Functional TRPV2 and TRPV4 channels in human cardiac c‐kit+ progenitor cells

The cellular physiology and biology of human cardiac c‐kit⁺ progenitor cells has not been extensively characterized and remains an area of active research. This study investigates the functional expression of transient receptor potential vanilloid (TRPV) and possible roles for this ion channel in regulating proliferation and migration of human cardiac c‐kit⁺ progenitor cells. We found that genes coding for TRPV2 and TRPV4 channels and their proteins are significantly expressed in human c‐kit⁺ cardiac stem cells. Probenecid, an activator of TRPV2, induced an increase in intracellular Ca²⁺ (Ca²⁺_i), an effect that may be attenuated or abolished by the TRPV2 blocker ruthenium red. The TRPV4 channel activator 4α‐phorbol 12‐13‐dicaprinate induced Ca²⁺_i oscillations, which can be inhibited by the TRPV4 blocker RN‐1734. The alteration of Ca²⁺_i by probenecid or 4α‐phorbol 12‐13‐dicprinate was dramatically inhibited in cells infected with TRPV2 short hairpin RNA (shRNA) or TRPV4 shRNA. Silencing TRPV2, but not TRPV4, significantly reduced cell proliferation by arresting cells at the G0/G1 boundary of the cell cycle. Cell migration was reduced by silencing TRPV2 or TRPV4. Western blot revealed that silencing TRPV2 decreased expression of cyclin D1, cyclin E, pERK1/2 and pAkt, whereas silencing TRPV4 only reduced pAkt expression. Our results demonstrate for the first time that functional TRPV2 and TRPV4 channels are abundantly expressed in human cardiac c‐kit⁺ progenitor cells. TRPV2 channels, but not TRPV4 channels, participate in regulating cell cycle progression; moreover, both TRPV2 and TRPV4 are involved in migration of human cardiac c‐kit⁺ progenitor cells.     

高产莫纳可林K低产桔霉素红曲霉菌株的筛选和发酵条件初步优化

采集全国各地红曲霉制品中的红曲霉资源,经分离获278株红曲霉菌株。高效液相色谱(HPLC)法测定各菌株发酵液中莫纳可林K(Monacolin K)和桔霉素含量,筛选获1株Monacolin K产量较高、桔霉素含量较低的红曲霉菌株编号为M-22。依据形态特征和ITS基因序列,参照红曲霉属分类检索表,鉴定M-22菌株为紫色红曲霉(Monascus purpureus)。通过摇瓶发酵对温度、初始pH、碳源、氮源、碳氮比(C/N)等因素进行优化,确定M-22菌株摇瓶发酵产Monacolin K适宜条件为发酵温度26℃、初始pH 5.0、转速160 r/min,甘油为碳源、蛋白胨为氮源、碳氮比(C/N)为5:1,Monacolin K产量显著提高,最高为107.16 mg/L。以优化的发酵条件对M-22菌株进行5 L发酵罐发酵,发酵液中Monacolin K产量最高为189.83 mg/L,桔霉素含量32.53μg/L,红曲色素色价为16.38 U/m L。     

蝉棒束孢菌子实体对大鼠肾小管上皮细胞的影响

对蝉棒束孢菌子实体（0.75g/kg）重复灌胃SD雄性大鼠90d及恢复28d的早期肾损伤生物标记物肾损伤分子-1（KIM-1）和中性粒细胞明胶酶相关载脂蛋白（NGAL）进行测定,评估蝉棒束孢菌子实体对肾小管上皮细胞的影响;研究不同剂量蝉棒束孢菌子实体（0.25g/kg、0.5g/kg、1.0g/kg）对肾小管上皮细胞增殖和增生能力的影响。给药30、60、90d及恢复28d时,SD大鼠血清中KIM-1浓度与对照组相比均无显著差异（P>0.05）,给药30d、60d时,SD大鼠血清中NGAL浓度与对照组相比均无显著差异（P>0.05）,给药90d及恢复28d时,SD大鼠血清中NGAL浓度低于对照组（P<0.05）,且给药90d组与对照组相比有显著性差异（P<0.01）;免疫组化检测增殖细胞核抗原法（PCNA）及四甲基偶氮唑盐微量酶反应比色法（MTT）表明：与对照组相比,蝉棒束孢菌子实体能使肾小管上皮细胞增生能力增强,未导致肾小管上皮细胞凋亡。     

Construction of an efficient genomic editing system with CRISPR/Cas9 in the vector mosquito Aedes albopictus

Aedes (Stegomyia) albopictus, also known as the Asian tiger mosquito, is a mosquito which originated in Asia. In recent years, it has become increasingly rampant throughout the world. This mosquito can transmit several arboviruses, including dengue, Zika and chikungunya viruses, and is considered a public health threat. Despite the urgent need of genome engineering to analyze specific gene functions, progress in genetical manipulation of Ae. albopictus has been slow due to a lack of efficient methods and genetic markers. In the present study, we established targeted disruptions in two genes, kynurenine hydroxylase (kh) and dopachrome conversion enzyme (yellow), to analyze the feasibility of generating visible phenotypes with genome editing by the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein 9 (Cas9) system in Ae. albopictus. Following Cas9 single guide RNA ribonucleoprotein injection into the posterior end of pre-blastoderm embryos, 30%-50% of fertile survivors produced alleles that failed to complement existing kh and yellow mutations. Complete eye and body pigmentation defects were readily observed in GI pupae and adults, indicating successful generation of highly heritable mutations. We conclude that the CRISPR/Cas9-mediated gene editing system can be used mAe. albopictus and that it can be adopted as an efficient tool for genome-scale analysis and biological study.     

镍钛形状记忆合金环抱器内固定术对多发性肋骨骨折患者术后疼痛、呼吸功能及肺部感染的影响

目的:探讨多发性肋骨骨折患者采用镍钛形状记忆合金环抱器内固定术治疗的临床效果。方法:选取2016年1月-2018年1月期间我院收治的97例多发性肋骨骨折患者。按照不同的治疗方法将患者分为对照组(n=47)和研究组(n=50),对照组患者给予胸部护板外固定术,研究组患者给予镍钛形状记忆合金环抱器内固定术。比较两组患者术后7 d的临床疗效,比较两组术前、术后7 d疼痛程度及呼吸功能情况,记录两组术后并发症发生情况。结果:研究组患者治疗后总有效率为92.00%(46/50),高于对照组患者的76.60%(36/47)(P0.05)。两组患者术后7 d视觉疼痛模拟评分量表(VAS)评分均较术前降低,且研究组低于对照组(P0.05)。两组患者术后7 d最大自主通气量占预计值百分比(MVV%)较术前升高,且研究组高于对照组(P0.05);两组患者术后7 d第1秒用力呼气肺活量占预计值百分比(FEV1%)比较无差异(P0.05)。研究组患者术后并发症总发生率8.00%(4/50),低于对照组的23.40%(11/47)(P0.05)。结论:多发性肋骨骨折经镍钛形状记忆合金环抱器内固定术治疗安全、有效,可明显减轻患者术后疼痛及改善呼吸功能,临床应用价值较高。     

乳腺癌患者心理韧性相关影响因素分析及心理韧性与焦虑的关系

目的:分析乳腺癌患者心理韧性的影响因素,并分析心理韧性与焦虑的关系。方法:于2017年2月～2018年4月期间,选择蚌埠医学院第二附属医院收治的乳腺癌患者287例为研究对象,分别采用心理韧性量表(RS)、焦虑自评量表(SAS)评估研究对象的心理韧性及焦虑状态,并采用多因素Logistic回归分析法分析乳腺癌患者心理韧性相关的影响因素,应用多元逐步线性回归分析法分析乳腺癌患者心理韧性与焦虑的关系。结果:乳腺癌患者RS得分为(76.72±9.82)分。单因素分析显示,不同年龄、居住地、家庭月收入及费用支出形式患者的RS得分比较差异有统计学意义(P0.05);不同文化程度、婚姻状况、手术类型及肿瘤分期患者的RS得分比较差异无统计学意义(P0.05)。多因素Logistic回归分析结果显示,年龄为20～40岁、居住地为农村、家庭月收入3000元、费用支出形式为自费是乳腺癌患者心理韧性的影响因素(P0.05)。乳腺癌患者SAS得分为(5.02±1.42)分,以心理韧性为自变量,焦虑为因变量进行多元逐步线性回归分析,结果显示,乳腺癌患者心理韧性为焦虑的预测因子,其心理韧性水平越高,焦虑程度越低(P0.05)。结论:乳腺癌患者心理韧性水平偏低,且其对患者焦虑状态具有预测作用,年龄为20～40岁、居住地为农村、家庭月收入3000元、费用支出形式为自费是乳腺癌患者心理韧性的影响因素,临床治疗中应该根据以上影响因素进行相关干预。     

胃大部切除术后消化性溃疡患者残胃癌危险因素分析及其干预措施

目的:探究消化性溃疡患者胃大部切除术后发生残胃癌的危险因素,并提出相应干预措施。方法:选取我院收治的行胃大部切除术治疗后的消化性溃疡患者180例作为研究对象,将其随机分为两组,每组90例。对照组给予常规干预,观察组给予综合干预,观察两组的干预效果,同时将发生残胃癌患者与未发生残胃癌患者各项资料进行对比,分析影响残胃癌发生的危险因素。结果:观察组中没有患者发生残胃癌,对照组中有10例患者发生残胃癌,发生率为11.11%,显著高于观察组(P0.05);两组患者在首次手术年龄、性别、HP感染上进行对比差异无统计学意义(P0.05);两组患者在首次手术吻合方式、首次手术溃疡位置上对比具有统计学意义(P0.05),残胃癌组组采用毕Ⅰ式2例,毕Ⅱ式7例,Roux-Y式1例;未发生残胃癌组采用毕Ⅰ式82例,毕Ⅱ式74例,Roux-Y式14例,残胃癌组首次手术溃疡位置以十二指肠球部为主,未发生残胃癌组以十二指肠球部、胃体部为主;Logistics回归分析结果证实首次手术吻合方式、首次手术溃疡位置属于残胃癌发生的主要危险因素。结论:术后残胃癌发生危险因素包括首次手术吻合方式、首次手术溃疡位置,同时相关并发症的发生同样会增加术后残胃癌发生风险,应加强引流管干预、并发症干预,并将此作为基础制定综合干预方案。     

围手术期应用ω-3多不饱和脂肪酸对胃癌根治术患者术后恢复及生活质量的影响

目的:探讨围手术期应用ω-3多不饱和脂肪酸对胃癌根治术患者术后恢复及生活质量的影响。方法:选择2015年1月～2016年12月期间南充市中心医院收治并行胃癌根治术的胃癌患者100例,按照随机数字表法分为对照组和观察组,每组50例。对照组接受常规静脉营养支持治疗,观察组在此基础上接受ω-3多不饱和脂肪酸治疗。检测并比较两组患者术前、术后1d、术后2周血清视黄醇结合蛋白(RBP)、前白蛋白(PA)、白蛋白(ALB)、转铁蛋白(RTF)、CD3~+、CD4~+、CD8~+细胞水平,并于术前、术后2周应用生活质量核心量表(EORTCQ LQ-C30)评价患者生活质量。结果:两组患者术后1d以及对照组术后2周的RBP、PA、ALB、RTF、CD3~+、CD4~+、CD4~+/CD8~+均较术前显著降低(P0.05),而CD8~+均较术前显著升高(P0.05),术后1 d、术后2周观察组RBP、PA、ALB、RTF、CD3~+、CD4~+、CD4~+/CD8~+显著高于对照组(P0.05),而CD8~+显著低于对照组(P0.05);术后两组患者EORTCQ LQ-C30功能量表各项评分较术前均显著提高,症状量表各项评分较术前显著降低(P0.05),术后观察组功能量表各项评分显著高于对照组,症状量表各项评分显著低于对照组(P0.05)。结论:应用ω-3多不饱和脂肪酸能够促进胃癌根治术后患者机体免疫功能的恢复,改善患者的营养状况,提高患者的生活质量。