Objective To explore the relationship between serum placental growth factor (PLGF) and carotid atherosclerosis (CAS) in patients with type 2 diabetes mellitus (T2DM ). Methods Serum PLGF were determined by ELISA in 53 patients with T2DM ,who were divided into T2DM without carotid plague (T2DM group ,n= 27) and T2DM with carotid plague (T2DM + CAS group ,n= 26). Fasting plasma glucose (FPG) ,glycosylated hemoglobin A1c (HbA1c) and some other metabolic variables were also measured. The results were compared to those of 27 healthy controls (NC group). Results Serum PLGF levels were significantly higher in T2DM group and T2DM + CAS group than in NC group (P<0.01). T2DM+CAS group showed significantly higher serum levels of PLGF compared to that of T 2DM group(P<0.01).Correlation analysis showed that serum PLGF was positively correlated with TC ,LDL‐C , hsC‐RP and IL‐6 in T2DM patients (P<0.01). Similarly ,multiple regression analysis showed that LDL‐C and IL‐6 were the independent factors of serum PLGF level (P<0.05). Furthermore ,Serum PLGF was positively associated with carotid plague. Conclusion PLGF is positively related with chronic inflammation factors in T2DM patients. Serum PLGF may play an important role in the occurrence and development of carotid atherosclerosis and other macroangiopathy in T 2DM.

BACKGROUND:The research at home and abroad for appropriate immature myocardial cardioplegia has no breakthrough, and it may be a better mean to improve the protection effect of existing cardioplegia on immature myocardial by adding ingredients. Adenosine can reduce ischemia and neutrophil-mediated reperfusion injury, and salvia miltiorrhiza can reduce the content of ischemic myocardial lipid peroxide and increase the scavenging of myocardial cells to oxygen radical. OBJECTIVE:To investigate the protection effect of improved myocardial protection fluid containing adenosine and salvia miltiorrhiza on the heart, liver and kidney. METHODS:Sixty cases of infant open heart operation of the first time were randomly divided into three groups, 20 cases in each group. The adenosine group received the improved myocardial protection fluid added with adenosine, the combination group received the improved myocardial protection fluid added with adenosine and miltiorrhiza salvia, and the control group received the improved myocardial protection fluid in the same volume. The blood samples were taken at the time before anesthesia induction, 30 minutes after cardio pulmonary bypass beginning, 1 hour after cardio pulmonary bypass end and 24 hours after cardio pulmonary bypass end. The serum levels of the creatine kinase, alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, creatinine, tumor necrosis factor-α, interleukin-6, interleukin-8 and interleukin 10 were detected. RESULTS AND CONCLUSION:The serum levels of the alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, creatinine and creatine kinase in three groups were increased, but the control group was increased significantly (P<0.01), and there were significant differences between adenosine group and combination group (P<0.05);the levels of tumor necrosis factor-α, interleukin-6 and interleukin-8 were significantly increased after operation (P<0.05). The levels of tumor necrosis factor-α, interleukin-6 and interleukin-8 in the control group were significantly higher than those in the adenosine group and combination group before induction and at the same time point (P<0.01);at 3 minutes and 24 hours after cardio pulmonary bypass, the level of interleukin-10 was increased in three groups, but the level of interleukin-10 in the adenosine group was significantly higher than that in the control group (P<0.01). The results indicate that improved myocardial protection fluid has great protection effect on immature myocardial, and can significantly reduce the serum levels of tumor necrosis factor-α, interleukin-6 and interleukin-8, promote the secretion of interleukin-10 that can inhibit inflammatory cytokine and can significantly reduce the inflammatory response caused by cardiopulmonary bypass. So the improved myocardial protection fluid has protection effect on important organs, such as heart, lung, liver and kidney.

Objective To evaluate the effects of acitretin combined with clarithromycin on tumor growth in human oral epidermoid carcinoma xenografts in nude mice,and to investigate their antitumor mechanisms.Methods A cell line of human oral epidermoid carcinoma was subcutaneously inoculated into 31 Balb/c nude mice to establish a xenograft model of human skin tumor.Then,the nude mice were randomly classified into 6 groups according to a double blind protocol:control group (n =6) remaining untreated,placebo group (n =5) treated with wheat flour,acitretin group (n =5) treated with acitretin 7.2 mg/kg per day,clarithromycin group (n =5) treated with clarithromycin 100 mg/kg per day,acitretin + placebo group (n =5) treated with both acitretin (7.2 mg/kg per day) and wheat flour,and acitretin + clarithromycin group (n =5) treated with acitretin (7.2 mg/kg per day) and clarithromycin 100 mg/kg per day.All the drugs were intragastrically administrated once daily.After three weeks of treatment,mice were sacrificed and xenografts were removed.Then,the size and weight of xenografts were measured,and pathological analysis was conducted.Real time-PCR was performed to quantify the mRNA expressions of vascular endothelial growth factor (VEGF) and nuclear factor (NF)-κB,and immunohistochemistry was carried out to observe the expression of VEGF as well as to determine microvessel density (MVD) and Ki-67 proliferation index.By using the software SPSS 19.0,analysis of variance was performed for comparison of measurement data,and least significant difference (LSD) test for paired comparisons.Results Both the size and weight of xenografts in the acitretin + clarithromycin group were significantly lower than those in the other groups (all P ＜ 0.05).Real-time fluorescence-based PCR revealed weaker mRNA expressions of VEGF and NF-κB in the acitretin + clarithromycin group compared with the control group,clarithromycin group and acitretin group (all P ＜ 0.05).As immunohistochemistry showed,the acitretin + clarithromycin group displayed a decrease in the expression rate (all P ＜ 0.01) and staining intensity of VEGF,MVD (all P ＜ 0.01) with a sparse distribution of microvessels,Ki-67 proliferation index (all P ＜ 0.05) and proliferative activity of tumor cells compared with the control group,clarithromycin group and acitretin group.Conclusion Acitretin combined with clarithromycin can synergistically inhibit the growth of human oral epidermoid carcinoma xenografts in nude mice,downregulate VEGF expression,and suppress angiogenesis and tumor proliferation.

ObjectiveTo observe the clinical efficacy of relaxing needling plus joint needling from the Internal Medicine in treating post-stroke spastic hemiplegia.MethodTotally 121 eligible subjects were randomized intoan ordinary acupuncture group (61 cases) and a relaxing-joint needling group (60 cases). The two groups were treated for 2 weeks, 5 sessions each week. The spasm of the lower limb was evaluated by using Fugl-Meyer Assessment Scale (FMA), modified AshworthScale (MAS), Tardieu Scale, and Activities of Daily Living (ADL) before and after treatment. Besides, 4 patients from each group were randomly selected for the three-dimensional gait analysis, and the dorsi-extension, range of motion (ROM) of ankle, strephenopodia, and heel-raising height were observed.Result and ConclusionRelaxing-joint needling and ordinary acupuncture both can produce significant efficacies in treating post-stroke spastic hemiplegia. The relaxing-joint needling produced more significant efficacies in improving the body balance, ROM, muscular tension, and lower limb spasticity compared to ordinary acupuncture. It’s plausible that relaxing-joint needling improves the strephenopodia and hemiplagia gait through renovating body balance, ROM, muscular tension, and lower limb spasticity. Since the three-dimensional gait analysis can analyze the dorsi-extension, strephenopodia, and heel-raising height, thus it can be used to evaluate the change of the spastic gait of post-stroke hemiplegia in amore objective, elaborate, and correct way.

AIM: To screen the lentiviral vector carrying siRNA with higher efficiency of suppressing the sphingosine-1-phosphate receptor 2 (S1P2) gene expression in the primarily cultured corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).METHODS: SHR and SD rats (n=5 each) were used for primarily culturing corpus cavernosum smooth muscle cells.The cells were randomly divided into 6 groups: SHR siRNA-1, SHR siRNA-2, SHR siRNA-3, SHR GFP, SHR control (SHR non-transfection group), and SD control (SD rat control group).Each group had 5 samples with 1.0×105 cells of each sample.At 72 h after transfection (MOI=60) with lentiviral vectors carrying S1P2 siRNA into the SHR corpus cavernosum smooth muscle cells, the expression of GFP was observed under fluorescence microscope.The protein expression of S1P2, ROCK1, ROCK2 and eNOS in the corpus cavernosum smooth muscle cells, and the mRNA expression of S1P2, ROCK1 and ROCK2 were determined by by Western blot and RT-PCR.RESULTS: The transfection efficiency of the corpus cavernosum smooth muscle cells in SHR siRNA-1, SHR siRNA-2, SHR siRNA-3 and SHR GFP groups were>80%.Compared with SHR control group, the mRNA levels and the protein expression of S1P2, ROCK1 and ROCK2 in SHR GFP group showed no remarkable changes, while those in SHR siRNA-1, SHR siRNA-2, SHR siRNA-3 and SD control groups were significantly lower than those in SHR control group (P<0.05).The protein expression of eNOS in SHR siRNA-1, SHR siRNA-2, SHR siRNA-3 and SHR GFP groups were not significantly changed as compared with SHR control group, but that in SD control group was significantly higher than that in SHR control group.CONCLUSION: Three groups of siRNA lentiviral vectors targeting S1P2 inhibit the expression of S1P2 in the corpus cavernosum smooth muscle cells of SHR, and by silencing the S1P2 expression, the expression of ROCK1 and ROCK2 is inhibited.Among them, siRNA-1 has the highest inhibitory efficiency.

Objective To investigate the relationship between the serum apolipoprotein B (ApoB)/ ApoA-Ⅰ ratio and intracranial artery stenosis in young patients with ischemic stroke.Methods Patients with ischemic stroke aged from 18 to 45 were enrolled in the study.Brain CT angiography was used to evaluate the degree of intracranial artery stenosis,and the concentrations of serum total cholesterol,triglyceride,highdensity lipoprotein cholesterol,low-density lipoprotein cholesterol,ApoA-Ⅰ,and ApoB were detected.The ratio of ApoB/ApoA-Ⅰ was calculated.The Demographic and clinical characteristics of the intracranial artery stenosis group and the non-intracranial artery stenosis group were compared.Multivariate logistic regression analysis was used to identify the independent risk factors for intracranial artery stenosis in young patients with ischemic stroke.Results A total of 161 young patients with ischemic stroke were enrolled,including 89 in the intracranial artery stenosis group and 72 in the non-intracranial artery stenosis group.The constituent ratios of diabetes mellitus (20.2％ vs.6.9％;x2 =4.641,P =0.032),smoking (47.5％ vs.15.2％;x2 =15.121,P=0.001),hyperlipidermia (56.1％ vs.48.6％;x2 =4.197,P=0.040),as well as the radios in serum high-density lipoprotein cholesterol (1.29 ± 0.30 mmol/L vs.1.65 ± 0.34 mmol/L;t =7.131,P=0.002),ApoA-Ⅰ (1.49 ± 0.65 g/L vs.1.63 ± 0.23 g/L;t =2.751,P =0.001),ApoB (1.49 ± 0.65 g/L vs.1.63±0.23 g/L;t=2.751,P=0.001),and ApoB/ApoA-Ⅰ ratio (1.49±0.65 vs.1.63± 0.23;t =2.751,P=0.001) had significant differences between the two groups.Multivariate logistic regression analysis showed that diabetes (odds ratio [OR] 3.052,95％ confidence interval [CI] 1.186-7.856;P =0.021),smoking (OR 2.997,95％ Cl 1.456-6.172;P =0.003),hyperlipidemia (OR 4.745,95％ CI 2.108-10.668;P =0.001),ApoB (OR 4.861,95％ CI 3.029-7.802;P=0.001),and ApoB/ ApoA-Ⅰ ratio (OR 5.684,95％ CI 2.215-14.584;P=0.002) were the independent risk factors for intracranial artery stenosis in young patients with ischemic stroke,while HDL-C (OR 0.561,95％ CI 0.354-0.888;P=0.014) and ApoA-Ⅰ (OR 0.065,95％ CI 0.010-0.409;P=0.004) were the independent protective factors.After adjustment for hypertension,diabetes,smoking,hyperlipidemia,HDL-C,ApoA-Ⅰ,and ApoB,ApoB/ApoA-Ⅰ ratio was still an independent risk factor for intracranial artery stenosis in young patients with ischemic stroke (each increase of 1 standard deviation,OR 4.255,95％ CI 2.348-7.711;P=0.001).Conclusion ApoB/ApoA-Ⅰ ratio is an independent risk factor for intracranial artery stenosis in young patients with ischemic stroke.

Objective To clarify the role of MafA gene in development of MODY (maturity onset diabetes of the young) by studying insulin production,gene expression,and serum glucose level in heterozygous MafA gene knockout mice.Methods C57BL/6J mice were used as control animals,MafA gene heterozygous mice were analyzed.The distribution curve of blood sugar levels over time and serum insulin of heterozygous mice were determined by using IPGTT.The sensitivity of the mice to insulin was examined by injecting insulin assay.The expression levels of genes of MafA,insulin,pdX1,Beta2,and other genes of heterozygous mice were analyzed by semi-quantitative RT-PCR.Morphological changes in pancreatic tissue and α-and β-cell counts were obtained by using immunofluorescence/histological examination.Results (1) Two weeks after birth,MafA gene heterozygous mice began to show that the blood glucose level was increased,weight was reduced,and the amount of insulin secretion was clearly decreased (P＜0.05 or P＜0.01) while the insulin sensitivity did not change significantly.(2)The islet volume in MafA gene heterozygous mice was increased significantly as compared with the control group.However there were no significant changes in the number of pancreatic cells,distribution patterns,and the ratio of α and β cell.(3) Semi-quantitative RT-PCR detection showed that,compared with the control group,MafA gene level,the amount of insulin and Beta2 gene in MafA gene heterozygous mice were significantly reduced(all P＜0.05),while no changes in the amount of glucagons and level of Pdx1 were found.Conclusions The blood glucose level of MafA gene heterozygous mice was raised early after birth.MafA gene is likely to be a new disease ralated gene of MODY.

Objective:The immunological effects of HCV-DNA vaccine with different adjuvants were detected by ELISPOT in mice.Methods:Female BALB/c mice were primed with naked HCV-DNA, HCV-DNA encapsulated by liposome DDAB/EPC or DC-Chol/DOPE, HCV-DNA mixed with Montanide ISA 720 or aluminum hydroxide, respectively, and boosted twice accordingly in a four-week interval. Cytokine production by splenocytes was assessed by ELISPOT.Results:In most cases, splenocytes from mice vaccinated with DDAB/EPC liposome produced more IFN-?. These splenocytes also have significant higher IL-2 production compared with the other groups. In expansion with NS5b, splenocytes from alum group have significance in IL-4 production compared with other groups. The profile of cytokine production revealed that the INF-? overwhelmed IL-4 in naked DNA, DDAB/EPC, and DC-Chol/DOPE groups while IL-4 surmounted IFN-? in alum and Montanide groups.Conclusion:Encapsulation with liposome DDAB/EPC has the strongest adjuvant effect in inducing Th1 dominated immunity. Alum and Montanide can convert the Th1 nature of DNA vaccine to Th2-biased immunity.

Objective To evaluate the safety and efficacy of bipolar plasmakinetic system in exfoliative tran-surethral resection of bladder tumors .Methods Clinical data of 72 patients with non muscle invasive bladder cancer (NMIBC) were retrospectively analyzed.Transurethral bipolar plasmakinetic system was used ,30°viewer,F27 outer sheath was pushed off and bladder tumor was cut .When the bladder filling state ,pushed off bladder mucosa distance from tumor basal 2cm,then electricity cut the exfoliative bladder tumor .When bladder half filling state electricity cut the base of the bladder muscle layer of bladder tumor .Results This group of 72 cases were successfully completed surgery,surgery time 37~93 min,without intraoperative bladder perforation ,slight obturator nerve reflex in 5 cases. The keeping intact pathologic specimens was good for pathological staging .Conclusion Exfoliative transurethral resection of bladder tumors with bipolar plasma is a safe , practical and effective way of operation , which can avoid severe obturator reflex occurred in the operation , and greatly reduce the occurrence of bladder perforation , without TURS,surgical removal of the pathological specimens is specification .

Objective To investigate the neuroprotective effect of lateral intracerebroventricular injection of Apelin-13 on the YAP expression and the cerebral ischemia/reperfusion (I/R) injury in Wistar rats.Methods Healthy adult male Wistar rats were randomly divided into the sham group, cerebral I/R group and Apelin-13 treatment group.The middle cerebral artery occlusion model was established with ischemia for 1 hour and reperfusion for 24 hours after restricting food and water intake for 12 hours.Apelin-13 was injected into rats' lateral ventricles of Apelin-13 treatment group after reperfusion.Neural function defects was assessed.The volume of infarction was evaluated by TTC staining.The expression levels of YAP were detected by western blot.Results Compared with the cerebral I/R group,the rats in the Apelin-13 treatment group had abetter neurologic score ((2.67±0.33) vs (1.67±0.33) , P＜0.05), the infarction volume was decreased ((30.60± 1.42) ％ vs (23.70± 2.20) ％,P＜0.05) , and YAP expression level was increased in each part of the cerebral tissue(P＜0.05).Conclusion Apelin-13 has a neuroprotective effect,which plays the therapeutic effect by regulating the expression of YAP on cerebral ischemia/ reperfusion (I/R) injury in Wistar rats.