黄河下游湿地芦苇形态变异研究

 以水盐为主导生态因子,分析了黄河下游湿地芦苇(Phragmites australis)种群形态变异的水平与格局。对基径、株高、叶长、叶宽、节间长、节间数和穗长7个形态特征在15个种群中的变异分析表明：种群内个体间形态差异极显著,7个形态特征在种群内的变异系数从大到小依次为节间长（0.284 6）、叶宽（0.253 6）、穗长（0.244 9）、叶长（0.208 5）、基径（0.187 5）、节间数（0.176 3）、株高（0.165 7）; 7个形态特征在种群间的变异度从大到小依次为株高、叶长、节间长、叶宽、基径、节间数、穗长,但种群间形态的显著差异主要存在于滨州种群（BZH）、淡水种群（ZHG、DPH、NYH,土壤总盐度<0.1%）、盐生种群（HS01、HS02、HS03、HS04、HS05、DWL、KD、GD、LZH、LJ、DLH,土壤总盐度>0.3%）之间,在来自相似生境的种群间形态差异普遍不显著;7个形态特征均与水盐联合影响因子显著相关;以形态特征为基础的聚类分析将研究的15个种群分为显著不同的３类。因此,根据黄河下游湿地芦苇形态变异规律和分化特点,建议将该地区芦苇分为盐生芦苇、淡水芦苇、巨型芦苇3个形态类型。     

口服补液减轻40%血容量失血大鼠肠组织缺血性损伤

目的：研究口服葡萄糖-电解质液（GES）对大鼠40%血容量失血时肠组织缺血性损伤的影响。方法：雄性SD大鼠,用氯胺酮-速眠新Ⅱ肌注复合麻醉后,行右颈动脉插管。随机分为3组（n=24）：GES对照组（GES）,失血性休克组（HS）和失血性休克＋口服GES液组（HS＋GES）。GES组：不放血,手术后口服GES;HS和HS＋GES组按全身血容量的40%分两次间隔15min放血制作失血性休克模型。HS＋GES组于失血后0.5h、1h和6h分3次给予3倍失血量的GES灌胃。用激光多谱勒血流仪测定失血后2h、4h和24h肠组织血流量（IBF）后,处死动物后取肠组织检测二胺氧化酶（DAO）和Na^＋-K^＋-ATP酶活性,测定肠组织含水率（WG）,并做病理学检查。结果：HS＋GES组IBF和Na-K-ATP酶活性均显著高于HS组（P〈0.05）,但显著低于GES组（P〈0.05）;失血各组肠WG显著高于GES组（P〈0.05）,24h时HS＋GES组WG显著低于HS组（P〈0.05）;HS＋GES组24hDAO活性均显著高于HS组（P〈0.05）,但显著低于GES组（P〈0.05）;失血后24h病理检查HS＋GES组肠组织水肿和充血改变比HS组明显减轻。结论：口服GES液能显著增加大鼠失血性休克早期IBF,提高Na^＋-K^＋-ATP酶和DAO活性,减轻肠组织水肿和缺血性损伤。     

瘢痕疙瘩及增生性瘢痕中MMP-2、MMP-9的表达

目的探讨基质金属蛋白酶-2、基质金属蛋白酶-9(MMP-2、MMP-9)在瘢痕疙瘩(keloid,Ke)及增生性瘢痕中(hypertrophic scar,HS)的表达。方法免疫组织化学SP法检测MMP-2、MMP-9在20例瘢痕疙瘩、15例增生性瘢痕及10例正常皮肤中的表达,采用图像分析技术对免疫组化结果进行定量分析。结果Ke中MMP-2表达高于正常皮肤(t=2.366,P<0.05),高于HS(t=2.223,P<0.05);MMP-9表达高于正常皮肤(t=3.198,P<0.01),高于HS(t=2.110,P<0.05)。HS中MMP-2表达与正常皮肤无差异(t=0.218,P>0.05),MMP-9表达与正常皮肤无差异(t=1.873,P>0.05)。正常人皮肤仅见MMP-2、MMP-9蛋白的弱阳性或阴性表达。结论MMP-2、MMP-9蛋白的表达与皮肤损伤后的过度增殖及肿瘤化倾向有关。     

靶向敲除β-珠蛋白基因座控制区增强子HS2对K562细胞转录组的影响

人类β-地中海贫血的发病机制与β-样珠蛋白基因异常表达息息相关。人类β-样珠蛋白基因以5′-ε-Gγ-Aγ-δ-β-3′的顺序排列于β-珠蛋白基因座,受5′LCR (locus control region)中5个超敏位点(hypersensitive site,HS)5′HS5～5′HS1和3′HS1调控。其中5′HS2是最重要的增强子,能产生增强子RNA(enhancerRNA)并调控ε-globin、γ-globin和β-globin的表达。为了进一步探究K562细胞中增强子5′HS2的功能,本研究首先通过染色质构象捕获技术在人慢性髓原白血病K562细胞中探测到5′HS2介导的染色质相互作用集中在以包含CTCF(CCCTC-bindingfactor)位点的3′HS1和5′HS5为边界的拓扑结构域中,5′HS2在三维空间上与HBE1、HBG2和HBG1启动子区域相互靠近。其次运用CRISPRDNA片段编辑技术在K562细胞系中删除了增强子5′HS2。最后通过RNA-seq和CUT&Tag (cleavage under target&tagmentation)实验...     

高张NaCl溶液对失血大鼠血压代偿的作用

使麻醉大鼠在5min 内失血,致使动脉血压下降至25mmHg,然后静脉注射1/10失血量的高张 NaCl 溶液(Hypertonic solution,HS,7.5%NaCl)或生理盐水(NS)。静脉注射少量 HS 能明显促进失血后血压回升,这个作用能为6-羟多巴胺或巯甲丙脯酸显著减弱,若将这两个药物同时应用,则 HS 的升压作用完全被解除。HS 使血浆 Na~+浓度明显升高,而 NS使之下降。侧脑室微量注射 HS 后也明显促进动脉血压的恢复。这些实验结果表明失血后静脉注射少量 HS 可升高血浆 Na~+浓度,高 Na~+作用于中枢神经系统,激活交感神经系统和肾素-血管紧张素系统,从而促使血压迅速恢复。     

一株嗜水气单胞菌HS01的偶氮还原脱色特性

[目的]研究嗜水气单胞菌HS01的偶氮染料还原脱色特性.[方法]建立HS01/偶氮染料/电子供体序批式厌氧反应体系,研究Fe(Ⅲ)/腐殖质还原菌HS01以偶氮染料为电子受体的厌氧呼吸特性及影响因素;并构建HS01/偶氮染料/电子供体/铁氧化物体系,探讨铁氧化物对HS01偶氮还原的影响.[结果]HS01可将金橙Ⅰ迅速还原,菌体增殖;柠檬酸、丙三醇、蔗糖和葡萄糖体系中,16h金橙Ⅰ的脱色率分别达87％、85％、88％、90％;不同pH和金橙Ⅰ初始浓度条件下的脱色率不同;在反应体系中加入α-FeOOH,脱色率从90％增加至95％,Fe(Ⅱ)生成量与无染料对照体系相当.[结论]HS01能以葡萄糖为电子供体,金橙Ⅰ为唯一电子受体,进行厌氧呼吸;蔗糖、柠檬酸、丙三醇也可作为有效的电子供体,脱色率依次递减;甲酸、乙酸、乳酸、乙醇及丙酸不能作为HS01厌氧呼吸的电子供体.金橙Ⅰ脱色的最佳pH范围为6.0-8.0;高浓度(2.0 mmol/L)金橙Ⅰ负荷下,HS01仍保持高脱色率(＞85％).在HS01/α-FeOOH/金橙Ⅰ体系中,异化铁还原作用与偶氮呼吸作用同时发生,异化铁还原能促进偶氮脱色,而脱色对Fe(Ⅲ)还原没有明显影响.这可为铁/腐殖质还原菌在环境修复和废水处理等领域的应用提供研究积累.     

Preclinical transplantation and safety of HS/PCs expanded from human umbilical cord blood

AIM: To expand hematopoietic/progenitor stem cells (HS/PCs) from umbilical cord blood (UCB) and prepare the HS/PC product, and analyze preclinical transplantation and safety of HS/PC product. METHODS: Human bone marrow-derived mesenchymal stem cells (MSCs) were used as feeder cells to expand HS/PCs from UCB in a serum-free culture system. The proliferation potential of HS/PCs was analyzed. The expanded HS/PCs were suspended in the L-15 medium to prepare the HS/PC product. The contamination of bacteria, fungi and mycoplasmas, the infection of exogenous virus, the concentration of bacterial endotoxin, and the SCF residual in HS/PC product were determined. Finally, cells from the HS/PC product with or without bone marrow-derived mesenchymal stem cells (BM-MSCs) were transplanted into the irradiated NOD/SCID mice to determine the in vivo engraftment potential. RESULTS: After co-culture for 10 d, the total nuclear cells (TNCs) increased 125-fold, and CD34 + cells increased 43-fold. The granulocyte-macrophage colonyforming cells (GM-CFCs) and erythroid colony-forming cells (E-CFCs) increased 3.3and 4.7-fold respectively. The expanded cells were collected and prepared as the expanded product of HS/PCs by re-suspending cells in L-15 medium. For preclinical safety, the HS/PC product was analysed for contamination by bacteria, fungi and mycoplasmas, the bacterial endotoxin concentration and the SCF content. The results showed that the HS/PC product contained no bacteria, fungi or mycoplasmas. The bacterial endotoxin concentration was less than the detection limit of 6 EU/mL, and residual SCF was 75 pg/mL. Based on clinical safety, the HS/PC product was qualified for clinical transplantation. Finally, the HS/PC product was transplanted the irradiated mice where it resulted in rapid engraftment of hematopoietic cells. CONCLUSION: HSPC product prepared from UCB in the serum-free culture system with hMSCs as feeder cells should be clinically safe and effective for clinical transplantation.     

抑癌因子miR-10a抑制Tiam1表达对胃癌细胞凋亡和迁移的影响

目的:探讨miR-10a抑制Tiam1表达对胃癌细胞凋亡和侵袭的影响。方法:获取胃上皮组织细胞及胃癌组织细胞,利用q PCR及Western blot实验检测两种细胞中mi R-10a表达与Tiam1的m RNA及蛋白表达水平,同时检测胃癌细胞S746T及正常胃粘膜细胞RGM-1和NGEC中mi R-10a表达与Tiam1蛋白表达水平。通过将mi R-10a mimic和mi R-10a inhibitor转染HS746T细胞,利用流式细胞术检测HS746T的细胞周期和细胞凋亡,TranswellTM实验检测HS746T细胞的侵袭能力,qPCR及Western blot实验检测凋亡相关蛋白caspase3、caspase9和Bax以及周期相关蛋白P21表达水平;荧光素酶活性分析实验检测Tiam1是mi R-10a的作用靶点。已构建的Tiam1高表达的Tiam1-pcDNA3.1质粒和敲除Tiam1基因的PX458质粒分别转染HS746T细胞,通过流式细胞术及TranswellTM实验检测HS746T细胞的凋亡及侵袭能力。结果:与胃上皮组织细胞相比,早期胃癌临床组织细胞中mi R-10a表达降低,Tiam1的m RNA及蛋白表达升高;mi R-10a的表达与早期胃癌患者的肿瘤转移密切相关,与年龄、性别和肿瘤分期无关;与正常胃粘膜细胞RGM-1和NGEC相比,胃癌细胞HS746T中的mi R-10a表达降低,而Tiam1蛋白表达升高;mi R-10a可抑制HS746T细胞侵袭,促进细胞凋亡,使其停滞于G0/G1期;mi R-10a靶向作用于Tiam1基因的3'非翻译区(3'UTR),减少Tiam1的蛋白表达;Tiam1可抑制HS746T细胞凋亡,促进HS746T细胞侵袭。结论:mi R-10a靶向作用于Tiam1基因的3'UTR,抑制HS746T细胞的增殖及侵袭,促进HS746T细胞凋亡。     

苏云金芽孢杆菌HS66的转座因子分析

移动遗传因子普遍存在于苏云金芽孢杆菌(Bt)基因组中,而且大部分位于Bt毒素基因两侧附近,与Bt毒素基因进化和转移密切相关。本研究完成了一株对鳞翅目昆虫表现出很强的杀虫活性的Bt菌株HS66的基因组序列草图测定,并利用本地BLAST软件和ISFinder软件,进行了插入序列和转座子序列的查找及基本信息的汇总,结果表明Bt HS66基因组序列含有丰富的插入序列和转座子序列。转座因子分析是整个Bt HS66基因组分析过程中的重要的一环,后续工作中若定位移动因子在染色体及质粒序列的位置,对解析转座因子功能,帮助预测相关杀虫基因信息,以及全面理解Bt HS66基因组将有极大的帮助。     

肝素硫酸转移酶优化表达及其在动物源肝素硫酸化中的应用

肝素是一种重要的凝血药物,目前主要依赖于动物小肠粘膜的提取。动物源肝素含有的抗凝血活性五糖单位GlcNS6S-GlcA-GlcNS6S3S-Ido2S-GlcNS6S少,抗凝血活性低下。文中提出并验证了一种基于酶法催化动物源肝素,提高其硫酸化程度和抗凝血活性的方法。通过比较3种硫酸转移酶肝素-2-硫酸转移酶(Heparan sulfate-2-O-sulfotransferase,HS2ST)、肝素-6-硫酸转移酶(Heparan sulfate-6-O-sulfotransferase,HS6ST)、肝素-3-硫酸转移酶(Heparan sulfate-3-O-sulfotransferase,HS3ST)在重组大肠杆菌及重组毕赤酵母中表达,确定了毕赤酵母作为3种硫酸转移酶的表达宿主;进一步通过N端融合麦芽糖融合蛋白MBP和硫氧还蛋白Trx A,HS2ST和HS6ST的酶表达水平分别提高至(839?14) U/L和(792?23) U/L。通过3种硫酸转移酶HS2ST、HS6ST和HS3ST共同催化动物源肝素,其抗凝血活性由(76?2) IU/mg提高至(189?17) IU/mg。     

Evaluation of the in vitro differential protein adsorption patterns of didanosine-loaded nanostructured lipid carriers (NLCs) for potential targeting to the brain

The preferential in vitro adsorption of apolipoprotein E (Apo E) onto the surface of colloidal drug carriers may be used as a strategy to evaluate the in vivo potential for such systems to transport drugs to the brain. The aim of this research was to investigate the in vitro protein adsorption patterns of didanosine-loaded nanostructured lipid carriers (DDI-NLCs), using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), in order to establish the potential for NLCs to deliver DDI to the brain. NLC formulations were manufactured using high-pressure homogenization using a lipid matrix consisting of a mixture of Precirol(?) ATO 5 and Transcutol(?) HP. The 2-D PAGE analysis revealed that NLCs in formulations stabilized using Solutol(?) HS 15 alone or with a ternary surfactant system consisting of Solutol(?) HS 15, Tween(?) 80, and Lutrol(?) F68, preferentially adsorbed proteins, such as Apo E. Particles stabilized with Tween(?) 80 and Lutrol(?) F68 did not adsorb Apo E in these studies, which could be related to the relatively large particle size and hence small surface area observed for these NLCs. These findings have revealed that DDI-loaded NLCs may have the potential to deliver DDI to the brain in vivo and, in addition, to Tween(?) 80, which has already been shown to have the ability to facilitate the targeting of colloidal drug delivery systems to the brain. Solutol(?) HS 15-stabilized nanoparticles may also achieve a similar purpose.     

Preparation and Optimization Lipid Nanocapsules to Enhance the Antitumor Efficacy of Cisplatin in Hepatocellular Carcinoma HepG2 Cells

This work aimed to develop and optimize several lipid nanocapsule formulations (LNCs) to encapsulate cisplatin (CDDP) for treatment of hepatocellular carcinoma. By comparing the effect of oil/surfactant ratio, lecithin content, and oil/surfactant type on LNC characteristics, two LNCs were selected as optimal formulations: HS15-LNC (Solutol HS 15/MCT/lecithin, 54.5:42.5:3%, w/w) and EL-LNC (Cremophor EL/MCT/lecithin, 54.5:42.5:3%, w/w). Both LNCs could effectively encapsulate CDDP with the encapsulation efficiency of 73.48 and 78.84%. In vitro release study showed that both LNCs could sustain the release CDDP. Moreover, cellular uptake study showed that C6-labeled LNCs could be effectively internalized by HepG2 cells. Cellular cytotoxicity study revealed that both LNCs showed negligible cellular toxicity when their concentrations were below 313 μg/mL. Importantly, CDDP-loaded LNCs exhibited much stronger cell killing potency than free CDDP, with the IC50 values decreased from 17.93 to 3.53 and 5.16 μM after 72-h incubation. In addition, flow cytometric analysis showed that the percentage of apoptotic cells was significantly increased after treatment with LNCs. Therefore, the prepared LNC formulations exhibited promising anti-hepatocarcinoma effect, which could be beneficial to hepatocellular carcinoma therapy.     

Evaluation of the in vitro differential protein adsorption patterns of didanosine-loaded nanostructured lipid carriers (NLCs) for potential targeting to the brain

The preferential in vitro adsorption of apolipoprotein E (Apo E) onto the surface of colloidal drug carriers may be used as a strategy to evaluate the in vivo potential for such systems to transport drugs to the brain. The aim of this research was to investigate the in vitro protein adsorption patterns of didanosine-loaded nanostructured lipid carriers (DDI-NLCs), using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), in order to establish the potential for NLCs to deliver DDI to the brain. NLC formulations were manufactured using high-pressure homogenization using a lipid matrix consisting of a mixture of Precirol^® ATO 5 and Transcutol^® HP. The 2-D PAGE analysis revealed that NLCs in formulations stabilized using Solutol^® HS 15 alone or with a ternary surfactant system consisting of Solutol^® HS 15, Tween^® 80, and Lutrol^® F68, preferentially adsorbed proteins, such as Apo E. Particles stabilized with Tween^® 80 and Lutrol^® F68 did not adsorb Apo E in these studies, which could be related to the relatively large particle size and hence small surface area observed for these NLCs. These findings have revealed that DDI-loaded NLCs may have the potential to deliver DDI to the brain in vivo and, in addition, to Tween^® 80, which has already been shown to have the ability to facilitate the targeting of colloidal drug delivery systems to the brain. Solutol^® HS 15–stabilized nanoparticles may also achieve a similar purpose.     

Heat Shock Preconditioning Reduces the Formation of 8-hydroxy-2′-deoxyguanosine and 4-hydroxy-2-nonenal Modified Proteins in Ischemia-reperfused Liver of Rats

Heat shock preconditioning (HSPC) is a promising strategy for providing ischemic tolerance. The objective of this study is to investigate the effectiveness of HSPC in preventing oxidative damage of cellular proteins and DNA during ischemia-reperfusion of the liver. Male Wistar rats were divided into a heat shock group (group HS) and control (group C). Forty-eight hours prior to ischemia, rats in group HS received HSPC at 42°C for 15 &#117 min. All rats received hepatic warm ischemia for 30 &#117 min and subsequent reperfusion. The formation of 8-hydroxy-2'-deoxyguanosine (8-OHdG), 4-hydroxy-2-nonenal (HNE) modified proteins in liver tissue, survival rate of the animals, and changes in biochemical and histological parameters were compared between groups. Heat shock protein 72 was produced only in group HS. The 7-day survival of rats was significantly better in group HS (10/10) than in group C (5/10) ( p <0.01). The serum release of alanine aminotransferase ( n =10, p <0.01) and the concentration of adenosine triphosphate in liver tissue ( n =10, p <0.01) 40 &#117 min after reperfusion was significantly better in group HS than in group C. The formation of 8-OHdG in liver tissue measured by high-performance liquid chromatography was suppressed in group HS ( p <0.01). The production of HNE-modified proteins as determined by Western-blot analysis was also decreased in group HS. These results were also confirmed by immunohistochemical analysis. As determined by levels of 8-OHdG and HNE-modified proteins produced during ischemia-reperfusion of the liver, HSPC reduced the oxidative injury of cellular proteins and DNA in the liver tissue.     

Cooling therapy upregulates protein C activation in heat stressed rats: An experimental study

Protein C (PC) pathway homeostasis is implicated in heat stress (HS). This study determines whether cooling could improve the PC pathway in HS. Fifty-six anesthetized rats were warmed to achieve HS (rectal temperature [Tr] 42°C). These rats were divided into seven groups: (a) control group:sacrifice immediately 15 min after HS; (b) HS+I:sacrifice immediately after 15 min ice-water treatment or (c) 3 hr after HS; (d) HS+C:sacrifice immediately after 15-min cold-water treatment or (e) 3 hr after HS; (f) HS: sacrifice immediately 15 min after HS or (g) 3 hr after HS. Plasma PC, activated protein C (APC), and soluble thrombomodulin (sTM) levels were tested at both time points. After cooling, Tr in the HS+I and HS+C groups significantly decreased, when compared with the HS group, and Tr was significantly lower in the HS+I group than in the HS+C group ( p < 0.05). Furthermore, sTM levels were highest in the HS group among the groups at both time points. Plasma PC and APC levels increased after HS. In the HS+I and HS+C groups, plasma APC levels and the APC/PC ratio significantly increased at both time points. The proportions were significantly higher in the HS+I group than in the HS+C group, and there was no significant increase in APC/PC ratio in the HS group. Cooling exerts an anticoagulant effect following HS by increasing APC levels. Ice-water blanket therapy is more effective than cold-water blanket therapy in increasing APC levels.     

Influence of heat shock on chlorophyll fluorescence of white oak (<Emphasis Type="Italic">Quercus pubescens</Emphasis> Willd.) leaves

Chlorophyll fluorescence parameters of Quercus pubescens Willd. as response to heat shock (HS) by immersing leaves for 5 and 15 min in water of temperatures between 38 and 59 °C were examined. Fluorescence was measured after different periods of recovery (15, 30, 90, 210, and 1 440 min at 24/26 °C night/day temperature and 100 % humidity). The effective quantum yield of photosystem 2 (Y) in control and HS-treated leaves was always measured after previous 15 min irradiation. Under a 5 min HS, Y did not change after using temperatures below 44 °C, was rapidly restored after HS of moderate temperatures (44–48 °C), and progressively decreased and recovered eventually to the initial value after HS of high temperatures (48–52 °C). Y did not recover after HS with temperatures higher than 52 °C. Increase in the duration of HS from 5 to 15 min lead to change of the initial Y at each HS temperature, but the recovery processes were similar to those characteristic after 5 min incubation. The processes of recovery may depend mainly on the specificity of injuries caused by different heat shock temperatures. Thus Q. pubescens is able to preserve and recover the functional potential of its photosynthetic apparatus in response to HS up to 52 °C.     

The Role of Formulation Additives in Increasing the Potency of Cydia pomonella Granulovirus for Codling Moth Larvae,in Laboratory and Field Experiments

Studies were undertaken to improve the biological efficacy of the granulovirus (CpGV) of the codling moth, Cydia pomonella , by evaluating the performance of some formulation additives that might improve virus persistence and/or virus uptake by first instar larvae. Laboratory studies, using a leaf disc bioassay, demonstrated that 15% cane molasses incorporated within a formulation of purified CpGV dramatically reduced the median lethal exposure time (LET 50 ) to CpGV for neonate larvae at a CpGV dosage rate of 10 7 occlusion bodies (OBs) ml -1 . Screening of a range of other compounds showed that sucrose, fructose and sorbitol (at 10% concentrations) and extracts of apple flesh and skin also gave significant reductions in the LET 50 of CpGV formulations containing these ingredients. Pectin, malic acid and &#102 -farnesene did not significantly reduce the LET 50 . In a field trial, molasses included at 15% (v/v) in a CpGV formulation, containing a dosage rate of 10 12 OBs ha -1 , gave as good control of codling moth damage as virus formulations containing the 'sticker' 0.2% skimmed milk at higher dosage rates of 10 13 and 10 14 OBs ha -1 . Studies of CpGV persistence on foliage revealed no significant improvement of virus persistence on apple foliage using 10% or 15% molasses formulations. A second field trial demonstrated that 10% molasses, 10% sorbitol or 0.08% &#102 -farnesene significantly reduced codling moth deep damage to fruit when these ingredients were added to formulations of pure CpGV. Substantial sooty-mould growth ( Cladosporium spp.) was observed on apple foliage treated with formulations containing molasses, indicating that this formulation additive has secondary consequences that would need to be taken into account if molasses was to be used in commercial CpGV formulations. Nonetheless, these studies clearly demonstrate that major biological improvements in CpGV performance can be achieved by the incorporation of formulation additives, including molasses and several other compounds, that probably function as attractants and/or feeding stimulants for codling moth larvae.     

Poly(ethyleneglycol) column for the determination of acetaminophen,phenylephrine and chlorpheniramine in pharmaceutical formulations

New polar reversed-phase stationary phases in HPLC provide specific selectivities which can help to solve traditional chromatographic problems related to the development of chromatographic methods with widely different retention times for the sample components. One such case is the analysis of pharmaceutical formulations against the common cold. Acetaminophen, phenylephrine and chlorpheniramine, compounds with different polarities, are frequently associated in these drugs. An isocratic and rapid HPLC method for the simultaneous determination of the three compounds, acetaminophen, phenylephrine and chlorpheniramine, in capsules as pharmaceutical formulations, including the separation of impurities (4-aminophenol and 4-chloracetanilide) and excipients, has been developed and validated. The final chromatographic conditions employed a Supelco Discovery HS PEG column poly(ethyleneglycol) 15x0.46 cm, 5 microm. The mobile phase was 20 mM phosphate buffer, pH 7.0-acetonitrile (90:10, v/v) at a flow-rate of 1 ml/min. UV detection was performed at 215 nm for all the compounds except acetaminophen, which was measured at 310 nm. Validation parameters permit us to consider this method suitable.     

Secretion of PGF2alpha and oxytocin during hyperthermia in cyclic and pregnant heifers

The effects of acute heat stress (HS) and oxytocin (OT) injection on plasma concentrations of PGF2alpha and OT were examined in cyclic (C; n = 15) and pregnant (P; n = 11) dairy heifers. On Day 17 of synchronized estrous cycles, animals were randomly assigned to either thermoneutral (TN; 20 degrees C, 20% RH) or HS (42 degrees C, 60% RH) chambers. The jugular vein of each heifer was cannulated and blood samples collected hourly for 4 h, then every 15 min for an additional 3 h. Oxytocin (100 IU) was injected (IV) 5 h after the start of blood collection. Plasma samples were assayed subsequently for concentrations of 13,14-dihydro-15-keto PGF2alpha (PGFM) and OT. During the 7-h experiment, body temperature of HS heifers reached 41.2 degrees C as compared to 38.5 degrees C in control heifers. Plasma concentrations of PGFM increased (P<0.05) and peaked 30 min after OT injection in C (890 pg/ml) and P (540 pg/ml) heifers. In C heifers, heat stress failed to alter PGFM concentrations either before or after OT injection. In the P group, PGFM concentrations following OT injection tended to be higher in HS heifers were further TN heifers (peak values of 690 vs. 410 pg/ml). Pregnant TN and HS heifers were further classified as responders or non-responders to OT challenge according to a cutoff value for PGFM of 193 pg/ml (overall mean of C heifers minus 1 SD). Five of six HS and one of five TN pregnant heifers were classified as responders (P<0.06). Oxytocin concentrations in plasma prior to injection of exogenous OT were not affected by HS or pregnancy status. It is concluded that in C heifers, acute HS in vivo does not cause any further rise in PGF2alpha secretion. However, in P heifers, HS appears to antagonize suppressive effects of the embryo on uterine secretion of PGF2alpha, as indicated by the larger proportion of P heifers responding to OT challenge.