肾炎益气液对肾毒血清性肾炎大鼠肾脏超微结构的影响

目的:观察肾毒血清性肾炎大鼠肾组织超微结构变化及肾炎益气液对该变化的影响。方法: 采用大鼠肾毒血清性肾炎模型,常规电镜制片、染色,观察肾组织超微结构的变化。结果: 注射肾毒血清(NTS)后,病理组大鼠电镜下可见明显的系膜细胞增生,系膜基质增多,毛细血管腔闭塞及上皮下、内皮下电子致密物沉积等多种病理损伤性变化。而肾炎益气液组上述病变有所减轻。结论: 肾炎益气液有不同程度减轻肾小球电镜下病变的作用。     

GPC-II-3液间歇低温灌流保存兔肺的形态学观察

目的探讨对移植供肺具有长期保护作用的新技术。方法用自制的GPC-Ⅱ-3液,以间歇低温灌流的方法保存兔肺24～192h,观察其组织结构的变化。结果保存120h以内的肺,其组织结构与对照组无明显区别,肺泡、各级支气管、血管的结构清楚、支气管上皮结构完整清楚,上皮细胞胞核清晰,染色质分布均匀、肺泡完整,肺泡上皮和毛细血管内皮清楚。部分肺泡腔中,可见结构清楚的尘细胞及其吞噬的粉尘颗粒;保存144h的肺,细胞成分的染色似乎有所加深,其余结构无明显变化。保存168h的肺,肺泡、各级支气管、血管的结构依然清楚,但细胞成分的染色有所加深,部分支气管上皮细胞有脱落现象。保存192h的肺,细胞成分的染色明显加深,有固缩迹象,支气管上皮有脱落现象加重。结论用GPC-Ⅱ-3液以低温冷藏的方法能够保存兔肺的组织结构120h。     

Ultrasonic nebulization of hypertonic solution: a new method for obtaining specimens from nasal mucosa for morphologic and biochemical analysis in allergic rhinitis

Various techniques are used to collect specimens from the nasal mucosa for morphologic and biochemical analysis. The purpose of this study was to devise a method that overcomes some of the disadvantages (e.g., invasive procedure, samples not suitable for cytologic and biochemical analysis, lack of standardization, and poor reproducibility) of these techniques. The new method requires subjects, with neck extended, to inhale an ultrasonic nebulization of a hypertonic (3% NaCl) solution (UNHS) for 5 min. They then blow their nose into a Petri dish, one nostril at a time with the other one blocked. The secretions are dispersed with 0.1% dithiothreitol in phosphate buffer solution for 20 min. Total cell count (TCC) is evaluated, and the cellular suspension is divided into two aliquots: one is centrifuged and the supernatants are collected for eosinophil cationic protein (ECP) measurements; the other is cytocentrifuged and the slides, stained with Diff-Quik, are used for differential cell count. The results obtained with the UNHS and nasal lavage (NL) methods were compared. Eleven nonatopic healthy subjects and 19 allergic rhinitic patients were studied. Total cell count (×10⁵) was significantly higher with UNHS than with NL (13.0±12.3 vs 1.911.6; P<0.0]) The differential cell count was similar with the two procedures. ECP levels (μg/l) were higher with UNHS than with NL (39.1+38.2 V.S 16.7±41.2; P<0.01). For evaluation of reproducibility, four healthy and six rhinitic subjects underwent UNHS on two occasions within 5 days, and the results of two samples (sample 1 vs sample 2) were analyzed. Reproducibility was good as to TCC, differential cell count, and ECP     

Induction of immunoglobulin-producing human peripheral blood lymphocytes in serum-free medium

Induction of immunoglobulin-secreting cells from human peripheral blood lymphocytes in a serum-free culture medium was studied. Albumin, transferrin, insulin and fibronectin can replace serum entirely for support of pokeweed mitogen (PWM)-stimulated B lymphocytes, measured by a reverse hemolytic plaque assay using protein A-coated red cells. In this serum-free system, growth and maturation to IgM and IgG secretion occur at the same or higher efficiency as in conventional serum-containing medium, with maximum numbers of plaque-forming cells on day 6 at optimal dose of PWM, 0.5 ～ 5 μg/ml. This system can be used to avoid the interference from undefined serum components.     

A monoclonal antibody (RH1-38) which inhibits multiple systems of cell-mediated cytotoxicity--I. Identification of a novel epitope on a killer cell surface bimolecular complex important in the cytotoxic response

This paper presents the initial characterization of a mouse monoclonal antibody (RH1-38) which blocks, in the absence of complement, three different systems of cell-mediated cytotoxicity. This monoclonal antibody markedly inhibits cytotoxicity mediated by human natural killer cells, a monocyte-like cell [phorbol myristate acetate (PMA) stimulated HL-60], and cytotoxic T-lymphocytes generated in a mixed leukocyte reaction. RH1-38 is not nonspecifically toxic to cells since antibody-dependent cellular cytotoxicity was not inhibited and viability as assessed by trypan blue exclusion was not affected. Inhibition is specific since control hybridoma culture supernatants, parent (NS-1) ascites supernatant, monoclonal anti-HLA and normal mouse IgG were not significantly inhibitory. In the NK system, the inhibitory effect appears to be due to binding of monoclonal antibody to effector cell surface since exposure of targets to antibody followed by washing yielded no inhibition of killing. Inhibition requires the antigen-binding portion of the antibody molecule and thus appears to be related to steric hindrance of an effector cell surface molecule which is important in the expression of cell-mediated cytotoxicity. Immunoprecipitation of surface-radioiodinated membranes from PMA-stimulated HL-60 cells and analysis on sodium dodecyl sulfate-polyacrylamide gels revealed a bimolecular complex (195,000 and 125,000 daltons) without significant change under reducing conditions. Control immunoprecipitates yielded no peaks of activity. This monoclonal antibody should serve as a useful probe of the function and biochemistry of a killer cell surface antigen important in the expression of cell-mediated cytotoxicity. Since RH1-38 inhibits cytotoxicity mediated by at least three apparently unrelated effector cells, the relevant antigen may be part of a common mechanistic step. As the companion paper demonstrates, this monoclonal antibody does not affect the conjugation step, but appears to block a late step in the NK cytolytic mechanism. Thus, RH1-38 recognizes either an epitope district from previously-described anti-LFA-1 antibodies or alternatively recognizes a distinct functional killer cell surface molecule.     

Early-life epileptiform discharges exert both rapid and long-lasting effects on AMPAR subunit composition and distribution in developing neurons

The perinatal period of brain is characterized by dynamic changes in structure and high propensity for epilepsy. Animal models have shown that alterations of AMPA receptor (AMPAR) assembly or function may be related to seizure-induced cell damage, long-lasting impairments in brain development and seizure threshold. However, effects of earlier epileptiform discharges on AMPAR composition and sub-cellular distribution remain understudied. In this study, we analyzed age-dependent variation of relative GluR1 and GluR2 protein levels in primary cultured rat cortical neurons at 7 DIV, 12 DIV, 17 DIV and 21 DIV. By inducing a single event of epileptiform activity at 6 DIV, we tested the effects of early-life seizure-like insults on AMPAR subunit distribution. We found a significant increase in synaptosomal membrane GluR1 expression in magnesium-free (MGF) medium-treated neurons at each time point detected (p < 0.05), while GluR2 expression increased at 7 DIV, and declined at 17 DIV and 21 DIV respectively (p < 0.05). That is, a trend of high GluR1 with much lower GluR2 expression on the surface membrane of epileptiform discharges experienced neurons over time in culture was presented. These findings in an in vitro model of early-life seizure may inform rodent models of epilepsy, as well as the cellular mechanism involved in epilepsy-associated brain dysfunction.     

The Follow-Up Study of Skin Reactivity to Recall Antigens and E- and EAC-RFC Profiles in Blood in Asbestos Workers

We have determined cutaneous DTH reactions to SK-SD and PPD and peripheral blood lymphocyte profiles in a group of asbestos workers in two consecutive surveys. It was found that asbestosis and, to a lesser extent, the presence of ANA are significantly correlated with the lack of response to the above antigens. 83% of asbestos workers when tested at a 4 year interval fell into the same two categories of responsiveness (lack of response or response at least to one antigen).The asbestosis cases had lower total lymphocyte count as well as proportions and absolute number of E-RFC as compared to asbestos workers without asbestosis and/or ANA. Furthermore, the latter group showed the lower percentages and absolute number of E-RFC than the matched controls. The presence of ANA is also correlated with lower proportions of E-RFC. However, this is related at least in part to asbestosis.     

Influence of antibody and complement components on phagocytosis and chemiluminescence of macrophages

Macrophages are known to release reactive oxygen species (O₂^?, ¹O₂, H₂O₂, OH·) in response to various membrane stimuliHowever, our studies show that phagocytic stimulation of macrophages is not necessarily accompanied by a stimulation of the oxidative burstWhereas IgG-opsonized erythrocytes were capable to induce phagocytosis and a chemiluminescence response, both being dependent on the number of IgG bound per erythrocyte, C3b-bearing erythrocytes were well ingested but failed to induce any chemiluminescence reactionFurthermore, stimulation of macrophages, via the Fc-receptors, seems to alter their functional state in regard to the activation of a receptor, which enables them to recognize membrane lesions on the target erythrocyteThe presence of IgG and membrane lesions, e.gthe C5b-9-complex of complement, induced a marked increase in chemiluminescence compared with stimulation by IgG-bearing particles aloneThe augmented response of macrophages was at least in part due to an additional release of H₂O₂, which was not liberated in response to IgG-bearing erythrocytesThis «Alesion recognizing receptor» in the macrophage membrane could not be activated by stimulation of C3b-receptors, indicating its functional linkage to the Fc-receptors.     

Diblock Copolymers Based on 1,4‐Dioxan‐2‐one and ε‐Caprolactone: Characterization and Thermal Properties

Summary: Various poly(ε‐caprolactone‐block‐1,4‐dioxan‐2‐one) (P(CL‐block‐PDX)) block copolymers were prepared according to the living/controlled ring‐opening polymerization (ROP) of 1,4‐dioxan‐2‐one (PDX) as initiated by in situ generated ω‐aluminium alkoxides poly(ε‐caprolactone) (PCL) chains in toluene at 25 °C. 1 ¹H NMR, PCS and TEM measurements have attested for the formation of colloids attributed to a growing PPDX core surrounded by a solvating PCL shell during the polymerization of PDX promoted by ω‐Al alkoxide PCL chains in toluene. The thermal behavior of the P(CL‐block‐PDX) copolymers was studied by DSC; showing two distinct melting temperatures (as well as two glass transition temperatures) similar to those of the respective homopolyesters. Finally, the thermal degradation of the P(CL‐block‐PDX) block copolymers was investigated by TGA simultaneously coupled to a FT‐IR spectrometer and a mass spectrometer for evolved gas analysis (EGA). The degradation occurred in two consecutive steps involving a first unzipping depolymerization of the PPDX blocks followed by the degradation of the PCL blocks via both ester pyrolysis and unzipping reactions.

TEM observation of P(CL‐block‐PDX) block copolyesters ( = 11 600 and = 22 100) as formed by vaporization starting from a diluted suspension in toluene/TCE mixture solvent (50/50 v/v).     

Involvement of a Na−Ca exchange mechanism in contraction induced by low-Na solution in isolated guinea-pig aorta

The possibility of involvement of a Na–Ca exchange mechanism in the contractile responses induced by a reduction of external Na concentration ([Na]₀) has been studied in isolated guinea-pig aorta. Low-Na (11.9 mM) solution (Lisubstituted) produced a contraction in ouabain-treated muscles in the presence of phentolamine (10^–6 M). The magnitude of the contraction was dependent on the duration of the pretreatment with ouabain (2×10^–5 M). Ca-free solution, but not verapamil (10^–6 M), abolished the contraction induced by low-Na solution. The muscles were loaded with various amounts of Na by incubating the tissue with ouabain and varying [Na]₀ (11.9–148.7 mM) in the absence of Ca. The magnitude of the contractions induced in these muscles by low-Na solution containing Ca (2.5 mM) was dependent on the cellular Na content. Loss of cellular Na into low-Na solution followed a single exponential time course and the rate coefficient of Na-loss in the presence of external Ca was about twice as great as in the absence of Ca. Cellular⁴⁵Ca uptake in low-Na solution was significantly greater in Na-loaded tissues (pretreated with ouabain for 3 h) than in normal tissues. The⁴⁵Ca uptake in low-Na solution was not inhibited by verapamil. These results suggest that the contraction induced by low-Na solution is caused by a Ca influx which is dependent on internal Na (a Na–Ca exchange mechanism).