AIM: To evaluate the effectiveness of small interference RNA on inhibiting bcl-2 gene expression. METHODS: With Ambion's software and kit, we designed and synthesized siRNA targeted bcl-2, which were transfected into astrocytoma cell line U251 with lipofectamine. The non-transfected cells and treatment with antisense drug G-3139 were taken as controls. MTT was used to detect the inhibitory rate of cell growth. Flow cytometric method was used to detect the change in cycle of the cells. The inhibitory effect of siRNA on mRNA level was detected by RT-PCR and on protein level was by immunohistochemical method. RESULTS: For the living rate of cell, siRNA 2, 3, 5, 6 groups were significantly lower than siRNA 1, 4 groups, lipofectamine and control group at 24, 48, and 96 h. siRNA 1-6 groups only had statistic difference with antisense group at 24, 48 h. As for PCR and immunohistochemical method, the expression of bcl-2 on siRNA 2, 3, 5, 6 groups were significant lower than other groups. The results of flow cytomytric method showed the cells transfected with siRNA 1-6 and antisense were blocked at S stage. CONCLUSION: siRNA inhibited bcl-2 gene expression more than 50%.

ObjectiveIn order to confirm the relationship of pathogenic factors and clinical manifestation of cerebral palsy. Methods185 children were divided into the prematurity group(91 cases) and maturity group(94 cases). The μ-test was applied to analyze the incidence of clinical manifestation of 185 children with cerebral palsy for different pathogenic factors. ResultsThe sever symptoms occurred more frequently in prematurity group than in maturity group. ConclusionThe earlier the careful follow-up for children with prematurity was performed, the earlier the diagnosis and treatment of cerebral palsy were achiened.

Objective To investigate the construction of compound membrane with corneal stromal cells and collagen-chitosan by tissue engineering technique and its biocompatibility.Methods Rabbit and human corneal stromal cells were separated and seeded into collagen-chitosan membrane.The compound membrane was transplanted into rabbit corneal stroma.Then the growth condition of keratocytes,the effect on normal keratocytes and degradation of compound membrane were detected by corneal confocal microscope,anterior OCT and histological and immunohistochemical methods ex vivo 1,2,4 weeks after grafting.Results The rabbit and human corneal stromal cells grown well in collagen-chitosan scaffold.The compound membrane degradated gradually after grafting.There was no necrosis and dissolvation.Corneal epithelium,stroma and endothelial cells were all normal.Conclusion Collagen-chitosan can be used as a biological scaffold for construction of corneal stroma.Corneal confocal microscopy and anterior OCT are new methods to observe the biological activity of constructed corneal stroma.

Objective To observe hypoglycemic efficacy of continuous glucose monitoring system (CGMS)combined with continuous insulin infusion(CSII)in the elderly diabetic patients aged 60 years and over.Methods A total 0f 100 elderly diabetic patients were randomly divided into two groups.50 were treated with CGMS or self-monitoring blood glucose(SMBG)and CSII.and whose insulin dose was adjusted based on results of monitoring,and the other 50 with SMBG and insulin pump as controls.whose insulin dose was adiusted based on the results of SMBG for three days.After two weeks.all the patients were monitored by CGMS for blood glucose.Results Mean blood level of glucose and mean amplitude of glycemic excursions(MAGE)were significantly lower in CGMS+CSII group than those in controls[(6.6±2.3)mmol/L vs.(7.5±2.1)mmol/L,and(3.9±0.9)mmol/L vs.(5.1±0.6)mmol/L,respectively,P＜0.05 1.Average insulin dose used was significantly lower in CGMS+CSII group than that in controls [(0.64±0.21)U/kg vs.(0.82±0.41)U/kg,P＜0.05],and duration of hypoglycemia was less in the former than that in the latter[(20±3)min vs.(40± 9)min,P＜0.05 ].Conclnsions CGMS combined with CSII can decrease blood glucose.glucose excursions.average dose of insulin and duration of hypoglycemia in the elderly diabetic patients,as well as prevent occurrence of hypoglycemia.

Objective:To investigate the rationality of prophylactic antibiotics application during perioperative period in the pa-tients with type Ⅱ incision operation in gynecology and obstetrics in our hospital after the intervention. Methods:Totally 274 medical records of obstetrics and gynecology patients with typeⅡincision operation in our hospital from April 2012 to March 2013 were collect-ed as the control group, another 321 medical records of obstetrics and gynecology patients with typeⅡincision operation in our hospital from April 2013 to March 2014 were collected as the intervention group. The rationality of prophylactic antibiotics application before and after the intervention was compared. Results:Compared with those in the control group, the average administration time, the hos-pitalization time and the cost of antimicrobial agents were significantly decreased in the intervention group (P<0. 05). The incidence of adverse drug reactions and operation incision infection between the two groups was not significant difference (P>0. 05). Compared with those in the control group, the incidence of irrational choice of antibacterial drugs, the combined use without indication, irrational medication time,over-time use, inappropriate use and improper choice of solvents in the intervention group was significantly reduced ( P<0. 05). Conclusion:After the intervention, the rationality of prophylactic antibiotics application during perioperative period in the patients with type Ⅱ incision operation in gynecology and obstetrics in our hospital is improved significantly.

Objective To understand the relationship among junior college students' anxious status in testing and studying behaviors and studying result. Methods TAS anxiety scale was used for the evaluation of anxious status, and data were analyzed by SPSS software. Results Statistical significance was observed among different specialties. Sex, socioeconomic status and living environment did not affect TAS score. The higher the anxious status was, the worse the studying results will be. The anxious status was associated with some studying behaviors. Conclusions Anxiety was correlated with specialities and studying behaviors, and affected studying result.

Objective To distinguish the increase of white blood cells after glucocorticoid is the adverse reaction of glucocor‐ticoid or infection aggravated .If we can analyse it properly ,the abuse of antimicrobials and waste of medical could be avoid re‐source .Methods 74 patients which were in our hospital from 2011 .1 to 2012 .12 were recruited into this study .They were divided into three groups ,including non‐infection with glucocorticoid(n=15) ,infection without glucocorticoid(n=29) and infection with glucocorticoid(n=30) according to disease and therapies .After being treated 0 day ,3 days ,7 days ,venous blood of patients were collected to detect white blood count ,CRP and morphology of leukocyte .Results The group of non‐infection with glucocorticoid applied glucocorticoid after 3 days and 7days ,WBC was increased significantly(P<0 .01 ,respectively) .And compare with the group of infection without glucocorticoid ,more band neutrophils could be seen in peripheral blood smear of the other two(P< 0 .05) . While there was no difference on level of CRP between the groups of infection without glucocorticoid and infection with glucocorti‐coid(P>0 .05) .Conclusion Glucocorticoid rise the count of WBC in peripheral blood especially the neutrophil .We can evaluate in‐fectious status by the level of CRP and the count of band neutrophils in peripheral blood smear .

Objective:The study focuses on the distinct distributions of γδT cells in various tissues and the changes after Sal-monella typhimurium infection,and attempts to explore the physiological significance of γδT cell distribution and the role of γδT cells in infectious diseases .Methods:Flow cytometry and PCR technique were used to detect the proportion of different γδ T cell subsets among thymus,spleen,lymph nodes,liver,skin,and intestinal intraepithelial lymphocytes .Flow cytometry was applied to detect the secretion of IFN-γand IL-17a.The changes of various γδT cell subsets in liver and intestinal intraepithelial lymphocytes were analyze after Slamonella typhimurium infection.Res ults: γδ T cells were rich in the intestinal epithelium , skin and liver, but poor in the thymus,spleen and lymph nodes .The distribution of different subsets was quite dissimilar .Vγ5+γδT cells chiefly existed in skin ,and Vγ1+,Vγ4+,Vγ7+γδ T cells largely existed in small intestine.γδ T cells in liver mainly secreted IL-17a;however,γδ T cells in intestinal intraepithelial secreted IFN-γ.After infection by Salmonella typhimurium , the proportion of γδ T cells in intestinal intraepithelial increased significantly ,particularly Vγ1 +γδT cells.In Liver,there was no significant change of total γδT cell ratio,but the ratio of Vγ1 +γδT cells reduced ,Vγ4 +γδT cells raised.Conclusi on:γδT cells are rich in the intestinal epithelium ,skin and liv-er.The distribution of different subgroups has specificity .There are large differences in the ability of cytokine secretion among various subgroups of γδT cells.The distribution of γδT cell subgroups in small intestine and liver changes during Salmonella typhimurium in-fection.

Objective To optimize purification technology of total lignans in Myristicae Semen; To study its anti-inflammatory effects. Methods Macroporous adsorption resin was used for enrichment and purification of total lignans in Myristicae Semen, with adsorption rate, desorption rate and content of total lignans as indexes. The resin type, concentration of eluent, sample weight and dosage of eluen were investigated. Anti-inflammatory effects were studied by determining swelling degree and inhibition rate of mice with ear swelling induced by xylene. Results Purification technology of total lignans was processed on type of AB-8 resin. 1 g crude extract was dissolved as sample; 40 mL preprocessed resin including 16 g resin was added and static adsorpted for 12 hours; 30% ethanol was used firstly to elute to colorless and then 300 mL 70% ethanol was used to elute; the eluent of 70% ethanol was collected. The purity of total lignans was 45.8%. There was statistical significance in ear swelling degree of total lignans in Myristicae Semen high-, medium-, and low-dosage groups compared with model group (P<0.05). Conclusion The purification technology of total lignans is stable and reproducible, and total lignans in Myristicae Semen has significant anti-inflammatory activity.

Objective To investigate the effects of chronic renal failure rabbit serum on proliferation and nuclear factor kappa B (NF-κB) activation of rabbit arterial smooth muscle cells (ASMCs) and to explore the possible mechanism. Methods Rabbit model of chronic renal failure was established by the ligation of renal arterial branches. ASMCs were incubated in the media with various concentrations of chronic renal failure serum cultured in vitro. Cell proliferation was assessed by MTT. Cell apoptosis was detected by Hoechst33342 staining. NF-κB p65 nuclear translocation was analyzed by immunofluorescence. Expression of proliferating cell nuclear antigen (PCNA) and NF-κB p65 proteins in response to chronic renal failure serum in ASMCs was determined by Western blotting. Results Lower concentrations of chronic renal failure serum (≤ 10%) could significantly promot the proliferation of ASMCs in a dose- and time-dependent manner. Higher concentrations of chronic renal failure serum (＞10%) could significantly inhibit the proliferation and induce apoptosis of ASMCs compared to the normal control (P＜0.05). Under the stimulation of lower concentrations of chronic renal failure serum, the expression of PCNA and NF-κB p65 increased significantly compared to the normal control (P＜0.01), while decreased markedly under the stimulation of higher concentrations of chronic renal failure serum compared to the normal control (P＜0.01). Under the stimulation of 10% chronic renal failure serum, nuclear translocation of NF-κB p65 in ASMCs was found. Conclusions Different concentrations of chronic renal failure rabbit serum can effectively induce ASMCs proliferation or apoptosis. The mechanism of promoting proliferation may be mediated by activating NF-κB, which will be useful for the treatment of accelerated atherosclerosis in chronic renal failure.