Objective To investigate the effects of insulin on the mRNA expression of insulin receptor substrate-2 (IRS-2) in MG-63 cells. Methods Semi-quantitative RT-PCR was used to study the action of insulin on the mRNA expression of IRS-2 in MG-63 cells. Results Insulin regulated the mRNA expression of IRS-2 in a dose- and time-dependent manners in MG-63 cells. Insulin up-regulated the expression of IRS-2 mRNA at 10~ -10～10~ -6mol/L(P

Objective To determine the expression of insulin receptor substract(IRS) family in human osteosarcoma cell line MG-63 and cultured normal human osteoblast-like cells (HOB). Methods The mRNA and protein expression of IRS family was measured using semi-quantitative RT-PCR and western blot analysis, respectively. Results MG-63 cells and HOB had the mRNA and protein expressions of IRS-1,-2,-3, and -4. Among IRS family, the expressions levels of IRS-1 mRNA and protein were the highest, and those of the IRS-4 mRNA and protein were the lowest in MG-63 cell line and HOB. Conclusion MG-63 and HOB can express the mRNA and protein of IRS family members, and the expressional levels of IRS family members were different.

Objective To observe the clinical sedativelefficacy of Tianwang-Buxin Dan treating chronic obstructive pulmonary disease patients undergoing mechanical ventilation.Methods 97 patients with chronic obstructive pulmonary disease undergoing mechanical ventilation were randomly recruited into a treatment group (47 cases) and a control group (50 cases).The control group was treated with midazolam and fentanyl,and the treatment group were treated by Tianwang-Buxin Dan on the basis of the control group.The amount of medazolam and fentanyl,duration of MV,and incidence of side-effects such as hypotension,bradycardia,delirium,nausea,etc were observed in both groups.Results The expected sedative and analgesia scores were obtained in patients of both groups.Compared with the control group,the patients in the treatment group were easier to be aroused and be kept in sedation and analgesia state,besides the dose of midazolam (μg/kg·h-1) (9.51 ±5.87 vs 20.01 ±6.24,P＜0.01)and fentanyl (μg/kg·h-1) was significantly fewer (0.22 ±0.13 vs 0.32±0.12,P＜0.05),duration of MV (days) was shorter(7.13±6.25 vs 12.85±9.13,P＜0.01),duration of disable sedatives for extubating (hours)was shorter (4.35 ± 2.57 vs 8.79±4.02,P＜0.01),the rates of hypotensionand brdycarcardia were significantly lower,the rates of delirium and nausea were clearly lower(both P＜0.05)in the treatment group.Conclusion Sedative effect of Tianwang-Buxin Dan is satisfactory for patients with chronic obstructive pulmonary disease undergoing mechanical ventilation,with the property of easier arousal,lower hypotension rates,lower brdycarcardia rates,lower delirium rates nausea rates,and fewer dosage of midazolam (by 50％) and fentanyl (by 30％) administration.

Objectives The aim of the study was to detect the frequency of B regulatory cells (Breg)and the correlation between Breg and T regulatory cells (Treg) in pancreatic cancer patients,and to investigate its role in pathogenesis of pancreatic cancer.Methods 50 pancreatic cancer patients and 21 healthy controls were enrolled.CD19+CD24hiCD38hi Breg and CD4+CD25high Treg was also determined via flow cytometry.The correlation between Breg and Treg was analyzed by Spearman correlation test.Results Upregulation of CD19+CD24hiCD381hi Breg was associated with pancreatic cancer progression.Furthermore,this B cell subset was positively correlated with the frequency of CD4 + CD25high Treg cells.Conclusions Together,CD19 + CD24hiCD38hi Breg cells are significantly elevated in pancreatic cancer patients,indicating this B cell subset might play an vital role in clinical progression of pancreatic cancer.The significant positive correlation between Breg and Treg may suggest CD 1TCD24hiCD38hi Breg are affecting tumor progression through Treg cells.

Objective:Investigation about immunological pathogenesis of thrombocytopenic purpura was made.Methods:Th1 and Th2 cell number between thrombocytopenic purpura patients and normal people was detected by flow cemetry.Results:Th2 cell number in thrombocytopenic purpura patients was significantly higher than that in control group,whereas Th2 cell number in thrombocytopenic purpura patients was significantly lower than that in control group.Conclusion:The imbalance between Th1 and Th2 could partly explained the mechanism of thrombocytopenic purprua.

Objective To explore the effects of erythropoietin (EPO) on the neural function recovery and the expressions of nuclear factor-κB (NF-κB) in rabbit spinal cord after ischemia-reperfusion injury. Methods 24 rabbits were randomized into 3 groups: EPO treatment group, ischemia-reperfusion (I/R) group and sham group with 8 rabbits in each group. The rabbit model of spinal cord ischemia-reperfusion injury was established with Tetik's method. The changes of neural functional recovery of rabbits of 3 groups were observed through Tarlov's scale at different time points post-operation. The expressions of NF-κB were tested with immunohistochemistry. Results The grade of nerve function improved distinctly in EPO treatment group (P<0.01).The expression of NF-κB in EPO group was lower than that of I/R group (P<0.01).Conclusion EPO can significantly improve the motor function of hind limbs in rabbits after spinal cord ischemia-reperfusion injury, which might be related with inhibition of the expressions of NF-κB.

This paper introduced the measures to construct the hierarchical medical system in Yichang city. The authors identified problems found and proposed the city to take the following countermeasures. Such steps included service capacity building at primary level, motivating primary medical workers, attracting patients to primary care, and improving policies of hierarchical medical system.

Hierarchical medical system empowers the current healthcare reform on the medical supply side in China, a key to resolving the contradictions in the healthcare sector. This study analyzed the necessity and obstacles for such a system with synergy between hospitals, insurance and pharmacy, putting forward suggestions for building a scientific and rational hierarchical system.

BACKGROUND:Most bone marrow mesenchymal stem cel s are infused intravenously and have very low efficiency of homing to the bone marrow. However, cel infusion via the femoral approach is little reported. OBJECTIVE:To explore the distribution of luciferase gene modified red fluorescent protein transgenic bone marrow mesenchymal stem cel s in vivo through different infusion routes. METHODS:Luciferase gene modified bone marrow mesenchymal stem cel s at different gradients (5×106, 1×106, 1×105, 1×104) were seeded or injected into the in vitro pore plate or free femurs to observe the fluorescence imaging and select the best concentration of cel s. Luciferase gene modified bone marrow mesenchymal stem cel s at the best cel concentration were injected into the mice via the femur and the tail vein, respectively. The distribution of fluorescence and cel number in the mice were explored by using bioluminescence, pathological examination, flow cytometry and quantitative PCR. RESULTS AND CONCLUSION:Ex vivo fluorescence intensity of luciferase gene modified bone marrow mesenchymal stem cel s was positively correlated with the cel concentration;fluorescent cel s in vivo appeared in the femur first and then quickly spread to the lungs in the femur group, while fluorescent cel s in the tail vein group spread to the lungs quickly after cel infusion. Fluorescent cel s could be seen in the spleen, liver and other organs 24 hours later in the two groups. The distribution and migration of cel s in mice could be observed successful y by bioluminescence;5 minutes after cel infusion, the lungs of mice in the two groups began to emit fluorescence that could spread to the liver, spleen and other tissues 24 hours later, and the fluorescence intensity reached its peak after 15 minutes. The distribution of bone marrow mesenchymal stem cel s in mice had no significant difference between the femur group and the tail vein group. To conclude, cel injection through the bone marrow cavity and tail vein fails to promote the homing of bone marrow mesenchymal stem cel s to the bone marrow.

Objective:To prepare monoclonal antibody ( mAb ) against human testis-specific conserved gene ( hTSC29 ) peptides and characterize its immunological and biological features.Methods:According to bioinformatics analysis and prediction of the antigenicity, surface property, hydrophilicity and flexibility of hTSC29, a 18-amino acid residue partial peptide of hTSC29 was synthesized,then immunized the BALB/c mice for preparing antiserum.The mAb against hTSC29 was produced using the routine hybridoma technique.The properties of the mAb against hTSC29 were identified by ELISA, Western blot and immunohistochemistry staining.Results:After cell fusion and subcloning, one hybridoma cell lines secreting specific mAb against hTSC29 protein were obtaind.The Ig subclass of the mAb was IgG2b(κ).ELISA detection showed that the titer of mAbs in cultured was 1∶104.Western blot analysis proved that the mAb could specifically recognize Mr 60 000 protein in human testis total protein.The hTSC29 protein main located at circumference of spermatocyte and spermatid in human testis tissue by immunohistochemistry staining and immunofluorescence assay.Conclusion:One hybridoma cell lines which can secrete specific mAb against hTSC29 protein with high titers and specificity have been established successfully.The mAb will provide efficient tools for functional studies of hTSC29 expressed in spermatogenesis.