经阴道三维超声联合CA125、CA199、NLR及PLR检测对绝经后子宫内膜癌的诊断效能

摘要 目的：探讨经阴道三维超声联合糖类抗原125（CA125）、糖类抗原199（CA199）、中性粒细胞/淋巴细胞比值（NLR）及血小板/淋巴细胞比值（PLR）检测对绝经后女性子宫内膜癌（EC）的诊断效能。方法：回顾性分析本院2019年5月至2022年5月收治的因绝经后出血就诊的92例疑似子宫内膜肿瘤患者的病历资料,经手术病理证实,EC患者48例（EC组）、子宫内膜良性病变患者44例（良性组）。比较两组CA125、CA199、NLR及PLR水平,对比两组子宫内膜血管指数（VI）、血流指数（FI）、血管-血流指数（VFI）、子宫内膜容积（EV）变化及血流信号分布情况。绘制受试者工作特征（ROC）曲线分析经阴道三维超声联合CA125、CA199、NLR及PLR检测对绝经后EC的诊断效能。结果：EC组CA125、CA199、NLR及PLR水平均高于良性组（P＜0.05）。EC组三维超声参数VI、FI、VFI及EV均高于良性组（P＜0.05）。EC组血流信号Ⅱ级占比25.00%,Ⅲ级占比70.83%;良性组血流信号Ⅱ级占比88.64%,Ⅲ级占比2.27%。EC组血流信号Ⅱ级占比低于良性组,血流信号Ⅲ级占比高于良性组（P＜0.05）。两组血流信号0级、Ⅰ级比较无差异（P＞0.05）。ROC结果显示五项检查联合检测曲线下面积（AUC）值为0.944（95%Cl：0.902~0.985）,具有更优的诊断效能（敏感度为89.10%、特异度为91.80%）。结论：CA125、CA199、NLR及PLR检测联合经阴道三维超声检查诊断绝经后EC效果显著,可为临床诊断提供参考。     

Binding, internalization and intracellular processing of 125I-epidermal growth factor purified by isoelectric focusing

When epidermal growth factor (EGF) which had been extensively purified by HPLC was subjected to iodination with sodium ¹²⁵iodide, 5 major species of differing isoelectric points were produced. Some of these species bound to rat fibroblasts with different affinities but were internalized with equal efficiency. Examination of the internalized ¹²⁵I-labelled molecules revealed processing of all the ¹²⁵I-EGF species to macromolecules with more acidic isoelectric points. The ¹²⁵I-EGF species with a pI of 4.5 corresponded in electrofocusing behavior with intact non-iodinated EGF. Other EGF species probably represented molecules which were covalently modified as a result of the iodination procedure.     

Characterization and determination of titratable groups of proteins by linearization of titration curves. II. Application to lysozyme

The potentiometric acid-base titration curve of fully protonated lysozyme at ionic strengths of 0.10 and 1.0 m has been performed. The stoichiometry and the pK_a values of each titratable group have been determined through the linearization of titration curves. Two types of carboxylic groups with pK_a values of 3.76 and 5.02, the imidazole group with pK_a 7.37 and the amine group with pK_a 9.63, have been identified at an ionic strength of 0.10 m at 25.0°C. The number of titratable groups found per mole of protein has been 5.12 and 5.60 for the two types of carboxylic groups, 1.13 for the imidazole group, and 3.19 for the amino groups. The endpoint of the titration of the protein obtained by this method accords quite well with the endpoint obtained by the use of Gran function applied to the excess of strong base.     

Molecular changes in cell surface membranes resulting from trypsinization of sarcoma 180 tumor cells

Sarcoma-180 tumor cells in culture or grown as an ascites form in the CD-1 mouse have been subjected to mild trypsinization procedures in order to study morphological and molecular changes resulting from proteolysis. The cells attached to a substratum become rounded within 20 min and most undergo cell division, but they do not detach from the substratum. Removal of trypsin permits the cells to go back to their original spindle shape over an 8–20 h period.Surface membranes were isolated from trypsinized ascites and cultured cells and subjected to dodecyl sulfate-acrylamide gel electrophoresis. Both cell types showed the same two kinds of changes in electrophoretic patterns. First, there was a loss of glycoproteins from both cell types, even though they show different complements of cell surface glycoproteins. Second, there is a loss of high molecular weight polypeptides, which have previously been suggested to play a role in membrane stabilization and cell shape. These results further implicate these polypeptides in the control of cell morphology and offer circumstantial evidence for transmembrane interactions of surface glycoproteins with the high molecular weight polypeptides as a factor in controlling cell morphology.     

Stimuli for cuticle formation and ecdysis in vitro of the infective larva of Anisakis sp. (Nematoda: Ascaridoidea)

Third-stage larvae of the genus Anisakis from the fish Leionura atun (Trichiuroidei: Perciformes) form a new cuticle and moult in vitro in about 72 h. If the culture medium is Krebs-Ringer under 5% carbon dioxide in air at 37°C, relatively few moult and survival is poor. But more moult and survival is enhanced if worms are incubated in tissue culture medium 199, even if the gas phase is air, although they moult more quickly if it contains 5% carbon dioxide. In both Krebs-Ringer and 199 the benefits of high concentrations of carbon dioxide only accrue if the gas is present during the first 40 h of incubation. Worms do not feed in these media until they have moulted.     

胰腺癌患者血清CEA、CA242、CA199水平变化及联合诊断的ROC曲线分析

目的:探讨胰腺癌患者血清癌胚抗原(CEA)、糖类抗原242(CA242)、糖类抗原199(CA199)水平变化,并分析上述指标对胰腺癌的联合诊断价值,为胰腺癌的临床诊断提供参考。方法:选择2014年2月至2018年2月我院收治的186例胰腺癌患者(胰腺癌组)、89例胰腺炎患者(胰腺炎组)作为研究对象,并取同期来我院检查的268例健康人作为对照组。比较三组受试者的血清CEA、CA242、CA199水平变化,对比分析血清CEA、CA242、CA199的单一以及联合诊断的准确度、特异度以及灵敏度,并绘制ROC曲线以分析上述指标的诊断价值。结果:三组受试者血清CEA、CA242、CA199水平差异具有统计学意义(P<0.05)。且胰腺炎组和胰腺癌组的血清CEA、CA242、CA199水平明显高于对照组,胰腺癌组患者的血清CEA、CA242、CA199水平明显高于胰腺炎组,差异均有统计学意义(P<0.05)。ROC曲线结果显示,CEA诊断价值最大,CA199诊断价值最小。CEA是胰腺癌单项肿瘤标志物中敏感度最高的,为85.48%;特异度最高的为CA242(96.72%);三项肿瘤标志物联合诊断的准确度增加至92.27%,敏感度增加至95.16%,特异度相比略有下降。结论:与单一肿瘤标记物诊断胰腺癌相比,CEA、CA242、CA199联合诊断的敏感度和准确度均明显升高,可以明显改善胰腺癌的漏诊率,提高患者的生存率,具有较好的临床应用价值。     

miR-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为研究

目的:探讨mi R-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为。方法:qRT-PCR法检测并比较肺癌组织、癌旁正常组织、肺癌细胞、正常肺上皮细胞中的mi R-199a-3p m RNA相对表达量。比较远处转移肺癌组织、未转移肺癌组织中mi R-199a-3p m RNA相对表达量。qRT-PCR法、Western Blot法检测并比较肺癌组织、癌旁正常组织中的CBX7 m RNA及蛋白的表达水平。荧光素酶活性法检测mi R-199a-3p与靶基因CBX7的结合。比较mi R-199a-3p模拟物转染组与阴性对照组的肺癌细胞中的CBX7 m RNA相对表达量及CBX7蛋白表达水平。CCK8实验检测mi R-199a-3p对肺癌细胞增殖的促进作用。Tranwell实验检测mi R-199a-3p对肺癌细胞侵袭与迁移能力的影响。结果:肺癌组织中mi R-199a-3p明显高于癌旁正常组织,发生远处转移的肺癌组织中mi R-199a-3p m RNA的表达量明显高于未发生转移的肺癌组织,差异有统计学意义(P<0.001)。肺癌组织中CBX7m RNA、CBX7蛋白表达水平均明显低于癌旁正常组织,差异有统计学意义(P<0.001)。荧光素酶活性法证实mi R-199a-3p可与靶基因CBX7结合抑制CBX7的表达。肺癌细胞中mi R-199a-3p m RNA的相对表达量明显高于正常肺上皮细胞,CBX7 m RNA相对表达量明显低于正常肺上皮细胞(P<0.05)。对于肺癌细胞,mi R-199a-3p模拟物转染组的CBX7 m RNA相对表达量及CBX7蛋白表达水平均明显低于阴性对照组(P<0.001)。CCK8实验证实mi R-199a-3p能够促进肺癌细胞的增殖,Tranwell实验证实mi R-199a-3p对肺癌细胞侵袭与迁移具有积极的促进作用。结论:mi R-199a-3p在肺癌的发生发展过程中发挥重要作用,能够通过抑制CBX7基因的表达,促进肺癌细胞的增殖、侵袭和转移。     

磁共振成像检查联合血清HE4、TK1、CA199检测在卵巢癌诊断中的应用价值

摘要 目的：研究磁共振成像（MRI）检查联合血清人附睾蛋白4（HE4）、细胞质胸苷激酶（TK1）、糖类抗原199（CA199）检测在卵巢癌诊断中的应用价值。方法：将2017年4月～2019年12月我院收治的卵巢癌患者54例作为卵巢癌组,卵巢良性肿瘤患者49例作为卵巢良性肿瘤组,所有受试者均进行MRI检测,并采集受试者血清检测HE4、TK1、CA199水平,以受试者工作特征（ROC）曲线分析MRI检查联合血清HE4、TK1、CA199检测诊断卵巢癌的效能。结果：卵巢癌MRI特点包括以下几点：①囊实性肿块;②形态不规则;③呈长T2/T1等信号;④病灶边界模糊;⑤增强扫描结果呈病灶显著强化;⑥腹膜转移者存在显著腹腔积液以及散在分布的结节状种植病灶。卵巢良性肿瘤MRI特点包括以下几点：①囊壁光滑;②呈长T1以及长T2信;③病灶边界清晰;④增强扫描结果提示病灶无强化/稍有强化。卵巢癌组血清HE4、TK1、CA199水平分别为（87.13±15.32）pmol/L、（2.15±0.13）pmol/L、（95.39±15.25）U/mL,明显高于卵巢良性肿瘤组的（66.42±10.19）pmol/L、（0.85±0.20）pmol/L、（37.94±1.05）U/mL（均P＜0.05）。经ROC曲线分析可得：MRI检查联合血清HE4、TK1、CA199检测诊断卵巢癌的灵敏度、特异度以及曲线下面积均高于上述各项单独诊断。结论：卵巢癌患者血清HE4、TK1、CA199水平较高,MRI检查联合血清HE4、TK1、CA199检测诊断卵巢癌的价值较高,具有一定的临床应用价值。     

African Swine Fever Virus Protein E199L Promotes Cell Autophagy through the Interaction of PYCR2

African swine fever virus(ASFV), as a member of the large DNA viruses, may regulate autophagy and apoptosis by inhibiting programmed cell death. However, the function of ASFV proteins has not been fully elucidated, especially the role of autophagy in ASFV infection. One of three Pyrroline-5-carboxylate reductases(PYCR), is primarily involved in conversion of glutamate to proline. Previous studies have shown that depletion of PYCR2 was related to the induction of autophagy. In the present study, we found for the first time that ASFV E199 L protein induced a complete autophagy process in Vero and HEK-293 T cells. Through co-immunoprecipitation coupled with mass spectrometry(CoIP-MS)analysis, we firstly identified that E199 L interact with PYCR2 in vitro. Importantly, our work provides evidence that E199 L down-regulated the expression of PYCR2, resulting in autophagy activation. Overall, our results demonstrate that ASFV E199 L protein induces complete autophagy through interaction with PYCR2 and down-regulate the expression level of PYCR2, which provide a valuable reference for the role of autophagy during ASFV infection and contribute to the functional clues of PYCR2.     

Biocatalytic Synthesis of Dihydroxynaphthoic Acids by Cytochrome P450 CYP199A2

CYP199A2, a bacterial P450 monooxygenase from Rhodopseudomonas palustris, was found to exhibit oxidation activity towards three hydroxynaphthoic acids. Whole cells of the recombinant Escherichia coli strain expressing CYP199A2 efficiently catalyzed the regioselective oxidation of 1-, 3-, and 6-hydroxy-2-naphthoic acids to produce 1,7-, 3,7-, and 6,7-dihydroxynaphthoic acid respectively. These results suggest that CYP199A2 might be a useful oxidation biocatalyst for the synthesis of dihydroxynaphthoic acids.