Objective:To develop and apply a method for real-time quantifying IL-6 mRNA.Methods:By Ficoll-Hypaque density gradient centrifugation, human peripheral blood mononuclear cells(PBMC) were isolated from whole blood treated with ethylenediaminetetraacetic acid(EDTA). Total RNA extracted from the PBMC in trizol solution was reverse transcribed to cDNA, which used as templates for fluorecent real-time quantitative PCR amplification of IL-6. PCR system consists of IL-6 primer, SBY Green Ⅰ and so on.Results:The method is wide linear range, sensitive, specific, reproducible and simple.Conclusion:The method can accurately quantify IL-6 mRNA.

Objective To study the differential proteome of kidney between lupus nephritis mouse and normal mouse.Methods The proteins of kidney were separated by two-dimensional gel electrophoresis(2-DE).The gels stained by silver were scanned by ImageScanner and analyzed by PDQuest software.Results About 573?52 and 658?43 protein spots were found in the three maps of control group and LN group respectively;the match ratio was 83% and 87% respectively.One hundred and fourteen spots were found increased that showed a two fold increase as comparing to control group.Conclusion A significant difference in protein expression of LN mouse kidney was found and may be related to the pathogenesis of LN.

Objective To assess the methylation patterns in CpG islands of SPARC genes and its relationship with clinicopathological parameters. Methods Bisulfite treatment of genomie DNA and sequencing analysis was used to study methylation patterns in the CpG islands of SPARC genes in fresh tissues from 6 cases of chronic pancreatitis, 6 normal pancreatic tissues, 17 pancreatic adenocarcinoma and the cancer adjacent tissues, as well as 6 normaI blood samples for normal control, and compared the results with clinicopathological parameters. Results WBC DNA showed no methylation of SPARC gene CpG islands. The methylation rates in CpG islands of SPARC genes in pancreatic adenocarcinoma, the cancer adjacent tissues, chronic pancreatitis and normal pancreatic groups (2, 3, 4, 5, 6, 7 CpG sites) were 61.6%, 47.1%, 37.5%, 24.7%, respectively. The methylation rates in CpG islands (1, 8, 9, 10, 11, 12 sites) were 52.0%, 28.7%, 16.7% and 0. The difference were statistically significant between the pancreatic adenocarcinoma and chronic pancreatitis as well as normal pancreas groups (P＜0.001), and the difference were not statistically significant between the pancreatic adenocarcinoma and the cancer adjacent tissues. CpG hypermethylation were not related to risk factors such as smoking, alcohol, history of CP, the tumor size, differentiation and TNM staging, lymph node metastasis. Conclusions CpG in SPARC gene extron 1 was hypermethylated in pancreatic cancer, and this may be an early event in the development of pancreatic cancer.

OBJECTIVETo study the relationship between genetic polymorphism of tumor necrosis factor-alpha (TNF-alpha, sites at -238 nt and -308 nt) and susceptibility to chronic benzene poisoning.

METHODThe polymorphism of TNF-alpha gene were detected by nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique from 50 benzene poisoning patients and 60 control individuals exposed to benzene.

RESULTSThe frequency of the TNF-alpha-238G/G, A/G and A/A in controls (95%, 5% and 0%, respectively), were not significantly different from those in patients (92%, 6% and 2%, respectively, P > 0.05). The frequency of the TNF-alpha-308G/G, A/G and A/A in patients were 76%, 24% and 0% respectively, and that in control individuals were 93%, 7% and 0% respectively. Chi(2) test showed that frequency of TNF-alpha-308A/G in patients was significantly higher than that in controls (chi(2) = 6.22, P = 0.013). Logistic analysis showed that TNF-alpha-308A/G was the independent risk factor of benzene poisoning (OR = 5.3, P < 0.05).

CONCLUSIONIndividuals with TNF-alpha-308A/G gene polymorphism are susceptible to benzene poisoning.

Adult ; Benzene ; poisoning ; Chronic Disease ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Occupational Diseases ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo explore the recording method of the electrophysiological signals in corticospinal tract (CST) of adult rats by plugging microelectrodes and analyze the characteristics of these signals. These could provide some valuable and basic neural electrophysiological information for further research of recovering and refunctioning after spinal cord injury.

METHODSThe microelectrodes were plugged into the corticospinal tract at the T8 spinal section of Sprague-Dawley rats and the neuro-electrical signals were identified and recorded from CST by means of the Cerebus System. The characteristics of the recorded signals were described with the help of the Offline sorter and Neuroexplorer softwares, including the wavelength, amplitude, discharging frequency, the synchrony among the multi-discharging units from the same electrode and two different electrodes, analysis of interspike interval (ISI), etc.

RESULTSThe continuous and steady spontaneous electrophysiological signals were recorded from CST. Three or four types of discharging signals originated from different discharging units were collected with each electrode. The waveform of the signals appeared bidirectional. The wavelengths were 0.6 - 1.3 ms with wave amplitudes at a grade of hundred microvoltage and high signal-noise ratios. The LFB staining proved that the electrodes were accurately plugged into the corticospinal tract.

CONCLUSIONThe neuro-electrical signals at a grade of hundred microvoltage could be recorded stably from the corticospinal tract of rats by the Cerebus System with the microelectrodes, which provided valuable and basic neural electrophysiological information for further research on recovering and refunctioning after spinal cord injury (SCI) by analyzing the characteristics of electrophysiological signals.

Animals ; Electrodes, Implanted ; Electrophysiological Phenomena ; physiology ; Evoked Potentials, Motor ; physiology ; Male ; Microelectrodes ; Pyramidal Tracts ; physiology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; physiology ; Spinal Cord Injuries ; physiopathology

No abstract available.
Animals ; Calcium ; Interstitial Cells of Cajal ; Jejunum ; Mice

AIM: To investigate the antitumor activity and mechanism of Paecilomyces tenuipes polysaccharide(PTPS).METHODS: PTPS-I was obtained by water extraction and alcohol precipitation,and purified by DEAE-cellulose and Sephadex G-100 chromatography.Human erythroleukemia cell line K562,laryngocarcinoma cell line Hep2 and hepatic carcinoma cell line SMMC-7721were co-cultured with PTPS-I or the conditioned medium which prepared with PTPS-I-stimulated human mononuclear cells(PTPS-I-MNC-CM),and the proliferation of tumor cells was determined.The cell counting kit-8(CCK-8) was used to determine the proliferation of MNCs.The FQ-RT-PCR was applied to investigate the expression of TNF-? and IL-6 mRNA in MNCs.RESULTS: PTPS-I-MNC-CM inhibited the proliferation of K562,Hep2 and SMMC-7721 cells in vitro(P

OBJECTIVETo determine the risk factors involved in myelodysplastic syndromes (MDS).

METHODSA 1:2 case-control study was conducted in 20 Shanghai' hospitals over a 3-year period, covering 266 "de novo" MDS cases corresponded to FAB criteria, and 532 age- and gender-matched controls from same hospitals with MDS cases. Subjects were all surveyed using the same standard questionnaire including histories of medications (Chloramphenicol, Sulfonamides, Meprobamate, Phenytoin, Colchicine, Cyclophosphamide, Propylthiouracil, Anti-TB medication, Tolbutamide, Primaquine and Chinese traditional herbs such as Bezoar, Angelica, Arsenic, Thunder cloud vine) at least 5 years prior to the onset of the disease, tumors, exposure to benzene, heavy metal, organic phosphates, pesticides, petrol/diesel, organic solvents, dye and hair dye products, radiation, house decorating, alcohol and smoking.

RESULTSOccupational exposure to benzene increased significantly the risk of MDS (OR: 8.52, 95% CI: 2.30 - 31.10). Living near high voltage power lines (100 m) increased significantly the risk of MDS (OR: 1.60, 95% CI: 1.10 - 2.32). House decorating (one year prior to the onset of the disease) increased significantly the risk of MDS (OR: 2.40, 95% CI: 1.38 - 4.14). Other investigated occupational poisons did not increase significantly the risk of MDS. Hair dye products, alcohol and smoking did not increase significantly the risk of MDS.

CONCLUSIONOccupational exposure to benzene, living near high voltage power lines and house decorating are the risk factors of MDS.

Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; etiology ; Risk Factors

Objective To study the genotypes of Mycobacterium tuberculosis(M.tuberculosis)strains isolated from Jiangsu province and to explore the relationship between the 'Beijing family' and the drug resistance of M.tuberculosis.Methods Two hundred and sixty M.tuberculosis strains were isolated from 30 drug surveillance sites in Jiangsu province.Susceptibility of the isolates to the first-line antituberculosis drugs(isoniazid,streptomycin,rifampicin and ethambutol)was tested by using the proportion method.Molecular typing of M.tuberculosis strains was determined by Spoligotyping and analyzed with BioNumerics software.Results Based on Spoligotyping fingerprint,260 strains showed 34 different genotypes,including 27 exclusive genotypes and 7 shared genotypes.These strains could be clustered into two groups:the Beijing family(80.4％,209/260)and the Non-Beijing family(19.6％,51/260).Data from logistic regression analysis revealed that infection by the Beijing family was related to an increased risk of multi-drug resistant M.tuberculosis,with the OR(95％ CI)of 11.07(1.45-84.50).Non-Beijing families including T1,T2,H3,H4,CAS,LAM,U and MANU2 families were also found.Among them,the CAS,LAM and MANU2 families were first reported in Jiangsu province.Conclusion It was revealed that the marked gene polymorphisms did exist in M.tuberculosis strains.The Beijing family had been the predominant strain circulating in Jiangsu province,which might be related to multi-drug resistant M.tuberculosis strains.

Objective: To analyze the characteristics of the healthy state of TCM (Traditional Chinese Medicine) of female, and preliminarily constructs the model of TCM health status identification of female with menstrual cycle. Methods: The text of disease diagnosis and syndrome differentiation of menstrual-cycle-linked disorders from Guidelines for diagnosis and treatment of common disease of gynecology in Traditional Chinese Medicine was analyzed, and the symptoms related with viscera functions, the waxing and waning of qi and blood, and pathogenic properties were extracted. The questionnaires of diseases and syndrome elements differentiation of female by calculating the weight relationship between symptoms and health status were designed to estimate the interior state of waxing and waning of yin-yang by figuring out the different periods of menstrual, and in the last step, Constitution in Chinese medicine Questionnaire was used to identify the constitution. Through the above four identification methods, a comprehensive identification of the female's health status of TCM was formed. Results: Through the disease identification model, we distinguished the state of disease and yet disease. And through the syndrome elements differentiation model, we knew the viscera functions, the waxing and waning of qi-blood and yin-yang, and pathogenic properties. And by identifying the internal qi-blood and yin-yang status, we initially built the model that can identify health status of female integrally with the constitution. Conclusion:The model identification results can reflect the comprehensive health status of female in multiple dimensions. It can make a fine classification of women.s TCM health status. It can not only help to know the previous health status but also help to speculate the future health trend, which contributes to the formulation of personalized health interventions of follow-up.