurons by zinc-deficient superoxide dismutase.
Mutations in copper, zinc superoxide dismutase (SOD) have been implicated in the selective death of motor neurons in 2 percent of amyotrophic lateral sclerosis (ALS) patients. The loss of zinc from either wild-type or ALS-mutant SODs was sufficient to induce apoptosis in cultured motor neurons. Toxicity required that copper be bound to SOD and depended on endogenous production of nitric oxide. When replete with zinc, neither ALS-mutant nor wild-type copper, zinc SODs were toxic, and both protected motor neurons from trophic factor withdrawal. Thus, zinc-deficient SOD may participate in both sporadic and familial ALS by an oxidative mechanism involving nitric oxide.
Amyotrophic Lateral Sclerosis↗
Id1 and Id3 are required for neurogenesis, angiogenesis and vascularization of tumour xenografts.
Id proteins may control cell differentiation by interfering with DNA binding of transcription factors. Here we show that targeted disruption of the dominant negative helix-loop-helix proteins Id1 and Id3 in mice results in premature withdrawal of neuroblasts from the cell cycle and expression of neural-specific differentiation markers. The Id1-Id3 double knockout mice also display vascular malformations in the forebrain and an absence of branching and sprouting of blood vessels into the neuroectoderm. As angiogenesis both in the brain and in tumours requires invasion of avascular tissue by endothelial cells, we examined the Id knockout mice for their ability to support the growth of tumour xenografts. Three different tumours failed to grow and/or metastasize in Id1+/- Id3-/- mice, and any tumour growth present showed poor vascularization and extensive necrosis. Thus, the Id genes are required to maintain the timing of neuronal differentiation in the embryo and invasiveness of the vasculature. Because the Id genes are expressed at very low levels in adults, they make attractive new targets for anti-angiogenic drug design.
Animals↗
A novel role for HEB downstream or parallel to the pre-TCR signaling pathway during alpha beta thymopoiesis.
TCR gene rearrangement and expression are central to the development of clonal T lymphocytes. The pre-TCR complex provides the first signal instructing differentiation and proliferation events during the transition from CD4-CD8-TCR- double negative (DN) stage to CD4+CD8+ double positive (DP) stage. How the pre-TCR signal leads to downstream gene expression is not known. HeLa E-box binding protein (HEB), a basic helix-loop-helix transcription factor, is abundantly detected in thymocytes and is thought to regulate E-box sites present in many T cell-specific gene enhancers, including TCR-alpha, TCR-beta, and CD4. Targeted disruption of HEB results in a 5- to 10-fold reduction in thymic cellularity that can be accounted for by a developmental block at the DN to DP stage transition. Specifically, a dramatic increase in the CD4low/-CD8+CD5lowHSA+TCRlow/- immature single positive population and a concomitant decrease in the subsequent DP population are observed. Adoptive transfer test shows that this defect is cell-autonomous and restricted to the alpha beta T cell lineage. Introduction of an alpha beta TCR transgene into the HEBko/ko background is not sufficient to rescue the developmental delay. In vivo CD3 cross-linking analysis of thymocytes indicates that TCR signaling pathway in the HEBko/ko mice appears intact. These findings suggest an essential function of HEB in early T cell development, downstream or parallel to the pre-TCR signaling pathway.
Adoptive Transfer↗
Imidazole substituted biphenyls: a new class of highly potent and in vivo active inhibitors of P450 17 as potential therapeutics for treatment of prostate cancer.
3- And 4-imidazol-1-yl-methyl substituted biphenyl compounds (named as meta- and para-substituted compounds) were synthesized bearing additional substituents in 3'-/4'-position as inhibitors of P450 17 (17alpha-hydroxylase-C17,20-lyase). P450 17 is the key enzyme of androgen biosynthesis. Its inhibition is a novel therapeutic strategy for treatment of prostate cancer (PC). Twenty-nine compounds were synthesized by Ar-Mg-Br, Negishi or Suzuki aryl-aryl cross coupling and tested toward human and rat enzyme. Most of the compounds showed moderate to excellent activity against one of the enzymes (0.087 microM < or = IC50 < or = 7.7 microM (ketoconazole: 0.74 microM) for the human enzyme, 0.63 microM < or = IC50 < or = 32 microM (ketoconazole: 67 microM) for the rat enzyme). Interestingly, strong species differences were observed. In addition compounds were tested for inhibition toward P450 arom. The 3-imidazol-1-yl-methyl substituted compounds showed good inhibitory activity of P450 arom, while for the 4-substituted compounds negligible inhibition was found. For the most active group of P450 17 inhibitors, (i.e. the 4-imidazol-1-yl-methyl substituted compounds) a QSAR study was performed for inhibition of the human enzyme leading to the result that a hydrophilic substituent in 3'-/4'-position is very important. The most promising compounds (with respect to activity toward both enzymes) were tested in vivo using SD-rats for reduction of plasma testosterone concentrations 2 and 6 h after single i.p. application. The fluorine substituted compound 8c decreased the testosterone plasma concentration to castration level (after 2 h; 5 mg/kg) showing a biological half live of about 6 h.
Animals↗
A novel superoxide dismutase-based trap for peroxynitrite used to detect entry of peroxynitrite into erythrocyte ghosts.
Peroxynitrite (ONOO-) is a relatively stable oxidant produced by activated macrophages and neutrophils. To detect peroxynitrite, a novel human superoxide dismutase (SOD) trap was developed by substituting a tyrosine near the copper in the active site. The copper can catalyze nitration of this tyrosine by peroxynitrite. The nitrated tyrosine can serve as a reporter for peroxynitrite by measuring the extent of nitration with Western blots developed with a nitrotyrosine antibody. The new SOD mutant differs from bovine SOD whose sole tyrosine is far removed from the active site. Nitration of bovine SOD was second-order with respect to SOD concentration, whereas nitration of the new mutant SODs followed first-order kinetics with respect to peroxynitrite. The tyrosine SODs were used to assess whether peroxynitrite crosses erythrocyte membranes through the band 3 anion exchange protein. Tyrosine-containing SOD entrapped within normal human erythrocyte ghosts became nitrated in proportion to peroxynitrite concentration. The band 3 anion exchange protein inhibitors, phenyl isothiocyanate (PITC) and 4, 4'-diisothiocyanatostilbene-2,2'-disulfonate (DIDS), inhibited up to 90% of the nitration. The erythrocyte membrane proteins, spectrin, band 3 anion exchange protein, and proteins 4.1 and 4.2, were also nitrated. Nitration of erythrocyte membrane proteins was also inhibited by PITC and DIDS. These data suggest that the band 3 anion exchange protein is the major route for the entry of peroxynitrite into erythrocytes. The ability of peroxynitrite to cross cell membranes can contribute to its toxicity by allowing access to intracellular target molecules.
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
Controlling lymphopoiesis with a combinatorial E-protein code.
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Animals↗
Targeted disruption of Smad3 reveals an essential role in transforming growth factor beta-mediated signal transduction.
The Smads are a family of nine related proteins which function as signaling intermediates for the transforming growth factor beta (TGF-beta) superfamily of ligands. To discern the in vivo functions of one of these Smads, Smad3, we generated mice harboring a targeted disruption of this gene. Smad3 null mice, although smaller than wild-type littermates, are viable, survive to adulthood, and exhibit an early phenotype of forelimb malformation. To study the cellular functions of Smad3, we generated Smad3 null mouse embryonic fibroblasts (MEFs) and dermal fibroblasts. We demonstrate that null MEFs have lost the ability to form Smad-containing DNA binding complexes and are unable to induce transcription from the TGF-beta-responsive promoter construct, p3TP-lux. Using the primary dermal fibroblasts, we also demonstrate that Smad3 is integral for induction of endogenous plasminogen activator inhibitor 1. We subsequently demonstrate that Smad3 null MEFs are partially resistant to TGF-beta's antiproliferative effect, thus firmly establishing a role for Smad3 in TGF-beta-mediated growth inhibition. We next examined cells in which Smad3 is most highly expressed, specifically cells of immune origin. Although no specific developmental defect was detected in the immune system of the Smad3 null mice, a functional defect was observed in the ability of TGF-beta to inhibit the proliferation of splenocytes activated by specific stimuli. In addition, primary splenocytes display defects in TGF-beta-mediated repression of cytokine production. These data, taken together, establish a role for Smad3 in mediating the antiproliferative effects of TGF-beta and implicate Smad3 as a potential effector for TGF-beta in modulating immune system function.
Animals↗
Impaired immune responses and B-cell proliferation in mice lacking the Id3 gene.
B-lymphocyte activation and proliferation induced by the B-cell receptor (BCR) signals are important steps in the initiation of humoral immune responses. How the BCR signals are translated by nuclear transcription factors into cell cycle progression is poorly understood. Id3 is an immediate-early gene responding to growth and mitogenic signals in many cell types including B cells. The primary function of the Id3 protein has been defined as that of inhibitor of basic-helix-loop-helix (bHLH) transcription factors. The interaction between Id3 and bHLH proteins, many of which are essential for cellular differentiation, has been proposed as a key regulatory event leading to cellular proliferation instead of differentiation. To further investigate the role of Id3 in tissue and embryo development and the mechanism of Id3-mediated growth regulation, we generated and analyzed Id3-deficient mice. While these mice display no overt abnormality in tissue and embryo development, their humoral immunity is compromised. The amounts of immunoglobulins produced in Id3-deficient mice immunized with a T-cell-dependent antigen and a type 2 T-cell-independent antigen are attenuated and severely impaired, respectively. Further analysis of lymphocytes isolated from Id3-deficient mice reveals a B-cell defect in their proliferation response to BCR cross-linking but not to lipopolysaccharide or a combination of BCR cross-linking and interleukin-4. Analyses of cultured lymphocytes also suggest involvement of Id3 in cytokine production in T cells and isotype switching in B cells. Finally, the proliferation defect in Id3-deficient B cells can be rescued by ectopic expression of Id1, a homologue of Id3. Taken together, these results define a necessary and specific role for Id3 in mediating signals from BCR to cell cycle progression during humoral immune responses.
Animals↗
[Genetic polymorphisms of serum orosomucoid in Han population in Nanjing of China].
OBJECTIVE: To determine the genetic polymorphisms of serum orosomucoid (ORM) in Han population in Nanjing of China and provide genetic data for studying the individual variance in the plasma protein binding of weak alkaline drugs. METHODS: The desialylated sera from 220 unrelated subjects were analyzed by isoelectric focusing on polyacrylamide gels followed by immunoblotting. RESULTS: The band patterns of ORM showed that the polymorphism of the structural locus ORM1 was controlled by three codominant autosomal alleles, ORM1*F1, ORM1*F2 and ORM1*S, which presented five phenotypes, ORM1 F1(47.27%), ORM1 S(5.46%), ORM1 F1F2 (3.18%), ORM1 F1S(43.64%), and ORM1 F2S(0.45%). The allele frequencies were: ORM1*F1=0.7068, ORM1*F2=0.0182, ORM1*S=0.2750. CONCLUSION: The ORM1 locus is polymorphic and the ORM2 locus is monomorphic in sera from the Han people in Nanjing of China.
Alleles↗
[The effect of inhibition of nitric oxide synthesis on vasoregulatory factors in pregnant rats].
OBJECTIVE: To study the effect of the inhibition of nitric oxide(NO) synthesis on plasma endothelin (ET), prostaglandlin I2(PGI2), thromboxane (TXA2) and angiotensin (ANGII) in pregnant rats, and to investigate the role of these factors in the pathogenesis of pregnancy-induced hypertension(PIH). METHODS: Pregnant rats were divided into two groups randomly. Saline solution or L-nitro arginine methyl ester (L-NAME) 125 mg/day was given subcutaneously from day 14 of gestation till delivery. Systolic blood pressure, urine protein, platelet count, and weight of pups and placentae were determined. Plasma NO, ET, PGI2, TXA2 and ANGII were measured by RIA. RESULTS: Pregnant rats which were given L-NAME produced physical signs similar to those of PIH, such as increase in systolic blood pressure (140.6 +/- 3.8) mmHg and urine protein (801.38 +/- 57.12) mg/L, and decrease in platelet count (533 +/- 42) x 10(9)/L and weight of pups and placentae. Compared with controls, plasma NO, PGI2 and ANG II were decreased significantly while plasma ET and TXA2 were increased significantly in rats given L-NAME. CONCLUSION: Inhibition of NO synthesis can cause imbalance of several vaso-regulatory factors, which may be responsible for the pathogenesis of PIH.
6-Ketoprostaglandin F1 alpha↗
[Effects of acute mild and moderate hypoxia on human short memory].
OBJECTIVE: To investigate the effects of acute mild and moderate hypoxia on human short memory. METHOD: Tests of pattern memory, scanning memory, continuous recognition memory and space memory were conducted on 18 healthy young male subjects during exposure to 300 m (control), 2800 m, 3600 m and 4400 m altitude in hypobaric chamber. RESULT: Compared to control group, only the performance of continous recognition memory decreased significantly after exposure to 2800 in for 1 h(P < 0. 05); total reaction time in all tests increased significantly (P < 0.0 1) and performance decreased after exposure to 3600 in for 1 h, but the error rates in memory scanning and space memory test were unchanged (P > 0.05); during exposure to 4400 in performance of all tests decreased further to about 90.4%, 93.7%, 83.3% and 80.7% of the control values (P < 0.01), respectively,and error rate increased significantly too (P < 0.05). CONCLUSION: Performance of human short memory decreased after exposure to acute mild and moderate hypoxia for 1 h, and these effects were aggravated with the increase of altitude. Characteristic and threshold of the effect on the various performance tests were different, but increase of reaction time was the first to appear in all cases.
Aerospace Medicine↗
Selection and determination of specific and protective antigens of Mycobacterium tuberculosis.
OBJECTIVE: To select and identify the specific and protective antigens of Mycobacterium tuberculosis (M. tuberculosis) for searching a new approach to diagnose and treat tuberculosis. METHODS: Extract and culture filtrates of M. tuberculosis were obtained by ultrasonic treatment and millipore membrane filtration, respectively. The protein samples were tested with monoclonal antibody (MAb) and patient sera. The proteins showing positive reaction were sequenced on Beckman/LF 3200/peptide amino acid sequencer. RESULTS: Proteins of M. tuberculosis with molecular weight of 31 ku and 30 ku showed positive results when reacted with anti-M. tuberculosis MAb and sera of tuberculosis patients, but not with normal mouse serum and healthy human sera. N-terminal sequences of the 31 ku and 30 ku antigen were Ala Glu Val Asp Trp Leu Val Phe Ala Val and Phe Ser Arg Pro Gly Leu Pro Val Glu Try respectively. CONCLUSION: 31 ku and 30 ku proteins of M. tuberculosis are immune protective proteins. They play important or dominant roles in determination of immunorecognization.
Amino Acid Sequence↗