Effects of Lactobacillus plantarum CJLP55 on Clinical Improvement,Skin Condition and Urine Bacterial Extracellular Vesicles in Patients with Acne Vulgaris: A Randomized,Double-Blind,Placebo-Controlled Study

Lactobacillus plantarum CJLP55 has anti-pathogenic bacterial and anti-inflammatory activities in vitro. We investigated the dietary effect of CJLP55 supplement in patients with acne vulgaris, a prevalent inflammatory skin condition. Subjects ingested CJLP55 or placebo (n = 14 per group) supplements for 12 weeks in this double-blind, placebo-controlled randomized study. Acne lesion count and grade, skin sebum, hydration, pH and surface lipids were assessed. Metagenomic DNA analysis was performed on urine extracellular vesicles (EV), which indirectly reflect systemic bacterial flora. Compared to the placebo supplement, CJLP55 supplement improved acne lesion count and grade, decreased sebum triglycerides (TG), and increased hydration and ceramide 2, the major ceramide species that maintains the epidermal lipid barrier for hydration. In addition, CJLP55 supplement decreased the prevalence of Proteobacteria and increased Firmicutes, which were correlated with decreased TG, the major skin surface lipid of sebum origin. CJLP55 supplement further decreased the Bacteroidetes:Firmicutes ratio, a relevant marker of bacterial dysbiosis. No differences in skin pH, other skin surface lipids or urine bacterial EV phylum were noted between CJLP55 and placebo supplements. Dietary Lactobacillus plantarum CJLP55 was beneficial to clinical state, skin sebum, and hydration and urine bacterial EV phylum flora in patients with acne vulgaris.     

合体细胞滋养层细胞外囊泡阻止母体恶性肿瘤侵袭及转移至胎儿相关机制研究

目的探讨合体细胞滋养层细胞外囊泡(STB-EV)阻止母体恶性肿瘤侵袭、转移至胎儿的相关机制。 方法选择2015年6月,于四川大学华西第二医院进行剖宫产术分娩的1例妊娠合并宫颈癌(CCP)患者被肿瘤侵犯的胎盘组织为研究对象。宫颈癌细胞系SiHa细胞和人滋养层细胞系HTR-8细胞,均由本院西部妇幼医学研究院-分子与转化医学实验室馈赠。其中,本例CCP患者被肿瘤侵犯的胎盘组织制作组织切片后,于光学显微镜(×400)和电子显微镜(×2 000)下,观察其组织形态学、STB-EV及合体细胞滋养层(STB)旁宫颈癌细胞自噬性死亡情况等;而SiHa细胞与HTR-8细胞培养后,用于Transwell迁移实验与细胞划痕实验。将SiHa与HTR-8细胞共培养,纳入研究组;单独培养的SiHa细胞,纳入对照组。对2组细胞进行Transwell迁移实验及细胞划痕实验,2组Transwell小室穿膜细胞数、细胞迁移率等比较,采用成组t检验。本研究遵循的程序符合2013年新修订的《世界医学协会赫尔辛基宣言》要求。 结果①对本例CCP患者被肿瘤侵犯的胎盘组织制作组织切片后,于光学显微镜(×400)下观察结果显示,与STB相邻宫颈癌细胞自噬性死亡、凋亡和固缩性坏死显著。电子显微镜(×2 000)下观察结果显示,宫颈癌细胞累及胎盘绒毛组织时,STB分泌的STB-EV水平显著高于正常胎盘组织。②2组细胞Transwell迁移实验结果显示,研究组穿膜细胞数为(597.6±87.7)个/视野,显著低于对照组的(1 358.4±203.0)个/视野,并且差异有统计学意义(t＝14.490、P<0.001)。③2组细胞划痕实验结果显示,划痕后48 h时,研究组细胞迁移率为(26.6±3.8)%,显著低于对照组的(45.9±3.7)%,并且差异有统计学意义(t＝3.122、P＝0.035)。 结论STB分泌的STB-EV可能参与母体肿瘤细胞广泛自噬性死亡,从而阻止母体肿瘤细胞侵袭胎盘、影响胎儿发育。     

整合素血小板膜糖蛋白IIb、IIIa在系膜增殖性肾炎中的表达

目的 :观察整合素血小板膜糖蛋白Ⅱb、Ⅲa(GPⅡb、GPⅢa)在人类系膜增殖肾炎 (MsPGN)中肾内表达 ,探讨GPⅡb Ⅲa在MsPGN的作用。方法 :18例肾活检标本均行光镜HE、PAS、PASM Masson染色 ,电镜及免疫荧光检查 ,以SABC法测定肾组织中GPⅡb、GPⅢa、P 选择素 (P 140 )、纤维连接蛋白 (FN)、粘连蛋白 (LN)表达与正常对照组分别比较 ,并行计算机图像分析。结果 :PAS阳性物 (PAS+ )、PASM阳性物 (PASM+ )及FN、LN明显增生 ;GPⅡb、GPⅢa、P 140表达上调 ;肾小球细胞数 (n)、平均肾小球体积 (AVG)及肾小球硬化指数显著增加 ;肾小管、间质轻度受损。结论 :GPⅡb、GPⅢa在肾小球及肾小管间质表达上调与系膜细胞基质增生、细胞浸润、肾小球硬化相关 ;可能与肾小球疾病中凝血功能紊乱、血小板在肾脏的致病作用相关     

氨甲酰胆碱对多巴胺诱导的小脑颗粒细胞凋亡的保护机制

【目的】研究多巴胺诱导小脑颗粒神经元凋亡的分子机制,以及胆碱受体激动剂氨甲酰胆碱对多巴胺诱导凋亡的作用。【方法】在培养的小脑颗粒神经元建立多巴胺凋亡模型。用相差显微镜观察形态学,DNA凝胶电泳和Hoechst33258核染色分析神经元凋亡,细胞的存活率用二乙酸荧光素（FDA）染色法检测。采用Westernblot分析细胞外信号调控的蛋白激酶（ERK）激活情况。【结果】多巴胺可诱导小脑颗粒神经元凋亡,并可持续激活ERK,二者均可被氨甲酰胆碱和PD98059抑制。氨甲酰胆碱对神经元的保护作用及对ERK激活的抑制作用可被阿托品阻断。【结论】多巴胺在小脑颗粒神经元诱导凋亡可能是通过持续激活ERK介导的。氨甲酰胆碱通过激活M胆碱受体,继而抑制了ERK的激活,从而起到对神经元的保护作用。     

Bivalent non-typhoidal Salmonella outer membrane vesicles immunized mice sera confer passive protection against gastroenteritis in a suckling mice model

Invasive non-typhoidal Salmonella (iNTS) serovars, especially Salmonella Typhimurium (ST) and Salmonella Enteritidis (SE), cause gastroenteritis worldwide. Due to the emergence of multi-drug resistance in iNTS, a broad-spectrum vaccine is urgently needed for the prevention of iNTS infection. Currently, there is no effective licensed vaccine against iNTS available in the market. We have formulated an outer membrane vesicles (OMVs) based bivalent immunogen as a vaccine candidate to generate broad-spectrum protective immunity against both recently circulating prevalent ST and SE. We have isolated OMVs from ST and SE and formulated the immunogen by mixing both OMVs (1:1 ratio). Three doses of bivalent immunogen significantly induced humoral immune responses against lipopolysaccharides (LPSs) and outer membrane proteins (OMPs) as well as a cell-mediated immune response in adult mice. We also observed that proteins of OMVs act as an adjuvant for generation of high levels of anti-LPS antibodies through T cell activation. We then characterized the one-day old suckling mice model for both ST and SE mediated gastroenteritis and used the model for a passive protection study. In the passive protection study, we found the passive transfer of bivalent OMVs immunized sera significantly reduced ST and SE mediated colonization and gastroenteritis symptoms in the colon of suckling mice compared to non-immunized sera recipients. The overall study demonstrated that OMVs based bivalent vaccine could generate broad-spectrum immunity against prevalent iNTS mediated gastroenteritis. This study also established the suckling mice model as a suitable animal model for vaccine study against iNTS mediated gastroenteritis.     

Extracellular vesicles released from irradiated neonatal mouse cheek tissue increased cell survival after radiation

Alopecia is one of the common symptoms after high-dose radiation exposure. In our experiments, neonatal mice that received 7 Gy X-ray exhibited defects in overall hair growth, except for their cheeks. This phenomenon might suggest that some substances were secreted and prevented hair follicle loss in the infant tissues around their cheeks after radiation damage. In this study, we focused on exosome-like vesicles (ELV) secreted from cheek skin tissues and back skin tissues, as control, and examined their radiation protective effects on mouse fibroblast cell lines. We observed that ELV from irradiated cheek skin showed protective effects from radiation. Our results suggest that ELV from radiation-exposed cheek skin tissue is one of the secreted factors that prevent hair follicle loss after high-dose radiation.     

Type IV collagenases in invasive tumors

Summary The matrix metalloproteinases appear to be elevated in tumors with metastatic potential, and may well be involved in penetration of the basement membrane and degradation of extracellular proteins including type IV collagen. An imbalance between the 72 kDa and 92 kDa type IV collagenases and the associated tissue inhibitors of these metalloproteinases (TIMPs) may therefore have a role in the invasive phenotype. Cultured tumor cells with invasive potential secrete both type IV collagenases, though in tumors there is some evidence that the 72 kDa form at least may be produced by stromal cells at the invading tumor front rather than primarily by the tumor cells themselves, while the 92 kDa form may be synthesized in macrophages near the front. These collagenases are elevated in invasive as compared within situ tumor components, but their specific roles and prognostic significance are not yet established.     

Efflux of taurine from renal brush border membrane vesicles: Is it adaptively regulated?

The uptake of the -amino acid taurine by rat renal brush border membrane vesicles (BBMV) adapts to changes in dietary sulfur amino acid intake. Initial rateV _max upregulates after ingestion of a low methionine and taurine diet (LTD) and downregulates after a high taurine diet (HTD). This is reflected in vivo by hypotaurinuria after a LTD (90% reduction in excretion) and an 18-fold increase in urine taurine after a HTD. This study was performed to determine whether taurine efflux from BBMV is adaptively regulated by external taurine concentration or by diet. Vesicles were preloaded with varying concentrations of radiolabelled and unlabelled taurine and a 150 mM concentration of various salts. Efflux conditions were: taurine and 150 mM salt inside and 150 mM salt outside. The efflux of five concentrations of taurine (10–500 M) was linear over 6 min, reached equilibrium by 21 min, and was dependent upon intravesicular taurine content. The kinetic characteristics of efflux (E) were significantly different from influx (I):K _m=109.8±5.8 (E) versus 23.6±4.2 (I),P<0.001 [time of linearity=360 s (E) vs. 20 s (I)]. Efflux of taurine was dependent on the presence of both sodium and chloride in the system, but neither external taurine content (0,100 M, 1,000 M) nor external -alanine altered initial efflux. Feeding rats a normal diet, LTD, or fasting altered taurine uptake but not efflux. Efflux does not appear to play a role in the adaptive regulation of taurine transport found in all mammalian species.     

A comparison of the haemodynamic and hormonal effects of low and conventional dose cyclopenthiazide in normal volunteers

Summary In this study we compared low (125 g) and conventional (500 g) doses of cyclopenthiazide on the renin angiotensin system, plasma and extracellular fluid volumes and the pressor responsiveness to angiotensin II since we have previously shown that the two doses have the same antihypertensive effect but different effects on plasma renin activity.Following a two week placebo run-in period, 8 healthy male volunteers received 125 g or 500 g of cyclopenthiazide for 2 treatment periods of 4 weeks as part of a double blind, 2-part crossover study with treatment periods separated by a 4-week placebo washout phase. Measurements were made on two study days at the beginning and end of the active treatment periods. On the first day serum potassium, plasma renin activity and plasma angiotensin II levels were measured after a 1 h period of supine rest. Plasma and extracellular fluid volumes were also measured after appropriate equilibration times. The blood pressure responses to angiotensin II were assessed on day 2.The 500 g dose of cyclopenthiazide had a greater effect than the 125 g dose on plasma renin activity, serum potassium, angiotensin II levels and extracellular fluid volumes. Neither drug had any effect on plasma volume or the responsiveness to infused angiotensin II.Low dose cyclopenthiazide failed to increase angiotensin II levels, contract body fluid volumes or attenuate vascular reactivity in normotensive volunteers.     

Nebulization of Liposomes. I. Effects of Lipid Composition

A series of multilamellar liposome dispersions was prepared from lipids of soy phosphatidylcholine or hydrogenated soy phosphatidylcholine containing from 0 to 30 mol% of either cholesterol, steary-lamine, or dipalmitoyl phosphatidylglycerol. The liposome dispersions were aerosolized with a Collison nebulizer for 80 min at an output flow rate of 4.7 liters of air/min. The effects of nebulization on the vesicles were determined by monitoring the release of encapsulated 5,6-carboxyfluorescein (CF) from dispersions containing 200 µg of total CF, of which 93.1 ± 2.4% (N = 18) was initially encapsulated. In all experiments CF was released from the liposomes while being aerosolized, and this ranged from a mean of 12.7 ± 3.8 to 60.9 ± 1.9% of the encapsulated CF, depending upon the lipid composition. The lipid concentration in the dispersions did not affect the rate or percentage release of CF over a range of 0.5 to 50 mg per nebulized dispersion. If liposomes are to be used as drug carriers in an inhalation aerosol a lipid composition should be employed which will minimize the release of encapsulated drug caused by nebulization.