Tissue-associated self-antigens containing exosomes: Role in allograft rejection

Exosomes are extracellular vesicles that express self-antigens (SAgs) and donor human leukocyte antigens. Tissue-specific exosomes can be detected in the circulation following lung, heart, kidney and islet cell transplantations. We collected serum samples from patients who had undergone lung (n?=?30), heart (n?=?8), or kidney (n?=?15) transplantations to isolate circulating exosomes. Exosome purity was analyzed by Western blot, using CD9 exosome-specific markers. Tissue-associated lung SAgs, collagen V (Col-V) and K-alpha 1 tubulin (Kα1T), heart SAgs, myosin and vimentin, and kidney SAgs, fibronectin and collagen IV (Col-IV), were identified using western blot. Lung transplant recipients diagnosed with bronchiolitis obliterans syndrome had exosomes with higher expression of Col-V (4.2-fold) and Kα1T (37.1-fold) than stable. Exosomes isolated from heart transplant recipients diagnosed with coronary artery vasculopathy had a 3.9-fold increase in myosin and a 4.7-fold increase in vimentin compared with stable. Further, Kidney transplant recipients diagnosed with transplant glomerulopathy had circulating exosomes with a 2-fold increased expression of fibronectin and 2.5-fold increase in Col-IV compared with stable. We conclude that circulating exosomes with tissue associated SAgs have the potential to be a noninvasive biomarker for allograft rejection.     

Circulating histocompatibility antigen (HLA) gene products may help differentiate benign from malignant indeterminate pulmonary lesions

Background

This study explores the potential diagnostic utility of soluble Human Leukocyte Antigen (sHLA) molecules differentially released by lung adenocarcinoma and benign lung lesions.

Occurrence,integrity and functionality of AcaML1–like viruses infecting extreme acidophiles of the Acidithiobacillus species complex

General knowledge on the diversity and biology of microbial viruses infecting bacterial hosts from extreme acidic environments lags behind most other econiches. In this study, we analyse the AcaML1 virus occurrence in the taxon, its genetic composition and infective behaviour under standard acidic and SOS-inducing conditions to assess its integrity and functionality. Occurrence analysis in sequenced acidithiobacilli showed that AcaML1-like proviruses are confined to the mesothermophiles Acidithiobacillus caldus and Thermithiobacillus tepidarius. Among A. caldus strains and isolates this provirus had a modest prevalence (30%). Comparative genomic analysis revealed a significant conservation with the T. tepidarius AcaML1-like provirus, excepting the tail genes, and a high conservation of the virus across strains of the A. caldus species. Such conservation extends from the modules architecture to the gene level, suggesting that organization and composition of these viruses are preserved for functional reasons. Accordingly, the AcaML1 proviruses were demonstrated to excise from their host genomes under DNA-damaging conditions triggering the SOS-response and to produce DNA-containing VLPs. Despite this fact, under the conditions evaluated (acidic) the VLPs obtained from A. caldus ATCC 51756 could not produce productive infections of a candidate sensitive strain (#6) nor trigger it lysis.     

Collagen gel contraction as a measure of fibroblast function in an animal model of subsynovial connective tissue fibrosis

Carpal tunnel syndrome (CTS) is a peripheral neuropathy characterized by non‐inflammatory fibrosis of the subsynovial connective tissues (SSCT). A rabbit model of CTS was developed to test the hypothesis that SSCT fibrosis causes the neuropathy. We used a cell‐seeded collagen‐gel contraction model to characterize the fibrosis in this model in terms of cellular mechanics, specifically to compare the ability of SSCT cells from the rabbit model and normal rabbits to contract the gel, and to assess the effect of transforming growth factor‐β1,which is upregulated in CTS, on these cells. SSCT fibrosis was induced in six retired breeder female rabbits which were sacrificed at 6 weeks (N = 3) and 12 weeks (n = 3). An additional two rabbits served as controls. SSCT was harvested according to a standard protocol. Gels seeded with SSCT cells from rabbits sacrificed at 6 weeks had significantly higher tensile strength (p < 0.001) and Young's modulus (p < 0.001) than gels seeded with cells from rabbits sacrificed at 12 weeks or control animals. TGF‐β1 significantly increased the decay time constant (p < 0.001), tensile strength (p < 0.001), and Young's modulus (p < 0.001) regardless of the cell source. This model may be useful in screening therapeutic agents that may block SSCT fibrosis, identifying possible candidates for CTS treatment. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 33:668–674, 2015.     

左氧氟沙星眼用凝胶的制备

目的左氧氟沙星眼用凝胶的制备及质量控制方法。方法卡波姆为增粘剂,磷酸盐为缓冲液,制备左氧氟沙星眼用凝胶。采用紫外分光光度法测定左氧氟沙星的含量。稳定性实验以60℃~90℃之间每5℃为1个梯度进行考察。结果该眼用凝胶pH值稳定在6.5左右。左氧氟沙星浓度在2~9 mg.L-1范围内和吸收度呈良好的线性关系C(mg.L-1)=10.7908A-0.0012(r=0.9998,n=7);平均回收率为100.20%,RSD=0.48%(n=5)。预测有效期为19 mo。结论该眼用凝胶延长了药物对眼睛的作用时间,稳定无刺激,质控可靠。     

FAME‐04: A Phase 1 trial to assess the safety,acceptability, pharmacokinetics and pharmacodynamics of film and gel formulations of tenofovir

Introduction

Fast‐dissolving vaginal film formulations release antiretroviral drugs directly into vaginal fluid and may be as efficient at drug delivery yet more acceptable to women than gels. In this Phase 1 vaginal film study, the safety, acceptability, pharmacokinetics and pharmacodynamics of two doses of tenofovir (TFV) film and TFV 1% gel were compared to corresponding placebo formulations.

Methods

Seventy‐eight healthy HIV negative women were randomized to self‐insert daily vaginal film (10 mg TFV, 40 mg TFV or placebo) or 4 mL of vaginal gel (TFV 1% [40 mg] or placebo) for seven days. Grade 2 and higher adverse events (AEs) related to study product were compared across study arms using Fisher's exact test. Plasma TFV concentrations were measured before and 2 hours after last product use. Paired cervical and vaginal tissue biopsies obtained 2 hours after the last dose were measured to determine tenofovir diphosphate (TFV‐DP) concentrations and exposed to HIV in an ex vivo challenge assay. Acceptability was assessed through questionnaire.

Results

There was only one grade 2 or higher related AE, the primary endpoint; it occurred in the placebo gel arm. AEs occurred in 90% of participants; the majority (91%) were grade 1. AEs were similar across study arms. TFV concentrations in plasma and TFV‐DP concentrations in cervical and vaginal tissues were comparable between 40 mg TFV film and the TFV gel groups. There was a significant relationship between reduced viral replication and TFV‐DP concentrations in cervical tissues. Film users were less likely to report product leakage than gel users (66% vs. 100%, p < 0.001).

Conclusions

Films were safe and well tolerated. Furthermore, films delivered TFV to mucosal tissues at concentrations similar to gel and were sufficient to block HIV infection of genital tissue ex vivo.

     

Efficacy of topical silicone gel in scar management: A systematic review and meta‐analysis of randomised controlled trials

To assess the efficacy of topical silicone gel in the management of scars, we conducted this meta‐analysis. The systematic search was performed on PubMed, Web of Science and Embase, and six randomised controlled trials with a total of 375 patients were involved. The outcome data of Vancouver Scar Scale were extracted from the studies and their effect sizes were calculated using Review Manager 5.3. As a result, topical silicone gel significantly reduced pigmentation, height, and pliability scores postoperatively compared with placebos or no treatment (Pigmentation: standard mean difference [SMD] = −0.55 [−0.83 to –0.26], P = .0002; Height: SMD = −0.73 [−1.02 to –0.44], P < .00001; Pliability: SMD = −0.49 [−0.95 to –0.03], P = .04). Topical silicone gel and silicone gel sheet were comparably effective (P > .05). The performance of topical silicone gel and other non‐silicone topical treatment was also similar (P > .05). In summary, topical silicone gel was effective in post‐operative scar prevention.     

Use of concentrate growth factors gel or membrane in chronic wound healing: Description of 18 cases

Treating chronic skin wounds in patients with diabetes, bed sores, or stasis dermatitis is typically a time‐consuming and costly process, and the outcome is not always promising. Concentrated growth factor (CGF) obtained from the autologous venous blood of patients via fractional centrifugation is employed for producing a CGF gel or membrane that can be applied to expedite self‐regeneration of skin wounds. In this case report, we presented the results from 18 patients with chronic skin wounds treated with a CGF gel or membrane produced from autologous venous blood. Noticeable granulation tissue and regenerated epidermal coverage were observed in 16 patients who received CGF treatment over various time courses, thereby demonstrating the significant therapeutic effects of CGF treatment in overall wound healing. The other two patients with stasis ulcers in their calves failed to respond to the treatment because of the comorbidity of iliac vein thrombosis. In addition, by culturing HaCaT keratinocytes using CGF membrane as the foundation, we observed that HaCaT cells attached to the CGF membrane migrated and proliferated to form an epithelium‐like structure. Comprehensively, the clinical results infer that CGF gel can expedite the regeneration of the soft tissue at the wound, whereas CGF membrane may facilitate its marginal re‐epithelialisation. The combination of the two can promote autologous regeneration of both deep and superficial wounds effectively and safely.     

盐酸奥布卡因凝胶在经鼻肠梗阻导管置入术中的应用价值

目的探讨肠梗阻病人行经鼻肠梗阻导管置入术中应用盐酸奥布卡因凝胶的价值。方法将60例拟行经鼻肠梗阻导管置入术的肠梗阻病人按随机数字量表法分为研究组和对照组,每组各30例,研究组病人术前应用盐酸奥布卡因凝胶10 ml(30 mg)进行鼻腔麻醉及导管前端润滑,对照组病人术前于鼻腔及导管前端涂抹石蜡油进行润滑,比较两组病人的一次性置管成功率、置管时间及术后并发症发生率,并采用视觉模拟评分法(visual analogue scale,VAS)进行评分。结果研究组病人的一次性置管成功率为96.7%(29/30),对照组病人的一次性置管成功率为70.0%(21/30),差异有统计学意义(P<0.05)。与对照组比较,研究组病人置管所用时间明显减少,术中疼痛VAS评分降低,差异均有统计学意义(均P<0.05)。两组病人均未出现穿孔、出血等并发症。结论盐酸奥布卡因凝胶在肠梗阻病人行经鼻肠梗阻导管置入术中应用,提高了一次性置管成功率,减少置管所用时间,明显地减轻了病人术中的疼痛不适,未见并发症,效果满意。     

自体肋软骨联合硅胶假体鼻整形效果探讨

目的:探讨自体肋软骨联合硅胶假体在鼻整形中的应用效果。方法:回顾性分析2015年5月-2017年5月笔者医院接受鼻整形的105例就医者临床资料,通过治疗方式不同分为观察组55例和对照组50例,对照组使用硅胶假体隆鼻,观察组使用自体肋软骨联合硅胶假体隆鼻,比较两组手术情况、鼻部塑形效果、二次手术率及并发症发生情况。结果:两组手术成功率、愈合时间比较,差异无统计学意义(P>0.05);观察组手术时间明显长于对照组,差异有统计学意义(P<0.05)。和术后3个月时比较,观察组术后12个月时鼻根高、鼻面角、鼻尖角、鼻额角差异无统计学意义(P>0.05);对照组术后12个月时鼻根高、鼻面角、鼻尖角均明显降低(P<0.05),且术后12个月时,观察组鼻根高、鼻面角、鼻尖角均明显高于对照组(P<0.05)。观察组二次手术率为3.64%(2/55)明显低于对照组的16.00%(8/50),差异有统计学意义(χ^2=4.646,P=0.031)。观察组术后并发症总发生率为14.55%明显低于对照组的32.00%,差异有统计学意义(P<0.05)。结论:和单用硅胶假体相比,自体肋软骨联合硅胶假体用于鼻整形中效果更加显著,具有鼻部塑形效果满意、二次手术率低、并发症少等特点,临床应用价值高。