1.Sequencing analysis on rpoB gene mutation of rifampin-resistance in Mycobacterium tuberculosis
Xin GAN ; Ling CHEN ; Jianhua WANG ; Nana LI ; Yang CHEN ; Jianyong ZHANG ; Hong ZHANG
Basic & Clinical Medicine 2010;30(3):268-271
Objective Genetic evaluation of relationship between rpoB gene mutation and rifampin-resistance of Mycobacterium tuberculosis (MTB).Methods This study was carded out with 81 clinical isolates and sensitivity test and genotypic analysis by PCR amplification and sequencing of rpoB gene were performed.Results Among these isolates,there were 27 rifampin-resistant and 54 sensitive strains.Of the 27 drug-resistant isolates,20 (74.1%) carried mutations on the amplified fragment of the rpoB gene with 10 mutation types at seven codons including 531 and 526,and two new mutation patterns were recognized.On the uther hand,one mutation (1.9%) appeared in 54 drug-sensitive strains.Conclusion The study shows geographical variation in the mutation types of rpoB gene in M.tuberculosis isolates from Guizhou,China.The result is valuable for the development of fast diagnostic methods.
2.Relationships of Vascular Endothelial Growth Factor and Interferon-?,Interleukin-4 in Asthmatic Rats and Effect of Budesonide on Their Expression
xiao-hong, JIN ; chang-chong, LI ; cun-guo, CHEN ; shao-bo, LI ; feng-xian, LI ; ling-hong, GAN
Journal of Applied Clinical Pediatrics 1994;0(04):-
Objective To explore the relationship of vascular endothelial growth factor(VEGF),IFN-? and IL-4 and effect of Bude-sonide on their in asthmatic rats.Methods Thirty-six male SD rats were randomly divided into 3 groups:asthma group,Budesonide-treatment group and control group.On the first day of the experiment and the 8th day,the rat models of the asthma group and Budesonid treatment group were allergized by the OVA/Al(OH)3 through intraperitoneal injection,respectively.And starting from the 15th day,they were challenged by the OVA through atomization for 2 weeks.Control group was allergized and challenged by NS atomization.Budesonid treatment group was interfered in Budesonide inhalation before suscitation in 0.5 h.After 12 h the same inhal done was again in Budesonide group.Twenty-four hours after the last challenge,the rats in 3 groups were sacrificed,and blood and bronchoalveolar lavage fluid(BALF) were collected.The concentrations of IL-4,IFN-? and VEGF in serum and BALF were measured by enzyme-linked immunosorbent assay.Results The concentrations of IL-4 in serum and BALF in asthma group and Budesonide treatment group were significantly increased than those in control group(P
3.Application of PCR-DGGE technique in G-6-PD deficiency.
Chang-Gan LI ; Xiao-Wen CHEN ; Yun-Sheng CHEN ; Ying WANG ; Wei-Ling ZHAO ; Hong-Song SHI ; Cheng-Rong LI
Chinese Journal of Contemporary Pediatrics 2007;9(6):529-532
OBJECTIVETo detect gene mutations of children with glucose-6-phosphorate dehydrogenase (G-6-PD) deficiency and of carriers of G-6-PD deficiency gene with the technique of polymerase chain reaction and denatured gradient gel electrophoresis (PCR-DGGE), and to explore the value of the technique in the diagnosis of G-6-PD deficiency and G-6-PD deficiency gene carrying.
METHODScDNAs were harvested by reverse transcription method after RNAs had been extracted from peripheral blood of 43 children with G-6-PD deficiency and of their family members (36 lineages). Electrophoresis behaviors of the fragment from exons 11-12 of G-6-PD cDNA were detected with the technique of PCR-DGGE. Gene sequencing was then performed for the abnormal electrophoresis bands.
RESULTSAbnormal electrophoresis bands were found in the 1304-1520 fragment of G-6-PD cDNA in 33 out of 36 family lineages. The G-6-PD/6-PGD ratio was below 1.00 in 9 mothers of patients. Three of them had the G-6-PD/6-PGD ratio lower than 0.50. The PCR-DGGE bands were the same in the 3 mothers. Gene sequencing showed double heterozygote in the 3 mothers, but the maternal carriers of G-6-PD deficiency gene who had normal G-6-PD/6-PGD ratio showed mono-heterozygote in gene sequencing. Three mutational sites were found in the 1304-1520 fragment, i.e., C1311TG1376T and G1388A. The electrophoresis behaviors were different among the 3 gene mutational sites.
CONCLUSIONSPCR-DGGE is a sensitive and reliable technique in the screening of gene mutations. It is useful in the diagnosis of G-6-PD deficiency, especially in the diagnosis of female G-6-PD deficiency gene carrying.
Base Sequence ; Electrophoresis, Polyacrylamide Gel ; Female ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA
4.Effect of arsenic trioxide on VEGF/R and MMP-2, 9 expressed in K562 cells.
Yu ZHANG ; Jian-Dong ZHANG ; Jian GU ; Li MA ; Wei-Gan SHEN ; Xiao-Ling WANG ; Hong CHEN
Chinese Journal of Hematology 2007;28(2):107-110
OBJECTIVETo explore the effect of arsenic trioxide (As2 O3) on the level of VEGF, VEGFR and the activity of MMP-2, 9 in K562 cells.
METHODSThe inhibition ratio of K562 cell was detected by MTT assay, the level of VEGF by Enzyme-linked immunosorbent assay (ELISA), the expression ratio of VEGFR by flow cytometry (FCM), and the activity of MMP-2, 9 by gelatin zymography assay.
RESULTS(1) The IC50 of K562 cells was (2.12 +/- 0.11) micromol/L. Proliferation of K562 cells was significantly inhibited at the concentration of 0.4 - 6.4 micromol/L As2 O3 (P < 0.05). (2) The expression of VEGF was slightly up-regulated by 0.05 micromol/L As2 O3 (P > 0.05) and prominently inhibited by 0.4 micromol/L and 3.2 micromol/L As2 O3 (P < 0.05). As2 O3 had no influence on VEGFR. (3) The activity of MMP-2 and 9 was partly inhibited by 0.05 micromol/L As2 O3 incubated 72 hours and by 0.4, 3.2 micromol/L, As2 O3. With the increase of As2 O3 concentration and the incubation time, the inhibited effect on MMP-2 and 9 was enhanced.
CONCLUSIONSAs2 O3 may down-regulate the expression of VEGF and inhibit the activity of MMP-2 and 9.
Arsenicals ; pharmacology ; Cell Proliferation ; drug effects ; Humans ; K562 Cells ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Oxides ; pharmacology ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism
5.Identification of tetracenomycin X from a marine-derived Saccharothrix sp. guided by genes sequence analysis.
Bin LIU ; Yi TAN ; Mao-Luo GAN ; Hong-Xia ZHOU ; Yi-Guang WANG ; Yu-Hui PING ; Bin LI ; Zhao-Yong YANG ; Chun-Ling XIAO
Acta Pharmaceutica Sinica 2014;49(2):230-236
The crude extracts of the fermentation broth from a marine sediment-derived actinomycete strain, Saccharothrix sp. 10-10, showed significant antibacterial activities against drug-resistant pathogens. A genome-mining PCR-based experiment targeting the genes encoding key enzymes involved in the biosynthesis of secondary metabolites indicated that the strain 10-10 showed the potential to produce tetracenomycin-like compounds. Further chemical investigation of the cultures of this strain led to the identification of two antibiotics, including a tetracenomycin (Tcm) analogs, Tcm X (1), and a tomaymycin derivative, oxotomaymycin (2). Their structures were identified by spectroscopic data analysis, including UV, 1D-NMR, 2D-NMR and MS spectra. Tcm X (1) showed moderate antibacterial activities against a number of drug-resistant pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) pathogens, with the MIC values in the range of 32-64 microg x mL(-1). In addition, 1 also displayed significant cytotoxic activities against human cancer cell lines, including HL60 (leukemia), HepG2 (liver), and MCF-7 (breast) with the IC 50 values of 5.1, 9.7 and 18.0 micromol x L(-1), respectively. Guided by the PCR-based gene sequence analysis, Tcm X (1) and oxotomaymycin (2) were identified from the genus of Saccharothrix and their 13C NMR data were correctly assigned on the basis of 2D NMR spectroscopic data analysis for the first time.
Actinomycetales
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chemistry
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genetics
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Anti-Bacterial Agents
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chemistry
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isolation & purification
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pharmacology
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Antineoplastic Agents
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chemistry
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isolation & purification
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pharmacology
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Benzodiazepinones
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chemistry
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isolation & purification
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pharmacology
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Cell Line, Tumor
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Data Mining
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methods
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Drug Resistance, Bacterial
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Enterococcus faecalis
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drug effects
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Fermentation
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Genomics
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Humans
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Inhibitory Concentration 50
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Marine Biology
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Methicillin-Resistant Staphylococcus aureus
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drug effects
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Microbial Sensitivity Tests
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Molecular Structure
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Naphthacenes
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chemistry
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isolation & purification
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pharmacology
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Phylogeny
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Staphylococcus epidermidis
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drug effects
6.Expression of miR-125a/b-5p in Serum and Urine of Preeclampsia and Related Clinical Studies
Lu GAN ; ling Xi LIU ; Ting WANG ; ying Xiao LIU ; hong Li CHEN ; Ming WEI ; Xin ZHANG
Journal of Modern Laboratory Medicine 2017;32(6):32-34,38
Objective To detect miR-125a/b-5p expression in serum and urine of patients with preeclampsia,explore the role of miR-125a/b-5p in peripheral circulation on the pathogenesis of preeclampsia.Methods Samples of serum and urine of pa-tients with preeclampsia(n=20),and the normal pregnancy group(n=20)were collected and the expression of miR-125a/b-5p was detected by stem-loop-based real-time fluorescence quantitative reverse transcriptase polymerase chain reaction.Re-sults The expression of miR-125b-5p in the serum of patients with preeclampsia were significantly decreased compared with the normal pregnancy(Z=-2.272,P=0.023).There were no statistically differences of the expression of miR-125a-5p in serum between the two groups(Z=-0.622,P=0.547).The differences of urinary miR-125a/b-5p between patients with preeclampsia and normal pregnancy were not statistically significant(Z=-0.663,-0.189,P>0.05).Urinary miR-125b-5p was negatively correlated with diastolic blood pressure in preeclampsia group(r=-0.513,P=0.021).Serum miR-125b-5p was positively correlated with urea nitrogen in normal pregnancy group(r=0.472,P=0.036).Urinary miR-125a/b-5p were positively correlated with systolic blood pressure in normal pregnancy group(r=0.526,0.502,P=0.017,0.024). Conclusion The decrease of serum miR-125b-5p was associated with the pathogenesis of preeclampsia.
7.Effect of parental factors on junior high students′ physical activity in their leisure time outside school
Dan-dan YU ; Ling-ling ZHANG ; Li ZHANG ; Dong-fang HAN ; Chen-chen LI ; Hong-wei GAN ; Li-hua WANG
Shanghai Journal of Preventive Medicine 2020;32(9):751-
Objective To investigate the effects of parental support for the moderate-to-vigorous physical activity (MVPA) of their children and to promote physical activity for children and adolescents. Methods A total of 622 students were selected by stratified clustering sampling methods.Physical activity and parental support factors were assessed by self-report questionnaire survey.Logistic regression was used to analyze determinants of physical inactivity. Results Nearly 58.7% of the parents purchased sports equipment for their children and 57.6% of the parents accompanied their children to do exercise.The rate of parental explicit modeling was 22.8%.The students who received parental logistic support and explicit modeling did longer MVPA on weekends.The students with parental logistic support were prone to be in activity (
8.Therapeutic efficacy and prognostic factors of sorafenib treatment in patients with unresectable primary hepatocellular carcinoma.
Lan ZHANG ; Zheng-gang REN ; Yu-hong GAN ; Yan-hong WANG ; Bo-heng ZHANG ; Yi CHEN ; Xiao-ying XIE ; Nin-ling GE ; Sheng-long YE
Chinese Journal of Oncology 2010;32(8):630-633
OBJECTIVETo evaluate the efficacy and analyze the prognostic factors of sorafenib treatment in patient with unresectable primary hepatocellular carcinoma (HCC).
METHODSDuring the period from December 2005 to March 2009, 50 patients with unresectable primary HCC of Child-Pugh status A were treated with sorafenib (400 mg, Bid). The tumor response was evaluated with CT or MRI imaging every 6 - 8 weeks according to the RECIST criteria. The overall survival (OS) and time to progression (TTP) were defined as the time from administration of sorafenib to the death or the last follow up and were evaluated by Kaplan-Meier method.
RESULTSThere was no PR or CR, but 28 patients (56.0%) achieved stable disease. The median follow up time was 15 months with a median OS of 14 months and median TTP of 4 months. The common adverse events were dermal reaction (68.0%, 34/50), diarrhea (52.0%, 26/50), hypertension (4.0%, 2/50), hair loss (14.0%, 7/50), myelosuppression (16.0%, 8/50), and liver dysfunction (20.0%, 10/50). However, most of the drug-related adverse events were grade I-II and reversible. The patients with lower tumor burden and without distant metastasis had better prognosis.
CONCLUSIONSoafenib is effective for unresectable primary HCC with tolerable toxicity. Tumor stage is a predominant prognostic factor.
Adult ; Aged ; Alopecia ; chemically induced ; Antineoplastic Agents ; adverse effects ; therapeutic use ; Benzenesulfonates ; adverse effects ; therapeutic use ; Carcinoma, Hepatocellular ; drug therapy ; Chemoembolization, Therapeutic ; methods ; Diarrhea ; chemically induced ; Disease Progression ; Follow-Up Studies ; Humans ; Hypertension ; chemically induced ; Liver Neoplasms ; drug therapy ; Male ; Middle Aged ; Neoplasm Staging ; Niacinamide ; analogs & derivatives ; Phenylurea Compounds ; Pyridines ; adverse effects ; therapeutic use ; Skin Diseases ; chemically induced ; Survival Rate
9.Detection of DD3 mRNA in the urine of prostate cancer patients and its clinical significance.
Guang-xiang LIU ; Hong-qian GUO ; Xiao-gong LI ; Wei-dong GAN ; Ling-qi ZENG ; Hong-lei SHI ; Shi-wei ZHANG ; Tie-shi LIU
National Journal of Andrology 2007;13(6):511-513
OBJECTIVETo detect the differential display code 3 mRNA (DD3 mRNA) in the urine sample of patients with prostate cancer and to evaluate its clinical significance.
METHODSDD3 mRNA in the urine collected from 48 patients with prostate cancer, 23 patients with benign prostate hyperplasia (BPH) and 9 healthy male volunteers was measured by reverse transcriptase polymerase chain reaction (RT-PCR).
RESULTSDD3 products could not be detected in the urine samples of the BPH patients and healthy male volunteers, but could in 39/48 urine samples of the patients with prostate cancer. Significant difference was found between them (P < 0.01).
CONCLUSIONThe detection of DD3 mRNA in the urine promises to be a non-invasive, simple and sensitive method for the early diagnosis and post-treatment monitoring of prostate cancer.
Aged ; Aged, 80 and over ; Antigens, Neoplasm ; genetics ; urine ; Humans ; Male ; Middle Aged ; Prostatic Neoplasms ; diagnosis ; RNA, Messenger ; urine ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity
10.Evaluation of the efficacy and prognostic factors for colorectal liver metastases treated with transcatheter arterial chemoembolization.
Tao YE ; Yao-hui WANG ; Jing-lin XIA ; Bi-wei YANG ; Yi CHEN ; Ning-ling GE ; Yu-hong GAN ; Yan-hong WANG ; Zheng-gang REN
Chinese Journal of Oncology 2012;34(9):706-709
OBJECTIVEThe aim of this study was to evaluate the therapeutic efficacy and to determine the prognostic factors of TACE in patients with colorectal liver metastases (CRLM).
METHODSThe clinical data of 183 patients with unresectable CRLM treated with TACE from Jan. 2002 to Dec. 2008 were retrospectively reviewed. Log-rank method was used for univariate analysis and Cox proportional hazard model was used for multivariate analysis of the prognostic factors.
RESULTSThe median survival time was 22 months, and the 0.5-, 1-, 2-, 3-, 5-year survival rates were 93.9%, 81.1%, 39.8%, 18.2%, and 3.9%, respectively. Multivariate analysis showed that tumor involved more than one lobe of the liver, and elevated CEA and CA19-9 levels were independent risk factors for the overall survival (P < 0.01). Females, more times of TACE, combination with regional therapy and received phase II resection were related with a good survival (P < 0.01) in CRLM patients after TACE treatment.
CONCLUSIONSTranscatheter arterial chemoembolization is an effective therapy for unresectable colorectal liver metastases. Patients with tumor spread more than one lobe of the liver, high CEA and CA19-9 levels are independent poor prognostic factors. Females, patients received more times of TACE, combined with regional therapy and received phase II resection may have a good survival.
Adult ; Aged ; Aged, 80 and over ; Antigens, Tumor-Associated, Carbohydrate ; blood ; Carcinoembryonic Antigen ; blood ; Chemoembolization, Therapeutic ; Colonic Neoplasms ; pathology ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Humans ; Iodized Oil ; administration & dosage ; Liver Neoplasms ; blood ; secondary ; surgery ; therapy ; Male ; Middle Aged ; Mitomycin ; administration & dosage ; Organoplatinum Compounds ; administration & dosage ; Proportional Hazards Models ; Rectal Neoplasms ; pathology ; Retrospective Studies ; Survival Rate