Inhibitory effect of berberine on the activation and proliferation of T lymphocytes

AIM: To investigate the effect of berberine (Ber) on the activation and proliferation of T lymphocytes and its mechanism of action. METHODS: Whole peripheral blood from normal subjects was stimulated with phytohemagglutinin (PHA) or phorbol ester (PDB) plus ionomycin (Ion) and the expression levels of CD69 and CD25 were evaluated with flow cytometry after the staining with appropriate fluorescent monoclonal antibody. The distribution of cell cycles was analyzed by propidium iodide staining and dead cells by 7-aminoactinomycin live staining. RESULTS: 100 μmol/L and 50 μmol/L of Ber had significant inhibition of the expression of CD69 on T cells stimulated with PDB plus Ion or PHA, while effect of 25 μmol/L Ber was not significant. And as time of action extended, the extent of inhibition decreased. For the expression of CD25, Ber at the concentrations as above all exerted significant inhibitory effect in a dose-dependent manner. Moreover, Ber could block lymphocytes cell cycle progression from G₀/G₁ phase to S and G₂/M phase without phase specificity. Besides, live staining analysis revealed that Ber did not have significant cytotoxicity on lymphocytes. CONCLUSIONS: Ber significantly inhibits the expression of early and mid activation antigens of T cells and also blocks the progression of lymphocytes cell cycles. These results suggest that Ber exerts immunosuppression effect through inhibiting the activation and proliferation of T cells.     

Protein expression of cyclin D2 and p16 in proliferation and differentiation of cultured cardiac myocytes

AIM:To investigate the protein expression of cyclin D₂ and p16 in proliferation and differentiation of cultured cardiac myocytes.METHODS:One-day-old Sparague-Dawley rats were used. Cardiac myocytes(CM) were collected by a trypsin-dispersal method and cultured. Cell growth line and fluorescence activated cell sorting (FACS) were used to investigate the proliferation of CM. Ultra-thin sections were made to observe the ultrastructure of CM under transmission electron microscope. The expression of cyclin D₂ and p16 in CM were measured using immunocytochemistry and image analysis.RESULTS:①Results of cell growth line and FACS analysis showed that cultured CM could proliferate in the first 3 cultured days, but the ability decreased quickly, concomitant with differentiation. CM was obseved quiescent in cell cycle three days later. The ultrastructure of CM showed the large amount of myofilaments and mitochondrion. ②The protein expression of cyclin D₂ in 3,4,5 day CM group was 0.89 times(P＜0.05),0.80 times (P＜0.05) and 0.56 times (P＜0.01) of that in 1 day group, respectively. The expression of p16 in CM was increased during the culture process, 2,3,4,5 day group were 1.63 times, 1.72 times, 1.99 times and 2.84 times (P＜0.01) of that in 1 day group, respectively.CONCLUSION:Cultured neonatal rat cardiac myocytes could proliferate during the first 3 days after incubation, but the ability of proliferation decreased, from the fourth day, concomitant with differentiation. Cyclin D₂ and p16 play the key roles in CM postnatal development. Downregulation of cyclin D₂ and upregulation of p16 may induce CM differentiation.     

对虾的免疫机制及其疾病预防策略的研究

在对虾的细胞免疫中血细胞是主要的作用因素,而体液免疫是血淋巴中的一些酶和调节因子,机体还可以被诱导产生特殊的免疫保护反应.应用免疫增强剂、疫苗和基因工程技术为预防对虾病害提供了有效的途径.本文根据国内外的有关资料,就对虾的免疫机制和疾病预防策略进行了综述.     

富铁酵母对家兔红细胞免疫功能的影响

为探讨富铁酵母对家兔红细胞免疫功能的影响,将32只家免随机分成富铁酵母高、低剂量组,FeSO4组及空白对照组,分别灌服富铁酵母(Fe元素8 mg/kg、4 mg/kg)、FeSO4(Fe元素4 mg/kg)及空白生理盐水;连续5 d后,检测红细胞C3g受体花环率和复合物(IC)花环率.结果:富铁酵母较FeSO4及空白组能显著提高红细胞C3h受体花环和红细胞复合物(IC)花环的百分率,但高、低剂量间无显著性差异.结论:富铁酵母能显著提高红细胞免疫功能.     

Effect of EGFP gene transfection on the cell cycle distribution of primary cultured human chondrocytes

AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35.37％ at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage.     

尿素水平对青山羊血液生化指标的影响

应用饲养试验与氮平衡试验,研究了不同尿素水平的日粮对青山羊血清总蛋白(STP)、血清清蛋白(ALB)、血清球蛋白(GLB)、血清清球蛋白比值(A/G)、血清尿素氮(SUN)及血清谷丙转氨酶的含量变化的影响。试验采用4×4拉丁方试验设计,试验分4个阶段,在基础日粮一致的情况下,采用4、9、14、0(对照)g/kg·DM4种不同的处理,试验结果表明:不同尿素水平极显著影响ALB、SUN浓度和GPT活性浓度(P<0.01),对血清清球比(A/G)无显著差异(P>0.05),尿素水平与STP、GLB浓度存在负相关;血清含氮生化指标STP、ALB、GLB、A/G、GPT、SUN随时间呈动态变化,出现高峰的时间比瘤胃氨浓度出现的时间平均推迟1～2h。进食尿素后,血液生化指标由平稳逐渐缓慢升高到高峰,随后又下降并趋于平衡。     

丝兰皂甙对绵羊瘤胃发酵及血液生化指标的影响

选用12只装有永久性瘤胃瘘管的成年雄性东北细毛羊，采用随机区组试验设计，分别饲喂0、100、200、300mg／kg水平丝兰皂甙，研究其对瘤胃发酵及血液生化指标的影响。对试羊采食后的0、2、4、6、8h瘤胃内环境指标进行测定，各组在同一时间点pH无显著差异（P〉0．05），但总体上随着丝兰皂甙添加水平的上升，pH随时间变化有变缓趋势；300mg／kg组较对照组4h和6h乙酸浓度下降15．1％和19．8％，丙酸浓度则提高22．7％和22．9％，差异显著（P〈0．05），各组总挥发性脂肪酸浓度差异不显著（P〉0．05）；100、200、300mg／kg组对氨的抑制率分别为17．17％、29．84％和27．12％，200mg／kg组较对照组差异显著（P〈0．05）；28d后，测定试羊血液生化指标，各组均处正常生理范围内且差异不显著（P〉0．05）。     

铜中毒对雏鸭某些血液指标的影响

选用1日龄天府肉鸭210只,随机分为3组,分别喂以对照日粮(Cu12.16mg/kg)、铜中毒日粮(Cu850mg/kg)和铜中毒日粮(Cu1050mg/kg)。试验期7周。以血液病理学方法观察了铜中毒对雏鸭某些血液学指标的影响。结果显示铜中毒组、组雏鸭分别于2周龄和3周龄开始出现临床症状,死亡18只和5只。血液病理学变化表现为,红细胞大小不等、变形变性、坏死溶解;红细胞数量和血红蛋白含量显著降低(P<0.01),血清谷丙转氨酶、谷草转氨酶活性显著升高(P<0.05或P<0.01),血清铜蓝蛋白活性下降。结果表明,铜中毒对雏鸭的生长发育和上述血液指标影响明显。     

乙烯对匍匐翦股颖ISR抗病反应中AsA-GSH循环及胼胝质沉积的影响

以匍匐翦股颖品种“Penn-A4”为试验材料,侵染立枯丝核菌后,经丁二醇(BDO)诱导产生系统抗性(ISR),喷施不同浓度的乙烯合成抑制剂氯化钴(CoCl₂)和促进剂1-氨基环丙烷羧酸(ACC)后,检测抗坏血酸-谷胱甘肽(AsA-GSH)循环中抗坏血酸(AsA)和还原型谷胱甘肽(GSH)含量及相关酶活性的变化,并观察匍匐翦股颖ISR反应中胼胝质沉积的变化。结果表明,乙烯抑制剂处理下,匍匐翦股颖幼苗中AsA含量较低,抗坏血酸过氧化物酶(APX)活性下降,大量GSH被催化还原为氧化型谷胱甘肽(GSSG),GSSG大量积累,同时谷胱甘肽还原酶(GR)活性较低,GSSG经GR少量还原为GSH。乙烯促进剂处理下,AsA含量较高,APX活性升高,GSSG在高活性GR作用下催化还原为GSH,使得GSH大量积累。因此在匍匐翦股颖ISR抗病反应中,高浓度乙烯促进AsA和GSH的大量积累,它们不仅参与了活性氧的代谢平衡,同时也作为信号分子在ISR抗病反应中起着重要作用。匍匐翦股颖感染褐斑病后,胼胝质主要沉积在厚壁细胞、韧皮部、木质部和表皮组织,其中厚壁细胞胼胝质沉积最多,表皮组织沉积最少。此外,胼胝质沉积面积在不同乙烯信号分子处理间存在差异,但随着处理时间的延长差异不显著。一定浓度的乙烯对ISR反应中胼胝质的沉积具有一定的影响,随着处理时间的延长显著增加,在100 μmol·L^-1ACC处理5 d后,胼胝质沉积总面积仅为9.916 mm²,处理10 d后升至最高,为38.396 mm²,但在病害侵染后期,胼胝质数量减少,在100 μmol·L^-1ACC处理15 d后,胼胝质沉积总面积降至20.052 mm²,反映了乙烯信号分子对胼胝质沉积的影响是一种短期效应,短期内可提高匍匐翦股颖植株抗病性,具有信号分子的短时效特点。研究结果为探清匍匐翦股颖ISR抗病响应中ET信号分子如何调控抗病生理特性提供了理论基础。     

Efficacy of a mephentermine‐based product as a vasopressor and a cardiac performance enhancer when given intramuscularly to cattle

The goal of this study was to confirm the vasopressor and cardiac effects of POTENAY^® INJETÁVEL (POT), a mephentermine‐based product, given to cattle with induced vascular/cardiac depression. Ten healthy Holstein cattle (206 ± 13 kg) followed a randomized‐complete‐block design (RCBD) utilizing crossover study design. Each animal randomly received (1 ml/25 kg, IM) of either POT (n = 10) or volume‐matched placebo control (0.9%NaCl, CP, n = 10). A subset of animals (n = 5) received POT first (day 0) while the remaining (n = 5) received CP; after a six‐day washout period, cattle received the opposite compound. Animals were anesthetized and catheterized for systemic/left ventricular hemodynamic monitoring. Myocardial dysfunction/hypotension was induced by increasing the end‐tidal isoflurane concentration until arterial blood pressure was 20% lower than at baseline and remained stable. Once the animal was determined to be hypotensive and hemodynamically stable, steady‐state hypotensive baseline data (BL2) were acquired, and treatment with either POT or CP was given. Data were acquired post‐treatment at every 15 min for 90 min. POT improved cardiac output (+68 L/min, ±14%, p < 0.05), MAP (+14 mmHg, ±4%, p < 0.05), HR (+22 bpm, ±8%, p < 0.05), and peak rates of ventricular pressure change during both systole (dP/dt_max: +37 mmHg/s ±13%, p < 0.05) and diastole (dP/dt_min: +31 mmHg/s, ±7%, p < 0.05). No improvements were noted following placebo‐control administration. Results indicate that POT improves cardiac performance and systemic hemodynamics in cattle with induced cardiovascular depression when given as single intramuscular injection.