OBJECTIVE:To explore ways to resolve the quality problems of Chinese herbal slice so as to provide reference for producers,processors and users.METHODS:The records of check and accept prepared slice into storeroom in our hospital during 2008 were summed up and sorted out.The number of total batches,the number and proportion of unqualified batches were calculated to analyze the causes of unqualified slice after checking.RESULTS:In 2008 there were 4 519 batches of herbal slice while 94 batches were unqualified and accounted for 2.1% in total.53 batches were refused because of their quality problem,accounting for 56.4% of unqualified batches.CONCLUSION:It is urgent to improve the quality of prepared slice and standardize the processing of herbal slice.

The problems to offer HIS course in plateau medical colleges and universities were pointed out and corresponding reform measures were put forward in view of the requirement for medical professionals in the 21stcentury information society and the current situation in HIS course teaching.

Objective:To investigate the inhibitory effect of adenovirus-mediated recombinant Buthus martensii Karsch chloride toxin artifact (Ad-rBmK CTa) on human glioma U251 cells and its related mechanisms.Methods:Groups of 3 titer gradients of 3.5×10 9, 7.0×10 9 and 3.5×10 10 pfu/ml Ad-rBmK CTa were set up and applied to U251 cells for 24, 48 and 72 h, and a blank control group (no cells and Ad-rBmK CTa were added) and a negative control group (only U251 cells were added) were set up at the same time. The virus infection status was observed by laser confocal fluorescence microscopy. The cell proliferation in each group was detected by methyl thiazolyl tetrazolium (MTT) assay. The cell cycle and apoptosis in each group were detected by flow cytometry. The expressions of apoptosis-related proteins bax, bcl-2 and caspase-3 were detected by Western blot. Results:The infection rate of Ad-rBmK CTa was over 90% after acting on U251 cells for 24 h. As the titer of Ad-rBmK CTa increased, the proliferation inhibition rate of U251 cells treated for the same hours gradually increased (all P <0.01); with the extension of time, the proliferation inhibition rate of U251 cells treated with the same titer of Ad-rBmK CTa also gradually increased (all P < 0.01). After 7.0×10 9 pfu/ml Ad-rBmK CTa acted on U251 cells for 48 h, the proportion of cells in G 0/G 1 phase was (40.7±0.8)%, and cells in S phase and G 2 phase accounted for (35.7±0.6)% and (23.6±1.4)%, and the difference was statistically significant ( F = 225.119, P < 0.01). When 7.0×10 9 pfu/ml Ad-rBmK CTa acted on U251 cells for 24, 48 and 72 h, the apoptosis rates were (7.4±1.4)%, (19.2±1.7)% and (22.3±1.7)% ( F = 49.470, P < 0.01). After 7.0×10 9 pfu/ml Ad-rBmK CTa acted on U251 cells for 48 h, compared with the negative control group, the expressions of bax and caspase-3 proteins increased, and the expressions of bcl-2 decreased. Conclusions:Ad-rBmK CTa may act on the DNA damage-induced G 1/S detection site to arrest the cell cycle in G 0/G 1 phase, thus inhibiting the proliferation of U251 cells in vitro. However, its induction of apoptosis in U251 cells is not obvious. The mechanism may be related to the direct or indirect inhibition of chloride ion channels.

OBJECTIVETo detect tumor cells in the peripheral blood of patients with hepatocellular carcinoma (HCC) by using the mRNA of the MAGE-1 and MAGE-3 genes as specific tumor markers.

METHODSPeripheral blood was obtained from 25 HCC patients and 20 healthy volunteers. The mRNA of the MAGE-1 and MAGE-3 genes in the peripheral blood mononuclear cells (PBMCs) was detected by nested RT-PCR. The MAGE-1 and MAGE-3 transcripts in the tumor tissues of these HCC patients were also detected by RT-PCR.

RESULTSOf the 25 HCC patients, MAGE-1 and MAGE-3 mRNA were positive in 44% (11/25) and 36% (9/25) of PBMCs respectively, and in 68% (17/25) and 56% (14/25) of HCC tissues respectively. In the PBMCs of the 25 HCC patients, 16 (64%) samples were detected to express at least one type of MAGE mRNA. MAGE mRNA were not detected in the PBMCs from the patients whose tumors did not express the MAGE genes, nor in the PBMCs from the 20 healthy donors. The positive rate of MAGE mRNA in the PBMCs was closely correlated with the TNM stages and the diameter of tumors, but there was no correlation between the positive rate of MAGE mRNA in PBMCs and tumor differentiation degree or serum alpha-FP level. Of 9 HCC patients whose serum alpha-FP was normal or slightly elevated (< 50 ng/ml), 6 were MAGE-1 and/or MAGE-3 mRNA positive in their PBMCs.

CONCLUSIONMAGE-1 and MAGE-3 mRNA could be specifically detected with high percentage in the PBMCs of HCC patients by our method. They can be used as specific tumor markers for the detection of the circulating HCC cells, and the detection results may be helpful to evaluate the prognosis of HCC patients.

Antigens, Neoplasm ; Carcinoma, Hepatocellular ; genetics ; Humans ; Leukocytes, Mononuclear ; Liver Neoplasms ; genetics ; Melanoma-Specific Antigens ; Neoplasm Proteins ; RNA, Messenger ; genetics

Objective: To explore the influence of pVR-IL15 gene adjuvant on the immune responses of different immunization strategies. Methods: The sequential immunization strategies were used in BALB/c mice with a DNA vaccine, recombinant modified vaccinia virus Ankara and recombinant adenovirus expressing HBsAg respectively and combined with a gene adjuvant pVR-IL15. Cellular and humoral immune responses were evaluated by IFN-γ enzyme-linked immunospot assay (ELISPOT) and enzyme-linked immunosorbent assay (ELISA), respectively. Results: The levels of humoral and cellular immune responses in the immune groups combined with pVR-IL15 were significantly higher than those of the non-combined with pVR-IL15. Conclusions The pVR-IL15 gene adjuvant can enhance the immune responses induced by the recombinant viruses expressing HBsAg.

Objective To further investigate the expression of MAGE-1 gene in hepatocellular carcincma (HCC). Methods The tumors and adjacent liver tissue from 45 HCC patients and liver tissue from 28 non-HCC patients (16 with liver cirrhosis and 12 with normal liver) were characterized by RT-PCR. A 421 bp PCR product from a cDNA fragment spanning exons 1,2 and 3 was sequenced. The HLA type wes assayed by standard ELISA in 43 HCC patients. Results Thirty-two of 45 tumor tissues from HCC patients expressed MAGE-1 mRNA (71.1%).In contrast, MAGE-1 mRNA was not detected in adjacent tissues. Three were found to have point mutations at 3 idntical sites resulting in the substitution of two amino acid residues.The most frequent HLA types in 43 HCC patients were: HLA-A2, 53.5%; A11, 25.6%; A24, 20.9%; A33, 20.9%; HLA-B13, 28.3% and B35, 23.2%. Expression of HLA-A33 (20.9%) was higher in HCC patients than that predicted in the normal Chinese population (8.8%). There was no discemable correlation between MAGE-1 expression and α-FP level, tumor size and hepatitis B or C virus infection.The identification of peptides which are restricted by haploptypes other than A1 should increase the opportunity for paptide based immunotherapy. Conclusions This study shows that MAGE-1 mRNA is highly expressed in HCC tumor tissue in Chinese patients. Previously unreported point mutations in the MAGE-1 gene are described and may also provide additional opportunities for immunotherapy.

Objective To construct the recombinant modified vaccinia Ankara vaccine expressing HBsAg and investigate its antigenicity and immune effect by different strategies.Methods Target gene was cloned into plasmid pSC1l,the recombinant plasmid pSC11-S was transfected into BHK-21 cells that infected with MVA.Homologous recombination occurred between MVA and pSC1 1-S.The recombinant virus MVA-S was selected by several rounds of blue/clear plaques.Target gene was identified by PCR.The expression of target gene was analyzed by Western Blot.The BALB/c mice were immunized with single immunization and consecutive immunization strategy.The level of specific cellular response was measured by ELISpot assays.Results The recombinant virus expressing target gene was confirmed by PCR and Western Blot showed that MVA-S could elicit specific cellular response.There were no significant differences among two kinds of DNA vaccine and recombinant MVA-S.The prime/boost regimen could induce higher cellular response than single DNA vaccine immunization.High doses of MVA-S boost with the same DNA vaccine prime could induce highest response.There was no significant difference between different DNA vaccines (pVR-S,pcDNA-S) followed by same MVA-S boost.Conclusions MVA-S expressing target was constructed successfully,and it could induce specific cellular immune response which was higher with the strategy of DNA prime/MVA-S boost than that of single immunization.

Consensus ; Critical Illness ; Humans ; Serum Albumin ; Serum Albumin, Human