Macrovascular complications are the main death or disabling causes of diabetic patients.In recent years,the results of a number of clinical trials aimed at preventing diabetic macrovascular complications have been unveiled.Comprehensive and systemic analysis of these results may give great enlightenment to the clinicans,as well as promote the work in preventing diabetic macroangiopathy in China.

Objective:To clone the mice interleukin-23(mIL-23) and express it in Pichia efficiently.Methods:Two subunits of IL-23 were amplified by PCR from pcDNA3/mIL-23,and ligated together with adaptor by over-PCR.The IL-23 cDNA confirmed by sequencing was inserted into expressing vector pPICZαA and expressed in Pichia X-33 strain.IL-23 protein expression was induced by methanol and ammonia.The recombinant IL-23 was identified by immunoblot and its biological activity was analyzed.Results:DNA sequencing confirmed that cloned cDNA was identical to the published sequence of mIL-23.The recombinant plasmid pPICZαA/mIL-23 was electroprated into X-33.An expected 72 kD protein of mIL-23 was founded both in the induced pellets and supermatants by SDS-PAGE and coomassie blue staining.The 72 kD protein could be recognized in Western blot.While we could detect bands at 72 kD in cell pellets induced more than 24 h by Western blot.Detected by Western blot using anti-His antibody,we could see positive band at 72 kD from supernatants induced 24 and 36 h.While we could detect band at 72 kD in cell pellets induced between 24 h and 96 h.Meanwhile,we could see obviously proliferation of mice peripheral blood mononuclear cells(PBMC) treated with the induced pellets and supermanants.Conclusion:We have successfully expressed mIL-23 protein in Pichia and the expressed product has its bioactivity in promoting the proliferation on PBMC.

Objective To explore the influence of levothyroxine(L-T4) in the treatment of primary hypothyroidism on myocardial enzymes and lipids.Methods 78 patients with primary hypothyroidism were selected and treated with L-T4 for 12 weeks.The fasting serum levels promote thyroid hormone(TSH),free T3 (FT3),free T4 (FT4),total cholesterol (TC),triglycerides (TG),low-density lipoprotein cholesterol (LDL-C),aspartate aminotransferase enzyme (AST),creatine kinase (CK) and its isoenzyme (CK-MB),lactate dehydrogenase(LDH) were monitored and analyzed before and after treatment.Results After L-T4 treatment for 12 weeks,compared with before treatment,TSH,TC,TG,LDL-C,CK,CK-MB,LDH,AST were significantly decreased or restored (t =10.5223,26.8498,22.7699,16.2735,22.9329,13.1910,32.0907,22.9597,all P ＜ 0.01).The FT3 was negatively correlated with TG (r =-0.3782),LDL-C(r =-0.3506),AST(r =-0.2843),LDH(r =-0.2974),CK(r =-0.3726) (all P ＜ 0.01)and CK-MB(r =-0.2559) (P ＜ 0.05).FT4 was negatively correlated with TC (r =-0.2660),TG (r =-0.4661),LDL-C(r=-0.5119),LDH(r=-0.5936),CK(r=-0.4877),CK-MB(r=-0.5463) (all P＜0.01)and AST(r =-0.2328) (P ＜0.05).TSH was positively correlated with TC(r =0.5341),TG(r =0.7567),LDL-C(r =0.8240),AST (r =0.3923),LDH (r =0.8073),CK (r =0.9661),CK-MB (r =0.7336) (all P ＜0.01).TSH had,the best correlationship with CK (r =0.9661).Conclusion L-T4 can significantly improve the thyroid function and reduce the blood lipids,myocardial enzymes levels of patients with primary hypothyroidism.

Objective To study the changes of diamine oxidase (DAO)levels in neonates with hypoxic -is-chemic encephalopathy (HIE)treated with selective brain hypothermi,also to further evaluate Scores for Newborn Gastrointestinal Functior (SNGF).Methods 60 newborns with moderate and severe HIE who were in the NICU of our department from June 2013 to December 2014 were collected.The 60 newborns were randomly divided into hypo-thermia treatment group (HG)and conventional treatment group(CG).The serum was collected and ELISA method was used to test the consistency of DAO on admission and 7 days afterwards,respectively.Meanwhile,the SNGF level of the two groups at the two time points was compared.Results Neither the DAO and SNGF level of the two groups had statistical difference on admission(all P >0.05).Seven days later,both the DAO of the two groups and the SNGF decreased[(12.51 ±1.53)u/mL vs (7.88 ±1.87)u/mL,however,the variation of the hypothermia treatment group was apparently more significant than the change of the conventional treatment group(P =0.011).The SNGF scores of the two groups all decreased,while the hypothermia treatment group was significantly lower than the conventional treat-ment group,the difference were statistically significant (P =0.044,0.006,0.013).Besides,there was remarkably positive correlation between serum DAO level and SNGF (r1 =-0.825,r2 =-0.876,all P <0.05).Conclusion Hypothermia treatment could effectively reduce the injury of asphyxia neonatal gastrointestinal tract by inhibiting the level of DAO,thus improve the gastrointestinal function.

Serum p-amyloid peptide(Aβ)40 and Ap42 levels in patients with type 2 diabetes mellitus (T2DM)and atherosclerosis(AS)were detected by ELISA.The results showed that serum Ap40 level in T2DM group was significantly higher than that in the control group[(274.70±159.51 vs 162.63±87.58)pg/ml,P<0.05],especially in the diabetic patients accompanied with AS[(616.95±195.13)pg/m],P<0.01].Serum Ap40 level in simple AS group was also higher than that in control group[(318.52± 188.65)pg/ml,P<0.05].These results suggest that Ap40 is a risk factor of T2DM complicated with AS.However,there was no difference in serum Ap42 levels among various groups.

Objective To study the impairment and the expression of receptor of advanced glycation endproduct (RAGE) in cultured rat glomerular mesangial cells ( GMC ) induced by constant and intermittent high glucose, and to investigate the pathogenesis of diabetic nephropathy. Methods After being cultured under constant and intermittent high glucose with different concentrations for 24 and 48 hours, the morphological changes of rat mesangial cells were observed, the proliferation of GMC was detected by MTT assay, the activity of superoxide dismutase (SOD)and the level of malondialdehyde (MDA)in supernatant were measured by spectrophotometer,and the expressions of RAGE mRNA were evaluated by RT-PCR. Results ( 1 ) Compared with the control group,the cellular morphology was changed in case of constant and intermittent high glucose. The damage of GMC with intermittent high glucose concentrations was more serious. (2)The activity of SOD was decreased and the level of MDA was raised in case of intermittent high glucose concentrations compared with the constant high glucose concentrations (P＜0.05). (3)The expression of RAGE mRNA with intermittent high glucose concentrations was significantly higher than that with constant high glucose concentrations ( P＜0. 01 ). Conclusions The damaging effects and increased expression of RAGE in cultured rat GMC induced by blood glucose fluctuation was much worse than that with constant high glucose. The blood glucose fluctuation may be one of the causes that induce diabetic nephropathy.

Objective To evaluate the effect of therapeutic communication on negative emotion and quality of life of stoke patients. Methods A total of 120 stroke patients were divided into the control group and the observation group equally. The former received routine nursing and the latter routine nursing plus therapeutic communication. After 4 weeks, they were evaluated by self-rating anxiety scale (SAS), self-rating depressive scale (SDS), trait coping style questionnaire (TCSQ) and the short-form health survey-36 (SF-36). Results After the intervention, the scores on anxiety, depression and negative coping in the observation group were lower than those in the control group (P<0.01), while the scores on positive coping and quality of life were higher in the observation group than those in control group (P<0.01). Conclusion Therapeutic communication can effectively improve stroke patients′negative emotion and quality of life.

Objective To study the changes and significance of plasma atrial natriuretic peptide (ANP),endothehn-1 (ET-1),von Willebrand factor (vWF) levels in newborns with persistent pulmonary hypertension (PPHN) after the treatment of sildenafil.Methods Sixty-six cases with PPHN group and 40 cases with non-PPHN (control group) were enrolled.PPHN group was in the treatment of sildenafil.Collected the blood when before the treatment of sildenafil and 3,7 d after treatment,respectively.Arterial blood gas were done and pulmonary arterial systolic pressure (PASP) was measured before treatment and 3,7 d after treatment,and by the same time recording pulse oxygen saturation (SpO2).Plasma ANP,ET-1,vWF levels were measured by ELISA method.Results The levels of PASP,SpO2 arterial partial pressure of oxygen (PaO2),arterial partial pressure of carbon dioxide (PaCO2) and plasma ANP,ET-1,vWF in PPHN group before treatment [(66.5 ± 13.4)mm Hg (1 mm Hg =0.133 kPa),0.726 ±0.531,(46.3 ±7.2)mm Hg,(59.2 ± 7.4) mm Hg,(272.6 ± 20.3)ng/L,(221.3 ± 24.3) ng/L,(142.5 ± 20.3)％] compared with controlgroup [(25.0±6.2) mm Hg,0.896 ± 0.767,(88.3 ±7.6) mm Hg,(41.1 ±6.1) mm Hg,(68.4 ± 7.9) ng/L,(39.8 ± 6.5) ng/L,(95.3 ± 18.5)％] were statistically significant(P ＜ 0.05).Their levels in PPHN group 3 dafter treatment[(48.3 ± 3.2) mm Hg,0.841 ± 0.416,(73.6 ± 9.3)mm Hg,(50.5 ± 7.2) mm Hg,(102.6 ±20.3) ng/L,(79.6 ± 15.2) ng/L,(103.6 ± 14.1)％] were significantly improved,there was significantdifference compared with before treatment and control group(P ＜ 0.05).Their levels in PPHN group 7 d aftertreatment [(25.2 ± 3.6) mm Hg,0.882 ± 0.724,(85.4 ± 7.4) mm Hg,(40.2 ± 6.4) mm Hg,(64.4 ± 3.6)ng/L,(37.3 ± 5.4)ng/L,(92.9 ± 11.7)％] were significantly improved,there was significant difference compared with 3 d after treatment (P ＜ 0.05),the difference was no statistically significant compared with control group (P＞ 0.05).Linear correlation analysis showed that ANP,ET-1,vWF and PASP,PaCO2 were significantly positively correlated (P ＜ 0.01),ANP,ET-1,vWF and SpO2,PaO2 were significantly negatively correlated (P＜ 0.01).On the basis of cardiac ultrasound monitoring PASP,ANP evaluation of the efficacy of sildenafil sensitivity was 82.2％,specificity was 83.4％ ;ET-1 was 86.4％ and 87.6％; vWF was 85.1％ and 84.7％.Conclusion ANP,ET-1,vWF may play an important role in the mechanism of the treatment of PPHN by sildenafil,and could be used as an objective index to evaluate the effect of sildenafil on PPHN.

This study was designed to explone whether catalpol exerts a protective effects on kidney of diabetic rats and its possible mechanism.Insulin-like growth factor- I ( IGF- I ) and protein kinase B (PKB/Akt)mRNA and protein expression in kidney of diabetic rats were up-regulated as results of assessments by RT-PCR,immunohistochemistry and Western blot.Catalpol could partially reduce IGF- Ⅰ and Akt expressions in kidney of diabetic rats,and thus alleviate the damage of kidney.

AIM: Our study focused on investigation of tissue specific regulatory activity of the newly cloned promoter: PLUNC-p with driving enhanced green fluorescent protein ( EGFP ). METHODS: Transgenic Xenopus Laevis system was applied.RESULTS: The green fluorescence protein directed under PLUNC-p was expressed strictly in branchial arches and epidermis of Xenopus Laevis embryos while CMV promoter showed ubiquitous regulation characteristic.CONCLUSION: PLUNC-p is able to direct epithelia specific expression of EGFP . This property of PLUNC-p might raise the possibility that lead target genes to express tissue-specifically.