Objective To compare the four kinds of Yinqiao Powder prepared by archaic decoction,Japan standard decoction,medical institution decoction and decoction combined archaic with modern technology,select the best method and provide the important theory basis for standardization of decoction technics of diaphoretic recipes.Methods The major contents of traditional Chinese medicine were determined by HPLC and used as the evaluating indicator as well as the ratio of dry extraction.Results The method of archaic decoction got the highest synthetic scores,and the validating experimental result was stable.Conclusion The preparation method of archaic decoction could be used as the reference standard for preparation technology of Yinqiao Powder,in order to further standardize the preparation of the standard decoction of diaphoretic recipes.

OBJECTIVE:To establish HPLC method for simultaneous determination of estazolam and diazepam in serum METHODS:The mobile phase consisted of methanol-water-triethylamine-acetic acid(55∶45∶0 5∶0 35)and the ?max was 254nm Phenytoin was adopted as the internal standard The drugs were extracted by diethyl ether under the alkaline condition RESULTS:The calibrating curves of estazolam and diazepam were linear in the ranges of 0 52～38 62?g/ml and 0 50～40 16?g/ml respectively CONCLUSION:The method was appropriate for determination of estazolam and diazepam qualitatively and quantitatively in intoxication accident

Objective To investigate the effect of insulin-like growth factor-1(IGF-1) and basic fibroblast growth factor (bFGF) on the differentiation of human dental pulp stem cells in vitro. Methods Mono-cell suspension was separated from the human adult dental pulp with collagenase Ⅰ and dispase digestion. Colony-forming efficiency of cells was calculated. Cells were observed under an invert microscope. Dentin sialoprotein (DSP), dentin matrix protein 1 (DMP-1) and dentin sialophosphoprotein (DSPP) mRNA of the 3rd passage cells induced with IGF-1 and bFGF in vitro were detected by immunohistochemistry and RT-PCR. Results Clonogenic cells were obtained from dental pulp and colony-forming efficiency of cells was 2.6-3.5 colonies/10 4 cells. Human specific DSP, DMP-1 protein and DSPP mRNA expressed in colony-forming cells after 7 d of induction with IGF-1 and bFGF in vitro. Conclusion Adult stem cells might exist in human dental pulp. Dental pulp stem cells could differentiate into odontoblast under the induction of IGF-1 and bFGF.

Objective To analyze the differentially expressed genes between the NCAM + c-Kit +RBE and NCAM-c-Kit-RBE of intrahepatic cholangiocarcinoma (ICC) cell lines,and to screen out the differentially expressed genes that are related to the stem cell signaling pathways.Methods Magnetic activated cell sorting was used to isolate the NCAM + c-Kit +/NCAM-c-Kit-subset cells,and then Agilent Whole Human Genome Microarray Kit was used to test the difference in gene expressions between the NCAM + cKit + and NCAM-c-Kit-subset cells.The difference in gene expressions related to the stem cell signaling pathways was analyzed by the SAS system.The result of the microarray was further confirmed by RT-PCR.Results The total differentially expressed genes which could be found through gene microarray were 7270 [foldchange(fc) ≥2 or fc ≤0.5].Compared with the NCAM-c-Kit-RBE,3572 genes were upregulated while 3698 genes were downregulated.The differences in gene expressions related to the stem cell signaling pathways were 421 (fc ≥2 or fc ≤ 0.5),among which 231 genes were upregulated while 190 genes were downregulated.Conclusions High-flux microarray could be used to screen out lots of differentially expressed genes between the NCAM + c-Kit + and NCAM-c-Kit-RBE cells.The differences in gene expression in the stem cell signaling pathways could also be further analyzed using the SAS system.

In comparison with their in vivo counterparts, the in vitro produced mammalian embryos had markedly lower rates of morula/blastocyst development and pregnancy after transfer to the recipients. Things became even worse in the cloned embryos. This necessitates improvement of the embryo culture system. Co-culture of embryos with different types of somatic cells was found beneficial for embryo development in vitro and many studies have been conducted in this area in recent years. In this paper, recent developments and the authors' own work in studies of co-culture of early mammalian embryos with somatic cells were reviewed, with emphasis on the effects of cell type, stage of estrous cycle and number of passages of somatic cells and supplement of serum on embryo development, and the mechanisms by which co-culture promote embryo development. The recent developments are summarized as follows: 1. Somatic cells of both homogeneous and heterogeneous origins can be used for co-culture of mammalian embryos, with similar developmental rates. 2. Supplementation of animal serum at appropriate concentrations improved the somatic cell growth and consequently the development of embryos in co-culture. 3. The estrous cycle stages of oviduct epithelial cells used for co-culture had no effect on the development of embryos. 4. Over-passaging of somatic cells reduced their efficiency in promoting development of the co-cultured embryos. In conclusion, studies have shown that co-culture overcame the block of embryo development in vitro and improved embryo quality with increased rates of implantation and pregnancy, but many problems remain to be solved on its influencing factors and mechanisms of action.
Animals ; Coculture Techniques ; methods ; Embryo Culture Techniques ; methods ; Embryo, Mammalian ; physiology ; Humans

One new dicyclopeptide cyclo-(L-N-methyl Glu-L-N-methyl Glu) (1), together with one new natural dicyclopeptide cyclo-(L-methyl Glu ester-L-methyl Glu ester) (2), and two known dicyclopeptides cyclo-(L-methyl Glu ester-L-Glu) (3), and cyclo-(L-Glu-L-Glu) (4), were isolated from the aerial parts of Dianthus chinensis L. Their structures were determined by spectroscopic analyses and chemical methods.
Dianthus ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry

Objective To investigate the single-nucleotide polymorphisms of PTPN22 gene rs2476601 ,rs3811021 and rs2488457 in patients with primary immune thrombocytopenia(ITP) .Methods Totally 100 patients with ITP and 100 cases as con-trol from Department of Hematology ,the Affiliated Baiyun Hospital of Guiyang Medical College and the Affiliated Hospital of Guiyang Medical College were collected .PTPN22 gene + 1858 loci (rs2476601) and 3′UTR region rs3811021 loci were detected by PCR-RFLP ,the promoter-1123 loci (rs2488457) were detected by PCR-SSP ,and the results were statistically analyzed .Results PTPN22 gene + 1858 locus in ITP patients and control group were all C allele ,T allele was detected ,and there was no single nucle-otide polymorphisms (R620W) exist .The frequency of PTPN22 gene rs3811021 locus TT ,CT ,CC three genotypes in ITP patients and control group had no significant difference(χ2 = 3 .686 ,P= 0 .158) .The frequency of T allele ,C allele in ITP patients and con-trol group had no significant difference(χ2 = 2 .828 ,P = 0 .093) .The frequency of PTPN22-1123 gene (rs2488457)GG ,GC ,CC three genotypes in ITP patients and control group had no significant difference(χ2 = 1 .802 ,P = 0 .406) .The frequency of C allele and G allele in ITP patients and control group had no significant difference(χ2 = 0 .003 ,P = 0 .954) .According to the gender fac-tors ,in females ,the genotype and allele frequency of SNP loci rs3811021 and rs2488457 in ITP patients and control group had no significant difference(P< 0 .05) ,so as in males(P < 0 .05) .Conclusion PTPN22 gene rs2476601 this SNP site does not exist in Guizhou Han population ,The addition of two SNP loci of PTPN22 gene (rs3811021 ,rs2488457) exists polymorphism ,but the two SNP loci has no sex difference ,the onset and ITP in Guizhou Han population had no significant correlation .

One new dicyclopeptide cyclo-(L-N-methyl Glu-L-N-methyl Glu) (1), together with one new natural dicyclopeptide cyclo-(L-methyl Glu ester-L-methyl Glu ester) (2), and two known dicyclopeptides cyclo-(L-methyl Glu ester-L-Glu) (3), and cyclo-(L-Glu-L-Glu) (4), were isolated from the aerial parts of Dianthus chinensis L. Their structures were determined by spectroscopic analyses and chemical methods.

Objective To identify the features of the NCAM+ c-Kit+ subset of hepatic progenitor cells in the intrahepatic cholangiocarcinoma (ICC) cell line RBE.Method Magnetic activated cell sorting was used to isolate NCAM+ c-Kit+/NCAM-c-Kit-subset cells,which were tested for hepatic progenitor cell properties and proliferation,colony formation,and invasive abilities in nude mice.Resuits The cell proliferation ability of NCAM+c-Kit+ subset cells was stronger than that of NCAMc-Kit-subset cells (P＜0.01).In serum-free condition,the number of colonies formed by NCAM+c-Kit+ subset cells was more than that of NCAM-c-Kit-cells (P＜0.01).1 × 104 NCAM+c-Kit+ cells were enough to form tumors in nude mice after subcutaneous inoculation for two weeks,while 1 × 106 NCAM-c-Kit-cells were necessary to form tumors for three weeks.The tumor formation rate of NCAM+c-Kit+ cells was higher than that of NCAM-c-Kit-cells (P=0.04).Conclusions It is possible that NCAM+c-Kit+ subset cells in RBE have the properties of hepatic progenitor cells,and NCAM combined with c-Kit might be a valuable marker for isolating and purifying ICC stem/progenitor cells.

Objective:To construct a p53-fused dual luciferase reporter and to test whether this reporter can mimic wild-type p53 activities in a high-throughput screen.Methods:A restriction endonuclease site was added to each terminus and the stop codon of the wild-type full-length p53 open reading frame (ORF) was removed by PCR. A restriction endonuclease site was added to each terminus and the start codon of the ifrelfy luciferase ORF was removed by PCR. The two modified ORFs were inserted upstream of the IRES-induced renilla luciferase ORF in a CMV-derived vector. hTe p53 fusion protein was expressed in cells to test its MDM2-mediated degradation, subcellular localization, and induction of p53-responsive promoter.
Results:hTe p53-fused dual luciferase reporter was successfully constructed. Atfer transfection into the host cells, the reporter expressing the p53 fusion protein that was degraded by oncoprotein MDM2, was mainly located inside the nucleus, and induced the p53-responsive promoter, respectively.
Conclusion:hTe p53-fused dual luciferase reporter (p53FL/IRES/RL) can identify modulators of P53 protein level in a high-throughput screen of genetic or chemical libraries.