Objective To investigate whether HBx alone is sufficient to directly transform the nontransformed immortalized human liver cell line QSG7701 and induce hepatocellular carcinoma in vivo,and to explore preliminarily the pathogenic mechanism of transplantation tumor in nude mice.Methods The pCMVX/QSG7701 cells were vaccinated into subcutaneous tissue of nude mice.The pRcCMV2/QSG7701 and QSG7701 Cells were used as control.At the fifth weeks after vaccination,the nude mice were killed to observe whether a tumor was formed.The activity state and food intake of the nude mice was recorded.The texture,volume,and metastasis of transplantation tumor were observed grossly.The transplantation tumor was observed microscopically on the hematoxylin and eosin (HE) staining section.Immunohistochemical surfactant protein (SP) method was used to analyze the protein expression of mutant p53 and C-Myc genes.Results The transplantation tumor was occurred in all of the six nude mice vaccinated with pCMVX/QSG7701 cells at the second week after vaccination.No metastatic tumor was found in other organs.Transplanted tumor was not formed in all of the negative control groups.HE staining analysis confirmed that the character of transplanted tumor was hepatocellular carcinoma.p53 and C-Myc proteins were expressed in pCMVX/QSG7701,pRcCMV2/QSG7701,and QSG7701 cells,and their expression levels in the pCMVX/QSG7701 cells were significantly higher than those in the pRcCMV2/QSG7701 and QSG7701 cells,respectively(P ＜0.01).Conclusions HBx alone is sufficient to directly transform the non-transformed immortalized human liver cell line QSG7701 and induce hepatocellular carcinoma in vivo through up-regulating the expression of mutant p53 and C-Myc genes.

Objective To explore the curative effects of repetitive transcranial magnetic stimulation ( rTMS) therapy on subcortical ischemic vascular disease( SIVD) patients with mild cognitive impairment.Methods Sixty-one SIVD patients with mild cognitive impairment but did not meet the diagnostic criteria for vascular dementia were enrolled and randomly divided into the treatment group (n=31) and the control group (n=30) in accordance with international common randomization form,TMT-A、TMT-B、VFT、AVLT score and P300 latency and amplitude changes were compared between the two groups before and after treatment.Results TMT-A、TMT-B、VFT、AVLT showed no difference before treatment between the two groups, but improved significantly after treatment in the treatment group ( P=0.040;P=0.041;P=0.034;P=0.010 ) .the results were also significantly different from the control group after treatment( P=0.019;P=0.009;P=0.044;P=0.045 ) .In the treatment group, P300 latency after treatment was significantly reduced than that before treatment (P=0.045),which was also significantly reduced from the control group ( P=0.025 ) , but P300 amplitude did not reach statistically significant difference before and after treatment in the treatment group.In the control group, P300 latency and amplitude did not reach statistically significant difference before and after treatment.Conclusion TMS therapy can improve cognitive function in SIVD with mild cognitive impairment.

Objective To establish a real-time fluorescence quantitative methylation assay to investigate the methylation status of GSH-sulphur-transferase P1(GSTP1) gene promoter region in hepatocellular carcinoma(HCC) and to investigate whether which can be used as the early diagnostic indicator of HCC .Methods Ninety-five serum samples were collected from 40 patients with HCC ,30 patients with liver cirrhosis and 25 individuals with healthy physical examination as controls .The methylation level of GSTP1 gene in these serum samples were quantitatively determined by using the real-time fluorescence quantitative methylated spe-cific PCR technique .The receiver-operation characteristic(ROC) curves were adopted to evaluate its diagnostic value for HCC .Re-sults The methylation quantitative level of GSTP1 gene in HCC serum was significantly higher than that in the healthy controls (P<0 .05) .The ROC curve analysis demonstrated that the methylation quantitative analysis of GSTP1 gene could efficiently distin-guish HCC and cirrhosis from healthy controls (AUC=0 .8641) .With the methylation rate of 2% as the critical value for diagno-sing HCC ,its diagnostic specificity was 87 .5% ,the sensitivity was 69 .6% ;the combination detection of serum GSTP1 gene methy-lation and serum AFP could increase the detection rate of HCC to 75% .Conclusion The real-time fluorescence quantitative methyl-ation assay can accurately quantify the methylation level of serum GSTP1 gene ,which has certain application value for the early di-agnosis of HCC .

Objective To determine whether HBx gene can directly induce hepatocellular carcinoma in vivo, and to explore the mechanism of transplantation tumor in nude mice.Methods pCMVX/QSG7701 cell lines were vaccinated into subcutaneous tissue of nude mice. pRcCMV2/QSG7701 and QSG7701 cell lines were used as controls. The sections of transplantation tumor were observed microscopically by HE staining. RT-PCR was used to detect the expression of mutant p53 and c-Myc mRNA in transplant tumor and an other 3 cell lines. Results The transplant tumor occurred within the subcutaneous tissue of the nude mice inoculated with pCMVX/QSG7701 cell lines at 2nd week after the vaccination. No metastatic tumor was found in other organs. Transplant tumor was not formed in all the controls. HE staining confirmed that the transplant tumor was hepatocellular carcinoma. The mutant p53 mRNA and c-Myc mRNA expression level of transplant tumor and pCMVX/QSG7701 cells was significantly higher than that of pRcCMV2/QSG7701 and QSG7701 cells, respectively (P＜0.01).Conclusion HBx gene can up-regulate the expression of mutant p53 and c-Myc genes, and directly induce hepatocellular carcinoma in vivo.

Objective To observe the effects of hepatitis B virus(HBV)X gene muhifunctional protein(HBx)on the biological characteristics of QSG7701 and the transformational effects on QSG7701 cell.Methods QSG7701 ceils were stably transformed by recombinant plasmid pCMV/X and eukaryotic expressed plasmid pRc/CMV2 by liposome-based assay,respectively.Non-transfeeted QSG7701 cells were employed as controls.The expressions of HBx,c-Myc and Bel-2 proteins in QSG7701 cells were detected by Western blot.MTT colorimetric analysis,flow cytometry and soft agar clone-forming assay were performed tO detect the biolo~dcal activity of cells.Results HBx Drotein was highly expressed in pCMV X/QSG7701 cells.The expression level of c-Myc protein in the pCMV X/QSG7701 cells was much higher than those in the other tWO groups of cells.The expression of Bcl2 protein was detected in the three groups of cells,but the expression levels were similar.Percentage of S stage cells in pCMV X/QSG7701 ceils was significantly higher than those in pRcCMV2/QSG7701 and non-transfected QSG7701 cells E(28.80±2.32)%,(15.5±2.64)%and(21.5±3.66)%,LSD 0.05=3.95%,LSD 0.01=5.47%,P<0.01].While percentage of G1 stage cells in pCMV X/QSG7701 cells was significantly lower than those in pRcCMV2/QSG7701 and non-transfected QSG7701 cells[(62.30±3.85)%,(78.70±4.12)%and(78.10q±4.45)%,LSD 0.05=5.63%,LSD 0.01-7.79 %,P<0.01].The apoptosis rate of pCMV X/QSG7701 cells was much higher than those in pRcCMV2/QSG770t and non-transfected QSG7701 cells[(14.90±1.01)%,(8.91±0.48)%and (4.03±0.47)%,LSD 0.05=O.94%,LSD 0.01=1.31%,P<0.01].The population doubling time of pCMV X/QSG7701 cells was shorter than those in pRcCMV2/QSG and non-trarmfected cells(14 h,29 h,38 h,respectively).The cloning ratio in soft agar of pCMV X/QSG7701 cells WaS significantly higher than those of pRcCMV2/QSG and non-transfected QSG7701 cells[(19.83±1.96)%,(1.76±0.03)%and (1.33±0.18)%,LSD 0.05=1.53%,LSD 0.01=2.11%,P<0.01].Conclusion HBx may transform human non-tumor hepatoeyte QSG7701,which makes the cell growth malignizeA.

Fluorescence spectroscopy was used to investigate the efficiency of coupling peptides to gold nanoparticles via 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-N-hydroxysuccinimide (EDC-NHS).The experiment conditions including buffer solution, pH value and concentrations of buffer solution, concentrations of NHS and EDC, concentration ratios of NHS to EDC, and coupling reaction time on the coupling efficiency were investigated and optimized.The experimental results indicated that the optimized experimental conditions were as follows: 25 mmol/L HEPES buffer solution, pH 7.0;2∶1 of concentration ratio of NHS to EDC, 0.4 mol/L NHS, 0.2 mol/L EDC, and coupling reaction time of 24 h.This study may provide references for the relative research involving coupling peptide or protein with gold nanoparticles

Objective To investigate the diagnosis value of immunophenotype and karyotypes in newly diagnosed chronic lymphocytic leukemia (CLL).Methods To retrospect the flow cytometry (FCM) immunophenotype and karyotypes characteristics in newly diagnosed 70 CLL cases.Results In all cases,the positive rates of CD19,CD20,CD5,CD23,CD22 were 100 ％,88.5 ％ (54/61),77.1％ (54/70),67.6 ％ (23/34)and 51.9 ％ (27/52),respectively.And 6 were misdiagnosed,2 was CD+5CD+19(+),but CD20,CD22 were strongly positive,final diagnosed as mantle cell lymphoma (MCL) by FISH t(11;14) examination and CyclinDl; CD+5CD+19(-) CLL were 16 cases (22.9 ％),but 4 were misdiagnosed,the misdiagnosis rate was 25 ％,significantly higher than that of CD+5CD+19(-) CLL (P =0.030).59 cases were examined by conventional cytogcnetic (CC),and 13 cases were with abnormal karyotypes,positive rate was 22.0 ％,with complex karyotypes in 5 cases (8.5 ％); 10 cases combined with FISH abnormalities karyotype examination rate was 60 ％.Conclusion Typical CLL immunophenotypic characteristics were CD5,CD19,CD23 co-expression,and CD－5 CLL with higher misdiagnosis rate,combined with CD20 (,) CD22 expression and karyotype analysis helps to CLL and other B lymphoid proliferative diseases (B-LPD) identification.Conventional cytogenetic detection combined with FISH scan can improve the recognition ability of abnormal chromosome.

Objective To determine the value of acute phase protein in the early diagnosis of recurrence after curative gastric cancer surgery. Methods Acute phase serum protein level was measured before and after the gastric cancer surgery in 120 patients,and was compared with that of the control group. At 1, 3, 6, 9, 12 months after the operation in 87 patients undergoing curative gastric cancer surgery, the levels of acute phase proteins were measured. They were followed up for at least 12 months or until death. Results The levels of serum C reactive protein(CRP), ?1 antitrypsin (?1 AT) and ? acid glycoprotein (? AG) in gastric cancer group were significantly higher than those in the control group (P0.05). In patients who underwent curative gastric cancer surgery in a certain period after the operation, the serum levels of CRP, ?1 AT and ? AG were significantly lower than those before the surgery(P0.05).There were significant difference in CRP,?1 AT and ? AG between the recurrence groups and nonrecurrence group before and after the operation (P

Fresh roots of healthy four-year-old American ginseng were randomly divided into two groups and stored for 180 days in sand without washing (treatment A) and in plastic bag with preservative after washing (treatment B), and sampled after 45, 90, 135 and 180 days of storage, respectively. The incidence of disease was surveyed and the contents of ginsenoside, polysaccharide, and free amino acids of roots were determined. The results indicats that the disease of ginseng could be controlled better by the treatment B than by the treatment A within 45 days, but the better effect was achieved in the treatment A after storage for 45 days. Both storage methods had significant influence on the contents of ginsenoside, polysaccharide and free amino acids of roots. For the treatment A, the ginsenoside content of roots had remained relatively high during the storage period, and the crude polysaccharide and free amino acids changed slowly within 135 days and then increase significantly until 180 days. For the treatment B, the content of crude polysaccharide of roots decreased obviously after 90 days of storatge and then became stable until 135 days. The change of content of free amino acids was similar to the crude polysaccharide, but the decline was not significant. In general, the treatment A is more benefit to keep the quality of fresh roots of American Ginseng during 180 days of storage.
Amino Acids ; analysis ; Ginsenosides ; analysis ; Panax ; chemistry ; Plant Extracts ; analysis ; Plant Roots ; chemistry ; Technology, Pharmaceutical ; methods

In order to develop clinical diagnostic tools for rapid detection of the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) and to identify candidate proteins for vaccine development, the C-terminal portion of the nucleocapsid (NC) gene was amplified using RT-PCR from the SARS-CoV genome, cloned into a yeast expression vector (pEGH), and expressed as a glutathione S-transferase (GST) and Hisx6 double-tagged fusion protein under the control of an inducible promoter. Western analysis on the purified protein confirmed the expression and purification of the NC fusion proteins from yeast. To determine its antigenicity, the fusion protein was challenged with serum samples from SARS patients and normal controls. The NC fusion protein demonstrated high antigenicity with high specificity, and therefore, it should have great potential in designing clinical diagnostic tools and provide useful information for vaccine development.
Antigens, Viral ; immunology ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; Genome, Viral ; Humans ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism ; SARS Virus ; genetics ; immunology ; Yeasts ; genetics