Contents Determination of 5 Heavy Metals in Tibetan Medicine Ershiwuwei Shanhu Pills by Atomic Absorption Spectrometry / 中国药房

OBJECTIVE:To establish a method for the contents determination of 5 heavy metals in Ershiwuwei shanhu pills, and to investigate the contents of heavy metals in Ershiwuwei shanhu pills produced by 5 manufacturers from different districts of Tibet. METHODS:Ershiwuwei shanhu pills were digested by HNO3-HClO4(4:1,V/V).The contents of 3 heavy metals as Cu,Pb, Cd in samples were measured by flame atomic absorption spectrometry(FAAS). The contents of 2 heavy metals as As,Hg in samples were measured by hydride generation atomic absorption spectrometry(HG-AAS). RESULTS:5 kinds of heavy metals have good linear relationship in the corresponding mass concentration range(all r≥0.999 1). The limits of detection of Cu,Pb, Cd were 0.001 6,0.041 2,0.036 3 mg/L,and the limits of quantitation were 0.005 3,0.137 3,0.121 0 mg/L;the limits of detection of As,Hg were 0.325 7,0.692 3 μg/L,and the limits of quantitation were 1.085 7,2.307 7 μg/L. RSDs of precision tests were ≤5.54%(n=6);RSDs of stability tests were all≤3.79%;RSDs of reproducibility tests were ≤3.72%. The average recovery rates were 91.34%-110.11%(RSDs were 0.66%-6.80%,n=6). Results of contents determination showed that the contents of Cu in samples from 5 manufacturers were not out of limits,but the contents of Cd and Hg were all out of limits; the contents of Pb in samples from 4 manufacturers were out of limits,and the contents of As in samples from 2 manufacturers were out of limits. CONCLUSIONS:The established method has good accuracy,sensitivity,stability and reproducibility,and it is suitable for contents determination of 5 heavy metals in Ershiwuwei shanhu pills. To some extent,there is a problem of excessive heavy metals in samples from 5 manufacturers.

Study on Anti-tussive and Anti-inflammatory Effects of Aqueous Extract of Pegaeophyti Radix Et Rhizoma / 中国药房

OBJECTIVE:To study the anti-tussive and anti-inflammatory effects of aqueous extract of Pegaeophyti Radix Et Rhizoma(P. Radix Et Rhizoma). METHODS:In the experiment,there were a model group,a positive control group [30 mg/kg co-deine for mice,18.38 mg/kg codeine for guinea pigs,20 mg/kg indometacin for mice and 15 mg/kg indometacin for rats] and P. Ra-dix Et Rhizoma low-dose,medium-dose and high-dose groups [2.0,4.0 and 8.0 g(medicinal materials)/kg for mice;1.2,2.4 and 4.8 g (medicinal materials)/kg for guinea pigs;1.4,2.8 and 5.6 g (medicinal materials)/kg for rats],with 10 animals in each group. The above-mentioned animals were given corresponding drug ig for 4 consecutive days,and then strong ammonia spraying method and sulfur dioxide stimulation method were employed to induce cough in mice and citric acid spraying method to induce cough in guinea pigs to observe the incubation period and times of cough,auricular swelling method to observe xylene-induced au-ricular swelling in mice and pedal swelling method to observe carrageenin-induced pedal swelling in rats. RESULTS:Compared to the model group,Aqueous extract of P. Radix Et Rhizoma could prolong the incubation period and reduce times of cough induced by strong ammonia spraying and sulfur dioxide stimulation in mice of positive control group and P. Radix Et Rhizoma medium-dose and high-dose groups;it could prolong the incubation period of and reduce times of cough induced by citric acid spraying in guinea pigs of positive control group and P. Radix Et Rhizoma medium-dose and high-dose groups;it could alleviate xylene-induced auricu-lar swelling in mice of positive control group and P. Radix Et Rhizoma medium-dose and high-dose groups;it could alleviate carra-geenin-induced pedal swelling in rats of positive control group and P. Radix Et Rhizoma medium-dose and high-dose groups. There were statistically significances (P<0.01 or P<0.05). CONCLUSIONS:Aqueous extract of P. Radix Et Rhizoma has anti-tussive and anti-inflammatory effects on the experimental animals.

Research on the genotypes of clinical varicella-zoster virus isolates in Tibet / 中华微生物学和免疫学杂志

Objective To analyze the genotypes of clinical varicella-zoster virus(VZV) isolates in Tibet.Methods The samples of vesicular fluid from patients with chickenpox or zoster were collected to isolate VZV in human embryonic fibroblast cells.Furtherly,the isolated VZV were identified by indirect immunofluorescence assay and the positive ones were continuely cultured in vitro to get their genome DNAs.Then,PCR and nucleotide sequencing were performed on the part of open reading frames 1,21,50,54.To confirm the genotypes of isolated virus strains,the gene sequences obtained from PCR products were compared with that in GenBank of VZV Dumas strains,and their restriction enzyme sites were analyzed via Primer 5.0.Results Ten clinical isolates of VZV were obtained from sixteen specimens,the positive isolation rate was 62.5％.Gene analysis showed that among all the ten clinical isolates of VZV in Tibet,three strains are genotype A1,four are genotype A2 and the other three are genotype J1.Conclusion The VZV genotype distribution in Tibet was tightly related to their special geographic locations.

Experssion of human FGFR2IIIc and its S252W mutant in MDA-MB-231 breast cancer cells with adenovirus vectors / 生物工程学报

To study the functions of human Fibroblast growth factor receptor 2IIIc (FGFR2IIIc) gene in cancer cells, breast cancer cells MDA-MB-231 were infected by recombinant adenoviruses containing FGFR2IIIc and its S252W mutant, respectively. FGFR2IIIc gene was amplified from an existing plasmid and its S252W mutant was obtained by overlapping extension PCR. These two genes were separately cloned into the adenoviral shuttle plasmid pAdTrack-CMV, confivmed by DNA sequencing linearized, and co-transformed into Escherichia coli BJ-5183 with the adenoviral vector pAdEasy-1. The resulting recombinant expression vectors Ad-FGFR2IIIc and Ad-FGFR2IIIcS252W were linearized and transfected into HEK293A cells to get adenoviral particles. GFP was used to verify the gene expression. The recombinant adenoviral particles were harvested, titrated, and then infected MDA-MB-231 cells. The expression of FGFR2IIIc and its S252W mutant were examined by RT-PCR and Western blotting, and the effect of these recombinant adenoviruses on MDA-MB-231 cell proliferation was analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry. The results showed the recombinant adenoviral particles could infect MDA-MB-231 cells and express the target proteins. MTT showed that both FGFR2IIIc and its S252W mutant inhibited MDA-MB-231 cell proliferation, but the mutant was more effective. Flow cytometry showed that both FGFR2IIIc and its S252W mutant arrested MDA-MB-231 cell cycle at G0/G1 phase, resulting in low cell proliferation.