Objective: To clone human augmenter of liver regeneration (hALR) cDNA, construct the recombinant expression vector, express and purify its product. Methods: The hALR cDNA was obtained by using RT PCR method with total RNA extracted from the fetal hepatic tissue. Then it was cloned into the pGEM T vector, and subcloned into expression vector pGEX 4T 3.After proved to be correct by sequencing, recombinant expression plasmid pGEX 4T 3(hALR) was transformed into E.coli BL21(DE3). The fusion protein GST hALR was produced by IPTG induction, isolated by affinity chromatography glutathione Sepharose 4B and cleaved by Thrombin. Results and Conclusion: Recombinant expression plasmid pGEX 4T 3(hALR) is constructed. The hALR is highly expressed in E.coli . The fusion protein in the plasma is resoluble. The procedure of purification and cleavage of fusion protein is easy and simple.

Purpose To construct a recombined antitumor peptide and to analyze its bioactivity. Methods Constructing a recombined gene and inserting the pGEX-4T-3 vector. The recombined protein was expressed in E. coli BL21 and purified with Amylose Resin. Then, citrostatin was subjected to the following tests separately: inhibition of endothelial cell proliferation, MTT test of cytotoxicity and inhibition of endothelial cell tube formation on ECMatrix. Results Citrostatin significantly inhibited the proliferation of human endothelial cell ECV304(IC_(50) = 2.28 μmol/L) .It also significantly inhibited the proliferation of human tumor cell 1990 and NCI-H64O(IC_(50) = 9.24,2.74 μmol/L) ,and the inhibitory effect became more marked with the increase of citrostatin concentration. The inhibitory effects of citrostatin on endothelial cell tube formation was also confirmed . Conclusion An antitumor peptide, citrostatin, has been successfully constructed and purified, which showed anti-angiogenesis effect and direct cytotoxic effect on tumor cells.

To study the effects of puerarin on the aromatase P450 (P450(arom)) mRNA expression and the effects of low-dose puerarin on transcription factors of the P450(arom) gene (P II) 5'-flanking region.

To compare angiopoiesis ability of eutopic and ectopic endometrial tissue isolated from women with endometriosis and endometrium isolated from women without endometriosis (control), and to explore the inhibitory effects of medicated serum of Neiyi Recipe, a compound traditional Chinese herbal medicine.

OBJECTIVETo investigate the effect of the expression and mutation of c-kit gene and its relationship with clinical pathology and prognosis of gastrointestinal stromal tumor (GIST).

METHODSImmunohistochemical and PCR-SSCP techniques were used to detect c-kit protein expression and c-kit gene exon 11 mutation in 82 patients with GIST.

RESULTSThe positive c-kit protein expression and c-kit gene mutation rates were 97.6% (80/82) and 41.5% (34/82). Correlating the results of these two methods and clinicopathological factors, the c-kit expression and c-kit gene mutation rates were 95.0% (19/20) and 0 in benign GIST, and were 98.4% (61/62), 54.8% (34/62) in malignant GIST. Mutation positive GIST showed higher frequency of adjacent tissue invasion, metastasis and recurrence as compared with mutation negative ones.

CONCLUSIONc-kit protein is an important diagnostic marker of gastrointestinal stromal tumor. c-kit gene mutation may play a significant role in the pathogenesis of GIST and also may be a prognostic marker.

Adolescent ; Adult ; Aged ; Child ; Female ; Gastrointestinal Neoplasms ; diagnosis ; genetics ; pathology ; Humans ; Male ; Middle Aged ; Mutation ; Neoplasm Metastasis ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Prognosis ; Proto-Oncogene Proteins c-kit ; genetics

Objective:This study by using the method of network pharmacology to screen the active constituent and related targets of Epimedii Folium aims to explore the mechanisms of the reinforcing effect of Epimedii Folium on kidney in the treatment of PCOS. Methods:By retrieving data from TCMSP datebase, screened out the active constituent of Epimedii Folium and the information of the targets corresponding to each active constituent; by using the gene database of NCBI, translated the information of the targets into gene names; by retrieving data from GeneCards datebase, extracted the genes related to PCOS; related targets of Epimedii Folium in the treatment of PCOS were obtained by Venn tool; by using Cytoscape 3.7.2 software, constructed a network diagram of Epimedii Folium-active constituents-targets-PCOS; by using STRING database, constructed the protein interaction network; then carried out GO enrichment analysis of related targets by Geneontology database and carried out pathway enrichment analysis of related targets by KEGG database. Results:There were 23 active constituents of Epimedii Folium and 132 related targets treating PCOS. The Epimedii Folium could play the reinforcing effect on kidney mainly by regulating the biological processes like steroid hormone receptor activity, as well as KEGG pathways such as Estrogen signaling pathway, GnRH signaling pathway, GnRH secretion, HIF-1 signaling pathway and VEGF signaling pathway in treating PCOS. Conclusion:From the perspective of network pharmacology, this study preliminarily analyzed the related targets and pathways of reinforcing effect on kidney of Epimedii folium in the treatment of PCOS, providing reference for further experiments and application inclinics.