血清TM、P-selectin、HDL-C以及凝血功能指标与创伤性骨折患者术后深静脉血栓形成的关系研究

摘要 目的：探讨血清血栓调节蛋白（TM）、P-选择素（P-selectin）、高密度脂蛋白（HDL-C）以及凝血功能指标与创伤性骨折患者术后深静脉血栓形成（DVT）的关系。方法：选择2018年1月至2020年1月我院收治的术后发生DVT的创伤性骨折患者100例作为DVT组,同期术后未发生DVT的创伤性骨折患者100例作为无DVT组,比较两组血清TM、P-selectin、HDL-C、血浆凝血功能指标[凝血酶时间（TT）、凝血酶原时间（PT）、活化部分凝血酶时间（APTT）、纤维蛋白原（FIB）、D-二聚体（D-D）],应用Pearson相关性分析血清TM、P-selectin、HDL-C与凝血功能指标的相关性,应用多因素Logistic回归分析创伤性骨折患者术后DVT的影响因素。结果：DVT组血清TM、P-selectin、血浆D-D、FIB水平、年龄≥60岁比例高于无DVT组,血清HDL-C水平低于无DVT组（P＜0.05）。DVT组血清TM和P-selectin与血浆D-D、FIB呈正相关（P＜0.05）,血清HDL-C与血浆D-D、FIB呈负相关（P＜0.05）。多因素Logistic回归分析显示,年龄≥60岁、血清TM≥9.50 IU/mL、P-selectin ≥70.00 ng/mL、HDL-C<1.00 mmol/L、血浆D-D≥700.00 μg/L、FIB≥4.00 g/L是创伤性骨折患者术后DVT的危险因素（P＜0.05）。结论：创伤性骨折患者术后发生DVT患者血清TM、P-selectin较无DVT患者升高,HDL-C较无DVT患者降低,联合检测血清TM、P-selectin、HDL-C和凝血功能指标可能有助于降低DVT的发生风险。     

The influence of sex and strain on trace element dysregulation in the brain due to diet-induced obesity

BackgroundThe objective of this study was to identify interaction effects between diet, sex, and strain on trace element dysregulation and gene expression alterations due to diet-induced obesity (DIO) in the hippocampus, striatum, and midbrain.MethodsMale and female C57BL/6 J (B6 J) and DBA/2 J (D2 J) mice were fed either a low fat (10 % kcal) diet (LFD) or high fat (60 % kcal) diet (HFD) for 16 weeks, then assessed for trace element concentrations and gene expression patterns in the brain.ResultsIn the hippocampus, zinc was significantly increased by 48 % in D2 J males but decreased by 44 % in D2 J females, and divalent metal transporter 1 was substantially upregulated in B6 J males due to DIO. In the striatum, iron was significantly elevated in B6 J female mice, and ceruloplasmin was significantly upregulated in D2 J female mice due to DIO. In the midbrain, D2 J males fed a HFD had a 48 % reduction in Cu compared to the LFD group, and D2 J females had a 37 % reduction in Cu compared to the control group.ConclusionsThe alteration of trace element homeostasis and gene expression due to DIO was brain-region dependent and was highly influenced by sex and strain. A significant three-way interaction between diet, sex, and strain was discovered for zinc in the hippocampus (for mice fed a HFD, zinc increased in male D2 Js, decreased in female D2 Js, and had no effect in B6 J mice). A significant diet by sex interaction was observed for iron in the striatum (iron increased only in female mice fed a HFD). A main effect of decreased copper in the midbrain was found for the D2 J strain fed a HFD. These results emphasize the importance of considering sex and genetics as biological factors when investigating potential associations between DIO and neurodegenerative disease.     

Characterization of a cryptic plasmid from a Greenland ice core Arthrobacter isolate and construction of a shuttle vector that replicates in psychrophilic high G+C Gram-positive recipients

Over 60 Greenland glacial isolates were screened for plasmids and antibiotic resistance/sensitivity as the first step in establishing a genetic system. Sequence analysis of a small, cryptic, 1,950 bp plasmid, p54, from isolate GIC54, related to Arthrobacter agilis, showed a region similar to that found in theta replicating Rhodococcus plasmids. A 6,002 bp shuttle vector, pSVJ21, was constructed by ligating p54 and pUC18 and inserting a chloramphenicol acetyl transferase (CAT) cassette conferring chloramphenicol resistance. Candidate Gram-positive recipients were chosen among glacial isolates based on phylogenetic relatedness, relatively short doubling times at low temperatures, sensitivity to antibiotics, and absence of indigenous plasmids. We developed an electroporation protocol and transformed seven isolates related to members of the Arthrobacter, Microbacterium, Curtobacterium, and Rhodoglobus genera with pSVJ21. Plasmid stability was demonstrated by successive transformation into Escherichia coli and four Gram-positive isolates, growth without antibiotic, and plasmid re-isolation. This shuttle vector and our transformation protocol provide the basis for genetic experiments with different high G+C Gram-positive hosts to study cold adaptation and expression of cold-active enzymes at low temperatures.     

高压电场对A549细胞中ABCG2和V-ATPase的表达及其耐药性的影响

目的:探讨高压电场对A549细胞中ABCG2和V-ATPase表达量的影响;探讨高压电场对A549细胞耐药性的影响。方法:MTT法测细胞生长曲线,明确能导致细胞可逆性电穿孔的最高电场强度。慢病毒构建ABCG2和V-ATPase低表达的A549细胞系,并用电场处理,用q-RT-PCR和Western-blot法检测处理前后ABCG2和V-ATPase的m RNA和蛋白表达量的变化。最适强度的高压电场处理各组细胞,在处理前后的细胞中分别加入阿霉素,用高效液相色谱法检测各组细胞中阿霉素浓度。结果:当电场强度为1500 V/cm时,肿瘤细胞增殖最慢;电场强度为1500 V/cm时,肿瘤细胞中ABCG2和V-ATPase的m RNA和蛋白的表达量分别降至对照组的58%和61%,具有统计学差异;1500 V/cm强度的电场可以提高肿瘤细胞内阿霉素的浓度3-4倍。结论:高压电场可以显著降低肿瘤细胞中V-ATPase和ABCG2的m RNA和蛋白的表达量并降低肿瘤细胞的耐药性。     

High-dose neoadjuvant chemoradiotherapy versus chemotherapy alone followed by surgery in potentially-resectable stage IIIA-N2 NSCLC. A multi-institutional retrospective study by the Oncologic Group for the Study of Lung Cancer (Spanish Radiation Oncology Society)

BackgroundThe optimal induction treatment in potentially-resectable stage IIIA-N2 NSCLC remains undefined.AimTo compare neoadjuvant high-dose chemoradiotherapy (CRT) to neoadjuvant chemotherapy (CHT) in patients with resectable, stage IIIA-N2 non-small-cell lung cancer (NSCLC).MethodsRetrospective, multicentre study of 99 patients diagnosed with stage cT1-T3N2M0 NSCLC who underwent neoadjuvant treatment (high-dose CRT or CHT) followed by surgery between January 2005 and December 2014.Results47 patients (47.5%) underwent CRT and 52 (52.5%) CHT, with a median follow-up of 41 months. Surgery consisted of lobectomy (87.2% and 82.7%, in the CRT and CHT groups, respectively) or pneumonectomy (12.8% vs. 17.3%). Nodal downstaging (to N1/N0) and Pathologic complete response (pCR; pT0pN0) rates were significantly higher in the CRT group (89.4% vs. 57.7% and 46.8% vs. 7.7%, respectively; p < 0.001)). Locoregional recurrence was significantly lower in the CRT group (8.5% vs. 13.5%; p = 0.047) but distant recurrence rates were similar in the two groups. Median PFS was 45 months (CHT) vs. “not reached” (CRT). Median OS was similar: 61 vs. 56 months (p = 0.803). No differences in grade ≥3 toxicity were observed. On the Cox regression analysis, advanced pT stage was associated with worse OS and PFS (p < 0.001) and persistent N2 disease (p = 0.002) was associated with worse PFS.ConclusionsCompared to neoadjuvant chemotherapy alone, a higher proportion of patients treated with preoperative CRT achieved nodal downstaging and pCR with better locoregional control. However, there were no differences in survival. More studies are needed to know the optimal treatment of these patients.     

宫颈病变患者阴道微生态与高危型HPV感染及宫颈癌相关增殖基因表达的相关性

目的探讨宫颈病变患者阴道微生态与高危型HPV感染及宫颈癌相关增殖基因表达的相关性。方法选择2018年1月至2019年1月间在我院确诊为原发性宫颈癌的患者50例作为宫颈癌组,在我院诊断为宫颈糜烂的患者78例作为宫颈糜烂组,同期在我院进行体检的健康女性100例作为正常对照组。对比3组研究对象的阴道微生态失调率、高危型HPV感染率以及宫颈癌组、宫颈糜烂组患者宫颈病灶组织中宫颈癌相关增殖基因(Prdx4、Nek2、Fhit、BLCAP)mRNA表达量的差异。采用Pearson检验分析宫颈癌患者阴道微生态失调率与高危型HPV感染及宫颈癌相关增殖基因表达的相关性。结果宫颈癌组、宫颈糜烂组患者的阴道微生态失调率、高危型HPV感染率高于正常对照组,其中宫颈癌组患者这两项指标水平高于宫颈糜烂组(均P0.05)。宫颈癌组患者宫颈病灶组织中Prdx4、Nek2 mRNA表达量高于宫颈糜烂组,Fhit、BLCAP mRNA表达量低于宫颈糜烂组(均P0.05)。相关性分析发现,宫颈癌患者阴道微生态失调率与高危型HPV感染率呈正相关,与癌基因(Prdx4、Nek2)mRNA表达量呈正相关,与抑癌基因(Fhit、BLCAP)mRNA表达量呈负相关(均P0.05)。结论宫颈癌患者阴道微生态失调率较高,可能与高危型HPV感染及癌细胞增殖旺盛密切相关。     

An acidic and thermostable carboxymethyl cellulase from the yeast Cryptococcus sp. S-2: purification, characterization and improvement of its recombinant enzyme production by high cell-density fermentation of Pichia pastoris

The extracellular carboxymethyl cellulase (CSCMCase) from the yeast, Cryptococcus sp. S-2, was produced when grown on cellobiose. It was purified to homogeneity from the supernatant by ultrafiltration, DEAE-5PW anion exchange column and TSK-Gel G3000SW gel filtration. The purified enzyme was monomeric protein with molecular mass of approximately 34 kDa. The optimum temperature and pH for the action of the enzyme were at 40–50 °C and 3.5, respectively. It was stable at pH range of 5.5–7.5 and retained approximately 50% of its maximum activity after incubating at 90 °C for 1 h. Moreover, it could able to hydrolyze carboxymethyl cellulose sodium salt higher than insoluble cellulose substrate such as Avicel, SIGMACELL^® and CM cellulose. Due to its action at acidic pH and moderately stable at high temperature, the gene encoding carboxymethyl cellulase (CSCMCase) was isolated and improved the enzyme yield by high cell-density fermentation of Pichia pastoris. The CSCMCase cDNA contains 1023 nucleotides and encodes a 341-amino acid. It was successfully expressed under the control of alcohol oxidase I promoter using methanol induction of P. pastoris fermentation in a 2L ABLE bioreactor. The production of the recombinant carboxymethyl cellulases was higher than that from Cryptococcus sp. S-2 of 657-fold (2.75 and 4.2 × 10⁻³ mg protein L⁻¹, respectively) indicating that the leader sequence of CSCMCase has been recognized and processed as efficiently by P. pastoris. Furthermore, the recombinant enzyme was purified in two-step of ultrafiltration and hydrophobic interaction chromatography which would be much more convenient for large-scale purification for successful industrial application.     

血栓通对急性脑梗塞患者血清D-二聚体和超敏C反应蛋白的作用

目的:观察血栓通对急性脑梗死(actue cerebral infarction,ACI)患者血清D-二聚体和超敏C反应蛋白(high sensitivity C-reactive protein,hs-CRP)水平的影响。方法:将68例ACI患者随机分为观察组和对照组。两组进行常规治疗,观察组加用血栓通0.5 g/d,静滴。于治疗前、治疗后3 d、7 d和14 d检测患者血清D-二聚体和hs-CRP浓度变化,比较两组治疗前后改良爱丁堡-斯堪的纳维亚神经功能评分(MESSS)及日常生活能力Barthel指数(BI)的变化。结果:观察组在治疗后3 d、7 d和14 d,血清D-二聚体和hs-CRP浓度较对照组明显降低(P0.05),MESSS和BI评分在治疗后7 d和14 d较对照组明显改善(P0.05)。观察组的显效率和总有效率高于对照组(P0.05)。结论:血栓通可降低ACI患者血清D-二聚体和hs-CRP水平,改善神经功能,是一种有效治疗急性脑梗塞的方法。     

巨细胞病毒中性红斑定量试验与高滴度病毒的制备

在巨细胞病毒(CMV)的研究中常需对病毒定量。CMV需低滴度传代,否则会产生没有感染性的缺损病毒颗粒;CMV的抗原性受其感染量的影响;检测CMV中和抗体或纯化病毒都需具备病毒空斑定量基础。另外,制备高感染滴度的无细胞病毒(游离病毒)是对CMV进行分子生物学研究的前提。本文建立了CMV微量板法中性红斑定量技术并比较了几种制备无细胞CMV的方法。     

Localization and characterization of neuropeptide Y-like peptides in the brain and islet organ of the anglerfish (Lophius americanus)

Summary Results from a previous report demonstrate that more than one molecular form of neuropeptide Y-like peptide may be present in the islet organ of the anglerfish (Lophius americanus). Most of the neuropeptide Y-like immunoreactive material was anglerfish peptide YG, which is expressed in a subset of islet cells, whereas an additional neuropeptide Y-like peptide(s) was localized in islet nerves. To learn more about the neuropeptide Y-like peptides in islet nerves, we have employed immunohistochemical and biochemical methods to compare peptides found in anglerfish islets and brain. Using antisera that selectively react with either mammalian forms of neuropeptide Y or with anglerfish peptide YG, subsets of neurons were found in the brain that labelled with only one or the other of the antisera. In separate sections, other neurons that were labelled with either antiserum exhibited similar morphologies. Peptides from brains and islets were subjected to gel filtration and reverse-phase high performance liquid chromatography. Radioimmunoassays employing either the neuropeptide Y or peptide YG antisera were used to examine chromatographic eluates. Immunoreactive peptides having retention times of human neuropeptide Y and porcine neuropeptide Y were identified in extracts of both brain and islets. This indicates that peptides structurally similar to both of these peptides from the neuropeptide Y-pancreatic polypeptide family are expressed in neurons of anglerfish brain and nerve fibers of anglerfish islets. The predominant form of neuropeptide Y-like peptide in islets was anglerfish peptide YG. Neuropeptide Y-immunoreactive peptides from islet extracts that had chromatographic retention times identical to human neuropeptide Y and porcine neuropeptide Y were present in much smaller quantities. These results are consistent with the hypothesis that peptides having significant sequence homology with human neuropeptide Y and porcine neuropeptide Y are present in the nerve fibers that permeate the islet.