A tetracycline-regulatable adeno-associated virus vector for double-gene transfer

An increasing demand for polycistronic vectors that express multiple genes simultaneously has arisen in recent years to obtain an efficient gene therapy. Armed with the knowledge that the expression of transgene in mammalian cells often requires tight control, we constructed in this study a tetracycline-regulated double-gene adeno-associated virus (AAV) vector carrying green and red fluorescent protein genes and expressed it in PC12 cells. When detected by fluorescence microscope and fluorescence-activated cell sorting, gene expression was induced by 44-66-fold and could be reversibly controlled by doxycycline. This double-gene AAV vector may be useful for regulated expression of two genes or a marker to monitor transgene expression.     

Different ion channel mechanisms between low concentrations of capsaicin and high concentrations of capsaicin and nicotine regarding peptide release from pulmonary afferents

Vagal nerve stimulation (1 Hz for 1 min), capsaicin (10^-8 M and 10^-6 M), resiniferatoxin (3 × 10^-10 M) and nicotine (10^-4 M) evoked a non-cholinergic bronchoconstriction in the isolated perfused guinea-pig lung preparation. Simultaneously there was an increase in the perfusate levels of calcitonin gene-related peptide-like immunoreactivity, suggesting release from sensory nerves. Both the bronchoconstriction and peptide release evoked by a low concentration of capsaicin (10^-8 M) and that evoked by nerve stimulation were depressed by tetrodotoxin, suggesting involvement of Na⁺ channel dependent depolarization. Since the effects of capsaicin (10^-8 M) and vagal nerve stimulation were inhibited by ω-conotoxin but not influenced by nifedipine, the Ca²⁺-channel involved is probably of N-type. Furthermore, the capsaicin analogue resiniferatoxin also evoked ω-conotoxin sensitive peptide release and bronchoconstriction. At the higher capsaicin concentration (10^-6 M), the functional response was only slightly inhibited by wconotoxin or tetrodotoxin indicating that capsaicin at this concentration evoked peptide release and functional effects through other mechanisms, probably involving Ca²⁺ fluxes in the non-selective cation channel associated with the proposed capsaicin receptor. The nicotine (10^-4 M) evoked peptide release and bronchoconstriction were only marginally influenced by ω-conotoxin or tetrodotoxin. It is concluded that the ion-channel mechanisms underlying the peptide releasing properties of antidromic nerve stimulation and low concentrations of capsaicin are similar and depend on action potential propagation, whereas capsaicin in high, toxic concentration and nicotine mainly act via receptor operated channels.     

肝细胞增殖因子促进受损肝细胞增殖的可能机制

目的： 探讨肝细胞增殖因子（ALR）促进受损肝细胞增殖的可能机制。方法: 采用细胞增殖试验（MTT法）观察经ALR刺激的大鼠肝枯否细胞(KC)的条件培养液(KCCM+)对受损肝细胞增殖的影响，同时用免疫组织化学方法检测正常大鼠肝脏组织中ALR的分布、大鼠枯否细胞膜表面结合的ALR以及ALR对枯否细胞IL-6表达的影响。结果: 受损肝细胞经枯否细胞条件培养液刺激后增殖明显。正常大鼠肝细胞内可表达合成ALR，枯否细胞膜表面可见ALR免疫反应颗粒，经ALR刺激后枯否细胞IL-6免疫反应信号增强。结论: ALR可能通过首先刺激枯否细胞，从而促进受损肝细胞增殖。     

三维及多平面重建CT技术在鼻部整形中的应用价值分析

目的:分析在鼻部整形中三维及多平面重建CT技术的应用价值。方法:选择2018年2月~2019年7月本院收治的60例行鼻部整形的患者进行分析,所有患者均开展三维及多平面重建CT技术检测,结合检查结果,开展相应的隆鼻手术。分析术后不良反应、患者满意度。术后6个月,在CT测量下,检测前后鼻部解剖数据。结果:在鼻部整形中,三维及多平面重建CT技术的应用,可以准确了解鼻部结构,如观察侧鼻骨、鼻部皮肤、鼻根等情况。术后评价,患者对鼻部整形满意度较高,术后6个月,患者对鼻部整形满意度较高,术后与术后6个月患者鼻部满意度相比,无统计学意义,P>0.05。术后60例患者没有出现出血、切口感染等反应,同时也没有出现肋软骨外漏与倾斜等现象。经随访术后6个月发现1例患者的鼻背软骨出现轻度弯曲,但无明显表现,没有做特殊处理;其余患者,鼻部外形均表现为鼻尖挺拔、鼻背曲线流畅。结论:在鼻部整形中,应用三维及多平面重建CT技术,可以辅助提高隆鼻手术准确度,并在CT技术的引导下,准确植入自体肋软骨,可以为手术成功奠定基础,促使鼻唇与周围器官形态的协调性,进一步提高患者满意度。     

低分子肝素钙抗凝对多囊卵巢综合征所致不孕症患者来曲唑诱导排卵疗效的影响

目的分析低分子肝素钙抗凝对多囊卵巢综合征(PCOS)所致不孕症患者来曲唑诱导排卵疗效的影响。方法选取2014年1月—2019年1月九江市第三人民医院和彭泽县妇幼保健院收治的156例PCOS所致不孕症患者,采用随机数字表分成两组,每组78例。对照组口服来曲唑治疗,观察组采取口服来曲唑联合低分子肝素钙注射液治疗。比较两组的疗效、子宫内膜容受性、凝血功能指标、促排卵结局及不良反应。结果观察组总有效率[94.87%(74/78)]显著高于对照组[80.77%(63/78)],差异有统计学意义(χ²=7.252,P=0.007)。治疗后,观察组子宫内膜厚度、子宫动脉搏动指数(PI)及阻力指数(RI)分别为(9.75±1.03)mm、(1.30±0.26)及(0.58±0.12),对照组子宫内膜厚度、PI及RI分别为(7.48±0.95)mm、(1.71±0.33)及(0.66±0.11),两组比较差异均有统计学意义(t=14.308,8.619,4.340,均P<0.05)。治疗后,两组凝血功能指标比较差异均有统计学意义(均P<0.05)。观察组妊娠率[52.56%(41/78)]显著高于对照组[32.05%(25/78)],自然流产率[2.44%(1/41)]显著低于对照组[16.00%(4/25)],差异均有统计学意义(均P<0.05)。观察组不良反应发生率[16.67%(13/78)]与对照组[12.82%(10/78)]比较差异无统计学意义(χ²=0.459,P=0.498)。结论PCOS所致不孕症患者在应用来曲唑诱导排卵的同时联合低分子肝素钙抗凝治疗能有效改善患者的凝血-纤溶状态,明显提高患者的子宫内膜容受性,改善促排卵结局。     

胎儿脑灰质异位合并FLNA基因突变病例报告

报告1例产前超声联合胎儿颅脑磁共振成像（magnetic resonance imaging,MRI）诊断胎儿脑灰质异位合并细丝蛋白A（filamin A,FLNA）基因突变的病例。患者超声检测结果显示侧脑室外壁凹凸不平,颅后窝池宽,透明隔间腔略窄,小脑延髓池轻度增宽。MRI诊断结果为胎儿脑灰质异位合并大枕大池。羊水穿刺获得脱落细胞,采用全外显子测序和基因拷贝数变异（copy number variations,CNV）分析,显示与脑灰质异位疾病相关的FLNA基因外显子出现缺失,其为可能导致脑灰质异位的致病基因。因此,超声结合胎儿颅脑MRI能有效提高脑灰质异位的诊断率,其中染色体检查可筛查出部分致病原因,并对下次妊娠提供帮助。该病例可丰富临床医生对胎儿大脑灰质异位的产前超声检查、合并畸形、临床表现等的认识。     

自体骨膜游离移植修复关节软骨缺损进展

探讨自体骨膜游离移植修复关节软骨缺损的研究进展。从骨膜的解剖生理功能和骨膜再生软骨能力两方面进行分析,并阐述了关节内营养环境、关节活动、骨膜生发层不同朝向、年龄及手术技巧等因素对移植修复的影响。同时总结了该方法的临床应用状况、效果及存在的问题。实验及临床的研究证明自体骨膜游离移植是修复关节软骨缺损的重要手段,局部环境因素影响骨膜成软骨。     

急性白血病患者血清可溶性白细胞介素-2受体水平及临床意义

用双抗体夹心酶联免疫吸附试验检测 30例急性白血病患者血清可溶性白细胞介素 - 2受体 (SIL -2R)水平。结果　急性白血病均显著高于正常对照组 ,且和病情的严重程度密切相关 ,是判定预后的一个有用的指标。     

盔瓣耳叶苔化学成分的研究

目的 研究耳叶苔科植物灰瓣耳叶苔Frullania muscicola Steph.的化学成分。方法 用硅胶、SephadexLH－20柱层析,通过理化性质鉴定及光谱数据分化确定化合物的结构。结果 分离到3个甾体化合物和4个黄酮化合物,分别鉴定为豆甾醇二十烷酸酯（I）、谷甾醇（Ⅱ）、洋芹素－7,4′－二甲醚（Ⅲ）、黄芩素－6,4′－二甲醚（Ⅳ）、6－羟基本犀草素－6,3′－二甲醚（Ⅴ）、黄芩素－6－甲醚（Ⅳ）和胡萝卜苷（Ⅶ ）。结论 化合物Ⅰ为新化合物,其余6个化合物均为首次从该植物中分得。     

幽门螺杆菌感染与冠心病发病的关系

目的　探讨幽门螺杆菌(Hp)感染是否为增加冠心病发病率的危险因素。方法　采用病例对照研究,从健康体检者中随机挑选127名作为正常对照组。根据病史、心电图、冠脉造影等确诊为冠心病的111例患者作为冠心病组,以血清抗HpIgG抗体阳性为感染标志,并测定血浆纤维蛋白原及脂质过氧化物浓度。结果　冠心病组Hp感染率(61%)高于正常对照组(39%)(P＜0.001);Hp感染是独立于其它冠心病易患因素的危险因素(比值比2.785,x2=14.82,P＜0.001);冠心病组CagA阳性感染率(98.5%)高于正常对照组(67.4%)(P＜0.05);冠心病组Hp阳性者的血浆纤维蛋白原及LPO浓度高于Hp阴性者(P＜0.05)。结论　Hp感染可能是发生冠心病的危险因素。慢性炎症引起脂质过氧化物、血浆纤维蛋白原增高,促进血栓形成可能是Hp感染诱发冠心病的机制之一。