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.

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 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.

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 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 .

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.

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 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.