Objective To investigate the way of gene therapy on congestive heart failure and other heart diseases. Methods Recombinant adeno associated viruses (AAV) encoding the human ? 2 adrenergic receptor(AR) and enhanced green fluorescent protein (EGFP) were constructed and the ? 2 AR and EGFP cDNA were transfected in vitro to the neonate rat ventricular myocytes. The expression of ? 2 AR and EGFP on cultured neonate rat ventricular myocytes and the ability of ? 2 AR to potentiate the beta adrenergic signaling were assessed. Results The presence of human ? 2 AR mRNA was demonstrated by Northern blot, the expression of the ? 2 AR gene was demonstrated by Westernimmuno blot. The density of ? AR in the myocytes was greater than that in the control 〔(204.0 ? 6.4) fmol/mg protein vs (76.0 ? 2.8) fmol/mg protein〕 according to a ligand binding assay. Human ? 2 AR gene significantly increased isoproterenol stimulated cAMP as compared with the control〔(116.2?5.8) pmol/10 6 cell vs (58.4 ? 4.6) pmol/10 6 cell〕. EGFP was also expressed in the transfected cardiomyocytes. Conclusions Human ? 2 AR gene could be transfected into cardiomyocytes by AAV vector and could potentiate the beta adrenergic signaling.

Microglia play a crucial role in inflammation after cerebral ischemia.A large number of studies have shown that microglia are highly plastic cells that can assume different phenotypes and functions in response to specific microenvironmental signals.Microglia can be polarized into the classically activated proinflammatory M1 phenotype or the alternatively activated anti-inflammatory M2 phenotype,and play different roles in ischemic injury.Irnhibiting M1 while stimulating M2 may be a new approach for the treatment of ischemic stroke.

Objective To evaluate the additional efficiency of aldosterone blockers (AB) during angiotensin Ⅱ receptor blockers (ARB) treatment on hypertensive left ventricle hypertrophy in the elderly. Methods Forty-five cases of hypertensive left ventricular hypertrophy detected by Doppler ultrasonic cardiogram examination were enrolled,and were randomized to receive treatment with ARB plus AB (Group A) or ARB plus diuretics (Group B) for 48 weeks. The indicators of left ventricle hypertrophy were checked before and after the treatment. Results The both regimens could significantly decrease the blood pressure,and there was significant difference in the effects on lowering blood pressure between the regimens. Both regimens could alleviate left ventricle hypertrophy,but the regimen of ARB plus AB was superior to the regimen in Group B. Conclusion The addition of AB treatment during the ARB treatment was a good antihypertensive regimen for reducing left ventricle hypertrophy.

Inflammation plays an important role in the pathophysiological mechanism of acute ischemic stroke.CD200 expressed in neurons interacts with CD200 receptor (CD200R) on microglia cells.It can inhibit microglia activation and alleviate the inflammation after cerebral ischemic injury.This article reviews the roles of CD200 and CD200R in the activation of microglia after cerebral ischemia.

Objective To observe the effects of treadmill training on hindlimb motor recovery and noradrenaline (NA) fibers in rats with spinal cord contusion.MethodsSpinal cord contusions at the T10 level were induced in 3-month old female SD rats by dropping a 10 g rod from a height of 25 mm.The injured rats were then randomly divided into a control group and a treadmill training group ( n =15 in each group).Treadmill training began 1 week post-injury.No treatment was administered to the control group rats.Hindlimb motor function was evaluated weekly from the 1 st to the 9th week post-injury using Basso Beattie Bresnahan (BBB) scores and average combined scores (ACOs).The expression of dopamine beta hydroxylase (DBH) in the spinal cord caudal to the injured area was detected by immunohistochemical staining.ResultsThe treadmill training group exhibited significantly greater improvement in BBB scores from the 3rd week to the 7th week and in ACOs from the 2nd to the 9th week post-injury compared with the control group.The number of DBH positive fibers in the injured spinal cords and the number of DBH positive neurons in the injured frontal horns were also significantly higher in the training group.ConclusionsTreadmill training can promote the recovery of hindlimb motor function in rats after spinal cord contusion,perhaps by increasing the number of DBH positive fibers and DBH positive neurons.

Objective To explore the feasibility of using irreversible electroporation (IRE)mediating HPV16 E6 shRNA into cervical cancer cell line SiHa,and to clarify the influence of their co-effect on the proliferation of SiHa cells and its mechanism.Methods A HPV16 E6 gene specific interference sequence was inserted in pGenesil-1 to build a interference vector.10 pulses of IRE with 800 V,100 μs,and 1 Hz were applied to the suspension of SiHa cells and vectors.According to the treatment factors,control group,IRE group,pGenesil-N group,pGenesil-N+IRE group,pGenesil-E6 group and pGenesil-E6 + IRE group were set up.The expression of green fluorescent protein (GFP)and transfection efficiency were confirmed by inverted fluorescence microscope 24 h after the vector was transfected by IRE,and the expression efficancy of GFP was calculated.The expression levels of E6 mRNA and protein were detected by RT-PCR and Western blotting method which was also applied to detect the expressions of P53 and PCNA.The proliferative activity of SiHa cells was determined by CCK-8 assay.Results Enzyme digestion and DNA sequencing verified that the vectors were correctly constructed.GFP was seen under inverted fluorescence microscope 24 h after IRE transfection.Compared with IRE group,the expression levels of E6 mRNA and protein were decreased detected by RT-PCR and Western blotting method after the vectors were treated with IRE,the P53 protein expression level was increased (P < 0.05),and the PCNA expression level was decreased (P <0.05).The CCK-8 assay results showed the proliferative activity of SiHa cells in pGenesil-E6+IRE group was decreased more obviously than that in pGenesil E6 group (P <0.05).Conclusion IRE can play the role of gene transfection of mediating HPV16 E6 shRNA into SiHa cells, and their co-effect can significantly inhibit the proliferation of SiHa cells.

Objective To evaluate the in vitro antifungal activity of 4 antifungal agents alone or in combination against Exophiala dermatitidis (E.dermatitidis) biofilms.Methods E.dermatitidis biofilms were prepared by using a modified 96-well plate-based method.The in vitro antifungal activity of amphotericin B,voriconazole,itraconazole and caspofungin alone or in combination against E.dermatitidis biofilms were investigated via the broth microdilution checkerboard technique.Results The sessile minimum inhibitory concentration ranges resulting in 50％ (SMICS0) and 80％ inhibition (SMIC80) of E.dermatitidis biofilms were all ＞ 32 mg/L for itraconazole,voriconazole and caspofungin,and the SMIC50 and SMIC80 ranges of amphotericin B were 1-2 mg/L and 4-8 mg/L respectively.The combination of amphotericin B with voriconazole showed synergistic inhibitory effects against E.dermatitidis biofilms,while the combination of amphotericin B with itraconazole or caspofungin,as well as the combination of voriconazole with caspofungin,revealed no synergistic effects.No antagonistic effect was observed in any of the combinations.Conclusion Amphotericin B appears more active against E.dermatitidis biofilms,and the combination with voriconazole can enhance the anti-biofilm effects against E.dermatitidis biofilms.

Objective To evaluate the feasibility and efficacy of laparoscopic single-port transumbilical renal pedicle lymphatic disconnection (TRPLD) for treatment of chyluria. Methods Nine cases of chyluria underwent laparoscopic single-port TRPLD. In all cases a 2-3 cm single inverted Ushaped supraumbilical incision was made, two 5-mm and one 12-mm trocars were inserted, and a medical rubber glove was sutured surrounding the three trocars and incision was made for gas proofing.Conventional straight and flexible instruments were used for dissection. Results All laparoscopic operations were successfully completed without conversion to open surgery. The mean operative time was 135 (96-178) minutes, and the mean estimated blood loss was 126 (50-250) ml. Chyluria disappeared in all patients after operation and did not reoccur during the follow-up (1 - 6 months).Conclusions Laparoscopic single-port transumbilical TRPLD represents a feasible and novel mini-invasive option for patients with chyluria.

Objective To explore the effects of the down-regulating peroxiredoxin 2 (PRDX2) on invasion, migration and proliferation of human gastric cancer MGC803 cells by using RNA interference. Methods MGC803 cells were divided into 3 groups:blank control group,negative control group and PRDX2 siRNA group. Transwell assay was used to examine the invasive ability change of MGC803 cells after transfection. Scratch test was used to detect the change of MGC803 cells migration ability after transfection. Western blot was used to examine the expression changes of PRDX2, matrix metalloproteinase (MMP)-2 and MMP-9. The proliferation ability of MGC803 cells was assessed by using CCK-8 assay. Results The expressions of PRDX2(0.345±0.006,0.721±0.013,0.720±0.014),MMP-2(0.067±0.012, 0.391±0.015, 0.371± 0.016) and MMP-9 (0.073±0.013, 0.341±0.028, 0.346±0.024) in the PRDX2 siRNA group were lower than those in the blank control group and negative control group (all P < 0.05). The cell invasion, migration and proliferation were inhibited in MGC803 cells (all P < 0.05). Conclusion PRDX2 is overexpressed in MGC803 cells. Down-regulating the expression of PRDX2 could inhibit the invasion, migration and proliferation of MGC803 cells.

Objective To investigate the clinical efficacy of implementation of standardized enteral nutrition (EN) and its effects on prognosis in patients with severe traumatic brain injury (sTBI) undergoing mechanical ventilation (MV). Methods Eighty-eight patients with sTBI undergoing MV admitted to the Department of Critical Care Medicine of Yuyao People's Hospital from January 2016 to December 2017 were enrolled, they were divided into a control group (42 cases) and an experiment group (46 cases) depending on the demarcation timing of January 1, 2017, the beginning time of implementing standardized EN. All the patients received early EN and conventional treatment in the two groups. Additionally, the procedure of standardized EN was implemented in the experiment group. The differences in starting time of EN, the first defecation time, the rates of EN therapeutic energy and protein supply reaching their respective targets, duration of MV and ICU stay and 28-day mortality were compared between the two groups. Results The starting time of EN (hours: 25.61±8.74 vs. 32.79±8.63) and first defecation time (days: 3.03±0.79 vs. 3.61±0.89) were significantly earlier in the experiment group than those in the control group (both P < 0.05); the rates of energy and protein supply reaching the respective targets on the 5th day and 7th day after receiving EN were all significantly higher in the experiment group than those in the control group [rates of energy supply reaching target on the 5th day: (44.83±13.99)% vs. 37.59±10.88, and on the 7th day: (68.07±10.68)% vs. (62.69±9.87)%; rate of protein supply reaching target on the 5th day: (31.93±9.49)% vs. (27.06±8.08)%, and on the 7th day: (62.09±9.91)% vs. (54.55±11.27) %, all P < 0.05]; the durations of MV (hours: 9.24±2.91 vs. 10.67±3.41) and ICU stay (days: 12.09±3.37 vs. 13.93±4.98) in the experiment group were significantly shorter than those in the control group (all P < 0.05). No statistical significant difference in the 28-day mortality was observed between the experiment group and control group [21.74% (10/46) vs. 19.05% (8/42), P > 0.05]. Conclusion The efficacy of implementation of standardized EN in patients with sTBI undergoing MV is very significant, as it can significantly improve the rate of reaching EN target, and shorten the duration of MV and ICU stay.