Objective To investigate the clinical characteristics and pathogenesis of paroxysmal dyskinesias.Methods Retrospective analysis was performed in 5 patients suffering from paroxysmal kinesigenic choreoathetosis(PKC) and 2 patients with paroxysmal persistent exercise-induced dystonia(PED).Results The episodes of all cases of PKC were induced by sudden movements.3 cases showed rigidty and hypertonia.3 cases presented with twist of limbs and dystonia.2 PED cases were induced by persistent movement,manifested involuntary movements of limbs,and the duration of the attack usually last seconds to minutes.5 patients showed epileptic discharges in EEG or AEEG.2 patients had abnormal findings of brain CT or MRI.4 PKC cases responded well to carbamazepine and 1 PED patients to large dose of valproate sodium.Conclusions Paroxysmal dyskinesias are usually induced by sudden movement and present paroxysmal extrapyramidal symptoms.Most of the patients show epileptic discharges in EEG and responded well to antiepileptic drugs.This implies the underlying relationship of pathogenesis between paroxysmal dyskinesia and epilepsy.

Objective To investigate the effects of the therapy of invigorating the kidney and qi and dissipating blood stasis to removing the toxin on the hepatocyte apoptosis and associated genes in the liver fibrosis rats,and to explore the treatment mechanism of liver fibrosis with TCM.Methods liver fibrosis model rats were induced by CCl4,and the Chinese herbs(Ganlike)with invigorating the kidney and qi and dissipating blood stasis to removing the toxin was given by daily gavage,after the experiment,the pathological changes of liver were observed with optical microscope.Hepatocyte apoptosis was detected with TUNEL assay,apoptosis-associated genes were detected with immunohistochemistry assay.Results The Chinese herbs Ganlike could obvously improve the pathological changes,alleviate the hepatocyte apoptosis and modulate the expression of apoptosis-associated genes.Conclusions The anti-fibrosis mechanism of Ganlike may be to regulate the hepatocyte apoptosis of liver fibrosis rats and to restrain the expressions of hepatocyte Fas/FasL,Bcl-2/Bax.

Objective To investigate the surgical diagnosis and treatment of chronic stenosis of mechanical prosthetic valve complicated with acute dysfunction,so as to deepen our knowledge on chronic stenosis of mechanical prosthetic valve.Methods The clinical data of 5 patients with chronic stenosis of mechanical prosthetic valve complicated with acute dysfunction were retrospectively analyzed,and the relevant literatures were reviewed.Results Re-operation(mechanical prosthetic valve replacement) was performed once the diagnoses were confirmed.The patients recovered well;the cardiac function was obviously improved;and there were no early complications.Conclusion Chronic stenosis of mechanical prosthetic valve should be strongly suspected when they have symptoms indicating valvular stenosis.Complication of acute mechanical prosthetic valve dysfunction is not difficult to diagnose,and prompt operation is important to save the life of patients.

The relationship between the expression of resistin in polycystic ovary syndrome (PCOS) and insulin resistance was investigated. The plasma resistin concentrations in 35 patients with PCOS and 40 controls were measured by ELISA. Luteinizing hormone (LH), follicle-stimulating hormone (FSH), and fasting insulin (FIN) were tested by radioimmunoassay. Insulin resistance index (HOMA-IR) was calculated. Fasting plasma glucose (FPG) was determined by oxidase test. Western blot and reverse transcriptase PCR (RT-PCR) methods were used to detect the expression of resistin in adipose tissues. The levels of plasma resistin, LH, LH/FSH and FIN and HOMA-IR in patients with PCOS were significantly higher than those in control group (all P<0.05). Plasma resistin was correlated positively with FPG, FIN, HOMA-IR, LH and LH/FSH (r=0.56, 0.60, 0.65, 0.48, and 0.42 respectively). Resistin protein and mRNA expression levels in patients with PCOS were significantly higher than those in normal tissues (all P<0.01). It was concluded that resistin might be involved in the pathogenesis of insulin resistance of PCOS.

BACKGROUND: Hyperpolarization-activated cyclic nucleotide-gated (HCN) current plays an important role in regulating heart spontaneous pulsation.OBJECTIVE: To observe target gene expression and electrophysiological characteristics of pig bone marrow mesenchymal stem cells (BMSCs) transfected with hyperpolarization-activated cyclic nucleotide-gated channel 2 (HCN2) gene recombinant adenovirus.DESIGN, TIME AND SETTING: Cell-gene in vitro study was performed at the Laboratory of Thoracic and Cardiovascular Surgery,Second Military Medical University of Chinese PLA from July 2007 to March 2008.MATERIALS: Yorkshire pig was supplied by Animal Institute, Second Military Medical University of Chinese PLA. HCN2 plasmid was presented by Professor Dario DiFrancesco from Italy. Recombinant adenovirus Ad.HCN2 was constructed and stored using Ad5 in this laboratory.METHODS: Pig BMSCs were isolated with combination of gradient centrifugation of Percoll and adherent treatment in vitro.Ad.HCN2 was transfected at multiplicity of infection=50. We also set non-transfection and transfected Ad.Null groups.MAIN OUTCOME MEASURES: Expression of HCN2 mRNA was detected with RT-PCR, and expression of HCN2 channel protein was examined with immunofluorescent staining. Electrophysiology of HCN2 channel protein was measured with whole-cell patch clamp.RESULTS: No amplified fragments were found in the non-transfection and transfected Ad.Null groups, but amplified fragments were determined at 250-500 bp following Ad.HCN2 amplification, which was the same as plasmid carrying HCN2 gene. Staining strength of cell nuclei following transfection was significantly weaken compared with cell membrane and plasma, which showed identical distribution as HCN2 protein. No HCN2 protein was detected in the non-transfection and transfected Ad.Null groups.Pacemaker current could be recorded with a whole-cell patch clamp. It was fully activated around -140 mV with an activation threshold of -60 mV, presenting voltage dependence. CsCI (4 mmol/L) reversibly blocked the inward currents. No pacemaker current was detected in the non-transfection and transfected Ad.Null groups.CONCLUSION: The HCN2 recombinant adenovirus carrier was transferred into serial subcultivation porcine bone marrow mesenchymal stem cells. HCN2 channel protein has been expressing. Pacemaker current could be recorded with a whole-cell patch clamp.

Objective To summary the methods and results of first and second operations on patients with aortic valve disease and non-specific aortitis.Methods The total 34 patients including 23 cases with aortitis and 11 cases with Behcet disease were studied from Jan 2000 to Dec 2010.The first operation was Bentall procedure in 18 cases and AVR in 16 cases.Fourteen of 16 cases who had AVR developed severe paravalvular leakage,and undewent the second operation including 10 aortic root replacement (8 valve-conduit and 2 homograft) and 4 non-anatomic AVR.Results Eighteen patients who had first operation of Bentall procedure all survive without aortic pseudoaneurysm after the follow-up of 6 months to 11 years.Fourteen redo cases all survive except for one case died of repture of aortic pseudoaneurysm 1.2 years postoperatively.Conclusion Preoperative diagnosis in these patients is very difficulty.The first operation of root replacement is of choice.The second operation is very difficulty to handle,root replacement can achieve satisfactory results.Non-anatomic AVR is easy to perform,and good hemo stasis intraoperatively,and is a satisfied alternative method with good results.

Background Retinal vein occlusion is a common retinal vascular diseases.Thromblysis and anticoagulation therapies are main approaches.However,systemic thrombolysis is relatively inefficient,and it often enhances the risk of hemorrhage.Objective This study was to investigate the therapeutic effects of PLM-ΔK,a kringle deficiency mutant of plasmin,on photochemically induced branch retinal vein occlusion (BRVO) after intravitreal injection.Methods BRVO models were established by the combination of caudal vein injection of Rose Bengal with argon laser radiation of periphery area of retinal veins in SD rats.Forty model rats were randomized into balance salt solution (BSS) group and 0.01 U,0.02 U,0.03 U PLM-ΔK group,and 10 μl corresponding drug was intravtreally injected 12 hours after modeling.Ophthalmoscopy and fundus fluorescein angiography (FFA) were performed to observe the change of retinal veins.The animals were sacrificed 3 days after intravitreal injection,and hematoxylin and eosin staining was used for the histopathological and ultrastructural examination of retinas.The retina of the rats was isolated for the stretched preparation of retina.The expressions of fibronectin (FN) and laminin (LN) in eyeball wall were assayed by immunofluorescence technology.The use and care of the animals complied with Statement of the Association for Research in Vision and Ophthalmology.Results The revascularization of over 2 retinal veins was found in 0,3,6 and 8 rats in the BBS group and 0.01 U,0.02 U,0.03 U PLM-ΔK group 3 days after intravitreal injection,respectively,showing a significant difference among the groups (x2=9.635,P =0.022),and the rat number with revascularization in 0.01 U PLM-ΔK group was not significantly different from that in BSS group (Z=-1.558,P =0.119),but the difference between 0.03 U PLM-ΔK group and 0.01 U PLM-ΔK group was significant (Z=-2.762,P=0.006).In the third day after intravitreal injection,retinal vein thrombus were found in the BSS group under the light microscope,and angiogenesis was seen on the retinal flatmount nuclear.In the 0.03 U PLM-ΔK group,posterior vitreal detachment was exhibited under the light microcope,and no retinal new vessel and cell damage were seen.FN was strongly expressed in the inner limiting membrane (ILM) layer,photocyte layer,outer limiting membrane (OLM) layer,choroid and scleral layer,and LN was expressed mainly in the ILM,OLM and scleral layer in the BSS group.However,the expression intensities of FN and LN were obviously weakened in the 0.03 U PLM-ΔK group.Conclusions Intravitreal injection of PLM-ΔK can enhance the reperfusion of occluded branch retinal vein and serve as a potential therapeutic drug for BRVO.Also it can permeate into choroid after intravitreal injection to degradate FN and LN.

Objective To define the incidence and perioperative risk factors of new-onset atrial fibrillation for patients with preoperative sinus rhythm after successful mitral valve replacement.Methods Retrospective analysis was performed on 586 patients with preoperative sinus rhythm who underwent successful mitral valve replacement in our hospital from 1998 to 2008.The cases were classified into postoperative atrial fibrillation (AF group) or postoperative sinus rhythm (SR group).Twenty-nine risk factors including clinical and echocardiography data were selected into univariate analysis by using student' s t test or chi-squared test according to the data type.The factors with a value of P ＜ 0.1 in univariate analysis were assessed by multivariate logistic regression.A value of P ＜ 0.05 (two-sided) was considered to be statistically significant in multivariate logistic regression. ResultsOne hundred and eighteen patients had atrial fibrillation postoperatively. The incidence was 20.1％.Univariate analysis revealed that the factors including age,chronic lung disease,left ventricular mass,left atrial volume,right atrial volume,tricuspid valve regurgitation,heart failure,valvular pathology,postoperative prosthetic mitral effective orifice area index,postoperative mechanical ventilation time,serum levels of potassium and magnesium significantly increased the risk of postoperative atrial fibrillation.However,in multivariate logistic regression,age,left atrial volume and postoperative prosthetic mitral valve effective orifice area index and serum of potassium had significant statistically significances between AF group and SR group.Conclusion Age,left atrial volume,postoperative prosthetic valve effective orifice area index and serum of potassium were significant predictors of postoperative new-onset atrial fibrillation for patients with preoperative sinus rhythm after mitral valve replacement.

Objective To develop a sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of forsklin in rat plasma.Methods After extraction with methyl tert-butyl ether,chromatographic separation was performed on a C18 column with the mobile phase consisting of water ( 0.1％ formic acid)-acetonitrile in a gradient elution mode.A tandem mass spectrometer equipped with electrospray ionization (ESI) source was used as detector in the positive ion mode.Quantification was performed using multiple reaction monitoring (MRM) with the precursor product combination ions of m/z 411→375.3 and 285→193 for forsklin and diazepam.Results Good linearity was obtained in the 0.5-1000 ng/ml range for the analyte and the analytical method was validated in terms of specificity,precision,accuracy,recovery,stability and matrix effect.These assays gave RSD values always lower than 14.4％ and RE values between -3.5 ％ and 3.8％.In addition,the specificity,extraction recovery,stability and matrix effect were satisfactory.Conclusion Due to its high sensitivity,specificity and simplicity,the method could be used for pharmacokinetic studies of forsklin.

Objective To investigate whether toll like receptor ( TLR) signaling pathways can in-crease the expression of IL-17 R in neuralglial cells , and if they can whether the increased IL-17 R is func-tional.Methods Experimental autoimmune encephalomyelitis (EAE) was induced in B6 mice by immuni-zation with an emulsion of myelin oligodendrocyte glycoprotein 35-55 ( MOG35-55 ) in complete Freund's adju-vant (CFA).The expression of Il17ra and Il17rc in the brains and spinal cords of mice with EAE were de-tected by real-time PCR.Luxol fast blue ( LFB) staining was performed to the spinal cord sections to detect tissue demyelination.Immunohistological staining against IL-17RA and CD3 were undertook to visualize IL-17RA+and CD3 + cells.Same approaches were also applied to immunized Rag1 -/- mice to figure out whether T cells infiltration is necessary for increasing IL-17RA expression in the central nervous system ( CNS) .Then B6 mice were immunized with incomplete Freund′s adjuvant ( IFA) plus different TLRs ago-nists to measure the expression of Il17ra in the brains and spinal cords by qPCR .The purified astrocytes , microglia and oligodendrocytes isolated from neonatal mice brains were cultured in vitro for two weeks , and then treated with different TLRs agonists .The expression of Il17ra at mRNA and protein levels in the cells were determined by qPCR and Western blot respectively .The astrocytes were treated with IL-17A and LPS individually or in the combination to detect the level of CCL 2, CXCL8 and IP-10 in the supernatant by ELISA.Results B6 mice with induced EAE showed significantly increased Il17ra expressions in the brain and spinal cord , which was also detected in immunized Rag1 -/-mice.Although no spinal cord demyeliza-tion and CD3 cells infiltration were detected in Rag1 -/-mice, significantly increased number of IL-17RA positive cells could still be visualized .In vivo TLRs agonist participated immunization and in vitro treatment of purified neuroglial cells demonstrated that TLRs agonists could directly evoke IL -17RA expression in the CNS or cultured astrocytes , microglia and oligodendrocytes with high efficiency .Both IL-17 A and LPS could stimulate astrocytes to secrete CCL2, CXCL8 and IP-10, however, a combined use of IL-17A and LPS fur-ther augmented the production of these chemokines to a large extend .Conclusion Taken together , we con-cluded that TLRs agonists could directly stimulate neuroglial cells to express IL -17RA which functionally re-spond to IL-17A by secreting chemokines .