Differentiation of the plasma membrane of hepatic cells in monolayer cultures

Hepatocytes from rats were isolated by treatment with trypsin and cultured. Plasma membranes at different culture stages were observed by electron microscopy. The activities of 5' nucleotidase and adenosinetriphosphatase on the plasma membranes were examined. The cell coat was also studied by use of the concanavalin A-peroxidase technique. The surfaces of single cells, covered with microvilli, are the site of adenosinetriphosphatase activity only and are devoid of 5'-nucleotidase activity. After a few h of culture, the cells are grouped together in tight clusters or long trails and are separated by an intercellular space of 250 A, partially permeable to lanthanum nitrate. The juxtaposed plasma membranes on which 5'-nucleotidase and adenosinetriphosphatase activities occur also delimit spaces similar to bile canaliculi. The formation of junction complexes and their permeability to lanthanum nitrate was also studied. No enzymatic activity is observed at the junctions. The numerous tight junctions, impervious to the tracer, are always accompanied by a profusion of microfilaments. Mature desmosomes are rare, and are present only in the form of "maculae adhaerentes diminutae." The gap junctions, nearly always permeable to the tracer, form rapidly and assume a variety of shapes (trail, bulge and ring-like), the significance of which is open to discussion. The use of concanavalin A permits localization of the free sugar sites on the surface of the cells, in the pinocytotic vesicles and in the internal space of the gap junctions.     

Autoradiographic study on the localization of estradiol-17β in neonatal mouse uterus and cervix

Summary The uptake of ³H-estradiol-17 in the neonatal mouse uterus and cervix has been studied by an autoradiographic method. When the radio-active hormone is administered in vivo and in vitro, grains are found to be concentrated above the nuclei both in the uterine and cervical epithelium and stroma. Grain counts revealed that the nuclear concentration of grains is higher at 4 h than at 2 h after isotope injection. The cervical epithelium has a higher nuclear concentration than the uterine epithelium both in vivo and in vitro. In the stroma, this situation is reversed except after in vitro treatment of the tissues.In the cervix, more of the hormone seems to be located within the nucleus while in the uterus a higher proportion of the grains are found in the vicinity of the nuclear periphery.Although the nuclear concentration of grains is higher at 4 h than at 2 h, the number of grains above the sections is lower at 4 h. Both in vivo and in vitro, the number of grains is higher above the stromal than above the epithelial compartments of the uterus and cervix.Five days old animals showed the same labeling pattern. The differences in uptake and distribution of ³H-estradiol are discussed in relation to other known differences in the hormone responsiveness in these tissues.We are greatly indebted to Professor W.E. Stumpf and the Laboratories for Reproductive Biology, University of North Carolina Medical School for the opportunity to study the method of dry mount autoradiography. The work has been supported by the Norwegian Research Council for Science and the Humanities and by the Norwegian Cancer Society (Landsforeningen mot Kreft)     

Pericytes and perivascular microglial cells in the basal forebrain of the neonatal rabbit

Summary Three types of pericytes outline the vascular bed in Golgi preparations of the newborn rabbit brain. Elongate cells (Type I) are restricted to capillaries, elements resembling smooth muscle cells (Type II) surround vessels of intermediate size, and large flat forms (Type III) cover the surface of arterioles and venules. Electron microscopy shows all types to be located within a well defined perivascular basement membrane. It also reveals the presence of filaments in the cytoplasm of some pericytes resembling the myofilaments of smooth muscle cells. It suggests the possibility that some pericytes are capable of contraction and may participate in regulating blood flow in small vessels.Microglia cells bear no resemblance to pericytes in terms of their shape, distribution or staining characteristics. Microglia cells are located outside the vascular basement membrane (external basal lamina) in the brain parenchyma, and they vary in form according to their location and the character of the surrounding extracellular space. This study does not support the hypothesis that microglia cells arise from pericytes but indicates that they originate either by in situ division or from hematogenous elements that enter the brain by crossing the vessel wall.Support provided by N.I.H. Grants No. NS 10864 and NS 07938 from the U.S. Public Health Service.     

Safety of ethanolic kava extract: Results of a study of chronic toxicity in rats

BACKGROUNDS: Recently, potential liver toxicity was discussed with the intake of kava extract preparations (Piper methysticum) as anxiolytic drugs. The aim of this study was to test chronic toxicity in rats by oral application of an ethanolic kava full extract. METHODS: Wistar rats of both sexes were fed 7.3 or 73 mg/kg body weight of ethanolic kava extract for 3 and 6 months. The animals were examined for changes in body weight, hematological and liver parameters, and macroscopical and microscopical histological changes in the major organs. RESULTS: No signs of toxicity could be found. CONCLUSIONS: The results are in accordance with the medical experience regarding the use of kava preparations and the long tradition of kava drinking in the South Pacific island states. Specifically, the results do not back the suspicion of potential liver toxicity.     

Alterations in the phosphodiesterase type 5 pathway and oxidative stress correlate with erectile function in spontaneously hypertensive rats

To explore how alterations in the phosphodiesterase type 5 (PDE5) signalling pathway and oxidative stress correlate with changes in the expression of relaxation and contraction molecules and erectile dysfunction (ED) in the corpus cavernosum smooth muscle (CCSM) of spontaneously hypertensive rats (SHR). In this study, SHR and Wistar‐Kyoto (WKY) rats were used. Erectile function was determined by apomorphine test and electrical stimulation (ES) of cavernous nerve. Masson''s trichrome staining and confocal microscopy were performed. Nitric oxide synthase (NOS), PDE5, phosphorylated‐PDE5 and α1‐adrenergic receptor (α1AR) were determined by RT‐PCR and Western blotting while oxidative stress in CC was determined by colorimetric analysis. SHR exhibited obvious ED. CC of SHR showed less SM but more collagen fibres. The expression of NOS isoforms in SHR was significantly decreased while all α1AR isoforms were increased. In addition, PDE5 and phosphorylated‐PDE5 were down‐regulated and its activity attenuated in the hypertensive rats. Meanwhile, the SHR group suffered oxidative stress, which may be modulated by endoplasmic reticulum stress and NADPH oxidase up‐regulation. Dysregulation of NOS and α1AR, histological changes and oxidative stress in CC may be associated with the pathophysiology of hypertension‐induced ED. In addition, PDE5 down‐regulation may lead to the decreased efficacy of PDE5 inhibitors in some hypertensive ED patients and treatment of oxidative stress could be used as a new therapeutic target for this type of ED.     

Exogenous neuritin treatment improves survivability and functions of Schwann cells with improved outgrowth of neurons in rat diabetic neuropathy

Pathogenesis and treatment for diabetic neuropathy are still complex. A deficit of neurotrophic factors affecting Schwann cells is a very important cause of diabetic neuropathy. Neuritin is a newly discovered potential neurotrophic factor. In this study, we explored the effect of exogenous neuritin on survivability and functions of diabetic Schwann cells of rats with experimental diabetic neuropathy. Diabetic neuropathy was induced in rats. 12‐week diabetic rats contrasted with non‐diabetic normal rats had decreased levels of serum neuritin and slowed nerve conduction velocities (NCVs). Schwann cells isolated from these diabetic rats and cultured in high glucose showed reduced cell neuritin mRNA and protein and supernatant neuritin protein, increased apoptosis rates, increased caspase‐3 activities and progressively reduced viability. In contrast, exogenous neuritin treatment reduced apoptosis and improved viability, with elevated Bcl‐2 levels (not Bax) and decreased caspase‐3 activities. Co‐cultured with diabetic Schwann cells pre‐treated with exogenous neuritin in high glucose media, and diabetic DRG neurons showed lessened decreased neurite outgrowth and supernatant NGF concentration occurring in co‐culture of diabetic cells. Exogenous neuritin treatment ameliorated survivability and functions of diabetic Schwann cells of rats with diabetic neuropathy. Our study may provide a new mechanism and potential treatment for diabetic neuropathy.     

白蛋白静脉滴注联合茵栀黄颗粒治疗新生儿黄疸血清 AKP、TBA、 FFA、γ-GT、HS-CRP的影响

目的:研究白蛋白静脉滴注联合茵栀黄颗粒对新生儿黄疸血清碱性磷酸酶(alkaline phosphatase,AKP)、总胆汁酸(totalbileacids,TBA)、游离脂肪酸(free fatty acid,FFA)、γ-谷氨酰基转移酶(γ-glutamyltransferase,γ-GT)、超敏C-反应蛋白(hypersensitive C-reactive protein, HS-CRP)的影响。方法:选择2016年1月～2019年1月我院收治的95例新生儿病理性黄疸患儿,随机分为两组。对照组服用茵栀黄颗粒治疗,观察组在服用茵栀黄的基础上静脉滴注白蛋白治疗。检测两组的血清间接胆红素(indirectreacting bilirubin,,IBIL)、总胆红素(total bilirubin,TBIL)和AKP、TBA、FFA、y-GT、HS-CRP水平。结果:观察组的有效率明显高于对照组(P0.05);两组治疗后的血清IBIL、TBIL水平明显降低(P0.05),观察组的IBIL、TBIL水平明显低于对照组(P0.05);两组治疗后的血清AKP、TBA、FFA、γ-GT、HS-CRP水平明显降低(P0.05),观察组的血清AKP、TBA、FFA、γ-GT、HS-CRP水平明显低于对照组(P0.05)。结论:白蛋白静脉滴注联合茵栀黄颗粒对新生儿黄疸的治疗效果良好,有助于促进血清胆红素和其他血清学相关指标恢复正常,且安全性好。     

微创肺表面活性物质对呼吸窘迫综合征新生儿肝肾功能、氧合功能及呼吸功能的影响

目的:探讨微创肺表面活性物质(pulmonary surfactant,PS)对呼吸窘迫综合征新生儿氧合功能、肝肾功能及呼吸功能的影响。方法:选择2018年3月至2019年3月我院收治的66例新生呼吸窘迫综合征的患儿作为研究对象,并按照随机数字表法分为观察组(n=33)和对照组(n=33)。对照组患儿采取常规的治疗方式,观察组患儿则在对照组治疗基础上应用微创PS治疗。观察比较两组患儿的动脉血气指标、Ⅱ型肺泡细胞表面抗原(KL-6)、巨噬细胞移动抑制因子-1(MIF-1)、高迁移率族蛋白1(HMGB-1)、肝肾功能、氧合指数及呼吸机参数。结果:治疗后,两组的氧分压(PaO_2)、二氧化碳总量(TCO_2)、氧饱和度(SaO_2)均较治疗前显著增高;且观察组以上指标均高于对照组。两组的Ⅱ型肺泡细胞表面抗原(KL-6)、巨噬细胞移动抑制因子-1 (MIF-1)、高迁移率族蛋白1(HMGB-1)均较治疗前显著降低,且观察组的以上指标均低于对照组。两组的谷草转氨酶(AST)、谷丙转氨酶(ALT)、尿素氮(BUN)、肌酐(CRE)水平均较治疗前显著降低,且观察组的以上指标均明显低于对照组。观察两组的呼吸机参数和氧合指数,发现两组的吸入氧浓度(FiO_2)、吸气峰压(PIP)、呼吸末正压(PEEP)、氧合指数(PaO_2/FiO_2,OI)均较治疗前有所改善,且观察组的以上指标均要优于对照组(P0.05)。结论:应用微创PS治疗新生儿呼吸窘迫综合征的效果显著,能明显改善患儿的动脉血气指标、肝肾功能以及氧合功能,减轻炎症反应,并减少机械通气的参数。     

持续镇痛分娩对产妇分娩结局和新生儿评分的影响

目的:研究持续镇痛分娩对产妇分娩结局和新生儿评分的影响。方法:选择2018年7月～2019年7月中国医科大学航空总医院(本院)采取硬膜外分娩镇痛的101例产妇,将其随机分为两组。当产生确切的镇痛效果,进入第二产程后,观察组的51例产妇采用0.4μg/m L舒芬太尼以及0.08%罗哌卡因进行持续镇痛分娩;对照组的50例产妇则在宫口开全后,使用生理盐水替代泵内的局麻药物,直到分娩结束。比较两组产妇催产素的使用率,宫口扩张度和第一、第二产程按压硬膜外自控镇痛泵的次数,分娩方式,新生儿的体质量,脐动脉血pH值,出生后1 min和5 min Apgar评分,产妇修复会阴部时的视觉模拟评分(visual analogue scale, VAS)评分及产妇对于第二产程镇痛的满意度评分。结果:两组产妇催产素的使用率、宫口扩张度和第一、第二产程按压硬膜外自控镇痛泵的次数、分娩方式(剖宫产率、器械助产率、自然分娩率)、第一产程镇痛时间、第一以及第二产程时间相比均无显著差异(P0.05);两组新生儿的体质量,脐动脉血pH值,出生后1 min和5 min Apgar评分小于8分的新生儿所占的比例相比没有明显的差异(P0.05);观察组产妇修复会阴部时的VAS评分明显低于对照组(P0.05),产妇对于第二产程镇痛的满意度评分明显高于对照组(P0.05)。结论:持续镇痛分娩对产妇分娩结局和新生儿评分无明显的影响,但可显著提高产妇对第二产程镇痛和修复会阴部时镇痛的满意度。     

沐舒坦联合布地奈德雾化治疗新生儿胎粪吸入综合征的临床应用分析

目的:探讨沐舒坦联合布地奈德雾化治疗新生儿胎粪吸入综合征的临床效果及安全性。方法:选择2017年1月～2018年2月我院新生儿科收治的76例新生儿胎粪吸入综合征患儿,按照随机数字表法将其分成两组,每组38例。对照组患者采用布地奈德雾化治疗,观察组在对照组的治疗基础上加用沐舒坦治疗,分析和比较两组的治疗效果,患儿治疗前后动脉血气分析指标变化以及预后情况。结果:治疗后,观察组临床总有效率明显高于对照组,观察组呼吸困难缓解时间、肺部湿罗音消失时间、发绀消失时间、血氧饱和度恢复时间均显著较对照组短(P0.05)。两组患儿治疗后PaCO_2、FIO_2、OI均较治疗前降低,PaO_2均较治疗前上升,其中观察组PaCO_2、FIO_2、OI明显低于对照组,PaO_2高于对照组,上述差异均具有统计学意义(P0.05)。观察组总并发症发生率显著低于对照组(P0.05),患儿治愈率显著高于对照组(P0.05),两组死亡率比较差异无统计学意义(P0.05)。结论:与布地奈德雾化治疗相比,沐舒坦联合布地奈德雾化治疗新生儿胎粪吸入综合征患儿可以更有效缩短临床症状改善时间,改善患儿肺功能及预后,且安全性更高。