Whole-exome sequencing identifies novel mutations in genes responsible for retinitis pigmentosa in 2 nonconsanguineous Chinese families

AIM: To detect the pathogenetic mutations responsible for nonsyndromic autosomal recessive retinitis pigmentosa (RP) in 2 nonconsanguineous Chinese families. METHODS: The clinical data, including detailed medical history, best corrected visual acuity (BCVA), slit-lamp biomicroscope examination, fundus photography, optical coherence tomography, static perimetry, and full field electroretinogram, were collected from the members of 2 nonconsanguineous Chinese families preliminarily diagnosed with RP. Genomic DNA was extracted from the probands and other available family members; whole-exome sequencing was conducted with the DNA samples provided by the probands, and all mutations detected by whole-exome sequencing were verified using Sanger sequencing in the probands and the other available family members. The verified novel mutations were further sequenced in 192 ethnicity matched healthy controls. RESULTS: The patients from the 2 families exhibited the typical symptoms of RP, including night blindness and progressive constriction of the visual field, and the fundus examinations showed attenuated retinal arterioles, peripheral bone spicule pigment deposits, and waxy optic discs. Whole-exome sequencing revealed a novel nonsense mutation in FAM161A (c.943A>T, p.Lys315*) and compound heterozygous mutations in RP1L1 (c.56C>A, p.Pro19His; c.5470C>T, p.Gln1824*). The nonsense c.5470C>T, p.Gln1824* mutation was novel. All mutations were verified by Sanger sequencing. The mutation p.Lys315* in FAM161A co-segregated with the phenotype, and all the nonsense mutations were absent from the ethnicity matched healthy controls and all available databases. CONCLUSION: We identify 2 novel mutations in genes responsible for autosomal recessive RP, and the mutation in FAM161A is reported for the first time in a Chinese population. Our result not only enriches the knowledge of the mutation frequency and spectrum in the genes responsible for nonsyndromic RP but also provides a new target for future gene therapy.     

Eleutheroside B or E enhances learning and memory in experimentally aged rats

Eleutheroside B or E, the main component of Acanthopanax, can relieve fatigue, enhance memory, and improve human cognition. Numerous studies have confirmed that high doses of acetylcholine significantly attenuate clinical symptoms and delay the progression of Alzheimer’s disease. The present study replicated a rat model of aging induced by injecting quinolinic acid into the hippocampal CA1 region. These rats were intraperitoneally injected with low, medium and high doses of eleutheroside B or E (50, 100, 200 mg/kg), and rats injected with Huperzine A or PBS were used as controls. At 4 weeks after administration, behavioral tests showed that the escape latencies and errors in searching for the platform in a Morris water maze were dose-dependently reduced in rats treated with medium and high-dose eleutheroside B or E. Hematoxylin-eosin staining showed that the number of surviving hippocampal neurons was greater and pathological injury was milder in three eleutheroside B or E groups compared with model group. Hippocampal homogenates showed enhanced cholinesterase activity, and dose-dependent increases in acetylcholine content and decreases in choline content following eleutheroside B or E treatment, similar to those seen in the Huperzine A group. These findings indicate that eleutheroside B or E improves learning and memory in aged rats. These effects of eleutheroside B or E may be mediated by activation of cholinesterase or enhanced reuse of choline to accelerate the synthesis of acetylcholine in hippocampal neurons.     

512例胆管细胞癌的临床及预后特点

目的： 胆管细胞癌(cholangiocarcinoma，CCA)恶性程度较高，总体预后较差，初诊时分期较晚，治疗手 段有效率欠佳。本研究探索影响CCA发生、发展的临床特点及预后相关因素，以期为CCA的早期诊断及临床治疗提 供潜在的手段。方法： 回顾性分析中南大学湘雅二医院2002 年以来经病理确诊且临床资料完善的512 例CCA患者的 病历资料。利用Kaplan-Meier 法绘制生存曲线、log-rank 进行单因素分析，多元Cox回归法对有意义的变量进行多因 素分析。结果： CCA发病率≤60 岁者高于>60 岁者(61.13% vs 38.87%)，男性略高于女性(52.54% vs 47.46%)，糖类抗 原19-9(carbohydrate antigen 19-9，CA19-9)≥35 μg/L 者占66.21%，病理分期III 和IV 期患者居多(分别占49.22%和 17.58%)。单因素分析发现ALB，ALP，CA19-9 等因素与预后相关，多因素Cox 生存分析发现ALP，CA19-9，肿瘤 最大直径等是CCA预后独立影响因素。结论：CCA发病率≤60 岁人群较高，初诊时分期较晚，CA19-9 是较为敏感的 实验室指标。ALP，CA19-9，肿瘤最大直径，合并子瘤，肝硬化及TNM分期是影响CCA预后的独立因素。     

Tenofovir gel for HIV prevention for women: Perspectives of Key Opinion Leaders from India

ObjectivesCountry decision-making regarding adoption of new health technologies is a lengthy process that is informed by scientific, public health, and policy analyses. Key Opinion Leaders (KOLs), who include a range of influential individuals (e.g., Ministry of Health officials, leaders of medical and pharmacy associations, civil society representatives, donors), play a critical role in country decision-making around adoption and availability of new HIV-prevention tools. Their perspectives can shape public opinion and influence policy, programming, and procurement decisions.MethodsIn-depth face-to-face interviews with a dozen purposefully selected national KOLs in India were conducted to gain their perspectives around ARV-based prevention for women, focusing on microbicides in general and tenofovir vaginal gel specifically. Theme-based content emerged and was analyzed using ATLAS.ti software.ResultsWhile generally supportive, KOLs expressed caveats regarding the role tenofovir gel might play in the overall HIV-prevention program. Key outstanding issues that would need to be addressed include product effectiveness, potential for resistance, product cost, feasibility of HIV testing and re-testing, and effective targeting of key populations for public health impact.ConclusionsAs new HIV-prevention technologies advance through clinical trials, there is a concomitant necessity to engage in policy analysis exercises. Formative research with stakeholders early in the process is an important yet often overlooked step. The potential role that the tenofovir gel could play in India will depend in large part on epidemiological, public health, and economic factors. Understanding target populations' perspectives on the product will also be critical for its successful introduction.     

Sodium phenylbutyrate improved the clinical state in an adult patient with arginase 1 deficiency

An adult female patient was diagnosed with arginase 1 deficiency (ARG1-D) at 4 years of age, and had been managed with protein restriction combined with sodium benzoate therapy. Though the treatment was successful in ameliorating hyperammonemia, hyperargininemia persisted. After being under control with a strict restriction of dietary protein, severe fall of serum albumin levels appeared and her condition became strikingly worsened. However, after sodium phenylbutyrate (NaPB) therapy was initiated, the clinical condition and metabolic stability was greatly improved.Current management of ARG1-D is aimed at lowering plasma arginine levels. The nitrogen scavengers, such as NaPB can excrete the waste nitrogen not through the urea cycle but via the alternative pathway. The removal of nitrogen via alternative pathway lowers the flux of arginine in the urea cycle. Thereby, the clinical complications due to insufficient amount of protein intake can be prevented. Thus, NaPB therapy can be expected as a useful therapeutic option, particularly in patients with ARG1-D.     

改良胰管空肠黏膜端侧吻合胰十二指肠切除术临床效果研究

目的探讨改良胰管空肠黏膜端侧吻合的胰十二指肠切除术临床效果。方法回顾性分析2016年1月至2019年3月62例行胰十二指肠切除术(PD)患者临床资料,其中30例行改良胰管空肠黏膜端侧吻合术(改良组),32例行传统胰管空肠黏膜端侧吻合术(传统组)。选用SPSS 20.00统计学软件进行数据分析,围术期相关指标和术后第1、3、5、7、9 d的腹腔引流液淀粉酶水平用(±s)表示,采用独立t检验;并发症发生率采用χ^2检验,P<0.05差异有统计学意义。结果改良组出血量明显小于传统组(P<0.05),其余相关围术期指标差异均无统计学意义(P>0.05);改良组术后发生1例胰瘘明显小于传统组9例(P<0.05),其中改良组B级胰瘘发生率明显小于传统组(P<0.05),生化瘘、C级胰瘘发生率、淋巴瘘、胃排空发生率比较差异均无统计学意义(P>0.05);两组患者腹腔引流液淀粉酶含量在术后第1、3、5、7、9 d的检测期间内,传统组腹腔引流液淀粉酶含量小于改良组(P<0.05)。结论改良胰管空肠黏膜端侧吻合术相比于传统胰管空肠黏膜端侧吻合术在PD中能有效降低胰瘘的发生率,具有一定的临床推广价值。     

非小细胞肺癌患者外周血癌胚抗原、铁蛋白和神经元特异度烯醇化酶水平与肿瘤增殖、侵袭的关系

目的 分析非小细胞肺癌患者外周血癌胚抗原（carcinoembryonic antigen,CEA）、血清铁蛋白（serum ferritin,SF）和神经元特异度烯醇化酶 （neuronal specific enolase,NSE）水平与肿瘤增殖、侵袭的相关性。方法 回顾性分析医院2019年8月-2020年12月诊治的86例非小细胞肺癌患者作为研究对象,将其纳入病例组。于同期选择体检健康的受试者86例,将其纳入对照组。使用倾向性评分匹配(propensity score matching,PSM)法保证两组研究对象年龄和性别平衡后检测血清CEA、SF和NSE水平,根据患者病理结果将其分成增殖组和非增殖组,侵袭组和非侵袭组。比较病例组及对照组CEA、SF和NSE水平,分析增殖组和非增殖组,侵袭组和非侵袭组CEA、SF和NSE水平,探讨不同CEA、SF和NSE表达水平对肿瘤增殖、侵袭的影响。结果 病例组患者血清CEA、SF和NSE水平高于对照组（均P<0.05）。病例组中86例患者中34例患者发生增殖,26例患者发生肿瘤侵袭。增殖患者血清CEA、SF和NSE高于非增殖患者（P<0.05）。侵袭患者血清CEA、SF和NSE水平高于非侵袭患者（P<0.05）。血清CEA、SF和NSE高表达与肿瘤增殖及侵袭情况呈正相关（P<0.05）。CEA、SF和NSE水平越高,肿瘤增殖及侵袭风险越大。结论 非小细胞肺癌患者血清 CEA、SF和NSE水平高表达,且表达情况直接影响肿瘤增殖和侵袭。     

血管紧张素Ⅱ1型受体拮抗剂对早期糖尿病肾病患者尿液中足细胞相关蛋白nephrin、自噬基因Beclin-1 mRNA排泄的影响

目的 观察早期糖尿病肾病(diabetic nephropathy, DN)患者尿液中足细胞、足细胞相关蛋白nephrin、自噬基因Beclin-1 mRNA 排泄及氯沙坦对上述指标的影响。方法 入选2018年1月-2021年1月我院确诊的符合标准的早期DN患者48 例,给予氯沙坦50 mg或100 mg口服6个月,观察治疗前后患者尿液中微量白蛋白、足细胞、足细胞相关蛋白nephrin mRNA、自噬基因Beclin-1 mRNA排泄。结果 早期DN患者尿液中微量白蛋白排泄、足细胞及足细胞相关蛋白nephrin、自噬基因Beclin-1 mRNA排泄均较健康人明显增加(P<0.01)。给予氯沙坦治疗6个月后,与治疗前比较,患者尿液中微量白蛋白、足细胞排泄减少(P<0.05),nephrin mRNA排泄呈下降趋势(P<0.05),自噬基因Beclin-1 mRNA排泄较治疗前有一定增加,但差异无统计学意义(P=0.067)。结论 早期DN患者可通过尿液检查证实存在足细胞损伤。血管紧张素Ⅱ1型受体拮抗剂可以减少DN患者尿微量白蛋白排泄,保护足细胞,并可能增强足细胞自噬作用。     

水通道蛋白4对一氧化碳中毒后迟发性脑病大鼠神经损伤的影响

目的 探讨水通道蛋白4(Aquaporin 4,AQP4)对一氧化碳(Carbon monoxide,CO)中毒后迟发性脑病(Delayed encephalopathy,DEACMP)大鼠神经损伤的影响。方法 将210只雄性SD大鼠随机分为空白对照(Blank control,BC)组、CO中毒(CO)组、钠-钾-氯共转运体(Na⁺-K⁺-Cl^- cotransporter,NKCC)抑制剂处理(布美他尼)组、p38-丝裂原活化蛋白激酶(Mitogen activated protein kinase,MAPK)抑制剂处理(MAPK)组和AQP4特异性抑制剂处理(AQP4抑制剂)组,每组各42只; 根据造模后不同时间点将每组大鼠进一步分为染毒3、6、12、24、48、72 h和7 d后共7个亚组,每亚组各6只; 取大鼠脑前额叶皮质组织,计算脑皮质含水量,采用HE法观察脑皮质形态; 采用免疫组化链霉菌抗生物素蛋白-过氧化物酶(Streptavidin-perosidase,SP)染色法测定大鼠脑皮质AQP4,p38 MAPK,NKCC1、胶质纤维酸性蛋白(Glial fibrillary acidic protein,GFAP)和S100钙结合蛋白B(S100 calcium binding protein B,S100B)蛋白表达水平。结果 与BC组比较,CO组大鼠在染毒3、6、12、24、48、72 h后尾静脉COHB水平和脑皮质含水量显著升高,脑皮质AQP4,p38 MAPK,NKCC1,GFAP和S100B蛋白表达水平显著升高,染毒7 d后恢复正常(P<0.05); 与CO组比较,布美他尼组、MAPK组和AQP4抑制剂组大鼠脑皮质含水量显著降低,脑皮质AQP4,p38 MAPK,NKCC1,GFAP和S100B蛋白表达水平显著降低,且AQP4抑制剂组变化更明显(P<0.05)。结论 p38-MAPK/NKCC信号通路可能参与调控CO中毒DEACMP大鼠脑皮质AQP4表达,抑制AQP4表达可有效减轻大鼠脑水肿并改善预后,有望成为预防和治疗DEACMP的新靶点。