Phytoplankton pigments and adenosine triphosphate (ATP) in Lake Peipsi-Pihkva

The material for pigment analysis was collected 1–3 times a year from Lake Peipsi-Pihkva in 1983, 1987, 1988, 1991 and 1992–1995. Concentrations of chlorophyll a, b and c (Chla, Chlb, Chlc), pheopigment (Pheo) and adenosine triphosphate (ATP) were measured biweekly in 1985–1986. The mean of all Chla values was 20.2 mg m^–1 (median 13.3 mg m^–1) indicating the eutrophic state of the lake. Average Chlb, Chlc, Pheo and carotenoid (Car) contents were 3.7 mg m^–3, 4.1 mg m^–3, 3.0 mg m^–3 and 4.8 mg m^–3, respectively. The average Chlb/Chla ratio was 22.9%, Chlc/Chla 23.4%, Pheo/Chla 38%, Car/Chla 37% and ATP/Chla 3%, the medians being 14.3, 13.6, 17.5, 39.4 and 1.9%, respectively. The proportion of Chla in phytoplankton biomass was 0.41%, median 0.32%. There were no significant differences in temperature, oxygen concentration, Chla, and ATP between the surface and bottom water; the lake was polymictic during the vegetation period. The Chla concentration had its first peak in May followed by a decrease in June and July. In late summer Chla increased again achieving its seasonal maximum in late autumn. The ATP concentration was the highest during spring and early summer, decreasing drastically in autumn together with the decline of primary production. ATP/Chla was the highest during the clear water period in June and early July, which coincided also with the high proportion of Chla in phytoplankton biomass. The highest Chla occurred in November (average 37.2 mg m^–3) when Secchi transparency was the lowest (1.05 m). Concentrations of Chlb, Chlc and carotenoids were the highest in August, that of Pheo in June. Concentrations of Chla and other pigments were the lowest in the northern part of Lake Peipsi (mean 14.7 mg m^–3, median 12.5 mg m^–3) and the highest in the southern part of Lake Pihkva (mean 47.9 mg m^–3, median 16.3 mg m^–3). An increase of Chla and decrease of Secchi depth could be noticed in 1983–1988, while in 1988–1994 the tendency was opposite.     

Continuous,high capacity reconstitution of nutrient media from concentrated intermediates

We designed an Integrated Media Preparation System (IMPS) for continuous, on-line preparation of cell culture media and delivery to intermediate storage vessels or directly to a bioreactor. Key components of the IMPS include: a high precision, continuous fluid mixing device; formulation-specific liquid medium concentrates; validated process controls and membrane filtration; and automated dispensing into large volume flexible plastic containers. The IMPS system is designed to produce sterile, single-strength liquid medium from common raw materials at a delivery rate of 1000–3000 liters per hour and will manufacture homogenous batches from several thousand liters to over 60,000 liters. Fortified nutrient media prepared from multi-component 50X concentrates have been demonstrated to accelerate bioreactor seed chains, increase product yield, and reduce the overall manufacturing cost of nutrient medium. A productivity matrix will analyze the fully-loaded costs and contrast alternative methods for media preparation against projected biological yield.Abbreviations IMPS Integrated Media Preparation System - 50X Nutrient fluid components formulated at fifty-fold final use concentration - 1X Nutrient fluid formulated at final, single-strength use concentration - cGMP Current Good Manufacturing Practices - SCADA Supervisory Control and Data Acquisition - PLC Process Logic Controller - LTI Life Technologies, Inc. - WFI Water for Injection - CIP Clean in place - SIP Sterilize in place - HPLC High performance liquid chromatography - DMEM Dulbecco's Modified Eagle's Medium     

Characterization and co-culture of novel nontransformed cell lines derived from rat endometrial epithelium and stroma

Summary Normal and neoplastic growth of epithelial cells depends on mutual interactions between epithelial and stromal cells. As a tool for the study of the underlying molecular mechanisms, we have developed temperature-sensitive, nontransformed cell lines derived from rat uterine epithelium and stroma by transfecting primary cultures with a temperature-sensitive mutant of the SV40 large T antigen. The epithelial and stromal cell lines obtained shared relevant morphological characteristics with the primary cells from which they were derived. Immunocytochemical analysis showed that the epithelial cell lines expressed the intermediate filament cytokeratin, whereas the stromal lines expressed the intermediate filament vimentin. Alkaline phosphatase activity was present in all cell lines examined. All cell lines were anchorage dependent and did not form foci. One epithelial cell line expressed oxytocin mRNA, a gene product recently shown to be highly expressed in vivo in the uterine epithelium at term. If grown on Matrigel, this cell line formed domelike structures, a further characteristic of its differentiated phenotype. In an attempt to reconstitute an endometrium in vitro, epithelial cells were seeded on top of a layer of stromal cells. Paraffin cross sections showed that this in vitro system consisted of a bilayer structure. Four to five cuboidal epithelial cells were typically anchored atop one stromal cell, forming an endometriumlike tissue. The present in vitro system should provide a useful model for further studies on endometrial functions and epithelial/stromal cell interactions at a molecular level.     

Human HE2 (μB) and μA motifs show the same function as whole IgH intronic enhancer in transgenic mice

In the murine IgH gene intronic enhancer (ENH_iH), two major functional domains were reported. One is the E4/octomer region and another includes the A and B motifs. In the human ENH_iH, it was reported that the HE2, which corresponds to the murine B, and E6 motifs play an important role in an enhancer activity and a tissue-specificity at cellular level. Here we examined thein vivo function of the E6, A and HE2 motifs within the human ENH_iH by using the transgenic mice technique. The A and HE2 motifs together revealed almost the same enhancer function as the whole human ENH_iH, but the E6 motif had lesser enhancer acitivty and tissue-specificity.     

Properties of the ATP-sensitive K+ current activated by levcromakalim in isolated pulmonary arterial myocytes

Tension and patch clamp recording techniques were used to investigate the relaxation of rabbit pulmonary artery and the properties of the K⁺ current activated by levcromakalim in isolated myocytes. Under whole-cell voltage clamp, holding at –60 mV in symmetrical 139 mm K⁺, levcromakalim (10 m) induced a noisy inward current of –116 ± 19 pA (n = 13) which developed over 1 to 2 min. This current could be blocked by either glibenclamide (10 m) or phencyclidine (5–50 M) and was unaffected when extracellular Ca²⁺ was removed. Both these drugs inhibited the levcromakalim-induced relaxation of muscle strips precontracted with 20 mm [K⁺] _o . Application of voltage ramps in symmetrical 139 mm K⁺ confirmed that the levcromakalim-induced current was carried by K⁺ ions and was weakly voltage dependent over the potential range from –100 to +40 mV.The unitary current amplitude and density of the channels underlying the levcromakalim-activated whole-cell K⁺ current was estimated from the noise in the current record. We estimate that levcromakalim caused activation of around 300 channels per cell, with a single channel current of 1.1 pA, corresponding to a slope conductance of about 19 pS. Furthermore, cells dialyzed with an ATP-free pipette solution developed a large noisy inward current at –60 mV, which could subsequently be blocked by flash photolysis of caged ATP. Analysis of the noise associated with this current indicated that the single channel amplitude underlying the ATP-blocked current was 1.4 pA, a value similar to that estimated for the levcromakalim-induced current. We conclude that the conductance of this ATP-sensitive channel is likely to be small under physiological conditions and that it is present at low density.We thank SmithKline & Beecham for the gift of levcromakalim, ICI Pharmaceuticals for the gift of charybdotoxin and Prof. D. Colquhoun for the noise analysis programs. We also thank Mr. R. Davey for technical assistance with tension experiments. This work was supported by the British Heart Foundation and the Wellcome Trust. L.H.C. is a Wellcome Research Fellow and P.L. is an intermediate fellow of the BHF.     

颅内压参数联合血清caveolin-1、AQP-4对高血压脑出血患者术后预后不良的预测价值

摘要 目的：探讨颅内压参数联合血清小窝蛋白-1（caveolin-1）、水通道蛋白4（AQP-4）对高血压脑出血（HICH）患者术后预后不良的预测价值。方法：选择2020年1月至2022年1月河北省胸科医院收治的106例HICH患者,术后随访3个月,根据格拉斯哥预后（GOS）评分将患者分为预后良好组（55例）,预后不良组（51例）。术后监测颅内压参数[压力反应指数（PRx）、平均颅内压波幅（MWA）、20 mmHg阈值下颅内压剂量（Dicp20）],检测血清caveolin-1、AQP-4水平。多因素Logistic回归分析HICH患者术后预后不良的因素。受试者工作特征曲线（ROC）分析颅内压参数联合血清caveolin-1、AQP-4预测HICH患者术后预后不良的价值。结果：预后不良组PRx、MWA、Dicp20以及血清caveolin-1、AQP-4水平高于预后良好组（P＜0.05）。低术前格拉斯哥昏迷评分（GCS）评分、高PRx、高Dicp20、高caveolin-1、高AQP-4是HICH患者术后预后不良的危险因素（P＜0.05）。联合PRx、Dicp20、caveolin-1和AQP-4预测HICH患者术后3个月预后不良的的曲线下面积为0.823,大于PRx、Dicp20、caveolin-1和AQP-4单独预测。结论：高PRx、Dicp20、caveolin-1、AQP4是HICH患者术后预后不良的危险因素,联合颅内压参数PRx、Dicp20及血清caveolin-1、AQP4预测HICH患者术后预后不良具有较高的价值。     

外周血NETs、TP53、STAT3表达与弥漫性大B细胞淋巴瘤临床病理及预后的关系分析

摘要 目的：探究外周血中性粒细胞胞外诱捕网（NETs）、TP53、信号转导与转录因子3（STAT3）表达与弥漫性大B细胞淋巴瘤（DLBCL）临床病理及预后的关系。方法：选取2020年3月-2021年12月收治的71例DLBCL患者作为研究对象,抽取患者外周静脉血,采用R-CHOP方案进行治疗,记录患者外周血NETs、TP53、STAT3表达情况并分析DLBCL患者外周血NETs、TP53、STAT3表达与其临床病理及预后的关系。结果：髓细胞组织增生蛋白（MYC）阳性在TP53阳性中的占比显著高于TP53阴性,差异有统计学意义（x²=28.844,P＜0.001）;Hans分型生发中心B细胞（GCB）在STAT3阳性中的占比显著高于STAT3阴性（x²=4.331,P=0.037）,其余差异无统计学意义（P>0.05）;随访截止至2022年6月,随访时长8~28个月,71例患者中共53例缓解DLBCL患者,其余18例为R/R DLBCL患者;NETs阳性、TP53阳性、STAT3阳性患者无进展生存期（PFS）显著低于NETs阴性、TP53阴性、STAT3阴性患者,差异有统计学意义（P<0.05）且NETs阳性、TP53阳性、STAT3阳性患者存活率均低于NETs阴性、TP53阴性、STAT3阴性患者（P<0.05）;单因素分析结果显示Ann Arbor分期、NETs、TP53、STAT3为DLBCL患者的影响因素（P<0.05）;以患者预后情况（R/R DLBCL=1,缓解DLBCL=0）为因变量,将Ann Arbor分期、NETs、TP53、STAT3单因素分析有统计学意义的因素纳入COX回归模型中,结果显示：NETs、TP53、STAT3为DLBCL患者预后的危险因素（P<0.05）。结论：TP53、STAT3表达与DLBCL临床病理存在一定相关性,临床应对DLBCL患者TP53、STAT3表达情况引起重视;NETs、TP53、STAT3表达为DLBCL预后的危险因素,可作为DLBCL患者不良预后的预测指标。     

血清GDF-15、hs-cTnT对冠状动脉旁路移植术后新发房颤及近期主要心血管事件的预测效能研究

摘要 目的：分析血清生长分化因子15（GDF-15）、高敏心肌肌钙蛋白T（hs-cTnT）对冠状动脉旁路移植术后新发心房颤动（房颤）及近期主要心血管事件的预测效能。方法：选择自2020年1月至2022年1月在我院行冠状动脉旁路移植术的140例冠心病患者作为研究对象,根据术后是否新发房颤,分为房颤组（46例）和非房颤组（94例）。检测两组术前血清GDF-15、hs-cTnT水平,使用多因素Logistic回归分析血清GDF-15、hs-cTnT与术后新发房颤的关系;随访6个月,观察主要心血管事件发生情况,通过受试者工作特征曲线下面积（AUC）评价血清GDF-15、hs-cTnT对术后新发房颤及近期主要心血管事件的预测效能。结果：两组患者年龄、性别、体重指数等一般资料比较无差异（P＞0.05）;房颤组SYNTAX积分高于非房颤组,差异有统计学意义(P＜0.05);房颤组血清GDF-15、hs-cTnT水平均高于非房颤组（P＜0.05）;经多因素Logistic回归分析,SYNTAX积分、血清GDF-15、hs-cTnT均是冠状动脉旁路移植术后新发房颤的独立预测因素（P＜0.05）;经ROC曲线分析,血清GDF-15联合hs-cTnT预测冠状动脉旁路移植术后新发房颤的AUC为0.933,大于SYNTAX积分的0.790,预测近期主要心血管事件的AUC为0.925,大于SYNTAX积分的0.750（P＜0.05）。结论：血清GDF-15联合hs-cTnT对冠状动脉旁路移植术后新发房颤及近期主要心血管事件均具有良好的预测效能,值得临床予以重视。     

阿司匹林通过调节Hippo途径抑制大鼠颅内动脉瘤的形成和机制

摘要 目的：探讨阿司匹林通过调节Hippo途径抑制大鼠颅内动脉瘤形成的机制。方法：选择40只健康SD大鼠分为对照组（不做任何处理）、假手术组（暴露双侧肾动脉后支和左颈总动脉,但不结扎）、动脉瘤组（结扎双侧肾动脉后支和左颈总动脉后生理盐水灌胃）和阿司匹林组（动脉结扎后阿司匹林灌胃）,各10只。灌胃12周后检测血清炎性因子[肿瘤坏死因子-α（TNF-α）、单核细胞趋化蛋白-1（MCP-1）、白细胞介素-6（IL-6）、IL-10]、血管内皮损伤标志物[一氧化氮（NO）、内皮素-1（ET-1）、血管内皮生长因子（VEGF）]。处死大鼠后,检测动脉瘤大小、壁厚比、内腔面积和中膜变薄长度,检测动脉瘤血管组织中Yes相关蛋白（YAP）表达情况。结果：（1）动脉瘤组和阿司匹林组大鼠Willis环上有明显凸起,且阿司匹林组大鼠凸起明显小于动脉瘤组。阿司匹林组大鼠动脉瘤大小、内腔面积和中膜变薄长度均显著小于动脉瘤组,壁厚比显著大于动脉瘤组（P＜0.05）。（2）动脉瘤组和阿司匹林组大鼠血清TNF-α、MCP-1、IL-6水平显著高于对照组和假手术组,IL-10水平显著低于对照组和假手术组（P<0.05）;阿司匹林组大鼠血清TNF-α、MCP-1、IL-6水平显著低于动脉瘤组,IL-10水平显著高于动脉瘤组（P<0.05）。（3）动脉瘤组和阿司匹林组大鼠血清NO水平显著低于对照组和假手术组,ET-1和VEGF水平显著高于对照组和假手术组（P<0.05）;阿司匹林组大鼠血清NO水平显著高于动脉瘤组,ET-1和VEGF水平显著低于动脉瘤组（P<0.05）。（4）动脉瘤组和阿司匹林组大鼠YAP蛋白表达相对吸光值显著高于对照组和假手术组（P<0.05）;阿司匹林组大鼠YAP蛋白表达相对吸光值显著低于动脉瘤组（P<0.05）。结论：阿司匹林能够显著减轻颅内动脉瘤大鼠炎性反应,改善血管内皮功能,抑制颅内动脉瘤形成,这可能与阿司匹林调控Hippo信号通路有关。     

七氟醚联合瑞芬太尼维持麻醉对介入治疗颅内动脉瘤患者神经功能、认知功能、炎性因子的影响

摘要 目的：探讨七氟醚联合瑞芬太尼维持麻醉对介入治疗颅内动脉瘤（IA）患者神经功能、认知功能及炎性因子的影响。方法：选入2020年1月~2022年6月在我院接受介入手术治疗的IA患者80例,根据麻醉方式不同分为A组（七氟醚联合瑞芬太尼）和B组（丙泊酚联合瑞芬太尼）,每组40例。评价两组患者的神经功能、认知功能、炎症因子水平等指标,并进行统计比较。结果：A组T1~T3时心率（HR）和平均动脉压（MAP）明显优于B组（P＜0.05）;A组术后24 h和术后7 d美国国立卫生研究院卒中量表（NIHSS）评分明显低于B组（P＜0.05）,易智能精神状态检查量表（MMSE）评分显著高于B组（P＜0.05）;A组术后24 h、术后3 d血清肿瘤坏死因子-α（TNF-α）、C反应蛋白（CRP）和白细胞介素-6（IL-6）水平均明显低于B组（P＜0.05）;A组术后自主呼吸恢复时间、苏醒时间、拔管时间及拔管后躁动发生率较B组低（P<0.05）;不良反应无差异（P＞0.05）。结论：将七氟醚联合瑞芬太尼维持麻醉应用于介入治疗IA患者,可平稳血流动力学,改善术后神经功能及认知功能,降低炎症反应,提高麻醉苏醒质量。