To study the expression of HCV non-structure 5 antigen in vitro, a human HepG2 cell line was incubated with a HCV RNA positive serum. The S ABC i mmunological techniques and gold-labeled colloid electron microscopy method wer e employed to examine for the viral proteins in those cells. The HCV non-struct ure 5 antigen was first detected in the HepG2 cells at 72 hours post incubation. The antigen was continuously observed in the cytoplasm or on the membrane as we ll on the cell wall of the HepG2 cells even after 1, 2, 3 and 4 weeks post incub ation. The observation of HCV non-structure 5 antigen continuously expressed in the HepG2 cells strongly indicates that the cells may have been infected by HCV virus and the virus may have replicated in the cells. Therefore, the HepG2 cell line may be served as a potential host for establishment of HCV infection and p ropagation in vitro.

BACKGROUND:An abroad study repoRed the distribution and expression of augmenter of liver regeneration(ALR)in the central nervous system.There are few literatures on how to prepare and evaluate ALR protein polyclonal antibody in recombinant rats,and how to construct prokaryotic expression vector.There are no repots concerning ALR in the central nervous system in China.OBJECTIVE:TO express ALR fusion protein in E coli BL21 and prepare and identify polyclonal antibody.METHODS:RNA was extracted from the hippocampus of Sprague Dawley rats.The prokaryotic expression plasmid pET28a-ALR was constructed and the positive recombinant plasmid was transformed into BL21.Protein ALR was expressed by inducing transformed BL21 with Isopropyl-β-D-thiogalactopyranoside(IPTG)and purified by Ni~(2+)affinity chromatography column after immune the rabbit for 4 times.the serum of rabbits was extracted from hear as polyclonal antibody.The titer and specificity of the rabbit's antiserum was respectively measured by ELISA and Western blotting The following parameters were measured:construction of prokaryotic expression plasmid pET26a-ALR;pET28a-ALR recombinant enzyme digestion evaluation;results of ELISA and Western-blotting.RESULTS AND CONCLUSION:Expecting bands were obtained by double enzyme digestion electrophoresis,respectively 5.3 kb and 0.4 kb.Nucleotide sequence analysis verified that prokaryotic expression vector pET28a-ALR was successfully constructed.The 19 ku fusion protein was successfuIly expressed.The titer of the antiserum measured by ELlSA could achieve 1:2 000 This indicated that antibody and purified recombinant ALR had a good reaction.and high titer.could meet the experimental require.Western blotting analysis proved that the antibody could identify the prokaryotic expression product of ALR.Prokaryotic expression system expressed ALR fusion protein,prepared and purified polyclonal antibody of ALR protein,and could meet the experimental require of ALR immunoblotting.

Objective To investigate the rehabilitative effect of low frequency repetitive transcranial magnetic stimulation (rTMS) on convalescing patients with Broca's aphasia. MethodsTwenty-eight patients with Broca's aphasia recovering from cerebral infarction were randomly divided into a stimulation group and a control group with 14 subjects in each.Patients in the control group accepted conventional drugs,speech rehabilitation and sham stimulation,while patients in the stimulation group were in given low frequency rTMS in place of the sham stimulation.Their speech performance was evaluated using the China Rehabilitation Research Center's aphasia examination (CRRCAE) pre-stimulation,post-stimulation and 90 days later. ResultsCompared with before treatment and with the controls,the speaking scores of the stimulation group increased significantly after treatment and also 90 days later. ConclusionLow frequency rTMS can not only improve the speech performance of Broca's aphasia sufferers in the short term,but it also plays a lasting role.It may thus have clinical application for patients with Broca's aphasia.

ObjectiveTo discuss the experimental method and the mechanisms on treating diseases by acupuncturing the ST36(Zusanli).MethodsUsing Positron Emission Tomography(PET) and functional Magnetic Resonance Imaging(fMRI) to obtain the experimental data about glycometabolism and cerebral blood stream,using SPM and ROI image-analytical method to obtain the visual experimental evidence when acupuncturing the ST36. ResultsThere are certain increases of glycometabolism and cerebral blood stream in ipsilateral hypothalamus and bilateral temporal lobe, when acupuncturing the ST36. Conclusions Acupuncturing the ST36 can lead to the functional changes in vegetative nerve center and temporal lobe, which is close correlated with the therapeutical effects of ST36.

This article studied the chemical constituents from the aerial part of Vitis thunbergii var. taiwaniana. The 60% ethanol extract was eluted with 95% ethanol though HP-20 macroporous adsorption resin column. 12 compounds, including (1) betulinic acid, (2)2, 2, 2'-bis (4-hydroxyphenyl) propane bis (2, 3-epoxypropyl) ether, (3) eriodictyol, (4) trans-ε-viniferin, (5) (+)-cis-ε-viniferin, (6) kobophenol A, (7) ampelopsin A, (8) nepalensinol B, (9) cis-miyabenol C, (10) cis-vitisin B, (11) cis-gnetin H and (12) (+)-hopeaphenol, were separated by using normal phase silica gel, ODS, Sephdadex LH-20 column chromatographies and semi-preparative or preparative HPLC. Compounds 2, 5, 6, 8, 9, 10, 11 were separated from the genus Vitis for the first time and compounds 3, 7, 12 were separated from Vitis thunbergii var. taiwaniana for the first time. At a concentration of 50 μmol · L(-1), compound 6, 7 and 11 showed strong cytotoxicity against MCF-7 cell lines with the inhibition rate of 66.58%, 57.16%, 52.84%, respectively.
Antineoplastic Agents, Phytogenic ; pharmacology ; Humans ; MCF-7 Cells ; Plant Extracts ; analysis ; pharmacology ; Vitis ; chemistry

OBJECTIVETo explore the mechanism of occupational medicamentosa-like dermatitis (OMDT) induced by trichloroethylene (TCE) and some immunity indexes in workers occupationally exposed to TCE.

METHODSThe blood samples from 8 cases with medicamentosa-like dermatitis in 1st, 2nd, 3rd, 4th and 5th weeks after admitting to hospital were examined for liver function, immunoglobulin and some complement indexes. Thirty nine workers occupationally exposed to TCE were investigated for urinary TCE and some immuno-complement indexes. The TCE concentrations of air in workplaces were monitored.

RESULTSC3d-CIC and C3 of patients before admission were (92.86 ± 44.80) mg/L and 0.91 ± 0.19 mg/L, respectively. C3d-CIC and C3 of patients before discharge were (52.41 ± 17.75) mg/L and (1.14 ± 0.22) mg/L, respectively. There were significant differences between admission and discharge (P < 0.05). The average TCE concentration in 4 workplaces was (351.96 ± 36.72) mg/m(3), which was higher than the occupational exposure limits (OELs). The number of workers exposed to the TCE concentration-time weighted and TCA in urine over OELs were 28.21% and 56.41% of total subjects, respectively. The serum IgG and CIC levels of patients before discharge were (10.03 ± 1.21) mg/L and 103.50 ± 29.17 mU/L, which were significantly lower than those (17.21 ± 1.85) mg/L and (227.46 ± 111.67) mU/L of patients before admission (P < 0.01).

CONCLUSIONThe type II and III hypersensitivity may be associated with OMDT and the organ injure induced by TCE.

Adolescent ; Adult ; Complement System Proteins ; immunology ; Dermatitis, Occupational ; immunology ; Female ; Humans ; Male ; Occupational Exposure ; Trichloroethylene ; toxicity ; Young Adult

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; DNA Mutational Analysis ; DNA, Viral ; blood ; genetics ; Female ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Male ; Middle Aged ; Mutation ; Young Adult

OBJECTIVETo detect the expession of THY1 in ovarian serous cystadenocarcinoma tissues.

METHODSImmunohistochemistry was performed to detect the expression of THY1 gene in formalin-fixed, paraffin-embedded specimens of normal ovaries (n = 25), ovarian serous cystadenoma (n = 25), and serous cystadenocarcinoma (n = 53). The correlation of THY1 expression with clinicopathological parameters was statistically analyzed.

RESULTSThe positive expression rates of THY1 protein in normal ovaries, ovarian serous cystadenomas and ovarian serous cystadenocarcinomas were 60.0% (15/25), 72.0% (18/25) and 34.0% (18/53), respectively. The values of IOD of THY1 protein expression were 288,449.2 +/- 60,087.3, 271,655.6 +/- 66,588.7 and 252,087.6 +/- 45,559.4, respectively. The expression of THY1 protein was significantly down-regulated in ovarian serous cystadenocarcinoma tissues compared with that in normal ovarian tissues and ovarian serous cystadenoma tissues (P < 0.05). THY1 expression was negatively correlated with surgical-pathological staging, histological differentiation and lymph node involvement (P < 0.05).

CONCLUSIONThe decreased level of THY1 expression may be related with the occurrence and development of ovarian serous cystadenocarcinoma.

Adult ; Aged ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Cystadenoma, Serous ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Ovarian Neoplasms ; metabolism ; pathology ; Thy-1 Antigens ; metabolism ; Young Adult

OBJECTIVETo investigate the effects of gambogic acid (GA) on the proliferation inhibition and apoptosis induction in Human lung adenocarcinoma A549 cells in vitro, as well as the regulation of steroid receptor coactivator-3 (SRC-3) to explore the relationship between them.

METHODSThe effect of GA on the growth of A549 cells was studied by MTT assay. Apoptosis was detected by Hoechst 33258 staining. The localization of SRC-3 was determined by confocal laser scanning microscopy. Western blot and RT-PCR technique were applied to assess the expression of SRC-3.

RESULTSGA presented a striking proliferation inhibition potency on A549 cells in vitro, as well as apoptosis induction activity in a time- and dose-dependent manner. The IC(50) value for 24 h was (3.17 +/- 0.13) micromol/L. Overexpression of SRC-3 was found in A549 cells, whereas the SRC-3 protein and mRNA expression levels were significantly downregulated in A549 cells induced by GA in a dose-dependent manner. The location of SRC-3 was situated mainly in the cell nuclei.

CONCLUSIONGA exhibits a potent proliferation inhibition and apoptosis induction in human lung adenocarcinoma A549 cells, which might correspond to the downregulation of the expression of SRC-3. Thus, it promises to be a new target drug for lung cancer treatment.

Adenocarcinoma ; metabolism ; pathology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Down-Regulation ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Nuclear Receptor Coactivator 3 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Xanthones ; administration & dosage ; pharmacology

OBJECTIVETo establish a new and efficient method(IEDA) for detection of hemorrhagic fever with renal syndrome virus (HFRSV) antigen.

METHODSAn immune enzyme dot assay (IEDA) with mixture of three sorts anti-HFRSV-IgG, which was obtained from rabbit vaccinated with EHFV R22, Chen and Hubei strain was employed to detect HFRSV antigen in serum and urine from epidemic hemorrhagic fever (EHF) patients, and compared with indirect immune fluorescence assay (I-IFA), 76 serum samples and 40 urine samples were detected in this study.

RESULTSThe results showed that the total positive rate of HFRSV antigen detected by IEDA was 73.68% in serum and 65.00% in urine, while that detected by I-IFA was 75.00% and 70.00%, respectively. The positive rate in primary phase (within 5 days) of HFRSV antigen detected by IEDA was 94.34% in serum and 83.33% in urine, while that detected by I-IFA was 64.42% and 55.56%, respectively, there was significant difference in both serum and urine detections. Correlation study showed a high correlation in the result of IEDA and I-IFA.

CONCLUSIONThe results of this study suggested that the IEDA, as compared with I-IFA, was a more specific, sensitive, rapid and simple method with higher positive rate in primary phase. IEDA could be widely used for early diagnosis of HFRS in hospital at grassroots level.

Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; analysis ; Early Diagnosis ; Female ; Fluorescent Antibody Technique, Indirect ; Hantaan virus ; immunology ; Hemorrhagic Fever with Renal Syndrome ; diagnosis ; immunology ; Humans ; Immunoblotting ; methods ; Immunoglobulin G ; immunology ; Male ; Rabbits ; Rats ; Sensitivity and Specificity