Objective To explore the effects of high glucose on glucose transport activity, and phosphorylationandexpressionofinsulinsignalingproteins in primary cultured rat adipocytes. Methods Isolated rat adipocytes were cultured at different glucose concentrations (5, 10, 15, 25 mmol/L) for 24 h. Then the glucose uptake, the phosphorylations of insulin receptor (IR), insulin receptor substrate (IRS) 1 and 2 and protein kinase B (PKB) as well as the protein expressions of IRS1, IRS2, p85 subunit of phosphatitylinositol 3 kinase (p85) and PKB were measured. Results These adipocytes treated with different high glucose showed the impairment of the basal and insulin induced increase in glucose uptake and significant decrease of IR, IRS1 and PKB phosphorylations as well as IRS1 protein expression, but up regulation of IRS2 protein expression. p85 and PKB contents and IRS2 phosphorylation were unaffected. Conclusion The exposure to high glucose inhibits glucose uptake and induces insulin resistance in adipocyte. The mechanism may be involved in affecting the multiple step phosphorylations and the expressions of insulin signaling proteins.

Objective: To get a general estimation of the pneumatic system in ventilators and supply technical assistance for screening and removing the faults of pneumatic component. Methods: Pneumatic system of three kinds of ventilators, Sevro-i/s, Rapheal XTC and PB840 were analyzed and the differences of the three pneumatic systems were compared. Results:Known the construct and differences of pneumatic system. Conclusion: Pneumatic system is the major component of ventilator, and the analysis of the pneumatic component is benefited for the common troubleshooting of ventilators.

Objective　To evaluate the effects of vitrectomy associated with extraction of intraocular foreign bodies in the posterior segment.Methods　Retrospective analysis was done on clinical records of 11 patients (11 eyes) having undergone vitrectomy company with extraction of intraocular foreign bodies in the posterior segment.Results　The intraocular foreign bodies in the posterior segment were extracted successfully in all cases, the retinal detachment and endophthalmitis were cured. Postoperation visual acuity was improved in 8 eyes and did not change in 3 eyes. None suffered from postoperative secondary intraocular hemorrahge, decompensation of corneal endothelium and other serious complications.Conclusion　Vitrectomy combined with extraction of intraocular foreign bodies in the posterior segment is less damaging, accurate and safe, and enhances the cure rate. It is an effective method for treating intraocular foreign bodies.

Objectives To prepare the HL-60 cell vaccine expressing heat shock protein 70 (HSP70) of Bacille calmette-Guérin (BCG), so as to study its anti-tumor effect and mechanism. Methods The whole BCG HSP70 gene was amplified from BCG genome by polymerase chain reaction (PCR) and sub-cloned into the polyclone endonuclease sites in pDisplay. The recom-binant vector of pDisplay-HSP70 was verified by sequencing. Then the HL-60 cell vaccine expressing the protein onto the cell surface was prepared by lipofectamine transfection. To detect the immunogenicity of HL-60 cells expressing HSP70, the test groups were divided into three subgroups, HL60-wt, HL60-pDisplay, and HL60-HSP70 respectively. Each group was cultured with peripheral blood T cells for 72 h, then the proliferation indices of T cells were assayed by CFSE-staining method, and IFN-γwere tested by enzyme-linked immunosorbent assay (ELISA). The HL-60 cells of different groups were cultured with peripher-al blood T cells for 6d. The wild-type HL-60 cells were added and co-cultured for another 12h. Cytotoxicity assay was measured by LDH release. Results (1) The fragment of BCG HSP70 was consistent with the theoretical value. DNA sequencing showed that the recombinant vector of pDisplay-HSP70 was correctly constructed. (2) BCG HSP70 expressed onto the HL-60 cells sur-face. (3) Detection of the immunogenicity: ①The most significant T cell proliferation was observed in the group of HSP70-transfected HL-60 cells (P<0.05). There was no difference between the HL60-wt group and HL60-pDisplay group (P>0.05).②The contents of IFN-γof the HSP70-HL60 group was the highest.③The inhibiting activity of CTLs on HL-60 cells in the group of HSP70-transfected HL-60 cells was more significant than that of wide-type and pDisplay--transfected HL-60 cells. And with the increase of the E:T ratio, the inhibiting activity of CTLs in the HSP70-HL60 group was rising. Conclusions The recombinant eukaryotic expression vector (pDisplay-HSP70) of BCG HSP70 was successfully constructed. And the HL-60 cell vaccine expressing BCG HSP70 onto its surface was successfully prepared. The results showed that gene transfection of BCG HSP70 could significantly enhance the immunogenicity of HL-60 cells.

Objective:To explore the therapeutic effect of Chinese herbal medicine for prevention and treatment of acute radiation oropharyngeal inflammation of nasopharyngeal carcinoma.Methods:120 cases of nasopharyngeal carcinoma were randomly divided into an experiment group of 60 cases treated with Chinese herbal medicine and a control group of 60 cases with gargling of Dobell's solution.The incidence rate and the extent of radiation oropharyngeal inflammation,total therapeutic time and short-term therapeutic effect in both groups were investigated.Results:There was no oropharyngeal inflammation of grade 0 in both groups. The incidence rate of radiation oropharyngeal inflammation in the experiment group was significantly lower and the effective rate was significantly higher than those in the control group(P0.05).Conclusion:Chinese herbal medicine combined with radiotherapy can relieve acute radiotherapy oropharyngeal inflammation in the patient of nasopharyngeal carcinoma with no significant adverse reaction and do not influence the short-term therapeutic effect.

Objective:To establish the quality standards for Zhichuan adhesive plasters;Methods:Herba ephedrae、Bergenin arolisiae、Flos caryophylli、Semen sinapis in Zhichuan adhesive plasters were identified by TLC. Ephedrine hydrachloride in Herba ephedrea was determined by RP HPLC.Results:These methods were simple and accurate.Conclusion: The methods can be used for the quality control of Zhichuan adhesive plasters.

Objective To observe the effect of advanced glycosylation end products(AGEs) on gene expression of connective tissue growth factor(CTGF) in NIH Swiss mouse embryo fibroblasts(NIH/3T3),and to assess the intervention actions of aminoguanidine(AG) and puerarin(Pue) on CTGF mRNA expression in NIH/3T3.Methods AGEs were synthesized by coincubation of BSA with glucose.The AGEs content was measured by fluorescence spectroscopy.NIH/3T3 cells were treated with AGEs(prepared with 20,50,80 mmol?L-1 glucose) for 24 h.The NIH/3T3 cells were treated with AGEs(prepared with 50 mmol?L-1 glucose) for 0,6,12,24 and 48 h.The intervention actions of AG and Pue with different concentration(0.25,0.5,1.0 and 1.5 g?L-1) were evaluated.The CTGF mRNA expression in NIH/3T3 was determined by RT-PCR.Results Compared with BSA control,the CTGF mRNA expression levels in NIH/3T3 were increased by treatment with AGEs(prepared with 20,50,80 mmol?L-1 glucose) for 24 h(P

Objective To develop techniques based on polymerase chain reaction(PCR) which can detect 2 of most common deletional ? thalassemia ? 3.7 deletion and ? 4.2 deletion in China accurately and speedily. Methods Two groups of primers were designed and synthesized. PCR conditions were optimized. The PCR pro ducts were analysed by 1.0% agarose gel electrophoresis. The gel was stained by EB and photographed using an UVP gel documentation. Results PCR product of a 1 700 bp DNA fragment with primers A′, B′, C3 indicates the ? 3.7 deletion while a 1 900 bp fragment indicates a normal or wild type of ? globin gene. Occurrence of the 1 700 bp and 1 900 bp simultaneously indicates a heterozygous of the ? 3.7 deletion. Neither of the 2 bands was presented, indicating a homozygous of South East Asia type of deletion (-? SEA ). According to patterns of 1 580 bp and 1 180 bp hand amplified by a PCR with primer G′, E, F′, we detected the ? 4.2 deletion and distinquished its heterozygous and homozygous. Conclusions The 2 PCR based techniques developed in our laboratory are accurate, simple, well reproducible and easy to use for screening of the 2 deletion types of ? thalassemia determinants.

Objective To culture the acute leukemia cells in vitro,and to prepare cancer vaccine expressing heat shock protein 70 (HSP70) of Bacille Calemette-Geérin(BCG) onto the cell surface,so as to study its anti-tumor effect and mechanism.Methods Acute myeloid leukemia (AML) cells were cultured in a serum-free Stemspan(H) culture supplemented with cytokines [stem cell factor(SCF),flt-3 ligand (FL),interleukin (IL)-3 and IL-6] in vitro.And B-lineage acute lymphoblastic leukemia (B-ALL) cells were cultured in a Iscove modified medium(IMDM) culture supplemented with cytokines (SCF,FL,IL-3 and IL-7) in vitro.Cellular morphology was observed by the microscopy and immunophenotype determination was used to verify the biological characteristics of acute leukemia cells after culture.Lipofectamine 2000 was used to transfect the pDisplay-HSP70 plasmid into acute leukemia cells.The expression of HSP70 on the cell surface was detected by fluorescene microscope.Then the immunogenicity of the leukemia cells expressing HSP70 were detected.The experimental groups were divided into 3 subgroups:the wide-type acute leukemia cells (wt-LC group),the pDisplay-leukemia cells (pDisplay-LC group),and the pDisplay-HSP70-leukemia cells (HSP70-LC group),respectively.The leukemia cells in different groups were cultured with autologous peripheral blood T cells for 72 hours.The proliferation indices of T cells were assayed by carboxyfluorescein diacetate succinimidyl ester (CFSE)-staining method,and the contents of interferon-γ(IFN-γ) were tested by enzyme-linked immunosorbent assay (ELISA).The leukemia cells in different groups were cultured with autologous peripheral blood T cells,and after 6 days,the fresh acute leukemia cells were added [in the different ratios of cytotoxicity T lymphocyte (CTL):leukemia cells were 10 ∶ 1,20 ∶ 1,40 ∶ 1 and 80 ∶ 1] and continued to be cultured for another 12 hours.Cytotoxicity assay was measured by lactate dehydrogenase (LDH) release.Results After short term culture in vitro,the leukemia cells were in colony-like suspension and maintain the proliferation characteristics were maintained.The cell proliferation was rapidly cultured for about 10 days and then was gradually slowed down.But there was no difference between the day 10 and day 0 in the expressions of CD13 and CD33 in fifteen cases of AML cells (P ＞ 0.05).Equally,there was no difference between the day 10 and day 0 in the expressions of CD19,CD10 and CD22 in fifteen cases of B-ALL cells (P ＞ 0.05).After BCG HSP70 gene transfection,the yellow-green fluorescence on the leukemia cells surface was observed under the confocal microscope.Detection of the immunogenicity:(1) Autologous T cell proliferation:the most significant T cell proliferation was observed in the group of HSP70-transfected leukemia cells (t =17.89,19.58,all P ＜0.05).There was no difference between the wt-LC group and pDisplay-LC group (P ＞ 0.05).(2) The contents of cytokines:the IFN-γ level in the group of HSP70-transfected leukemia cells was higher than those of wide-type acute leukemia cells and the pDisplay-transfected ones (t =24.72,24.81,all P ＜ 0.05).(3) Cytotoicity of CTL:the killing rate in HSP70-transfected leukemia cells was significantly higher than those of wide-type acute leukemia cells and pDisplay——transfected ones(F =13.66,P ＜ 0.05).And with the increase of the ratio from 10 ∶ 1 to 80 ∶ 1,the inhibiting activity of CTL in the HSP70-LC group was raising(F =19.69,P ＜ 0.05).Conclusions Fresh acute leukemia cells can be successfully cultured in vitro.Short-term culture can significantly increase the number of leukemia cells,but has little effect on surface antigen expression.So,the biological characteristics of the leukemia cells can be maintained.The leukemia cells vaccine expressing BCG HSP70 onto its surface was successfully prepared,and gene transfection of BCG HSP70 can significantly enhance the immunogenicity of leukemia cells.

Objective To explore the clinical characteristics and the influence factors of prognosis of ad -vanced ovarian cancer patients after abdominal metastases .Methods We retrospectively analyzed 65 cases dur-ing January 2013 to January 2016 in the First People′s Hospital of Tianmen .The patients were diagnosed clearly with pathological ,diagnosed for the first time and has the complete clinical data including peritoneal metastases in 58 cases,which were analyzed for single factor and multiple factors of influencing factors survival analysis .Results 58 cases of patients with diagnosis of peritoneal metastasis in ovarian cancer that the average is (49.2 ±6.5), from ovarian cancer diagnosis to abdominal metastases for an average time of 11 months,ovarian cancer patients with peritoneal metastasis of the median survival time was 8 weeks,but 7 cases without abdominal metastases that the median survival time was 15 weeks.The single factor survival curves(Kaplan-meier)showed that marital sta-tus,reproductive history,history of breastfeeding,malignant ascites,neoadjuvant chemotherapy,comprehensive treatment(chemotherapy and tumor cells to destroy the loss and laparotomy abdominal hot perfusion chemothera -py) ,peritoneal metastasis tumor number ,residual lesion size ,quality of life in patients with residual lesions ( KPS scores),D-dimer level in plasma and urine trace albumin levels related to the prognosis of patients (P<0.05). Conclusion Patients with advanced ovarian cancer patients with peritoneal metastasis survival time is shorter , and also company with poor prognosis;Neoadjuvant chemotherapy combined tumor cells to destroy the loss and postoperative intraperitoneal hot perfusion chemotherapy ,short-term curative effect is good ,it can not only im-prove the advanced ovarian cancer treatment effectiveness ,but also improve the quality of life in patients of late phase.The D -dimer level in plasma and urine trace albumin levels before treatment have remained in the pa-tients with high level or not reduce ,treatment effect and prognosis was poor .These can be used as a new index of judging prognosis .