To simply and rapid authenticate Lonicera japanica. Rapid allele-specific PCR primer was designed base on trnL-trnF 625 G/T Single nucleotide polymorphism and the PCR reaction systems including annealing temperature was optimized; optimized results were performed to authenticate L. japanica and its 9 adulterants. When 100 x SYBR Green I was added in the PCR product of 87 degrees C initial denatured 1 min; 87 degrees C denatured 5 s, 68 degrees C annealing 5 s, 30 cycle; L. japanica visualize strong green fluorescence under 365 nm UV lamp whereas adulterants without. The results indicate rapid allele-specific PCR could authenticate L. japanica and its adulterants rapidly and simply.
Alleles ; DNA Primers ; genetics ; Drug Contamination ; prevention & control ; Lonicera ; classification ; genetics ; Polymerase Chain Reaction ; methods ; Quality Control

This study aimed to develop the calculation method of pre-hospital emergency physician allocation gap and apply it to Shanghai.In order to reduce the ambulance dispatch lag frequency, through the analysis of its da-ta in the Shanghai urban area, the research group obtained the gap and extended the data to Shanghai city.The peak method establishes the association between pre-hospital emergency physician increment and the ambulance dispatch lag decrement.Based on descriptive statistics, the peak method by which the Shanghai ambulance dispatch lag data were analyzed uses the SAS programming software.This method of using programming software provides it with good reliability and validity.After an increase of 40 duty vehicles (381 pre-hospital emergency physicians), the ambu-lance dispatch lag ratio would drop from 25.61 percent to 0.22.Therefore, the association between the pre-hospital emergency physician increment and the ambulance dispatch lag decrement was established and can provide a scientif-ic evidence for the policy formulation.

Objective:The research aimed to figure out the geographic availability of the residence in the aspect of receiving health service thereby studying the spatial distribution regulation and the distance attenuation coefficient of the inpatients.It also provides the decision-making basis for the regional health planning.Methods: Select 23 medical institutions and randomly pick up patients from each of them, calculate the distance between the inpatients home and the medical institutions, then simulate the corresponding probability patients using the Geographical Infor-mation System ( GIS) .Perform a variety of curve fitting models according to the determination and simulation graph-ics coefficient choosing the most appropriate model, and draw corresponding the distance attenuation coefficient.Re-sults:An exponential curve model is better than power curve model for the secondary medical institutions inpatients distribution and contrarily for those in the tertiary medical institutions.The mean distance attenuation coefficient was 0.51 for exponential curve model of secondary comprehensive medical institutions inpatients and its average was 0.50 for the secondary specialist medical institutions inpatients.The mean distance attenuation coefficient was 0.84 for the power curve model of the tertiary comprehensive medical institutions inpatient and its average was 0.53 for tertiary specialist medical institutions inpatients.Conclusion: The secondary medical institutions inpatient probable attenua-tion trends with distance are faster than in tertiary medical institutions, those for the comprehensive medical institu-tions inpatient are faster than in the specialist medical institutions.In the regional health planning, for different lev-els, categories of medical institutions should adopt different distance attenuation coefficients to calculate the health re-sources level.

Objective To observe the distribution of oligodexynucleotide (ODN)MT01 in main organ tissues of the rats at different time points and to discuss the regularity of the distribution of MT01 preliminarily. Methods 60 male Wistar rats were randomly divided into experimental group(n=30)and control group(n=30). The rats in experimental group was locally injected with Cy5 labeled MT01 in gingival mucosa,whereas the rats in control group were injected with MTO1.The samples of rat lung,liver spleen,kidney,heart,and brain tissues were collected at 15 min,1 h,4 h,8 h,16 h,1 d,2 d,3 d,4 d,and 5 d after injection,and the distribution of MT01 fluorescence was observed by laser scanning confocal microscope.The ratio of fluorescence positive cells indicated the amount of MT01 that had been taken up by different organs.Results No positive fluorescence cells were observed in control group.Whereas,in experimental group ,the positive fluorescence cells were detected in the tissue samples of lung,liver,spleen and kidney but not in the tissue samples of heart and brain.The positive fluorescence cells distributed focally in kidney tissue and presented primarily in the cytoplasm of renal tubular epithelial cells.The ratios of positive fluorescence cells changed regularly with time in liver, spleen and kidney tissues and the highest level was detected at 4,3 and 4 d after injection.No distinct regularity of the ratio of positive fluorescence cells was observed in lung tissue.Conclusion MT01 can be taken up by liver,spleen and lung tissue and primarily by kidney with regularity in distribution.

[ ABSTRACT] AIM:To evaluate the effects of 1α, 25-dihydroxyvitamin D3 on T helper cell 17 ( Th17 cells) and its related cytokines in a mouse model of corneal allograft transplantation.METHODS:C57BL/6 mice were transplanted with corneal grafts from BALB/c mice and treated intraperitoneally with 1.0μg 1α, 25-dihydroxyvitamin D3 or soybean oil every other day after operation.The transparency of the corneal grafts was evaluated for potential rejection signs by slit lamp biomicroscopy and histopathology.The expression levels of IL-17, RORγt and IFN-γin the spleen were measured by real-time PCR.Moreover, the protein expression of RORγt and IL-17 in the peripheral blood was analyzed by Western blotting. IL-17 and IFN-γin peripheral blood were measured by flow cytometry.RESULTS:1α, 25-dihydroxyvitamin D3 signifi-cantly inhibited the rejection of the corneal allograft and reduced the numbers of inflammatory infiltrates in the corneal graft. In the spleen, 1α, 25-dihydroxyvitamin D3 treatment reduced the expression levels of IL-17, RORγt and IFN-γ.In the pe-ripheral blood, 1α, 25-dihydroxyvitamin D3 treatment downregulated the expression levels of RORγt, IL-17 and IFN-γ. CONCLUSION:The effects of 1α, 25-dihydroxyvitamin D3 on suppressing corneal transplantation-induced allograft rejec-tion in mice are closely associated with its modulation on IL-17 and related cytokine RORγt.

OBJECTIVETo investigate the relationship of Fas mRNA and protein expression and apoptosis in human oral squamous cell carcinoma.

METHODSNorthern blot and flow cytometry (TUNEL method) were used to detect the expression of Fas mRNA and Fas protein, cell cycle and apoptotic level in oral squamous cell carcinoma. The relationship between Fas gene expression and OSCC apoptosis was analyzed statistically.

RESULTSFas mRNA and protein could be detected in all five normal oral mucosa specimens. There was positive correlation between expression of Fas mRNA/protein and cell differentiation as well as apoptosis in OSCC (P < 0.005).

CONCLUSIONThe expression of Fas gene was highly correlated with the differentiation and apoptosis in OSCC.

Apoptosis ; Carcinoma, Squamous Cell ; metabolism ; Humans ; Mouth Neoplasms ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo investigate the effect of fas gene transfection and monoclonal anti-fas antibody on tumorigenicity and proliferation of transplanted tumor of Tca8113 cell.

METHODSPlasmid including fas gene was transfected into Tca8113 cell by lipofectamine kit. Some transfected cells were treated by monoclonal anti-fas antibody after 48 hours since transfection. Untransfected cell (control), fas-tansfected cell and fas-transfected cell treated with antibody were transplanted to nude mice subcutaneously. Growth of transplanted tumor was observed and recorded regularly. Animals were sacrificed and tumor samples were harvested at the end of experiment. Fas expression in each neoplasm was assessed by RT-PCR. Apoptosis, proliferation and expression of fas protein in tumor tissue were measured by flow cytometry (FCM).

RESULTSTumor occurred much later in fas-transfected group and fas-transfected plus antibody treated group. Growth arrest was found in them. RT-PCR and FCM suggested that fas-transfection up-regulated the expression of fas mRNA and protein, increased apoptosis index (AI). But no effect on proliferation index (PI) was observed. Monoclonal anti-fas antibody did not effect the expression of fas mRNA and protein, but increased AI and decreased PI.

CONCLUSIONFas-transfection suppressing tumorigenesis of Tca8113 cell transplanted in nude mice might be caused by up-regulation of expression of fas gene and enhancement of apoptosis. However, anti-fas antibody suppressing tumorigenesis might be associated with activation of apoptosis and repression of proliferation.

Animals ; Antibodies, Monoclonal ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Fas Ligand Protein ; biosynthesis ; genetics ; pharmacology ; Humans ; Mice ; Mice, Inbred BALB C ; Mouth Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Transfection

OBJECTIVEThe purpose of this study was to establish transfergeneic Tca8113 cell and evaluate the expression of human endostatin (hES) gene in the cell colone in vitro.

METHODSTo transfect hES gene into Tca8113 cells, lipofectamin was complexed with plasmid encoding hES gene, and blasticidin S antibiotic was adopted to select Tca8113--hES cell clone. Immunohistochemistry S-P method was adopted to detect the expression of hES in the transfergenic Tca8113 cell in vitro.

RESULTSTransfected by hES, the transfergenic Tca8113 cells could grow and proliferate in RPMI--1640 culture medium containing blasticidin S antibiotic. The expression rate of hES reached 100%.

CONCLUSIONhES gene can express in hES-transfected Tca8113 cell in vitro.

Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Division ; Cloning, Molecular ; Endostatins ; biosynthesis ; genetics ; Humans ; Lipids ; pharmacology ; Tongue Neoplasms ; genetics ; metabolism ; pathology ; Transfection ; Tumor Cells, Cultured

Objective: To observe the clinical effect of acupoint sticking therapy with Mian Tan Gao (facial paralysis paste) plus electroacupuncture (EA) for treating peripheral facial paralysis and its influence on patients' facial nerve functions, facial disability index and clinical symptoms and signs. Methods: A total of 96 peripheral facial paralysis patients were allocated into an observation group, a medicine group and an EA group by simple randomization, with 32 cases in each group. Patients in the medicine group were treated with oral mecobalamine and prednisone acetate; patients in the EA group were treated with EA on the basis of the medicine treatment; while patients in the observation group were treated with acupoint sticking therapy with Mian Tan Gao (facial paralysis paste) plus EA. After 4-week treatment, the clinical efficacy, the adverse events, and the scores of House-Brackmann (H-B) facial nerve function grading scale, visual analog scale (VAS), clinical symptoms and signs, and facial disability index (FDI) were compared. Results: After 4-week treatment, the total effective rate was 96.9％ in the observation group, higher than 68.7％ in the medicine group and 75.0％ in the EA group (both P<0.05). After 4-week treatment, the scores of H-B grading scale, VAS and clinical symptoms and signs in the three groups dropped significantly compared with those before treatment, and the scores in the observation group were lower than those in the medicine group and EA group (all P<0.05). After 4-week treatment, the facial disability index-physical function (FDIP) in the FDI in the three groups increased significantly, with a higher value in the observation group compared with that in the medicine group and EA group (both P<0.05). The facial disability index-social function (FDIS) in the FDI dropped significantly, with a lower score in the observation group compared with that in the medicine group and EA group (both P<0.05). However, the comparisons of the items above between the medicine group and the EA group showed no statistical significance (all P>0.05). The between-group comparison of the adverse event across the three groups showed no statistical significance (P>0.05). Conclusion: Acupoint sticking therapy with Mian Tan Gao (facial paralysis paste) plus EA can decrease H-B grade, reduce pain severity and improve clinical symptoms and signs as well as the facial disability condition in peripheral facial paralysis patients. This method produced more significant efficacy compared with oral medicine and medicine plus EA.

The influences of E23K polymorphism of inwardly rectifying K~+channel 6.2 (Kir6.2) gene on the clinical phenotype of type 2 diabetes and glucose-lowering effect of gliclazide were investigated.The result showed that E23K polymorphism did not influence glucose-lowering effect of gliclazide,but serum creatinine level of patients with K/K genotype was higher than those of E/E and E/K genotypes (P