Objective To explore the clinical and molecular biological characteristics of spinocerebellar ataxia type 3(SCA3).Methods Clinical manifestation and brain MRI data of 12 patients with SCA in two families were analyged.The polymorphic CAG repeated time in the encode region of SCA3,SCA1 and SCA7 genes were compared in 15 family numbers without abnormal presentations,and 12 healthy persons of controls.Results Among 27 numbers of 4 generations in the two families had 12 patients,male and female were affected,average onset was 32 years old.The main clinic features included gait ataxia,ambiguity in speech and action clumsiness.Brain MRI showed remarkable atrophy on cerebellum and brain stem.In the two families,the CAG lengths of SCA1 and SCA7 were normal in all numbers.The repeated times of CAG of SCA3 were 11~39 in two control groups,65 ~87 in 10 cases,diagnosed as SCA3 patients.The child Ⅳ2 of family 1 was 8 years old,the repeated times of CAG of SCA3 were repeats 21 and 64 times,repectively.He might be a asymptomatic patient,because he was too young to onset the disease.Conclusions SCA3 is an autosomal dominant genetic disease.The clinical manifestations are ataxia and dysarthria.The detection of repeated times CAG can provide an effective way for the genetic and asymptomatic diagnosis.

Objective To investigate the single nucleotide polymorphism at G-395A site of the Klotho (KL) gene and to analyze its correlation with the coronary heart disease(CHD) and serum Cystatin C(Cys C) level in the elderly Chinese Han population in central China.Methods A case-control study was conducted in 278 elderly Chinese Han population in this department,who were divided into CHD group(138 cases)and control group(140 cases) according to bear angiography coronary or not.G-395A polymorphism of KL gene was determined by TaqMan Gene probe method,and the relationship between G-395A polymorphism and coronary heart disease and serum Cys-C level was analyzed.Results Compared with the control group,the frequency of GG genotype of G-395A in CHD group was significantly higher,and the frequency of AA genotype and AG genotype was not statistically significant.The levels of Cys-C in the GG genotype were higher than those in the AA and AG genotypes both in the control group and coronary heart disease group.Conclusion In the elderly Han population in central China,the risk of suffering coronary heart disease is higher among the GG genotype of the G-395A locus of the KL gene.KL gene G-395A site gene mutation may affect the development of atherosclerosis by affecting blood Cys-C levels.

Objective: To interview the role of CGRP to hepatic microcirculation in acute liver injury induced by coneanavalin A(Con A). Methods: Sixty Kunming rats were divided into three groups randomly(n=20), acute liver injury was established by injection with 20 mg/kg Con A through the tail vein. A saline control group was established by injection with saline. In CGRP-administered group, CGRP was given to rats 30 min before Con A injected. Ten mice in each group were used to observe the average liver blood flow volume and concentration by laser-doppler flow-instrument, and the other ten rats were observed the hepatic microcirculation velocity in vivo by an inverted microscope. Liver damage was assessed by histological evaluation after the rat had been killed. Results: Compared to the injury group, the average liver blood flow volume and velocity were significantly increased in CGRP-administered group, meanwhile, the pathological injury was markedly alleviated, whereas the blood concentration was almost the same. Conclusion: CGRP can decrease the dysfunction of hepatic microcirculation by means of improving the tissue perfusion, and alleviate the pathological damage during acute liver injury.

Objective To investigate the effect of swimming exercise on insulin sensitivity and interleukin-1 beta(IL-1β)mRNA expression in muscle tissue of insulin-resistant (IR) rats induced by high-fat diet.Methods The IR rat model was induced by high-fat diet feeding for 10 weeks and assessed by hyperinsulinemic-euglycemic clamp technique (HEC). The rats were divided into the fllowing three groups: normal group without exercise, IR groups with and without swimming exercise. After 4 weeks swimming excersie, insulin sensitivity and serum IL-1β levels were measured.Results Glucose infusion rate (GIR) of the IR rats was significantly elevated than that of non-exercise IR rats (P<0.01). Moreover, serum IL-1β levels were reduced markedly in IR rats (P<0.01). Conclusion High-fat diet feeding can partly reverse IR of rats to normal value. Swimming exercise improves insulin sensitivity and reduces the levels of IL-1β in IR rats induced by high-fat diet.

Objective:To explore the effect of sarpogrelate on platelet function in patients at the bridging stage before coronary artery bypass grafting (CABG).
Methods: A total of 40 consecutive patients with peripheral artery stent and scheduled for CABG in our hospital from 2011-05 to 2013-04 were enrolled in this study. The patients were randomly divided into 2 groups, Low molecular weight heparin (LMWH) alone group, n=19 and Sarpogrelate+LMWH group, n=21. The medications started at 5-7 days before CABG and stopped at 24 h before CABG. The platelet inhibition rates (platelet aggregation induced by collagen+ serotonin) were examined and compared between 2 groups at the baseline (before randomization), 24h and 1h before CABG respectively.
Results: The platelet inhibition rates were similar between 2 groups at the baseline (87.33 ± 6.82) % vs (86.11 ± 6.87) %, P=0.577 and 1h before CABG (62.60 ± 12.39) % vs (56.19 ± 14.99) %, P=0.148. At 24h before CABG, the platelet inhibition rate in Sarpogrelate+LMWH group was higher than that in LMWH alone group (83.87 ± 8.99)%vs (63.13 ± 10.88)%, P<0.001. Compared with the baseline, the falling range of platelet inhibition was lower in Sarpogrelate+ LMWH group at 24h before CABG, (3.46 ± 6.18) % vs (22.98 ± 9.43) %, P<0.001 and the falling range was similar between 2 groups at 1h before CABG (24.73 ± 14.19)%vs (29.92 ± 14.28)%, P=0.257.
Conclusion: Sarpogrelate + LMWH may result better platelet inhibition rate with quicker recovery of platelet function upon the medication stopping, which might be a feasible management in patients at the bridging stage before CABG.

Objective T o study the correlation of daily living activities with location and severity of trau-m atic brain injury (TBI) and to provide a theoretical basis for im proving the accuracy of expert opinion. Methods Five hundred and one cases of patients with TBI were selected. D etailed records included fol-lowing: pre-injury situation, location and severity of injury, treatm ent and education. D aily living activi-ties scale (B arthel index) was applied to test the subjects’daily living activities. T he relevance am ong location and severity of TBI and B arthel index was statistically analyzed. Results In m ild TBI group, there wasnosignificantdifferenceinBarthelindex among each location (P>0.05).Inmoderate TBIgroup, there were significant differences in B arthel index between subarachnoid hem orrhage and cerebral lobe injury, also between parietal, occipital lobes injury and frontal lobe injury, parietal, occipital lobes injury and temporallobe(P<0.05),respectively,whereasnosignificantdifference in Barthelindexbetween frontal lobe injury and tem poral lobe injury (P>0.05).In severe TBI,there were significant differences in Barthel index between every two different locations (P<0.05). Conclusion T here is som e correlation between the location of TBI and B arthel index, which provides an im portant reference value for analyzing and deter-m ining daily living activities after TBI.

OBJECTIVE:To establish a method for the 5 main components (original syringin,chlorogenic acid,eleutheroside E,isofraxidin and quercetin-3-rhamnoside) in the fruits and roots of wild Acanthopanax senticosus. METHODS:UPLC was per-formed on the column of Waters ACQUITY UPLC HSS T3 with mobile phase of acetonitrile-0.3% phosphoric acid (gradient elu-tion)at a flow rate of 0.2 ml/min. Detection wavelength was 300 nm,column temperature was 30 ℃,and injection volume was 10μl. RESULTS:The linear range was 24.56-184.2 μg/ml for syringin(r=0.9993),18.454-138.405 μg/ml for chlorogenic acid(r=0.9993),8.416-63.12 μg/ml for eleutheroside E (r=0.9997),3.286-24.645 μg/ml for isofraxidin (r=0.9993) and 2.522-18.915μg/ml for quercetin-3-rhamnoside(r=0.9998);RSDs of precision,stability and reproducibility tests were lower than 1%;recover-ies were 99.14%-100.50%(RSD=0.48%,n=6)for syringing in the fruits of A. senticosus、99.03%-100.45%(RSD=0.50%,n=6) for chlorogenic acid in the fruits of A. senticosus、99.22%-100.44%(RSD=0.44%,n=6)for eleutheroside E in the fruits of A. sen-ticosus、99.80%-100.80%(RDS=0.44%,n=6)for isofraxidin in the fruits of A. senticosus、99.76%-101.10%(RSD=0.51%,n=6) for quercetin-3-rhamnoside in the fruits of A. senticosus;99.21%-101.20%(RSD=0.73%,n=6)for syringing in the root of A. senti-cosus、99.81%-101.20%(RSD=0.52%,n=6)for chlorogenic acid in the root of A. senticosus、100.00%-101.50%(RSD=0.62%, n=6)for eleutheroside E in the root of A. senticosus、99.22%-100.40%(RSD=0.47%,n=6)for isofraxidin in the root of A. senti-cosus. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determi-nation of original syringin,chlorogenic acid,eleutheroside E,isofraxidin and quercetin-3-rhamnoside in the fruits and root of wild A. senticosus.

This study inquired into the mechanisms of co-immobilized cytokines and free cytokines-induced apoptosis on HeLa cells. With the use of photochemical fixed method, TNF-alpha/IFN-gamma were co-immobilized on a 24-well polystyrene culture plate. HeLa cells were stained with fluorescent probe JC-1 to detect the changes of mitochondrial membrane potential (deltapsim), and then were examined by flow cytometry. The results showed that co-immobilized cytokines could induce the apoptosis of HeLa cells in a dose-independent manner. When treated with low-dose of co-immobilized cytokines (20ng/ml), the mitochondrial membrane potential (deltapsim) of HeLa cells continually decreased in 6 days. These indicate that low dose co-immobilized cytokines have a long-term of apoptosis-inducing effect on HeLa cells. We assume that there is close relationship between the mitochondrial membrane potential decrease and the apoptosis of HeLa cells.
Apoptosis ; drug effects ; Dose-Response Relationship, Drug ; HeLa Cells ; Humans ; Immobilized Proteins ; pharmacology ; Interferon-gamma ; pharmacology ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondrial Membranes ; drug effects ; physiology ; Tumor Necrosis Factor-alpha ; pharmacology

Objective To evaluate the sensitivity, repeatability and accuracy of microarray digital PCR system in detecting JAK2 V617F mutation, which was closely related to myeloproliferative neoplasms (MPN).Methods All of the 31 MPN patients with JAK2 V617F mutation, including 18 cases of polycythemia vera(PVs),11 primary thrombocythemias (ETs) and 2 primary myelofibrosis (PMFs), were collected from Huashan Hospital, Fudan University during 2014 -2015, while 10 normal controls and 6 cases with abnormal increased hemoglobin were involved.Human erythroleukemia cell line ( HEL ) and colorectal cancer cell SW480 were used as the mutant and the wild type control, respectively.The sensitivity of microarray digital PCR were verified by detecting the gradient diluted mutation standard harboring 30%, 10%, 1%, 0.1%and 0.01%mutant allele burden, respectively .Repeatability was evaluated by detecting 1%and 10% mutated samples for 5 times, respectively.MGB probe real time PCR was selected as the reference method to verify the accuracy of the digital PCR.Results With digital PCR, the accurate quantitation of JAK2 V617F mutation was achieved down to 0.1%, which is approximate to 0.16 copies per microliter.The results obtained from the two kinds of technique showed a high correlation by linear regression analysis (R2 =0.998 3).The results of repeated samples showed CVs as 17.18% for 1%mutant allele burden and 7.50%for 10%.Among all cases, the 31 patients known mutated were detected as positive and 10 controls as negative by both digital PCR and Real time PCR.In another 6 cases, 2 were found JAK2 V617F mutation of low allele burdens of 0.37% and 0.18% by digital PCR but detected as negative by real time PCR.Conclusions Microarray digital PCR offers a higher sensitivity and better repeatability than real time PCR which could help detect rare JAK2 V617F mutations in MPNs accurately.

Objective: To explore a single center large cohort of patients with Takayasu’s arteritis for their clinical manifestation and long-term outcome in China. Methods: We retrospectively analyzed 566 patients with Takayasu’s arteritis admitted in our hospital from 2002-01 to 2013-11 for their clinical characteristics, laboratory ifndings, angiographic features, treatment and long-term outcomes. Results: The patient’s ratio for female to male gender was 1 to 3.8 and the average onset age was (28.9 ± 12.0) years. The most common non-speciifc symptom, initial symptom and complication were fever (52/566 patients, 9.2%), dizziness (214 patients, 37.8%) and hypertension (392 patients, 69.3%) respectively. The patients with pulmonary artery and coronary artery involvement were 83 (14.7%) and 66 (11.7%) respectively, and 131 (23.1%) patients had faster erythrocyte sedimentation rate. The major vascular damage was steno-occlusive lesion and the most common involvement was left sub-clavian artery, which was observed in 278 (49.1%) patients. The treatments were mainly included in medication, interventional therapy, autologous blood vessel transplantation, artiifcial blood vessel transplantation and aortic valve replacement. There were 32 patients died during the mean follow-up period of (5.0 ± 0.2) years. Hypertension, complication and the progressive stage of disease were the major factors affecting prognosis in relevant patients (regression coefifcients: 4.664, 1.959 and 1.870 respectively, allP<0.05). Conclusion: Hypertension was the leading reason for patients’ hospital visit. Takayasu’s arteritis was closely related to cardiovascular disease, the early diagnosis and treatment were really important in clinical practice.