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Objective To learn the present situation and bottlenecks of human resources at private hospitals in Sichuan province.Methods Questionnaires,discussions and in-depth interviews were used to learn human resources of 35 private hospitals in the province.Results The findings indicate significant differences among doctors at such hospitals,most of whom age 30-40 years old(45.13％) ; technical titles of the medical workers are low in general,as only 20.08％ of them have vice-senior titles or above; employees are dissatisfied with the remuneration,training setup and culture of their hospitals.Conclusion Private hospitals must enhance training of professionalism,raise payment,improve training setup,and create a positive hospital culture if they want sustained development.In the meantime,hospital regulators should enhance supervision and provide supportive policies.
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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.
Subject(s)
Female , Humans , Adenoviridae , Genetics , Metabolism , Antineoplastic Agents , Pharmacology , Breast Neoplasms , Metabolism , Pathology , Cell Line, Tumor , Genetic Vectors , Genetics , Mutant Proteins , Genetics , Receptor, Fibroblast Growth Factor, Type 2 , Genetics , Recombinant Proteins , Genetics , Pharmacology , TransfectionABSTRACT
BACKGROUND: Studies have shown that chitooligosaccharides have antitumor effect. However, the influence of chitooligosaccharides on cyclin D1, bcl-2 and bcl-xl remains unclear.OBJECTIVE: To observe the JnhJbJtJon effect of chitooligosaccharides on the proliferation of Hela cells, and the influence on cyclin D1, bcl-xl and bcl-2 mRNA expression.METHODS: Hela cells were stimulated by different concentrations of chitooligosaccharides (0.1, 0.5, 1, 2, 5 g/L). The effects of chitooligosaccharides on Hela cells were detected by CCK8 kit. Using real-time PCR methods, the gene expression of cyclin D1, bcl-xl and bcl-2 mRNA were determined.RESULTS AND CONCLUSION: Chitooligosaccharides inhibited the proliferation of Hela cells. With the concentrations of chitoolJgosaccharides increased from 0.1 g/L to 2 g/L, the inhibition effects on the gene expressJon of cyclin D1, bcl-xl and bcl-2 were enhanced, peaked at 2 g/L, and decreased at high concentration (5 g/L). Antitumor activity of chitooligosaccharides may exert through two aspects: it inhibits cyclin D1 mRNA expression to suppress the proliferation of tumor cells; on the other hand, chitooligosaccharides inhibits the expression of bcl-xl and bci-2 to promote the apoptosis of tumor cells. Moreover, the effects of the former are stronger than the latter.
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AIM: To obtain a high and stable expression analog of human basic fibroblast growth factor by genetic engineering. METHODS: The cysteins 78 and 96 of natural hbFGF polypeptide was substituted with serines by means of site-directed mutagenesis. Using pET- 3c as vector, the mutated polynucleotide was cloned and then transferred into BL21 (DE3)plysS. After induction by IPTG, the analog was obtained and analyzed by SDS - PAGE. RESULTS: After purification the form of soluble mutant increased remarkably but the forms of dimmer and higher multimer were reduced greatly to no more than 8% of the total recombinant protein. By MTT assay, the analog showed the same biological activity. This new analog represented a desirable complementation for native hbFGF to develop pharmaceutical drug in clinical use. CONCLUSION: Substitution of certain amino acids of polypeptide without altering native protein' s bioactivity to get the analog is an effective means to increase stability of foreign protein and its solubility in E. coli.
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Objective Non-small-cell lung cancer with ipsilateral mediastinal lymph node metastasis (N2) belongs to heterogeneous subgroup. We analyzed the prognosis of patients with resected N2 NSCLC to determine the clinical significance. Methods The present study comprised 146 consecutive patients whom underwent surgical resection of N2 NSCLC between January 1997 and January 2000. Histological type, location, T primary tumor status, operation mode, clinical N2 (cN2) factor, N2 level(single or multiple), number of positive nodes(N2-num), and the cycles of adjuvant chemotherapy were estimated from the date of operation using the Kaplan-Meier and Log Rank analysis. The Cox regression model evaluated the influence of factors on the survival. Results The 3-and 5-year survival rate of these N2 NSCLC patients were 19.86% and 14.56%, respectively. The patients with tumor in the right lower lobe showed a significantly longer survival than left lobes. The histological type, tumor location, cN2 factor, N2 level and N2-number were associated with survival. A multivariate analysis using Cox regression identified 4 factors of prognosis: tumor site, T status, N2 level and clinical N2 status. Conclusion This article has identified N2 NSCLC subgroups and found that patients with mN2, N2L1 and single N2 have better prognosis.
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This paper summerized the surgical treatment of primary bronchogenic carcinoma in 3568 cases between 1957-1991. The resectability rate was 90. 3%, postoperative morbidity rate 8.32% and operative mortality 0. 89%. Pathological diagnoses of the resected specimens included squa-mous cell carcinoma for 48.7%, adenocarcinoma 22.9%, small cell cancer 15. 4%, large cell cancer 1. 3% and squa-mous-adenocarcinoma in 10.1%. The follow-up rate was 93%. The 5-and 10-year survival rates were 34.6% and 22.79% respectively. Analyses of the data demonstratad that the histologic type, pathological stage and metastasis of mediastinal lymph node are the important factors affecting the prognosis. According to UICC P-TNM,42. 6% of the patients in this group were in stage III. The 5 year survival rate was 19% in IIIa patients and 6% in IIIb. According to authors experience, it is recommended that in IIIa patients with nonsmall cell cardinoma, active surgical treatment should be adopted; in patients with small cell carcinoma, chemotherapy and radiotherapy should be given pre-and postoperatively, in IIIb patients with small cell carcinoma, surgical treatment is generally not indicated.
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To improve the expression level of non fusion hbFGF in E coli , the coding sequence of human bFGF gene, which had been cloned from primarily cultured human fibroblast, was mutated according to the principle of lowering the GC content and increasing the codon preference After being ligated into pET 3c and transformed into BL21(DE3), the recombinant induced by IPTG Expression level was up to 30% of the total bacterial protein The result indicated that optimizing of the TIR would promote the expression level of recombinant protein