2.An epidemiological investigation of bats carrying SARS-CoV in Guangzhou and its vicinity.
Zhi-feng LI ; Yong HU ; Hui-chun ZHAN ; Xue-xia YUN ; Yu-ping DU ; Xue-mei KE ; De-xian YU ; Jian-dong LI ; Ying-chun DAI ; Qing CHEN ; Shou-yi YU
Journal of Southern Medical University 2006;26(7):949-953
OBJECTIVETo detect serve acute respiratory syndrome-associated coronavirus (SARS-CoV) and SARS-like-CoV in fruit bats captured in Guangzhou and its vicinity.
METHODSTotally 927 bats of 9 species (Cynopterus sphinx, Rousettus leschenaulti, Miniopterus schreibersi, Hipposideros pratti, Rhinolophusasinicus, Scotophilusakuhlii, Hipposideros Pomona, Rhinolophus affinis, and Rhinolophus pusillus) captured in Guangzhou and its vicinity from September 2004 to November 2005 were available for this investigation, from which 3,043 samples (813 throat swasb, 524 sera, 853 lung tissues and 853 colorectal tissue specimens) were obtained. SARS-Cov and SARS-like-CoV were detected in these specimens using diagnostic kit for novel coronavirus N protein (ELISA), SARS-CoV Virus RNA detection kit, fluorescence PCR, Genchip, RT-PCR and cell isolation culture methods.
RESULTS AND CONCLUSIONNo SARS-CoV and SARS-like-CoV were detected in the 3043 samples, indicating the current absence of SARS-CoV and SARS-like-CoV in the bats captured in Guangzhou and its vicinity.
Animals ; China ; epidemiology ; Chiroptera ; virology ; Disease Vectors ; Enzyme-Linked Immunosorbent Assay ; Humans ; Nucleocapsid Proteins ; metabolism ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; metabolism ; Severe Acute Respiratory Syndrome ; epidemiology ; transmission ; virology
3.Influence of hepatocyte cell adhesion molecule on gene expression profile of human bladder transitional cell carcinoma cell line.
Qiu-ju WANG ; Chang-kun LV ; Jia TAO ; Hong-fei DU ; Yan-ru FAN ; Xue-dong SONG ; Chun-li LUO
Acta Academiae Medicinae Sinicae 2013;35(2):190-198
OBJECTIVETo investigate the changes of gene expression file in transitional cell carcinoma of bladder after hepatocyte cell adhesion molecule(hepaCAM) overexpression.
METHODSAffymetrix Human Genome U133 Plus 2.0 Array was used to investigate the changes of gene expression profile between adenovirus-green fluorescent protein(GFP) -hepaCAM group and GFP group in transitional cell carcinoma of bladder EJ cells.Significant Analysis of Microarray(SAM) was used to screen the differentially expressed genes, DAVID software was used to conduct gene ontology analysis and wikiPathway analysis based on the differentially expressed genes. Reverse transcription-polymerase chain reaction and Western blot were applied to verify microarray data.
RESULTSCompared with the GFP group, a total of 2469 genes were up-regulated or down-regulated by more than 2 times in the GFP-hepaCAM group. Among these genes, 1602 genes were up-regulated and 867 were down-regulated.Most of the differentially expressed genes were involved in the function of cell proliferation and cell cycle regulation. The mRNA expressions of nibrin, liver kinase B1, and cyclin D1 detected by reverse transcription-polymerase chain reaction in three different bladder cancer cell lines were consistent with the microarray data.The protein expressions of nibrin and liver kinase B1 in these three cell lines measured by Western blot were consistent with the mRNA expression.
CONCLUSIONSHepaCAM can alter the gene expression profile of bladder cancer EJ cells. The well-known anti-tumor effect of hepaCAM may be mediated by regulating the gene expression via multiple pathways.
Carcinoma, Transitional Cell ; genetics ; pathology ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; physiology ; Humans ; Nuclear Proteins ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Proteins ; genetics ; physiology ; Urinary Bladder Neoplasms ; genetics ; pathology
4.Determination of nineteen organonitrogen pesticides in Paeoniae Radix Alba by capillary electrophoresis-mass spectrometry.
Xue-chun DU ; Yi-min XU ; Shen JI ; Xiu-hong MAO ; Ke WANG ; Shu-Fang WANG
Journal of Zhejiang University. Medical sciences 2012;41(1):25-31
OBJECTIVETo establish a capillary electrophoresis-mass spectrometry(CE-MS) method for the analysis of nineteen organonitrogen pesticides in Paeoniae Radix Alba.
METHODSCE-MS analysis was performed on a 70 cm X 50 μm fused-silica capillary. The optimal buffer was composed of 1 % formic acid and 15 % methanol(V/V, pH 2.2). The temperature of capillary was controlled at 25 degree. The separation voltage was +20 kV. The optimal MS parameters were as follows: ESI-MS analysis was performed in the positive mode; 90 % methanol containing 0.2 % formic acid with a flow rate of 8 μl·min(-1) was selected as the sheath liquid; the flow rate and temperature of drying gas were 6 L·min(-1) and 250 degree, respectively; The nebulizing gas pressure was set at 5 psig; The optimal values of fragmentor and ESI voltage were 100 V and 5 000 V, respectively.
RESULTSThe nineteen pesticides had good linearity over the testing ranges. The average recoveries were in the range of 80.1 %-108.4 % with RSDs less than 20 % (except ethoxyquin and spiroxamine, those of which were 29.2 % and 22.3 % at 0.01 mg.kg(-1) concentration level). The LODs of nineteen pesticides were 0.503 ≊10.1 μg.kg(-1).
CONCLUSIONThe method can be used effectively to analyze the nineteen organonitrogen pesticides residue in Paeoniae Radix Alba.
Electrophoresis, Capillary ; methods ; Mass Spectrometry ; methods ; Paeonia ; chemistry ; Pesticide Residues ; analysis
5.Chemical constituents of Geranium eristemon.
Shu-shan DU ; Wen-sheng ZHANG ; Chen WU ; Yan-chun XU ; Lu-xue WEI
China Journal of Chinese Materia Medica 2003;28(7):625-626
OBJECTIVETo study the chemical constituents of Geranium eristemon.
METHODChromatography and spectral analysis were used to isolate the constituents and elucidate their structures.
RESULTFive compounds were isolated from acetone extract of the whole grass of G. eristemon, and identified as beta-sitosterol, protocatechuic acid, myricetin, kaempferol-7-O-alpha-L-arabifuranoside and kaempferol-3-O-alpha-L-arabifuranoside.
CONCLUSIONkaempferol-7-O-alpha-L-arabifuranoside and kaempferol-3-O-alpha-L-arabifuranoside were isolated from G. genus for the first time.
Arabinose ; analogs & derivatives ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Geranium ; chemistry ; Hydroxybenzoates ; chemistry ; isolation & purification ; Kaempferols ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Sitosterols ; chemistry ; isolation & purification
6.Isolation, purification and characterization of endothelial cells from endometrial cancer and normal endometrial tissue.
Xue-lian DU ; Xiu-gui SHENG ; Chun-xiao YAN ; Ting LIU ; Chun-hua LU ; Cong WANG ; Hao YU ; Qing-shui LI
Chinese Journal of Oncology 2012;34(6):409-413
OBJECTIVETo immunopurify human endometrial endothelial cells (HEEC) from fresh surgical specimens of endometrial cancers and normal endometrial tissues, and investigate their biological characteristics.
METHODSEndothelial cells of endometrial cancers and normal endometrial tissues were isolated using anti-CD31 conjugated magnetic microbeads. The isolated endothelial cells were cultured in vitro and their origins were identified. Their angiogenic characteristics were observed by MTT, wound healing, Transwell cell invasion and tube formation assays.
RESULTSFlow cytometry revealed that the immunopurification technique yielded endothelial cell purity of > 95% in all samples. All purified HEEC were characterized as endothelial cells on the basis of expression of the classical endothelial markers vWF and CD31 as shown by immunofluorescence examination. Although the tumor-associated HEEC didn't show more rapid proliferation than normal HEEC, they exhibited enhanced migration ability (P = 0.006), potent invasiveness (P = 0.033), and elevated tube formation in vitro (P = 0.029).
CONCLUSIONSHuman endometrial endothelial cells can be efficiently isolated from endometrial cancer and normal endometrial tissues by immunomagnetic methods. Tumor-associated HEEC exhibit enhanced migratory ability, potent invasiveness, and elevated tube formation in vitro.
Adult ; Aged ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Endometrial Neoplasms ; metabolism ; pathology ; Endometrium ; cytology ; metabolism ; pathology ; Endothelial Cells ; metabolism ; pathology ; Female ; Humans ; Middle Aged ; Neoplasm Invasiveness ; Neovascularization, Pathologic ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; von Willebrand Factor ; metabolism
7.Bacterial carriage and influencing factors of mobile phones used by health care workers in municipal hospitals in a city
Chun-Bei ZHOU ; Bing ZHU ; Chun-Yan LIAO ; Jiang DU ; Ya-Ming HE ; Xue-Fan YANG ; Ning YAO ; Tong-Jian CAI
Chinese Journal of Infection Control 2017;16(12):1141-1146
Objective To understand the status of mobile phone use and bacterial carriage on surface of mobile phones used by health care workers(HCWs) in municipal hospitals in a city,explore the influencing factors of mobile phone use behavior and bacterial carriage status.Methods In April-June,2016,111 HCWs in 24 hospitals in a city were performed questionnaire survey,on-site observation,and sampling of mobile phone surface.Results A total of 111 (100.00%) available questionnaires were distributed and returned.The average age of the respondents were (32.00 ± 9.03)years old,female and nurses were predominant.95.50% of respondents used touch screen mobile phones,24.32% used mobile phones during diagnosis and treatment,65.77% used mobile phone >2 hours every day,93.69% cleaned and disinfected mobile phones,98.20% thought that pathogenic microorganisms exited on the surface of mobile phones.A total of 111 mobile phone surface specimens were collected,the qualified rate was 80.18%,contamination rate was 95.50%,average colony number was 2.90 CFU/cm2,the maximum bacterial content was 111.60 CFU/cm2.Among 44 specimens of mobile phone surface,55 strains of 18 species of pathogenic bacteria or opportunistic pathogenic bacteria were detected.Age,gender,and occupation were the influencing factors of mobile phone use behavior and attitude;qualified rates were all significantly different among mobile phones used by HCWs of different gender,occupation,and duration of mobile phone use (all P<0.05);bacterial contamination on the surface of mobile phones used by HCWs of different age,gender,occupation,duration of mobile phone use,and whether to use the phone shell/set were significantly different respectively(all P<0.05).Conclusion Potential pathogens on the surface of mobile phones may cause healthcare-associated infection through the use of mobile phones by HCWs during the process of medical diagnosis and treatment.
8.Effects of nitric oxide and hydrogen sulfide on the relaxation of pulmonary arteries in rats.
Yan-fei WANG ; Prabha MAINALI ; Chao-shu TANG ; Lin SHI ; Chun-yu ZHANG ; Hui YAN ; Xue-qin LIU ; Jun-bao DU
Chinese Medical Journal 2008;121(5):420-423
BACKGROUNDThe balance between vasodilation and vasoconstriction plays a major role in maintaining vascular homeostasis. However, the underlying mechanisms are unclear. More and more evidence suggested that there was an interaction in the regulation of vasorelaxation between nitric oxide (NO) and hydrogen sulfide (H(2)S). We explored the interaction between and effects of NO and H(2)S on the relaxation of pulmonary arteries in rats.
METHODSSeven male Sprague-Dawley rats were anaesthetized with chloral hydrate and the pulmonary arteries of each rat separated for the study of vascular activities. The vasorelaxing activities of pulmonary artery rings in response to different doses of a NO donor, sodium nitroprusside (SNP), or a H(2)S donor, sodium hydrogen sulfide (NaHS), were measured in vitro. When pulmonary artery rings were treated with a cystathionine-gamma-lyase inhibitor, DL-propargylglycine, in the presence of SNP or a nitric oxide synthase inhibitor, N(omega)-nitro-L-arginine methyl ester, in the presence of NaHS, the changes in relaxing activities were analyzed.
RESULTSThe relaxation of pulmonary artery rings was in a dose dependent manner in response to either SNP or NaHS. The relaxation rates of pulmonary artery rings increased from (30.90+/-4.62)% to (60.50+/-8.08)% when the concentration of SNP increased from 1 micromol/L to 3 micromol/L and from (26.13+/-4.12)% to (53.09+/-14.01)% when the concentration of NaHS increased from 25 micromol/L to 100 micromol/L. However, when appropriate inhibitor was added, the relaxation responses to SNP and NaHS decreased.
CONCLUSIONSThe results suggested that similarly to NO, H(2)S acted as a vasorelaxant either independently of, or synergistically with NO in the regulation of vasorelaxation. The interaction between NO and H(2)S played an important role in regulating relaxing activities of pulmonary arteries.
Animals ; Hydrogen Sulfide ; pharmacology ; In Vitro Techniques ; Male ; Nitric Oxide ; physiology ; Nitroprusside ; pharmacology ; Pulmonary Artery ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Vasodilation ; drug effects
9.Clinical significance of pancreatic beta-cell function in obese children with acanthosis nigricans.
Xue-jun LIANG ; Cheng ZHU ; Chun YAN ; Gui-chen NI ; Zhong-liang LIU ; Zhong-min DU ; Ming LI
Chinese Journal of Pediatrics 2004;42(6):405-407
OBJECTIVEThe strong relation between type 2 diabetes mellitus and obesity with acanthosis nigricans is widely concerned. This study investigated the pancreatic beta-cell function in obese children with acanthosis nigricans, so as to find out the role of insulin secretion and insulin resistance in obese children with acanthosis nigricans.
METHODSThirty-five obese children with acanthosis nigricans (19 males and 16 females with mean age 12.8 +/- 1.5 years) were enrolled in this study. Thirty-eight obese children (21 boys and 17 girls with mean age 11.9 +/- 2.6 years) and 39 normal children (20 boys and 19 girls with mean age 11.2 +/- 2.2 years) were recruited as obese and normal control groups. The levels of serum fasting insulin, C-peptide, proinsulin and true insulin were measured in all the subjects. The ratios of proinsulin/insulin and proinsulin/C-peptide were calculated. Homeostasis model assessment was applied to assess the status of insulin resistance and basic function of pancreatic beta-cell.
RESULTSThe levels of fasting insulin, C-peptide proinsulin, true insulin, the ratios of proinsulin/insulin and proinsulin/C-peptide, insulin resistance index and insulin secretion index of obese children with acanthosis nigricans, obese control children and normal control children were: 18.5 (5.0-60.5) pmol/L, 12.4 (6.1-35.8) pmol/L and 5.1 (2.0-32.8) pmol/L; 3.9 (1.3-14.0) microg/L, 2.4 (1.1-4.0) microg/L and 1.1 (1.0-4.2) microg/L; 28.8 (9.9-64.2) pmol/L, 9.5 (2.2-34.5) pmol/L and 4.2 (2.0-16.0) pmol/L; 33.0 (6.2-66.0) pmol/L, 10.6 (4.8-29.4) pmol/L and 4.5 (1.3-30.1) pmol/L; 1.2 (0.4-8.9), 0.9 (0.2-1.9) and 0.8 (0.4-2.0); 6.9 (2.5-36.6), 4.7 (1.2-12.3) and 3.6 (1.2-9.6); 5.0 (0.8-14.1), 2.6 (1.3-8.1) and 1.2(0.4-6.9); 303.3 (52.2-1,163.8), 213.6 (84.6-572.0) and 51.1 (19.1-561.4). The levels of fasting insulin, C-peptide, proinsulin, true insulin, the ratios of proinsulin/insulin and proinsulin/C-peptide, insulin resistance index and insulin secretion index in obese children with acanthosis nigricans were significantly higher than those in obese children (P < 0.001) and normal children (P < 0.001).
CONCLUSIONObese children with acanthosis nigricans had higher insulin resistance and pancreatic beta-cell dysfunction; acanthosis nigricans may be a skin sign of high risk of type 2 diabetes mellitus.
Acanthosis Nigricans ; complications ; Adolescent ; C-Peptide ; blood ; Child ; Diabetes Mellitus, Type 2 ; etiology ; Female ; Humans ; Insulin ; blood ; Insulin Resistance ; Islets of Langerhans ; physiopathology ; Male ; Obesity ; complications ; physiopathology ; Proinsulin ; blood
10.Construction and identification of RNA interference vector for human tissue factor gene.
Wen TANG ; Shi-long XIONG ; Hong-ling YANG ; Jie BAO ; Jiang DU ; Shao-dong HUA ; Xue-gang SUN ; Zhi-chun FENG
Journal of Southern Medical University 2007;27(7):1065-1067
OBJECTIVETo construct a RNA interference vector for human tissue factor (TF) gene.
METHODSHuman TF short hairpin RNA (shRNA) sequence was designed using online design software (Invitrogen) and synthesized into double-strand oligonucleotide (ds oligo), which was cloned into the pENTRTM/U6 plasmid, followed by transformation of the product into competent Top10 E. coli cells. After expansion of the transformed bacteria, the plasmid was extracted and sequenced, which was subsequently transfected into human umbilical vein endothelial cells (HUVECs). The interference effect of the vector on the target gene expression was detected by RT-PCR and immunofluorescence assay.
RESULTSThe sequencing result indicated that the plasmid pENTRTM/U6-RelB-shRNA was constructed correctly, which resulted in effective inhibition of TF expression in HUVECs after transfection.
CONCLUSIONThe RNA interference vector against human TF gene has been constructed successfully, which may provide a stable transfection vector for potential treatment of blood coagulation abnormalities.
Base Sequence ; Cell Line ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; Humans ; Molecular Sequence Data ; Oligonucleotides, Antisense ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Thromboplastin ; deficiency ; genetics ; Transfection