Impact of Ocular Residual Astigmatism on Visual Outcomes of Myopic Astigmatism Correction after Small Incision Lenticule Extraction

Objective: To calculate ocular residual astigmatism (ORA) by vector analysis and to evaluate the impact of ocular residual astigmatism on refractive and visual outcomes after correction of myopic astigmatism by small incision lenticule extraction (SMILE). Methods: In this prospective case-series study, 115 eyes with myopic astigmatism were treated with SMILE from Jauary 2019 to August 2019 atRefractive Center of Ophthalmology Department, Shengjing Hospital, China Medical University. ORA was calculated by vector analysis and patients were divided into two groups according to the magnitude of ORA: high ORA group (ORA≥1.00 D) with 42 eyes and low ORA group (ORA<1.00 D) with 73 eyes. The follow-up period was 3 months after the operation. Uncorrected visual acuity (UCVA) and pythagorean length were measured and compared after the operation. The following measurements were compared: refractive outcomes, total higher order aberrations (tHOAs), vertical trefoil, vertical coma, horizontal coma, oblique trefoil, oblique quadrafoil, oblique secondary astigmatism, spherical aberration, vertical secondary astigmatism, and vertical quadrafoil. Contrast sensitivity was compared between the high and low ORA groups under either 85 cd/m2 or 3 cd/m2 illumination from 1.5 c/d to 18 c/d. The data were analyzed by an independent-samples t test and Mann-Whitney U test to compare the differences between the two groups. Results: Three months postoperatively, refractive outcomes showed that postoperative residual astigmatism in the high ORA group was significantly higher than in the low ORA group (t=3.293, P=0.001). Vector analysis showed that the error vector (EV), absolute error of angle (absolute EA) and index of success (SI) were higher in the high ORA group (t=-3.235, P=0.001; t=-2.326, P=0.020; t=-2.587, P=0.010). After SMILE，all cases had achieved a desirable visual acuity: high ORA group: -0.15±0.05, low ORA group: -0.15±0.05. There were no statistically significant differences in visual acuity, pythagorean length, tHOAs, vertical trefoil, vertical coma, horizontal coma, oblique trefoil, oblique quadrafoil, oblique secondary astigmatism, spherical aberration, vertical secondary astigmatism, or vertical quadrafoil between the two groups. An obvious difference was found in contrast sensitivity, which showed a higher contrast sensitivity at a spatial frequency of 18 c/d with 85 cd/m2 in the low ORA group than in the high ORA group (t=-2.877, P=0.005), but this tendency was not present at 3 cd/m2 . Conclusions: Ocular residual astigmatism impacts the precision of SMILE in correcting myopic astigmatism. This leads to a tendency of more residual astigmatism in refractive outcomes, and reduces contrast sensitivity at high spatial frequencies. However, it does not affect the outcome of obtaining good visual quality in both visual acuity and higher order aberrations.     

角膜内皮基因转移

基因治疗的关键技术是基因转移。角膜内皮基因转移是角膜内皮基凶治疗的基础,其主要方法有病毒载体和非病毒载体介导法,他们各有其优缺点。对角膜内皮基因转移技术和载体的研究现状,及未来载体的研究方向进行综述。     

抗人膀胱癌单克隆抗体重链可变区基因的克隆及原核表达载体的构建

目的 克隆抗人膀胱癌单克隆抗体重链可变区基因(VH),并构建其原核表达载体.方法 从能分泌抗人膀胱癌单克隆抗体的杂交瘤细胞BDI-1中提取总RNA,通过RT-PCR扩增出VH cDNA,用HindⅢ和XhoⅠ酶切纯化的RT-PCR产物和原核表达载体pET28a( ),在T4 DNA连接酶作用下室温连接.重组质粒经酶切鉴定,阳性克隆测序并进行序列分析.结果 扩增出VH cDNA片段,大小约为370bp,重组质粒的酶切鉴定结果与预期一致.VH基因序列长度为366bp,编码122个氨基酸.VH基因属于鼠免疫球蛋白重链Ⅱ亚类.结论 成功克隆出抗人膀胱癌单克隆抗体重链可变区基因,并成功构建其原核表达载体.     

靶向非病毒载体一次及持续介导Ⅰ型单纯疱疹病毒胸腺嘧啶核苷激酶基因转导卵巢癌细胞的比较

目的比较靶向非病毒载体GE7系统介导Ⅰ型单纯疱疹病毒胸腺嘧啶核苷激酶(HSV1-tk)基因一次及持续转导卵巢癌细胞的转导效率及杀伤效应。方法靶向非病毒载体系统GE7分别与标志基因β-半乳糖苷酶(β-gal)基因和治疗基因HSV1-tk基因构成载体复合物,一次及持续体外转导卵巢癌细胞CaOV3,通过5-溴-4-氯-3-吲哚-β-D-半乳糖吡喃糖甙(X-gaI)染色比较GE7系统一次及持续转导外源基因的转导效率。将丙氧鸟苷(GCV)加入一次及持续转导HSV1-tk基因的卵巢癌细胞,通过细胞生长抑制曲线、流式细胞分析等,观察GCV对一次及持续转导GE7/HSV1-tk的卵巢癌细胞的杀伤作用。结果X-gal染色显示,GE7-次转导外源基因的平均转导效率可达80%,持续转导达85%。当GCV为1μg/mL时,GE7/HSV1-tk一次转导的生长抑制率达82%,凋亡指数为15,而持续转导可达90%,凋亡指数为30。在相同的GCV浓度下,持续转导GE7/pCMV-tk卵巢癌细胞生长抑制率显著高于一次转导(P<0.05)。结论GE7载体系统持续转导卵巢癌细胞较一次转导的平均转导效率高,持续转导的GE7/HSV1-tk/GCV基因治疗系统杀伤作用更大。     

短暂性脑缺血发作中医证候量化诊断标准研究

目的：建立短暂性脑缺血发作中医证候量化诊断标准。方法：根据文献研究、专家咨询、临床流行病学调查获得短暂性脑缺血发作四诊信息。将493例临床患者随机分为运算组（370例）和考核组（123例）,以基于熵的复杂系统分划方法提取运算组患者症状信息,分析证候之间组合规律,确立症状对证候贡献度,以诊断性试验ROC曲线分析方法建立诊断阈值,并依据考核组数据对其进行前瞻性检验。结果：短暂性脑缺血发作存在以络气郁滞、络气虚滞、痰湿、血瘀、阴虚、火热等基本证候,分别确立了不同症状对证候诊断的贡献度,建立了诊断阈值,前瞻性检验显示其具有良好的灵敏度、特异度及符合率。结论：基于熵的复杂系统分划方法可用于中医证候的量化研究,建立的短暂性脑缺血发作量化诊断标准具有良好的诊断效能,为中医证候规范化及进一步开展络病与血管病变相关性研究奠定了基础。     

巨噬细胞炎症蛋白1α修饰的小鼠肺癌疫苗的制备及体内抗肿瘤活性研究

目的以小鼠趋化因子巨噬细胞炎症蛋白1α(MIP-1α)的重组腺病毒载体(AdmMIP-1α)体外感染小鼠Lewis肺癌细胞株(LLC),观察其体内抗肿瘤活性,探讨其成为肺癌疫苗的可能性。方法AdmMIP-1α体外感染LLC细胞,通过绿色荧光蛋白(GFP)的表达检测感染率。连续14 d隔天进行细胞计数,观察细胞的体外生长曲线。取3×106修饰后的LLC细胞接种于C57BL／6小鼠,4周后在MIP-1α瘤苗组未荷瘤小鼠原接种部位的对侧再接种1×106野生型LLC或小鼠淋巴瘤细胞株(EL4)细胞。观察肿瘤体积。结果AdmMIP-1α能有效感染靶细胞LLC细胞,感染后LLC细胞的体外生长无明显改变。修饰后LLC细胞的体内成瘤性下降;4周后再接种LLC细胞,肿瘤生长速度明显减慢;而再接种的EL4细胞呈渐进性生长,与对照组无明显差异。结论表达MIP-1α基因的LLC细胞的体内成瘤性下降,并且能激发特异性免疫保护反应,有可能成为有效的肺癌疫苗。     

反向重复序列法构建神经病靶标酯酶RNA干涉表达质粒

目的构建基于通用真核表达载体的人神经病靶标酯酶双链RNA的稳定表达载体。方法在正义和反义引物两端分别加上EcoRⅠ和BamHⅠ的识别位点扩增得到神经病靶标酯酶活力域序列,构建含有目标基因的倒置重复序列的双链RNA表达载体pcDNA.NTE.dsRNA,酶切分析鉴定重组载体。脂质体法转染到Hela细胞中瞬时表达重组载体,酶活力测定其对细胞内神经病靶标酯酶活力的影响。结果成功构建了NTE双链RNA稳定表达载体pcDNA.NTE.dsRNA,表达该载体细胞内NTE活力明显下降至对照细胞的20%左右。结论采用反向重复序列法将靶标基因的编码序列正反向插入到通用真核细胞表达载体中,是构建哺乳细胞基因双链RNA稳定表达载体的有效方法。     

重组 GST-β-GT 融合蛋白的原核表达、纯化及活性鉴定

目的：构建噬菌体β‐GT蛋白的原核重组体系,诱导表达、纯化GST‐β‐GT 融合蛋白并对其进行酶活性测定。方法利用PCR技术从T4噬菌体中扩增β‐GT基因;经T‐A克隆,获得β‐GT基因片段,经酶切连接,构建原核表达载体pGEX‐6P‐1‐β‐GT ;经测序鉴定序列正确后,将所构建的载体转化进入大肠埃希菌BL21（DE3）中进行表达。利用IPTG诱导,经10％ SDS‐PAGE鉴定目的蛋白表达后,采用GST柱纯化目的蛋白;通过酶切和qPCR鉴定其活性。结果成功扩增了噬菌体β‐GT基因;重组质粒在大肠埃希菌BL21（DE3）中诱导表达出GST‐β‐GT融合蛋白;通过GST柱纯化后获得了GST‐β‐GT融合蛋白,纯度达95％;通过酶切和qPCR验证,此GST‐β‐GT融合蛋白具有T4‐β‐GT酶的活性。结论。原核表达GST‐β‐GT融合蛋白具有糖基转移酶活性。     

Integrated decision strategies for skin sensitization hazard

One of the top priorities of the Interagency Coordinating Committee for the Validation of Alternative Methods (ICCVAM) is the identification and evaluation of non‐animal alternatives for skin sensitization testing. Although skin sensitization is a complex process, the key biological events of the process have been well characterized in an adverse outcome pathway (AOP) proposed by the Organisation for Economic Co‐operation and Development (OECD). Accordingly, ICCVAM is working to develop integrated decision strategies based on the AOP using in vitro, in chemico and in silico information. Data were compiled for 120 substances tested in the murine local lymph node assay (LLNA), direct peptide reactivity assay (DPRA), human cell line activation test (h‐CLAT) and KeratinoSens assay. Data for six physicochemical properties, which may affect skin penetration, were also collected, and skin sensitization read‐across predictions were performed using OECD QSAR Toolbox. All data were combined into a variety of potential integrated decision strategies to predict LLNA outcomes using a training set of 94 substances and an external test set of 26 substances. Fifty‐four models were built using multiple combinations of machine learning approaches and predictor variables. The seven models with the highest accuracy (89–96% for the test set and 96–99% for the training set) for predicting LLNA outcomes used a support vector machine (SVM) approach with different combinations of predictor variables. The performance statistics of the SVM models were higher than any of the non‐animal tests alone and higher than simple test battery approaches using these methods. These data suggest that computational approaches are promising tools to effectively integrate data sources to identify potential skin sensitizers without animal testing. Published 2016. This article has been contributed to by US Government employees and their work is in the public domain in the USA.     

速度向量成像技术评价左房功能的可行性研究

目的 采用超声速度向量成像技术(VVI)评价左房功能,探讨其可行性及临床应用价值.方法 健康志愿者58例,应用Simpson法计算左心房被动射血分数(LAPEF)主动射血分数(LAAEF)和左房总排空分数(LATEF),采用VVI检测左心房长轴整体收缩期峰值应变(Ss)及峰值应变率(SRs),左心房长轴整体舒张早期峰值应变(Se)及峰值应变率(Sre)和左心房长轴整体舒张晚期峰值应变(Sa)及峰值应变率(Sra),并与左房容积参数进行比较.结果 左心房长轴整体收缩期峰值应变及峰值应变率与左房总排空分数相关,(r=0.398,P=0.005; r=0.365,P=0.007).左心房长轴整体舒张早期峰值应变及峰值应变率与左房被动射血分数显著相关(r=0.475,P<0.001; r=0.505,P<0.001).左心房长轴整体舒张晚期峰值应变及峰值应变率与左心房主动射血分数不相关(r=0.199,P=0.165;r=0.267,P=0.062).结论 速度向量成像技术对左房功能的评价具有可行性.