Aim To investigate the effects of TMCC on abnormal L-type calcium current (ICa,L) in rat ventric-ular cardiomyocytes during hypoxia-reoxygenation to find out the mechanism of antiarrhythmic effect. Methods Whole-cell patch clamp was used to record ICa,L in the ventricular cardiomyocytes during hypoxia-reoxygenation in rat under amiodarone and different concentrations of TMCC. Results In hypoxia-reoxy-genation model, peak ICa,L increased from ( 3. 35 ± 0. 50 ) pA/pF to ( 5. 69 ± 0. 25 ) pA/pF ( n =6 , P <0. 01 ) , which was restored to ( 5. 28 ± 0. 18 ) pA/pF (n=6, P>0. 05),(4. 41 ± 0. 22) pA/pF, (3. 82 ± 0. 21)pA/pF(n=6, P<0. 01) by TMCC(100, 200, 400 μmol·L-1 ) and amidodarone 24. 24 μmol·L-1 restored peak ICa,L to(3. 66 ± 0. 27)pA/pF (n=6,P<0. 01 ) . Compared to control group, hypoxia-reoxy-genation turned ICa,L steady-state activation curves to left and inactivation curves to right, which quickened activation and slowed inactivation, TMCC ( 200, 400μmol · L-1 ) and amiodarone could restore the left shift activation curves and right shift inactivation curves. Conclusion TMCC can concentration-de-pendently restore the increase of calcium current due to hypoxia-reoxygenation by promoting inactivation process and inhibiting activation process, and the effect is equal to that of amiodarone. TMCC blocks ICa,L of the ventricular cardiomyocytes, which may be one of its antiarrhythmic mechanisms.

Objective To evaluate the method, clinical efficacy and safety of one phase treat-ment of renal calculi associated with pyonephrosis by percutaneous nephrolithotripsy(PCNL) by pneu-matic combined with ultrasonic lithotriptor. Methods Sixty-six cases of renal calculi accompanied with pyonephrosis were treated with PCNL. The renal calyx was punctured under ultrasound gui-dance, then the tract was dilated from F8 to F16 by peel-away vascular access sheathes. After the in-sertion of the flexible sheath, metallic dilator was inserted and the flexible sheath was pulled out. The tract was dilated by metallic sheath to F21 and the operation sheath and nephroseope were placed into working tract. EMS III LithoClast Master was used. Ultrasonic powered lithotriptor probe with suc-tion was used to clear the liquor puris and calculus fragments with low-pressure or no-pressure. The combined pneumatic and ultrasonic powered lithotriptor was used to break and clear the calculi. Re-salts Of the 66 cases, there was no bacteremia or pyaemia intraoperatively and postoperatively. And there was no other severe complication occurred intraoperatively. One phase PCNL was successfully completed in 60 cases. Other 4 cases had residual calculi less than 1.5 em in diameter and received ESWL to break the calculi, 2 cases had bigger residual calculi and accepted second PCNL 1 week after the first intervention. In the follow-up period, the 3 month post-operative serum Cr was 56-203 μmol/L with an average decrease of 40 μmol/L, GFR was 5.0-56.2 ml/min with an average increase of 23.6 ml/min compared with the pre-operative data. At 6 months postoperative serum Cr was 56-158 μmol/L with average decrease of 31 μmol/L, GFR was 5.0-79.2 ml/min with an average in-crease of 30.2 ml/min. Conclusion Application of PCNL in the treatment of patients with renal cal-culi accompanied with pyonephrosis is safe, cost-effective and clinically efficient by pneumatic com-bined with ultrasonic lithotriptor.

Objective To prepare quality control samples for St.Louis encephalitis virus(SLEV)molecular detection by constructing pseudovirus containing target sequences of SLEV.Methods According to the principles of armored RNA technique, the prM gene sequence of SLEV was cloned into the prokaryotic expression vector to generate recombinant plasmid pSE380-MS2-SLEV.Then, recombinant E.coli transformed with the corresponding plasmid was induced with IPTG to produce recombinant pseudovirus particles.The particles were purified by chloroform and further characterized by double enzyme digestion and transmission electron microscopy.The temperature sensitivity experiments and quantitative RT-PCR were performed to validate the potential of these pseudovirus particles as quality control samples.Results PCR amplification and sequencing analysis confirmed that the prM gene sequence of SLEV was cloned into vector pSE380-MS2.Transmission electron microscopy showed that homogenous spherical particles with a diameter of about 25 nm were produced upon IPTG induction.The SLEV genomic RNA within the pseudovirus particles was resistant to DNaseⅠand RNase A digestion, and remained stable for 20 days at 37℃.These samples were validated with quantitative RT-PCR for SLEV.Conclusion The RNase-resistant and stable pseudovirus particles containing prM fragment of SLEV are constructed successfully, which can be used as positive quality control samples for RNA extraction and molecular detection.

Aim To investigate the antiarrhythmic mechanism of taurine-magnesium coordination compound on sodium current in single rat ventricular myocytes of arrhythmia induced by aconitine.Methods Whole-cell patch clamp was used to record INa in normal cardiomyocytes and single rat ventricular cardiomyocytes of arrhythmia induced by aconitine.Results In ventricular cardiomyocytes of rat,INa was blocked by 100~400 ?mol?L-1 TMCC in a concentration-dependent manner.INa was increasd from(45.56?1.96)pA/pF to(59.19?11.49)pA/pF by 1 ?mol?L-1 aconitine,while decreased to(34.23?1.33)pA/pF by 24.24 ?mol?L-1 amiodarone.TMCC(100,200,400 ?mol?L-1)could restore INa to(51.61?5.96)pA/pF,(40.91?6.73)pA/pF,(41.50?5.50)pA/pF respectively.Amiodarone could restore INa to(40.22?1.47)pA/pF.Conclusions TMCC can restore INa,which is increased by aconitine,and the effect is equal to that of amiodarone.TMCC blocks INa of ventricular cardiomyocytes,which may be one of its antiarrhythmic mechanisms.

Aim To investigate the effects of dioscin ( Dio) on rat myocardial contractility. Methods Left ventricular contractile function was measured using the Langendorff non-recirculating mode of isolated rat heart perfusion. Effects of low, middle and high concentra-tion of Dio were investigated by measuring left ventricu-lar systolic pressure ( LVSP ) and left ventricular end diastolic pressure ( LVEDP) . Also, peak rates of rise/fall of left ventricular pressure ( ± dp/dtmax ) of isolated rat heart were calculated. Effects of Dio on intracellu-lar free calcium concentration in rat H9 c2 cells were measured by using the confocal microscopy. Mitochon-drial membrane potential was detected with multifunc-tional microplate reader. Results With 0. 1, 1 μmol · L-1 Dio, LVSP were significantly enhanced from (11. 55 ± 0. 52), (10. 53 ± 0. 28) kPa to (13. 08 ± 0. 72), (12. 53 ±0. 64) kPa(P<0. 01); +dp/dtmax were dramatically increased from ( 0. 38 ± 0. 10 ) , (0. 40 ± 0. 07) kPa·ms-1 to (0. 42 ± 0. 11), (0. 43 ± 0. 02) kPa·ms-1(P<0. 05). With the 10μmol· L-1 Dio, LVSP and + dp/dtmax were both decreased from (12. 13 ± 0. 33) kPa and (0. 42 ± 0. 04) kPa· ms-1 to ( 9. 46 ± 0. 77 ) kPa and ( 0. 24 ± 0. 04 ) kPa ·ms-1 (P <0. 01). With 0. 1, 1, 10 μmol·L-1 Dio, the relative fluorescence intensity of intracellular free calcium concentrations was increased significantly from (16. 62 ± 0. 89) to (21. 48 ± 0. 80), (25. 68 ± 0. 69) and (19. 84 ± 0. 66)(P <0. 01)respectively. 0. 1, 1μmol·L-1 Dio showed no significant effects on the mitochondrial membrane potential of rat H9 c2 cells, while with effects of 10 μmol·L-1 Dio, the ra-tio of JC-1 monomer and J-aggregates was changed from (1. 14 ± 0. 03) to (1. 35 ± 0. 06)(P<0. 01), indica-ting a decrease in the mitochondrial membrane poten-tial. Conclusion Low and middle concentrations of Dio show a positive inotropic effect on isolated rat heart, as the LVSP and + dp/dtmax are enhanced, which may concern with the increase of the intracellu-lar concentration of Ca2+. It will not cause the calcium overload while the intracellular concentration of Ca2+ is increased by low and middle concentration of Dio in the myocytes except high concentration of Dio.

Aim To investigate the antiarrhythmic mechanism of taurine-magnesium coordination com-pound on abnormal sodium current channel ( INa ) in-duced by hypoxia-reoxygenation in ventricular myocytes of rats. Methods Single ventricular myocytes were i-solated from each rat heart using enzymatic dissociation through Langendorff retrograde aortic perfusion. Whole-cell patch clamp was applied in voltage clamp mode to record INa both in normal ventricular myocytes and single ventricular myocytes of arrhythmia induced by hypoxia-reoxygenation. Results The peak density of INa was changed from ( 56. 89 ± 2. 07 ) pA/pF to (35. 05 ± 1. 52) pA/pF( n=6, P <0. 01 vs control) by hypoxia-reoxygenation with the INa I-V curve shifting upward. TMCC(200, 400 μmol·L-1)was able to re-store the reduction caused by H/R to (35. 78 ± 1. 95) pA/pF, (41. 52 ± 0. 86) pA/pF, (n=6,P <0. 01) and (48. 34 ± 0. 99) pA/pF(n=6,P<0. 01) respec-tively, but not at 100 μmol·L-1(n=6, P>0. 05), in a concentration-dependent manner, while amioda-rone restored it to (39. 44 ± 1. 24) pA/pF (n=6,P<0. 01 ) . Both high concentration of TMCC and amioda-rone could shift the I-V curve downward. In addition, TMCC and amiodarone could restore the INa inactivation curve and slow down its inactivation, whereas the acti-vation curves showed no significant differences among groups. Conclusion TMCC(200,400 μmol·L-1) could restore the H/R induced INa reduction and shift the I-V curve downward by inhibiting steady-state inac-tivation, which is suggested to be one of the mecha-nisms of the antiarrhythmic effects of TMCC in hypoxia-reoxygenation model.

Aim To investigate the antiarrhythmic mechanism of taurine magnesium coordination compound(TMC)on the potassium current in single ventricular myocytes of guinea pig.Methods Whole-cell patch clamp was used to record IK,IK1 in single ventricular myocytes of guinea pig.Results In ventricular myocytes of guinea pig,IK was decreased from(8.67?1.04)pA/pF to(6.31?1.16)pA/pF at +70 mV.TMC had no effect on the IK1.Conclusions TMC had inhibitory effect on IK directly and this effect maybe resulted in prolonging the action potential duration(APD)and effective refractory period(ERP).It could be one of the basis of antiarrhythmic effect of TMC.

The important progress of the relationship between genomics and cardiovascular diseases has elucidated several abnormalities of genes leading to familial cardiomyopathy and ion channel diseases.Investigations of genomics show that the difference including resistance and susceptivity to diseases among human is determined by over 3 million base pairs of all 3,000 million.Single nucleotide polymorphism plays an essential role in elucidating hereditary cardiovascular diseases.

Objective: To investigate the effectiveness of Nice knot combined with elastic intramedullary nailing fixation in treatment of Robinson type 2B midshaft clavicular fracture in adults. Methods: Between March 2016 and January 2018, 20 patients with Robinson type 2B midshaft clavicular fractures were treated with reduction and internal fixation by Nice knot and elastic intramedullary nailing. There were 13 cases and 7 cases, with an average age of 43 years (range, 18-56 years). The causes of injury included the traffic accident in 6 cases, falling in 12 cases, and falling from height in 2 cases. The interval between injury and admission ranged from 1 hour to 2 days (mean, 3.2 hours). The fractures were classified as Robinson type 2B1 in 16 cases and type 2B2 in 4 cases. The length of incision, the operation time, the visual analogue scale (VAS) score on the 2nd day after operation, the fracture healing time, the postoperative shoulder function and the Disability of Arm Shoulder and Hand (DASH) score, the complications, and the time of second surgical removal of internal fixator and incision length were recorded. Results: The length of incision was 2-6 cm (mean, 4.7cm). The operation time was 45-120 minutes (mean, 77.2 minutes). The VAS score was 1-5 (mean, 3.2) on the 2nd day after operation. All incisions healed by first intention and no infection or nerve injury occurred. All patients were followed up 12-32 months (mean, 18.6 months). All fractures healed with the healing time of 10-15 weeks (mean, 12.1 weeks). The Constant score was 92-98 (mean, 96.3) and DASH score was 0-6.4 (mean, 3.1). The elastic intramedullary nailing bending and hypertrophic nonunion occurred in 1 case and the skin stimulated by elastic nail tail in 1 case after operation. The internal fixators were removed at 12-26 months (mean, 14.6 months) after operation. And the length of incision was 1-2 cm (mean, 1.3 cm) and the operation time was 5-15 minutes (mean, 9.0 minutes). Conclusion: For the midshaft clavicular fracture in adults, the procedure of the Nice knot combined with elastic intramedullary nail has advantages of small incision, light pain, rapid fracture healing, small secondary operation injury, and avoiding the risk of clavicular epithelial nerve injury, and can obtain good effectiveness.

Aim To study the protective effects of sphingosine 1-phosphate (S1P) postconditioning on rat myocardial cells injured by hypoxia/reoxygenation in reperfusion injury salvage kinase ( RISK ) signal path-way. Methods The cultured rat H9c2 cells were ran-domly divided into seven groups: ( 1 ) control group;(2) hypoxia/reoxygenation (H/R) group; (3) S1P group;(4) S1P+LY294002 group(S1P+LY); (5) LY group; ( 6 ) S1 P +PD98059 group ( S1 P +PD );(7) PD group. The viability of H9c2 cells was detec-ted using MTT method. The content of MDA in the cultured medium and the activity of T-SOD and Mn-SOD were measured with colorimetry. The concentra-tion of intracellular free calcium ion was detected by confocal microscopy. The rate of cell apoptosis was de-termined by flow cytometric analysis. Western blot was used to assess phosphorylation of Akt and ERK1/2 in H9c2 cells. Results Compared with the H/R group, S1P significantly increased vaibility of cells, lowered the rate of apoptosis, decreased the content of MDA in the culture medium, increased the activity of T-SOD and Mn-SOD, reduced concentration of intracellular calcium and increased the phosphorylation of Akt and ERK1/2 . When added LY294002 or PD98059 , the effects of S1P above were inhibited. Conclusion S1P protects H9 c2 cells against hypoxia/reoxygenation inju-ry. The protection of S1P was inhibited by LY294002, the inhibitor of PI3 K/Akt and PD98059 , the inhibitor of ERK1/2 . S1 P protects H9 c2 cells against hypoxia/reoxygenation injury via RISK pathway.