Objective To observe the expression of CD_ 105 in breast cancer, and explore the correlation between CD_ 105 and growth, invasion and metabasis of breast cancer. Methods 50 cases of breast infiltrating duct cancer, 20 cases of breast duct cancer and 20 cases of breast benign hyperplasia were enrolled in this study. The microvascular density(MVD) was marked with CD_ 105 and F-8RAg using immunohistochemical S-P method. Results The expression level of CD_ 105 in breast infiltrating duct cancer was significantly higher than that in breast duct cancer and benign hyperplasisa(P

Objective To compare the binding characteristics of insulin receptors (IR) on ovary,adrenal and hepatic plasma membrane from rats. Methods The number and affinity of IR was detected by radioligand binding assay. Results Scatchard plot analyses showed that the receptor numbers for high affinity sites are 7. 359?10/mg protein, 8. 029? 10/mg protein and 6. 440?10/mg protein of liver,ovarian and adrenal plasma membrane with KD of 6.147?107 M-1, 1. 528?10, M-1, and 1. 010?107 M-1 that. The receptor numbers for low affinity sites are 2. 403?10/mg protein,2. 212?10/mg protein,and 2. 257?10/mg protein respectively with KD of 2. 920?10M-1,2. 008?10 M-1 and 0. 433?10sM-1. Conclusion There are abundent IR on ovary and adrenal just as many as those on liver, although the KD are smaller. It suggests that insulin may play an important regulatory role in ovary and adrenal.

Cells, Cultured ; Endothelial Cells ; cytology ; Humans ; Stem Cells

Previous work from this laboratory has cloned a novel gene NOR1 and showed its extensive expression in normal tissues and down-regulation in carcinomas. To further investigate its downstream target genes and better understand its function, NOR1 was over-expressed in HepG2 hepatoma cells and global changes in gene expressions from a stable line were identified by cDNA microarrays. The results discovered 59 genes up-regulated in these cells compared with the original cells, including Grb2, HBP17,TNFRSF11B genes that have been implicated in tumorigenesis and cancer development. In addition, 103 down-regalated genes were also identified, including genes encoding Bik, MAP2K6 and ZFP95 proteins. The expression patterns of certain genes identified by microarrays were validated by quantitative real-time PCR and the results showed that expression difference were statistically significant (P< 0.05). These data suggest that NOR1 may influence the biology and cancerous behaviors of HepG2 cells by regulating expression of a set of genes involved in signal traasduction, cell cycle regulation, transcription and Wanslation controls.

The genetic analysis of herpesviruses has been a constant challenge, due to the large, complex genomes of herpesviruses and mutagenesis of viral genes by conventional recombination methods in cell culture. Recently, a completely new approach for full-length infectious clones of herpesviruses based on bacterial artificial chromosomes (BACs) has been developed. This technique allows the maintenance, propagation and genetic modification of the viral genome as a BAC plasmid in E.coli, thus making the procedures fast, safe and effective in prokaryotic cells. This technique also makes it possible for the reconstitution of viral progeny or mutants by transfection of the BAC plasmid into eukaryotic cells, thereby facilitating the analysis of viral gene functions in the context of genome. In this presentation, Epstein-Barr virus was used as an example to describe the principle, establishment of the technique and mutation introduction into the BAC plasmid, and to discuss the perspective in the use of BAC-cloned herpesviruses.

The effects of several factors on the chlamydospores production of Trichoderma aureoviride T-33 during the fermentation were researched.Based on the results above, the orthogonal test was made to screen out the best prescription.The results showed that, the best single-factor conditions for the chlamydospores production of T.aureoviride T-33 were, liquid culture of oat powder, 30℃, pH4.0, 120r/min, 24 hours oscillate incubating as well as liquid culture volume of 80mL/bottle when the 250mL size triangle bottle was used.The result of orthogonal test showed that, the best prescription for temperature, pH and oscillating speed was 30℃, pH4.0 and 140r/min.3.37?10~ 7 spore/mL chlamydospores were obtained at this combined condition.

Objective To investigate the short term effect of gastric bypass surgery for the treatment of nonobese type 2 diabetes mellitus and possible mechanisms. Methods The clinical data of 58 patients with nonobese type 2 diabetes mellitus who received gastric bypass surgery from March to August, 2009 were retrospectively analyzed. The levels of fasting plasms glucose (FPG), 2-hour postprandial plasma glucose (2h PG) and glycosylated hemoglobin (HbA1c) were dynamically monitored, and the insulin resistance index (HOMA-IR) and body mass index ( BMI) were calculated. All data were analyzed using variance of analysis and LSD test. Results Of the 58 patients, 48 (83% ) met the requirement of complete response criteria and stopped administration of hypoglycemic agents; 7( 12% ) had to use hypoglycemic agents, but the dose of the agents was lowered by 50% compared to that before surgery. The surgery was ineffective in 3 patients (5% ). The levels of FPG, 2h PG, HbAlc and HOMA-IR of the 58 patients showed a significant decreasing trend after surgery when compared to those before surgery (F = 67. 867, 50. 885, 78. 278, 572. 757, P <0.05), while there was no significant change of the BMI after surgery ( F = 3. 503, P > 0. 05 ). Conclusions Gastric bypass surgery has a good effect on nonobese patients with type 2 diabetes mellitus whose BMI was less than 25 kg/m2. The improvement of insulin resistance after the surgery might be the main reason.

Objective To dynamically observe the changes of cytokines of splenocytes in mice immunized with recombinant bifidobacteria bifidum (Bb)- Eg95-EgA31 vaccine of Echinococcus grauulosus (Eg). Methods Balb/c mice were vaccinated by 5× 108 colony forming unit(CFU) orally and 5 × 105 CFU intranasally, respectively.Mice were killed on week 0,2,4,6,8,10, 12,14,16, 18 and 20 after immunization, respectively, and spleens were separated for cell culture with the stimulation of EgAg, concanavalin A (ConA) or lipopolysaccharide (LPS). The splenocyte supernatants were collected to determine the levels of interferonγ(IFN-γ), interleukin(IL)-12, tumor necrosis factor α(TNF-o) and IL-l0 using enzyme linked immunosorbent assay(ELISA) with MRS as control. Results In the oral immunization group, the levels of IFN-γ, IL-12, TNF-α and IL-10 showed a significant increase from week 2 to week 8, week 2 to week 8, week 4 and week 6 to week 10 after vaccination, respectively, and reached the highest level on week 4, week 2, week 4 and week 6 after vaccination, respectively;in EgAg stimulation group, the levels of IFN-γ, IL-12, TNF-α and IL-10 were (700.0 ± 115.5), (45.0 ± 5.8), (350.0 ± 57.7), (112.5 ± 14.4)ng/L, respectively, compared with week 0[(35.0 ± 5.8), (12.5 ± 2.9), (190.0 ± 11.6), (25.0 ± 5.8)ng/L, P ＜0.05 or ＜ 0.01] and MRS control group[(37.5 ± 5.0),(13.8 ± 2.5), (195.0 ± 5.8), (27.5 ± 2.9)ng/L, P＜ 0.05or ＜ 0.01]. In the intranasal immunization group, the levels of IFN-γ, IL-12, TNF-α and IL-10 showed an obvious increase from week 2 to week 8, week 2 to week 8, week 2 to week 6 and week 6 to week 16 after vaccination,respectively, and reached the highest level on week 2, week 2, week 4 and week 8 after vaccination, respectively;in EgAg stimulation group, the levels of IFN-γ, IL-12, TNF-α and IL-10 were (700.0 ± 115.5), (55.0 ± 5.8),(275.0 ± 28.9), (140.0 ± 11.6)ng/L, compared with week 0[(35.0 ± 5.8), (12.5 ± 2.9), (190.0 ± 11.6), (25.0 ±5.8)ng/L, P ＜ 0.05 or ＜ 0.01] and MRS control group[(37.5 ± 5.0), (13.8 ± 2.5), (195.0 ± 5.8), (27.5 ± 2.9)ng/L, P ＜ 0.05 or ＜ 0.01]. The cytokine levels in the groups with EgAg, ConA or LPS stimulus were significantly higher than those in the corresponding splenocytes suspension groups(P ＜ 0.05 or ＜ 0.01) , and the cytokine levels in the groups with ConA or LPS stimulus were obviously higher than those in the corresponding groups with EgAg stimulation(P ＜ 0.05 or ＜ 0.01). Conclusion The mixed Th1 and Th2 type response can be induced in mice immunized with the recombinant Bb-Eg95-EgA31 vaccine of Echinococcus granulosus in the early stage of immunization(2 to 6weeks).

Objective To construct and express the recombinant plasmid pET28α-Sj26GST-Sj32 of Schistosoma japonicum(Sj) in Escherichia coli BL21 (DE3). Methods Total RNA was extracted from Sj adult worms by ultrasound-breaking, Sj26GST and Sj32 antigen gene was respectively amplified by RT-PCR from the total RNA; Sj26GST-Sj32 fusion gene obtained with gene splicing by overlap extension(SOEing) was cloned into prokaryotic expression plasmid pET28α and transformed into Escherichia coli BL2 (DE3) to construct pET28α-Sj26GST-Sj32;BL21 (pET28α-Sj26GST-Sj32) was induced with isopropyl-β-D-thiogalactopyranosid (IPTG), and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE)and Western blotting. Results The 1991 bp Sj26GST-Sj32 fusion gene was successfully amplified by gene SOEing and cloned into pET28α by restriction analysis and PCR identification, the recombinant plasmid pET28α-Sj26GST-Sj32 was successfully constructed; the relative molecular mass of the expressed recombinant protein was approximately 69 × 103 by SDS-PAGE, and the amount of the expressed protein was 25% of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pET28α-Sj26GST-Sj32 is successfully constructed and highly expressed in Escherichia coli in fused form with His-tag, and the expressed fusion protein shows specific antigenicity.

Objective To construct and identify recombinant Bifutobacteria (rBb)-Eg95 vaccine of Echinococcus granulosus (Eg). Methods The total RNA was extracted from hydatid cyst protoscoleces shattered by ultrasound, Eg95 antigen encoding gene was obtained by reverse transcription-polymerase chain reaction(RT-PCR) from the template of total RNA using the primer designed according to the DNA sequence of Eg95, the gene was cloned into Escherichia coli-Bifutobacteria(E.coli-Bb) shuttle plasmid pGEX-1λT and transformed into E.coli BL2 (DE3) competent cell to construct recombinant plasmid pGEX-Eg95 using BamH Ⅰ and EcoR Ⅰ, the recombinant plasmid was identified by restriction endonuclease digestion, then was electroporated into Bb to construct rBb-Eg95 vaccine, the vaccine was identified by PCR. Results Four hundred and seventy-one bp Eg95 gene was amplified by RT-PCR, the products of restriction endonuclease digestion were the same as expected(471 bp Eg95 gene and 4947 bp pGEX-1λT), 471 bp Eg95 gene fragment was amplified by PCR from the template of pGEX-Eg95 extracted from rBb vaccine. Conclusion rBb-Eg95 vaccine of Eg is successfully constructed, which lays the theoretical foundation for exploitation and utilization of this vaccine.