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.

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 develop a novel method for treating complete heart block by autotransplantation of simus node node cells to right ventricular anterior wall.Methods Twenty healthy mongrel dogs were involved in the present study.The dogs were randomly assigned to transplant group or control group(n=10).The sinus node (SN)was harvested and isolated in vitro after an electronic pacemaker was implanted and complete heart bolck was introduced.The SN cells from dogs of transplant group were injected to autogenic right ventricular wall.Commensurable culture medium was implanted to ghe same position of dogs in control group.Two weeks later,detailed electropohysiological study was performed.For investigating the variation of the rhythm,epinephrine was administrated through femoral vein to dogs of transplant group.Results Most of isolated SN cells from dogs were thin-spindle shape,and cell activity was fine.The SNs by VG stained displayed typical structural feature.2 weeks after cell autotransplantation,higher heart rates were achieved from transplant group than that in control group(P＜0.05).This rhythm was stable in 4 weeks and became faster remarkably after administration of eninephrine(P＜0.05).Conclusion SN cell of dogs tutorgrfted into right ventricular anterior wall can form new pacemaker site in ventrcle and improve ventricular rate of complete heart bolck.This pacemaker site can also be regulated by epinephrine.

Objective The results of Aortic valve replacement (AVR). Combined with ascending aortic replacement(group A) or aortoplasty (group B) in patients with aortic valve disease and ascending aortic dilatation were analysed to assess the clinical outcomes and respective indications. Methods Among the two groups, the age, gender, NYHA class, types of aortic valve lesions and left ventricular ejection fraction were not different statically. The ascending aortic diameters in group A[(50.41 ±3.71) mm] and group B [(48.29±2.18) mm] were not statically different. Ascending aortic replacement was performed in Group A. A Dacron tube(diameter 28 ～ 30mm) was routinely wrapped around the ascending aorta after aortoplasty in group B. Results There was 1 postoperative death in group B, blood transfusion volume and postoperative complications were not stasticaly different in the two groups. Cardiopulmonary bypass time [(110.52 ± 27.51) min] and aortic across clumping time [(71.70 ± 17.13)min] in group A were significantly longer than that of group B [(97.31 ± 19.46) min,P=0. 004; (57.13 ±19.46) min, respectively. P=0.025]. Conclusion Aortic valve disease, especially bicuspid valve disease often combines with ascending aortic dilatation or aneurysm. In younger patients, ascending aorta should be actively treated surgically when the diameter is equal or more than 40mm. Aortoplasty with external reinforcement of a Dacron tube is simpler and safer than aortic replacement in patient without aortic atherosclerosis or ulceration, and large aneurysm.

Objective To investigate whether rat grimace scale (RGS)could be used to assess pain in chronic pancreatitis,so as to provide evidence for pain research and clinical assessment of rat pain.Methods Twenty-eight adult male wister rats were evenly randomized into two groups (n =14):an experimental group and a control group.The experimental group was intravenously given 8 mg/kg body weight dibutyltin dichloride (DBTC)to induce chronic pancreatitis,and the control group was injected with ethanol and glycerin solution.Abdominal hypersensitivity,RGS scores and weight at different time points was detected.HE staining was used to detect the histological changes of pancre-atic tissue.Results Compared with the control group,the rats in the experimental group showed chro-nic inflammation in pancreatic tissue in two weeks.There was a significant increase in the number of abdominal withdrawals (P < 0.001 )and RGS in the experimental group.Conclusion Rat grimace scale might lead to a successful transition of basic science findings into clinical application.

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 .