Alport Disease: A Review of the Diagnostic Difficulties

In patients with familial hematuria, ultrastructural study of the renal biopsy has been the gold standard for the diagnosis of Alport disease, based on characteristic findings of glomerular basement membrane thickening due to reduplication of the lamina densa. But the diagnosis has difficulties as not all biopsies from Alport disease patients have these structural changes. In adult female patients or in children, extensive thinning of the basement membrane can be the major abnormality by electron microscopy. Until the genetic mutation of collagen IV responsible for Alport disease can be demonstrated in all patients, the diagnosis will continue to be a challenge at the clinical and at the ultrastructural levels.     

Cytologic Observations on Trypsinized Cells Obtained from Human Endometrial Epithelium in Tissue Culture

Abstract

Although epithelial cells have been successfully grown from tissue culture of endometrial explants, they survived for only a few weeks before fibroblasts overgrew and eliminated the epithelium. We describe a method for growing endometrial epithelium from menstrual blood and passing them using 0.25% trypsin in EDTA; cells survived in vitro for 24 weeks. The morphology of the processed cells differed from that observed in vitro: trypsinized cells were rounded with long surface processes and microvilli; nuclei were irregular and contained a macronucleus; the cytoplasm appeared very granular. Immunocytochemistry revealed 90–95% positivity for anti-keratin; EM confirmed the light microscopic findings. Nuclei were large and irregular with a macronucleus and cytoplasmic organelles. The most striking feature of these cells was the abundance of rough endoplasmic reticulum, surface projections, and microvilli.

Endometrial epithelium undergoing trypsinization is not irreversibly damaged and retains its epithelial characteristics although it may be mechanically disrupted as a tissue and separated into single cells or small groups of cells. (The J Hislotechnol 10:233, 1987.)     

Surface engineering of artificial heart valve disks using nanostructured thin films deposited by chemical vapour deposition and sol-gel methods

Pyrolytic carbon (PyC) is widely used in manufacturing commercial artificial heart valve disks (HVD). Although PyC is commonly used in HVD, it is not the best material for this application since its blood compatibility is not ideal for prolonged clinical use. As a result thrombosis often occurs and the patients are required to take anti-coagulation drugs on a regular basis in order to minimize the formation of thrombosis. However, anti-coagulation therapy gives rise to some detrimental side effects in patients. Therefore, it is extremely urgent that newer and more technically advanced materials with better surface and bulk properties are developed. In this paper, we report the mechanical properties of PyC-HVD, i.e. strength, wear resistance and coefficient of friction. The strength of the material was assessed using Brinell indentation tests. Furthermore, wear resistance and coefficient of friction values were obtained from pin-on-disk testing. The micro-structural properties of PyC were characterized using XRD, Raman spectroscopy and SEM analysis. Also in this paper we report the preparation of freestanding nanocrystalline diamond films (FSND) using the time-modulated chemical vapour deposition (TMCVD) process. Furthermore, the sol-gel technique was used to uniformly coat PyC-HVD with dense, nanocrystalline-titanium oxide (nc-TiO₂) coatings. The as-grown nc-TiO₂ coatings were characterized for microstructure using SEM and XRD analysis.     

Comparative performance of Thin Layer Agar and Löwenstein–Jensen culture for diagnosis of tuberculosis

Sputum smear microscopy for the diagnosis of tuberculosis (TB) is cheap and simple but its sensitivity is low. Culture on Löwenstein–Jensen (LJ) is more sensitive but it takes a long time to yield results. Thin-Layer Agar (TLA) culture was suggested as an equally sensitive and faster alternative. We evaluated the performance of TLA for diagnosing TB in Jogjakarta, Indonesia. People with suspected TB presenting from July 2010 to July 2011 to two chest clinics of the National TB Control Programme network of Jogjakarta were eligible for inclusion. A sputum sample was sent to the Gadjah Mada University microbiology laboratory for concentration, smearing, Ziehl–Neelsen staining and culture on LJ and TLA. Sensitivity of cultures was evaluated against a composite reference standard (any positive culture). Time to detection of Mycobacteria was recorded. Out of 1414 samples, 164 (12%) were smear positive, 99 (7%) were scanty and 1151 (81%) were negative. On TLA and LJ respectively, 168 (12%) and 149 (11%) samples were positive, 72 (5%) and 32 (2%) were contaminated (κ = 0.64; 95% CI 0.59–0.69, p <0.01). Using the reference standard, 196 (14%) TB cases were identified. The sensitivity of TLA was 0.86 (95% CI 0.80–0.90), significantly higher (p 0.03) than for LJ (0.76; 95% CI 0.69–0.81). The median time to detection in days was significantly shorter (p <0.01) for TLA (12; 95% CI 11–13) than for LJ (44; 95% CI 43–45). TLA is a rapid and sensitive method for the diagnosis of TB. Implementation studies to evaluate the cost-effectiveness and impact of its introduction into programmatic settings are urgently needed.     

Pathology of the endometrium in women with chronic hepatitis C and HIV infection as a cause of unsuccessful in vitro fertilization attempts

Abstract

Aim: The purpose of the study is to detect endometrial pathology in individuals with blood-borne infections (hepatitis C virus [HCV] and HIV) and to assess the impact of quality of the endometrium on the efficiency of assisted reproductive technologies (ART).

Materials and methods of research: The study involved 56 women of reproductive age, 33 women with HCV, 22 with HIV infection (stages 3 and 4A) and 1 patient with co-infection of HCV and HIV, which was excluded from the further study. Ultrasound investigation of the small pelvic organs, hysteroscopy with biopsy of the mucosa of the uterine cavity, histological examination of the endometrium, determination of HCV RNA and HIV RNA in serum and endometrial tissue by PCR method were conducted.

Results: Ultrasound and histological studies of the endometrium revealed a high incidence of development of hyperplastic and inflammatory processes of the endometrium in women with HCV (86% and 100%) and HIV (51% and 99%). When molecular biological examination of endometrial tissue of these patients was performed with the use of a set of reagents “AmpliSens HCV/HBV/HIV-FL” produced by Federal Budgetary Institution of Science (FBUN) Central Research Institute of Epidemiology of Rospotrebnadzor of Russia, HCV was detected in 16.7% and HIV in 14.3% of cases. In 5% of patients with HCV infection and 7% with HIV, pathogens were detected in the endometrium under the non-detectable viral load in the blood plasma. Lower effectiveness of IVF in women with HIV and HCV is defined compared to the women without blood-borne infections.

Conclusion: Detected HIV and HCV replications in the endometrium is the likely cause of hyperplastic and inflammatory processes of the endometrium leading to reduced efficiency of the ART programs in patients with chronic hepatitis C and HIV.     

子宫内膜癌组织微小RNA-373表达情况及其对子宫内膜癌细胞增殖、迁移的影响研究

背景微小RNA-373 (miRNA-373)在多种恶性肿瘤(如肝癌、肺癌)的发生发展过程中具有重要作用,但其在子宫内膜癌(EC)发生发展中的作用尚不完全清楚。目的探讨EC组织miRNA-373表达情况及其对EC细胞增殖、迁移的影响。方法选取2012年6月-2013年6月在邯郸市妇幼保健院行手术治疗的64例EC患者的EC组织作为试验组,另选取同期在邯郸市妇幼保健院行子宫切除术的30例良性子宫肌瘤患者的正常组织作为对照组。采用实时荧光定量聚合酶链反应(qRT-PCR)检测两组miRNA-373表达情况,绘制Kaplan-Meier生存曲线以分析不同miRNA-373表达情况EC患者术后5年预后;分别比较转染miRNA-373模拟物(miRNA-373 mimic)及其阴性对照物(mimic-NC)、miRNA-373抑制剂(miRNA-373 inhibitor)及其阴性对照物(inhibitor-NC)的HEC-1B细胞大型肿瘤抑制基因2 (LATS2)表达情况、转染96 h后吸光度值、迁移细胞数量;采用双荧光素酶报告基因系统验证miRNA-373与LATS2间的靶标关系。结果 (1)试验组miRNA-373相对表达量高于对照组(P<0.05)。根据miRNA-373相对表达量平均值将试验组患者分为低表达组(n=26)和高表达组(n=38),Kaplan-Meier生存曲线显示,高表达组患者术后5年累积生存率为52.6%,低于低表达组的84.6%(P<0.05)。(2)转染miRNA-373 mimic的HEC-1B细胞LATS2相对表达量低于转染mimic-NC的HEC-1B细胞(P<0.05);转染miRNA-373 inhibitor的HEC-1B细胞LATS2相对表达量高于转染inhibitor-NC的HEC-1B细胞(P<0.05)。转染96 h后,转染miRNA-373 mimic的HEC-1B细胞吸光度值高于转染mimic-NC的HEC-1B细胞(P<0.05);转染miRNA-373 inhibitor的HEC-1B细胞吸光度值低于转染inhibitor-NC的HEC-1B细胞(P<0.05)。转染miRNA-373 mimic的HEC-1B细胞中迁移细胞数量多于转染mimic-NC的HEC-1B细胞(P<0.05);转染miRNA-373 inhibitor的HEC-1B细胞中迁移细胞数量少于转染inhibitor-NC的HEC-1B细胞(P<0.05)。(3)双荧光素酶报告基因系统检测结果显示,LATS2含有miRNA-373的潜在结合位点。野生型LATS2中miRNA-373相对荧光强度低于空载对照(P<0.05);突变型LATS2中miRNA-373相对荧光强度与空载对照比较,差异无统计学意义(P>0.05)。结论 EC组织miRNA-373呈高表达,其可通过直接靶向调节LATS2的表达而促进EC细胞增殖、迁移,进而影响EC患者预后。     

滋癸益经汤联合滋肾育胎丸治疗卵巢早衰的研究

【目的】观察滋癸益经汤联合滋肾育胎丸治疗卵巢早衰(POF)的临床疗效。【方法】将66例POF患者随机分为治疗组和对照组,每组各33例。治疗组给予口服滋癸益经汤加滋肾育胎丸治疗,对照组给予口服滋癸益经汤加维生素E胶囊治疗,3个月为1个疗程,共治疗2个疗程。观察2组患者治疗前、治疗3个月及6个月后的各项证候积分、月经情况、血清性激素水平[促卵泡生成激素(FSH)、促黄体生成激素(LH)、雌二醇(E2)、抗苗勒氏管激素(AMH)]及彩超检查指标[子宫内膜厚度、卵巢体积、卵泡数量、卵巢最大血流速度(PSV)和卵巢阻力指数(RI)]的变化情况,并评价2组患者的疗效。【结果】(1)疗效方面:治疗6个月后,治疗组的总有效率为63.6%,对照组为39.4%,治疗组的疗效明显优于对照组(P<0.05)。(2)证候方面:治疗3个月后,治疗组在改善心烦失眠、潮热盗汗、咽干口燥方面明显优于对照组(P<0.05);而治疗6个月后,治疗组在改善咽干口燥、大便异常方面明显优于对照组(P<0.05)。(3)月经方面:治疗3个月后,治疗组在改善月经周期和经质方面明显优于对照组(P<0.05);而治疗6个月后,治疗组在改善经量和痛经方面明显优于对照组(P<0.05)。(4)彩超方面:治疗3个月后,治疗组在降低RI和增加PSV方面明显优于对照组(P<0.05);而治疗6个月后,治疗组在增加子宫内膜厚度方面明显优于对照组(P<0.05)。(5)性激素水平方面:治疗3个月后,2组患者血清FSH、LH、E2、AMH水平差值比较,差异均无统计学意义(P>0.05);而治疗6个月后,治疗组在提高患者血清AMH水平方面明显优于对照组(P<0.05)。【结论】滋癸益经汤联合滋肾育胎丸治疗POF疗效确切,不仅能改善患者症状和月经情况,还可提高患者血清AMH水平,增加子宫内膜厚度,改善双侧卵巢血供,降低卵巢血流阻力。     

双调蛋白在女性生殖系统作用的研究进展

在女性生殖功能实现的复杂过程中,双调蛋白(amphiregulin,Areg)参与调控颗粒细胞的黄素化、卵母细胞减数分裂的恢复及调控卵子成熟、卵丘复合体的扩张及排卵,而且其表达水平与卵子的发育潜能密切相关。Areg与子宫内膜的容受性密切相关,可能参与到胚胎的着床过程中。Areg诱导合体滋养层细胞分泌大量绒毛膜促性腺激素(h CG),参与孕早期妊娠的维持。