Objective To explore the related factors of atherosclerosis in senile obstructive sleep apnea hypopnea syndrome (OSAHS) patients.Methods From May 2012 to January 2014,129 elderly hypertension and type 2 diabetes patients were recruited.According to the presence or absence of OSAHS,they were divided into OSAHS (n =60) and non-OSAHS (n =69) groups.And 30 physical examination subjects were selected as control group.The parameters of age,gender,height,weight,high sensitivity C-reactive protein (Hs-CRP),homocysteine (Hcy),triglyceride (TG),total cholesterol (TC),high density lipoprotein-cholesterol (HDL-C),low density lipoprotein-cholesterol (LDL-C) and intimate-media thickness of carotid artery (CIMT) were measured with ultrasonography.And their levels were compared among OSAHS,non-OSAHS and normal control groups.And the correlations were analyzed between CIMT and Hs-CRP,Hcy,TG,TC,HDL-C,LDL-C,age and body mass index (BMI).Results The levels of Hs-CRP[(3.40 ± 0.91) mg/L,(2.57 ± 1.01) mg/L],Hcy [(16.50 ± 2.89) μmol/L,(11.20 ± 2.32) μmol/L],CIMT[(1.06 ±0.14) mm,(0.93 ±0.13) mm],TG[(2.52 ± 1.40) mmol/L,(2.51 ±2.02) mmol/L],TC[(5.74 ± 1.16) mmol/L,(5.49 ± 1.09) mmol/L] and LDL-C[(3.15 ±0.47) mmol/L,(3.05 ±0.70) mmol/L] in both OSAHS and non-OSAHS groups were higher than those in normal control group [Hs-CRP:(2.06 ± 1.22) mg/L,Hcy:(6.04 ± 1.85) μmol/L,CIMT:(0.65 ± 0.07) mm,TG:(0.99±0.24) mmol/L,TC:(3.63 ±0.71) mmol/L and LDL-C:(2.47 ±0.27) mmol/L] (OSHAS group vs.control group,t =5.88,18.15,15.62,5.91,9.17,7.47,non-OSAHS group vs.control group,t=2.20,10.73,11.51,4.09,8.56,4.45,all P＜0.05).The levels of Hs-CRP,Hcy,CIMT and plaque score were higher in OSAHS group than those in non-OSAHS group [(1.92 ± 1.03) vs.(1.35 ± 0.97) points] (OSHAS group vs.non-OSAHS group,t =4.84,11.72,5.54,3.91,all P ＜0.05).CIMT or carotid artery plaque were related with age,BMI,TG,TC,LDL-C,Hs-CRP and Hcy in OSAHS group.Conclusions OSAHS may elevate the level of inflammatory factors and cause atherosclerosis.And the changes of CIMT,Hs-CRP and Hcy are parallel.Hcy also has important clinical significance in the progression of atherosclerosis.

Lycopus lucidus is a herbal drug that can invigorate blood circulation,resolve blood stasis and improve microcirculation. In our further study of its pharmacological function and to identify its effective components,its effect on the hemorrheology of rat was used as a guide to evaluate the relative strength of four different extraction fractions of the drug. Results showed that fraction L. F04 and L. F02 were effective in reducing the blood viscosity,with L. Fo4 showing the best effect. This function of L. F04 was believed to be related to its ability to inhibit platelet aggregation and to improve the compostion of plasma

Objective To study the clinical efficacy of intra-arterial thrombolysis with recombinant tissue plasminogen activator (rt-PA) for the treatment of ischemic cerebrovascular disease caused by cerebral thrombosis. Methods A total of 245 patients accepted by our hospital during May 2013 and July 2015 were divided into the observation group (n=148) and the control group (n=97). All patients were given conventional process for controling blood pressure and blood lipids. Patients in observation group received intra-arterial thrombolysis with rt-PA, while patients in control group accepted conventional treatment. At the time of admission, the demographic characteristic, vascular influencing factors, baseline clinical findings, laboratory findings and neurological deficits were collected. The improvement of neurological function was evaluated by the modified Rankin scale 3 months after treatment. The levels of fibrinogen (FIB), D-Dimer, activated partial thromboplastin time (APTT) and thrombin time (PT) were measured before and 24 h after the treatment. Results There were no significant differences in demographic characteristic and general clinical data between the two groups ( P>0.05). The proportion of patients with improved neurological function was significantly higher in observation group than that of the control group (83.11％vs. 53.61％, P<0.05). There were no significant difference in coagulation index and fibrinolysis index before treatment between the two groups (P>0.05). Twenty-four hours after the treatment, the levels of FIB, D-Dimer, APTT and PT were significantly improved in the observation group compared with those before treatment. The level of FIB was significantly decreased, D-Dimer was significantly increased, APTT and PT were significantly prolonged in observation group compared with those of control group (P<0.05). Conclusion The rt-PA can effectively dissolve thrombosis and correct the coagulation system and fibrinolytic system.

BACKGROUND:The influence of Tol-like receptor 2 (TLR2), Tol-like receptor 4 (TLR4), Tol-like receptor 6 (TLR6) signal transduction pathway and active Treg in children with Henoch-Schonlein purpura has been unknown.
OBJECTIVE:To investigate the expression of TLR2, TLR4 and TLR6 in peripheral blood mononuclear cells and Treg immune response in patients with Henoch-Schonlein purpura, and to explore the role of TLR2, TLR4, TLR6 and Treg activation in the pathogenesis of Henoch-Schonlein purpura.
METHODForty-two hospitalized children with Henoch-Schonlein purpura were enrol ed in this study. Another 15 healthy children were selected as controls. TLR2, TLR4 and TLR6 at protein level in peripheral blood mononuclear cells were detected by flow cytometey;reverse-transcription PCR and real-time PCR were used to evaluate the level of MyD88;the levels of transforming growth factor-βand interleukin-10 were measured by enzyme-linked immunosorbent assay. t-test or t’-test was used to compare the levels of these genes and proteins. Pearson’s correlation test was done for correlation analysis.
RESULTS AND CONCLUSION:Compared with the control group, the protein levels of TLR2, TLR4, TLR6 and the relative expression level of MyD88 mRNA were significantly up-regulated (P<0.01). The serum levels of transforming growth factor-βand interleukin-10 were higher in the Henoch-Schonlein purpura children than the healthy children (P<0.05). There was a significant correlation between the protein levels of TLR2, TLR4, TLR6 and mRNA level of MyD88 (P<0.01), but no relationship was found between TLRs and interleukin-10, transforming growth factor-β(P>0.05). The excessive activation of TLR2, TLR4, TLR6 may be involved in the process of Henoch-Schonlein purpura via MyD88-dependent pathway, and the compensatory activation of Treg may participate in protective immunity.

AIM: To study the pharmacodynamics of Yajiao Hadun San(Vitex trifolia;streptocaulon griffibhii;Asparagus pseudofilicinus;Inula cappa;Benincasa hispida;Tacca esquirolli). METHODS: The pyrexia model rat induced with active yeast,the mice's auricular swelling test caused by croton oil;the mice's toe swelling test caused by albumen;the mice's twisting body test caused by the irritation of acetic acid and the mice's carbon clearance test were used to determine antipyretic,analgesic,anti-inflammation effect and immunity function of Yajiao Hadun San. RESULTS: Yajiao Hadun San could reduce the anus temperature of the pyrexia model rat,inhibit the mice's toe and auricular swelling,reduce the times of twisting body sharply and also increase the rate of eliminating colloid carbon in mice. CONCLUSION: Yajiao Hadun San has certain antipyretic,anti-inflammation,analgesic effect and enhance the phagocytosis of mono-karyon macrophage in mice.

Objective To explore the transfection efficiency of primary lymphocytes from human peripheral blood by different methods to acquire the method with higher transfection efficiency.Methods Mononuclear cells from human peripheral blood were isolated using Ficoll-Hypaque.Cell viability was detected by Trypan blue staining.Suspending lymphocytes were sucked out and were incubated in 24-well plate after cultured in 6-well plate for 2 h.Activated lymphocytes were transfected by electroporation with plasmid(PEGFP-N1).Resting or activated lymphocytes were transfected by lentivirus vector(LVGFP) single infection or repeated infection,respectively.Green fluorescence protein (GFP) was detected under the fluorescence microscopy and percentage of positive cells was checked by flow cytometry at different time points after infection.At the same time,the effectiveness of lentivirus infection was compared under different conditions.Results Purity of mononuclear cells isolated by Ficoll-Hypaque was 95 ％ and its viability was over 95 ％.The percentage of lymphocytes obtained with a uniform shape was 90 ％-95 ％.Scattered fluorescence was observed by electroporation under the conditions of voltage 2 100 V,pulse width 10 ms,pulse number 1 for lymphocyte,while fluorescent became weaker over time and no green fluorescent was observed after transfection for 72 h.After resting lymphocytes were infected once for 48 h by lentivirus vector,green fluorescent was not found and positive cells were less than 1％.1％-5 ％ of activated lymphocytes could express GFP after single lentivirus infection and the expression levels were enhanced with concentration increasing,while 5 ％-10 ％ of activated lymphocytes showed strong green fluorescent by repeated lentivirus infection.In contrast with electroporation,the fluorescent with lentivirus infection was stronger over time.Conclusion Repeated lentivirus infection could efficiently transfect exogenous genes into activated lymphocytes for stable expression.

Objective To establish a method for detection of gene mutations in β-thalassemia by high-resolution melting (HRM) and study its preliminary clinical application.Methods Two common mutations [ IVS-2-654 ( C ＞ T ) and -28 ( A ＞ G ) ]of β-thalassemia in Wenzhou city population were selected.The plasmid DNA fragments of these mutations were constructed by TA clone technology as PCR templates or genotyping controls.A method for detection of β-thalassemia gene mutations based on HRM analysis was established and its specificity,sensitivity and repeatability were methodologically evaluated.One hundred and seventeen patients with clinically suspected β-thalassemia from Second Affiliated Hospital and Yu ying Children's Hospital of Wenzhou Medical College were enrolled into this study.The genomic DNA was extracted from whole blood cells and detected by HRM method.The results were compared with the direct sequencing data.Results HRM method could detect the mutations [ IVS-2-654( C ＞ T) and -28 ( A ＞ G ) ]of β-thalassemia and the results did not show any non-specific amplified fragments.All within-run and between-run coefficients of variation for different DNA types' Tm were smaller than 0.1％.And minimum 103 copies of DNA of each assay and 10％ mutation could be determined by this method.One hundred and seventeen patients with clinically suspected β-thalassemia were detected with HRM and all the results were in accordance with direct DNA sequencing.There were 45 IVS-2-654 ( C ＞ T)heterozygous mutation and 9-28 ( A ＞ G)heterozygous mutation and none homozygous mutation.Conclusion The method of rapid identification of β-thalassemia gene mutations based on HRM analysis is successfully established,which is a convenient,rapid,specific,sensitive and accurate technique for screening gene mutations in β-thalassemia as well as a general technical platform to identify other gene mutations.

Objective To investigate the expressions of CXCR4,MMP-2 and VEGF in pancreatic carcinoma and the relationship with biological behavior of pancreatic carcinoma.Methods Immunohistochemical PV6000 technique was used to detect the expression of CXCR4,MMP-2 and VEGF in 47 cases of pancreatic carcinoma tissues,and the relationship between the expressions and pathologic parameters,and the relationship among the expressions of the 3 proteins were analysed.Results The expression rates of CXCR-4,MMP-2 and VEGF were 72.3%,66.O%and 61.7%in pancreatic carcinoma tissues,and all these proteins were correlated with metastasis,clinical staging and prognosis(X2=5.84～12.69;P＜0.05).The expression of CXCR-4,MMP-2 and VEGF was not correlated with sex,age,tumor size and differentiation(X2=0.03～4.27;P＞0.05).There was a positive relationship between the expression of CXCR-4 and MMP-2 or VEGF in pancreatic carcinoma(r=0.587,0.521;P＜0.01).Conclusions CXCR-4 could up-regulate the expression of MMP-2 and VEGF in pancreatic carcinoma,and they may contribute to metastasis and invasion of pancreatic carcinoma.

Objective:To investigate the sensitivity and the specificity of scorpions amplification refractory mutation system ( ARMS) in comparing with that of direct DNA sequencing in the detection of BRAF gene mutations in patients with papillary thyroid microcarcinoma.Methods:Direct sequencing and ARMS were used simultaneously to detect BRAF mutation status in 56 patients with PTMC.Results:BRAF mutations were identified in 46 cases with a mutation rate of 82.9%by ARMS,while in 18 cases with a mutation rate of 32.1%by direct sequencing.Besides,the sensitivity of ARMS was 100%and that of direct sequencing was 39.1%.There were significant differences of both mutation rate and sensitivity between two methods ( P<0.01 ).Conclusion: Compared to direct sequencing,ARMS gains a higher sensitivity in the detection of BRAF mutations in samples with tiny lesions.