Synthesis of lipophilic dimeric C-7/C-7-linked ciprofloxacin and C-6/C-6-linked levofloxacin derivatives. Versatile in vitro biological evaluations of monomeric and dimeric fluoroquinolone derivatives as potential antitumor, antibacterial or antimycobacterial agents

The synthesis of C-7/C-7-linked ciprofloxacin (CP) and C-6/C-6-linked levofloxacin (LV) derivatives with modulated lipophilicity is described herein. The synthesized compounds, along with the monomeric analogs described previously, were evaluated in vitro for (i) their growth inhibitory effect against five human cancer cell lines, (ii) their antibacterial activity against Gram-positive Staphylococcus aureus and Enterococcus hirae and Gram-negative Escherichia coli and Pseudomonas aeruginosa strains and (iii) their antimycobacterial activity. The most efficient derivatives as antiproliferative agents (C-7/C-7-linked CP 7e and C-6/C-6-linked LV 11f) displayed IC₅₀ values in the 0.1–8.7 and 0.2–0.7 μM ranges respectively while IC₅₀ values for parent CP and LV ranged from 89 to 476 μM and from 67 to 622 μM respectively depending on the cell line. A specific antibacterial activity against S. aureus was found for the monomeric and dimeric derivatives of CP. The most efficient derivative against S. aureus (monomeric oxoethyloctanoate CP derivative 3d) displayed MIC <1 nM. Monomeric alkanoyloxymethyl LV esters (9a,c,e,f) and C-6/C-6-linked LV derivatives (11f–h) were the most efficient derivatives as antimycobacterial agents with MIC and IC₅₀ values in the 2.5–5 μM and 1.3–≤2.5 μM ranges respectively while MIC and IC₅₀ values for parent LV were 2.5 and 0.8 μM, respectively.     

TFPR1 acts as an immune regulator and an efficient adjuvant for proteins and peptides by activating immune cells,primarily through TLR2

Triflin, a non-toxic protein found in the venom of the Habu snake, belongs to the CRISP (cysteine-rich secretory protein) family, which comprises two domains: a C-terminal cysteine-rich domain (CRD) and an N-terminal pathogenesis-related-1 (PR-1) domain. The function of the highly structurally conserved PR-1 domain is unknown. Here, we successfully expressed the PR-1 domain of triflin (hereafter called TFPR1) in E. coli. Animal experiments showed that TFPR1 augmented Th1-biased antibody- and cell-mediated immune responses in mice immunized with two protein antigens (OVA and HBsAg) or a peptide antigen (HIV-1 pep). A flow cytometry-based binding assay and in vitro stimulation with TFPR1 showed that it triggered Th1-biased proinflammatory and immunoregulatory cytokine secretion primarily by binding to B cells and macrophages within the mouse splenocyte population. Quantitative RT-PCR, antibody blocking assays using a specific anti-mTLR2 antibody, and stimulatory experiments in vitro using splenocytes from TLR2-KO mice demonstrated that TFPR1 activated murine immune cells, primarily by stimulating toll-like receptor 2 (TLR2). These results suggest that TFPR1 acts as a novel immune modulator and potent adjuvant primarily by activating TLR2. Thus, the PR-1-based core domain might play a role in immune regulation.     

Ang-1、Ang-2、MVD与复发性流产的相关性研究

目的:通过观察复发性流产(RSA)患者绒毛、蜕膜组织中血管生成素-1(Ang-1)、血管生成素-2(Ang-2)的表达及微血管密度(MVD),探讨Ang-1、Ang-2与血管生成及RSA的关系。方法:选择2013年5~11月在山西大医院妇产科门诊行流产清宫术的RSA患者30例(RSA组),既往有正常胎儿分娩史、自愿行人工流产的正常早孕妇女30例(正常组)。在人工流产的同时留取绒毛及蜕膜组织,经免疫组化SP法检测Ang-1、Ang-2、CD34在两组绒毛及蜕膜组织中的表达水平,计数抗-CD34单抗标染的MVD,分析它们与RSA的关系。结果:(1)RSA组绒毛组织中Ang-1、Ang-2的表达及Ang-1/Ang-2比值均高于正常组(P0.05);蜕膜组织中Ang-1、Ang-2的表达均低于正常组(P0.05)。两组蜕膜中Ang-1/Ang-2比值差异无统计学意义(P0.05);(2)RSA组中,绒毛Ang-1、Ang-2表达均高于蜕膜,Ang-1/Ang-2比值低于蜕膜(P0.05);正常组中,绒毛中Ang-1表达低于蜕膜(P0.001),Ang-2在绒毛、蜕膜组织中表达无统计学差异,绒毛Ang-1/Ang-2比值低于蜕膜(P0.05)。(3)RSA组绒毛组织及蜕膜组织中MVD值低于正常组(P0.05)。结论:Ang-1、Ang-2的过度表达可能是导致血管生成缺陷及自然流产的原因之一,Ang-1过度表达对血管生成缺陷的影响可能更大。     

Antifertility, spermicidal and ultrastructural effects of gossypol and derivatives administered orally and by intratesticular injections

Because there are problems, in men, associated with the use of gossypol related to reversibility and, infrequently, hypokalemia, several laboratories around the world have resorted to the synthesis and evaluation of experimental analogs and optical isomers of gossypol in an attempt to find a compound which retains its pharmacologically desirable antifertility effects while eliminating its suboptimal ones. The present study documents: (a) the effects of fourteen new, orally-administered synthetic analogs of gossypol on testicular ultrastructure and fertility in hamsters, (b) the in vitro effects of these compounds as well as of the optical isomers of gossypol against hamster and human sperm, and (c) the morphological and antifertility effects of intratesticular injections of gossypol-PVP and its optimal isomers in the rat. The results of the study demonstrate that these new analogs are not effective as male antifertility agents and that their in vitro activity is unrelated to their in vivo contraceptive potential. In addition, this report establishes the validity of the intratesticular injection model for the analysis of the mechanism of action of gossypol and its analogs by making these compounds directly available at the testicular site. The significance of these findings is discussed.     

尿激酶溶栓治疗ST段抬高型心肌梗死的时间窗与疗效的相关性研究

目的：研究尿激酶溶栓治疗ST段抬高型心肌梗死的时间窗与疗效的相关性。方法：将医院急诊收治的60例ST段抬高心肌梗死患者根据接受溶栓治疗的时间分为观察组（3 h内接受溶栓）和对照组（3-6 h接受溶栓）,检测心肌细胞坏死指标及心室舒缩功能指标,并分析与溶栓时间的相关性。结果：观察组患者的cTnT、LDH及CK-MB水平均低于对照组,与溶栓时间呈正相关,E峰、E/A值、LVEF及LVEDD均高于对照组,且与溶栓时间呈负相关。结论：心肌梗死发生后溶栓治疗时间越短,治疗后心肌细胞坏死越少、心室功能恢复越好,临床价值也越明显。     

Synthesis, antiarrhythmic, and antihypertensive effects of novel 1-substituted pyrrolidin-2-one and pyrrolidine derivatives with adrenolytic activity

A series of 1-substituted pyrrolidin-2-one and pyrrolidine derivatives were synthesised and tested for electrocardiographic, antiarrhythmic, and antihypertensive activity as well as for alpha(1)- and alpha(2)-adrenoceptors binding affinities. Among the newly synthesised derivatives several compounds with 3-(4-arylpiperazin-1-yl)propyl moiety displayed strong antiarrhythmic (7a-12a) and antihypertensive (7a-11a) activities. Compound 11a, 1-[2-acetoxy-3-[4-(2-methoxyphenyl)piperazin-1-yl]propyl]pyrrolidin-2-one, was the most potent in this series. The pharmacological results and binding studies suggest that their antiarrhythmic and hypotensive effects may be related to their alpha-adrenolytic properties, and that those properties depend on the presence of the 1-phenylpiperazine moiety with a methoxy- or chloro- substituent in the ortho position in the phenyl ring.     

Excretion of purine derivatives by ruminants: endogenous excretion, differences between cattle and sheep

The endogenous urinary excretion of the purine derivatives allantoin, uric acid and xanthine plus hypoxanthine were measured in twenty-nine lambs, ten cattle (six steers, one cow and three preruminant calves) and four pigs. The sheep and mature cattle were nourished by intragastric infusion and the calves were given a milk-substitute. The pigs were fed on a purine-free diet. The excretion of total purine derivatives was substantially greater by the cattle, being 514 (SE 20.6) mumol/kg live weight (W)0.75 per d compared with 168 (SE 5.0) mumol/kg W0.75 per d by the sheep and 166 (SE 2.6) mumol/kg W0.75 per d by the pigs. Plasma from normally fed sheep, cows and pigs was incubated with either xanthine or uric acid. Sheep and pig plasma had no xanthine oxidase (EC 1.2.3.2) activity whereas plasma from cattle did. Uricase (EC 1.7.3.3) was not present in plasma of cattle and pigs and appeared to be present in trace amounts only in sheep plasma. It is suggested that the species differences in endogenous purine derivative excretion were probably due to the different profiles of xanthine oxidase activity in tissues and particularly in the blood. This is because a high xanthine oxidase activity would reduce the chance to recycle purines, by increasing the probability of degradation to compounds which could not be salvaged.     

Anti-EGFR activation, anti-proliferative and pro-apoptotic effects of polyclonal antibodies induced by EGFR-based cancer vaccine

Up to now clinical experiences focusing EGF receptor, an attractive target for cancer therapy, have been limited to passive therapies, suggesting that therapeutic cancer vaccines inducing anti-epidermal growth factor receptor (EGFR) antibodies could also work. Here, the humoral immune response induced in mice with a vaccine formulation containing the human EGFR-extracellular domain and very small-sized proteoliposomes (VSSP), a novel nanoparticulated adjuvant was assessed. In vaccinated mice sera average of the specific polyclonal antibodies (PAb) titers was 10(-5). Anti-EGFR PAb were able to bind EGFR+ tumor cell lines, expressing different levels of the molecule. Noteworthy, the presence of Cetuximab only partially inhibited the vaccine-induced antibodies binding to H125 cells. Anti-EGFR PAb abrogated ligands-dependent EGFR phosphorylation, provoking tumor cells apoptosis. The described EGFR-based vaccine might be a superior therapeutic approach for patients with EGFR+ tumors.     

Toll样受体-2和Toll样受体-4、单核细胞趋化蛋白-1与绒毛膜羊膜炎的相关性研究

目的考察Toll样受体-2(Toll likereceptor-2,TLR-2)和Toll样受体-4(Toll likereceptor-4,TLR-4)、单核细胞趋化蛋白-1(Monocyte chemoattractant protein,MCP-1)在绒毛膜羊膜炎的表达及其意义。方法随机选取2016年1月-2018年1月海南医学院第二附属医院收治的绒毛膜羊膜炎早产患者45例,无绒毛膜羊膜炎早产患者45例,绒毛膜羊膜炎足月患者45例,无绒毛膜羊膜炎足月孕妇40例。采用定量双抗体夹心酶联免疫吸附(Enzyme-linked immunosorbent assay,ELISA)法测定宫颈分泌物MCP-1水平,免疫组织化学法测定羊膜组织TLR-2和TLR-4蛋白表达情况,RT-PCR法测定羊膜组织TLR-2和TLR-4 mRNA表达情况。结果 TLR-2和TLR-4表达于羊膜上皮细胞、绒毛膜细胞和蜕膜细胞中;早产绒毛膜羊膜炎组MCP-1和TLR-2高于早产无绒毛膜羊膜炎组(P<0.05),足月绒毛膜羊膜炎组MCP-1和TLR-2高于足月无绒毛膜羊膜炎组(P<0.05);早产绒毛膜羊膜炎组MCP-1 mRNA和TLR-2 mRNA高于早产无绒毛膜羊膜炎组(P<0.05);足月绒毛膜羊膜炎组MCP-1 mRNA和TLR-2 mRNA高于足月无绒毛膜羊膜炎组(P<0.05);TLR-2 mRNA和MCP-1 mRNA呈正相关(r=0.473,P<0.05)。结论 TLR-2和MCP-1参与了绒毛膜羊膜炎的发生、发展,病原微生物可能通过激活TLR-2,诱导MCP-1的产生。     

感染和炎症反应与冠心病的相关性研究

目的：探讨肺炎衣原体(Cp)、幽门螺杆菌(Hp)和巨细胞病毒(CMV)感染与全身炎症反应和冠心病的关系。方法：将疑有冠心病行冠脉造影检查者根据造影结果分为冠心病组(45例)和对照组(33例)。测定血清学Cp、Hp和CMV抗体及C反应蛋白(CRP)水平。结果：(1)Cp IgG、Hp IgG和IgA阳性率均与冠心病相关(P＝0．017、P＜0．001、P＝0．009)。去除冠心病传统危险因子影响后，仍保持相关性；(2)冠心病组血清CRP较对照组明显升高(P＜0．001)，经多因素分析后仍有统计学意义(P＝0．03)；(3)Cp IgG、HpIgG和CMV IgG阳性率与高基础CRP水平无关。三种病原体IgG阳性组和两种或两种以下病原体IgG阳性组高CRP比率分别为32．1％和14．9％，但经统计学分析无意义(P＝0．078)。结论：Cp和Hp慢性感染与冠心病相关，而CMV感染与冠心病无关。尚缺乏证据证明冠心病患者全身炎症反应的增强与Cp和Hp等感染之间的相关性。     

乳腺导管内原位癌的MRI特征与HER-2表达的相关性分析

目的 探讨乳腺导管内原位癌（DCIS）的MRI特征与HER-2表达的相关性。方法 回顾性分析2018年9月—2021年6月在德州市第二人民医院经手术病理证实的53例乳腺DCIS患者分为HER-2阳性组29例,HER-2阴性组24例,比较2组间MRI特征的差异。结果 2组间内部强化特征、微血管征、TIC类型差异有统计学意义（P < 0.05）;病灶形态学、非肿块样强化分布形式、ADC值差异无统计学意义（P > 0.05）。HER-2阳性组以集群状强化（65.5%）为主,TIC类型以Ⅱ型（48.1%）、Ⅲ型（29.6%）为主,微血管征（89.7%）;HER-2阴性组以簇环状强化（50.0%）为主,TIC类型以Ⅱ型（45.8%）、Ⅰ型（54.2%）为主,微血管征（54.2%）;集群状强化,微血管征、TIC Ⅲ型3个参数同时具备诊断HER-2阳性的特异度及阳性预测值均为100%。结论 MRI参数中集群状强化、微血管征及TIC Ⅱ型或Ⅲ型曲线在较大程度上能反映 乳腺DCIS中HER-2表达情况,三者联合应用可提高HER-2阳性的诊断效能。     

大肠癌组织中生长抑素与cyclins、CDKs表达的相关性研究

目的 研究大肠癌组织中生长抑素（SS）的表达与细胞周期调控因子——细胞周期素（cyclins）、细胞周期依赖性蛋白激酶（CDKs）表达的相关性，了解SS对大肠癌细胞增殖的具体调控机制，从而进一步完善大肠癌的发病机制，为激素依赖性大肠癌的内分泌联合基因治疗提供理论依据。方法随机选择79例大肠癌（结肠癌、直肠癌）患者的手术切除标本，用免疫组织化学方法中的SP法检测SS、cyclinD1、cyclinE、cyclinA、cyclinB1、cdc2、CDK2、CDK4的表达情况。结果CyclinE、CDK2在大肠癌组织SS高表达组的表达低于低表达组（P〈0．05）。CyclinD1、cyclinA、cyclinB1、cdc2、CDK4在大肠癌组织SS高、中、低表达组三组间相比较差异无统计学意义（P〉0．05）。结论在大肠癌组织中，SS表达的水平越高，cyclinE、CDK2的阳性表达率越低，因此，SS对大肠癌细胞周期的调控可能是：SS抑制大肠癌细胞cyclinE、CDK2基因的表达，使cyclinE-CDK2复合物的水平降低，进而抑制细胞从G1期到S期的转化，使进入S期的细胞数减少，以诱导细胞周期阻滞，抑制细胞增殖。     

大肠癌组织中生长抑素与cyclins、CDKs表达的相关性研究

目的研究大肠癌组织中生长抑素(SS)的表达与细胞周期调控因子———细胞周期素(cyc lins)、细胞周期依赖性蛋白激酶(CDKs)表达的相关性,了解SS对大肠癌细胞增殖的具体调控机制,从而进一步完善大肠癌的发病机制,为激素依赖性大肠癌的内分泌联合基因治疗提供理论依据。方法随机选择79例大肠癌(结肠癌、直肠癌)患者的手术切除标本,用免疫组织化学方法中的SP法检测SS、cyc linD1、cyc linE、cyc linA、cyc linB1、cdc2、CDK2、CDK4的表达情况。结果Cyc linE、CDK2在大肠癌组织SS高表达组的表达低于低表达组(P<0.05)。Cyc linD1、cyc linA、cy-c linB1、cdc2、CDK4在大肠癌组织SS高、中、低表达组三组间相比较差异无统计学意义(P>0.05)。结论在大肠癌组织中,SS表达的水平越高,cyc linE、CDK2的阳性表达率越低,因此,SS对大肠癌细胞周期的调控可能是:SS抑制大肠癌细胞cyc linE、CDK2基因的表达,使cyc linE-CDK2复合物的水平降低,进而抑制细胞从G1期到S期的转化,使进入S期的细胞数减少,以诱导细胞周期阻滞,抑制细胞增殖。     

Examination of the structure of an unknown green fluorescent compound, compound G2, accumulated in non-growing cells of Eremothecium ashbyii by the addition of dimeric diacetyl

The addition of dimeric diacetyl to Eremothecium ashbyii caused simultaneous accumulation of two green fluorescent compounds with the inhibition of riboflavin formation in non-growing cells. One compound, referred to as Compound G1, was identified as 6, 7-dimethyl-8-ribityllumazine as reported previously and the other is referred to as Compound G2. The latter compound was considered to be 6-methyl-7-(2-hydroxy-2-methyl-3-oxobutyl)-8-ribityllumazine because a violet fluorescent compound, 6-methyl-7-hydroxy-8-ribityllumazine was derived from Compound G2 in the presence of p-quinone and because the known action mechanism of dimeric diacetyl as a trapping agent of possible intermediates in the biosynthetic pathway. The results indicate that an immediate intermediate to 6, 7-dimethyl-8-ribityllumazine in the biosynthetic pathway of riboflavin is 4-ribitylamino-5-amino-2, 6-dihydroxypyrimidine, whose pyrimidine portion is derived by the elimination of a dimeric diacetyl fragment from the molecule of the isolated lumazine.     

静态平衡仪与儿童运动评估成套工具评定儿童平衡功能的相关性分析

目的 分析Tetrax静态平衡仪与儿童运动评估成套工具(MABC-2)在评定7~10岁儿童平衡功能中的相关性,为临床上儿童平衡评估方法的合理运用提供客观依据。方法 2015年1月-2017年5月共选取61例7~10岁门诊儿童,由专人采用Tetrax静态平衡仪和儿童运动评估量表对每位受试儿童平衡功能进行评定,并将每位儿童静态平衡仪测试结果和MABC-2量表中平衡项目评分结果进行Pearson相关性分析。结果 MABC-2量表动静态平衡项目总分结果与Tetrax静态平衡仪检测结果间具有显著相关性(r=0.50,P<0.01),其中MABC-2量表平衡项目中的静态平衡项目得分与Tetrax平衡仪垫上睁眼站立评分具有高度相关性(r=0.73,P<0.01);MABC-2量表静态平衡项目得分与Tetrax平衡仪睁眼站立评分具有中度相关性(r=0.36,P<0.01)。结论 MABC-2中平衡测试与静态平衡仪在评定儿童平衡功能方面具有一致性;MABC-2量表法简单易行可用于筛查儿童整体的平衡功能,而静态平衡仪可量化、客观地评定儿童在站立位静态平衡状态并分析其平衡机制,两者可相互补充,在临床应用时需综合运用从而能全面评定儿童的平衡能力。     

宫颈分泌物CT、UU、Mh检测与不孕症的相关性研究

目的研究宫颈分泌物沙眼衣原体(CT)、解脲脲原体(UU)及人型支原体(Mh)检测结果与不孕症间的相关性。方法将2017年1月-2018年1月收治的316例女性不孕症患者纳为不孕组,同期于妇科门诊要求终止妊娠的孕早期146例患者纳为早孕组,94例输卵管性不孕患者纳为输卵管性不孕组,103例腹腔镜手术者纳为手术组。比较不孕组与早孕组宫颈分泌物CT、UU及Mh培养阳性率;比较输卵管不孕组与手术组宫颈分泌物及盆腔积液CT、UU及Mh培养阳性率。根据患者输卵管功能将输卵管性不孕者分为低分组(24例)、中分组(26例)与高分组(44例),观察3组宫颈分泌物及盆腔积液CT、UU及Mh培养阳性率。结果不孕组CT培养阳性率为12. 97%,支原体感染率为41. 14%,均显著高于早孕组(均P<0. 05);输卵管性不孕组宫颈分泌物CT阳性率为11. 70%,支原体感染率为36. 17%,盆腔积液CT感染率为5. 32%,均显著高于手术组(均P<0. 05);低分组、中分组及高分组输卵管性不孕患者宫颈分泌物CT培养阳性率及支原体感染率依次升高,组间差异均有统计学意义(均P<0. 05)。结论女性不孕症及输卵管性不孕患者宫颈分泌物CT、UU及Mh感染率均明显高于正常水平,且患者输卵管功能越差,宫颈分泌物CT、UU及Mh感染率越高,提示CT、UU及Mh感染与不孕间关系密切,可能是诱发并加重输卵管障碍的原因之一。     

胃癌患者血清miR-204、HSP-70及IL-2与Hp感染及病情关系研究

目的探讨胃癌患者血清中微小RNA-204(MicroRNA-204,miR-204)、热休克蛋白-70(Heat shock protein-70,HSP-70)及白细胞介素-2(Interleukin-2,IL-2)的变化与幽门螺杆菌(Helicobacter pylori,Hp)感染、TNM分期及淋巴结转移的关系。方法选取2015年1月-2018年10月郑州大学附属肿瘤医院经病理学或胃镜确诊的初诊胃癌患者96例作为胃癌组、胃炎患者96例作为对照组,检测对比两组的血清miR-204、HSP-70及IL-2与Hp感染水平,并按照Hp感染结果、TNM分期、淋巴结转移结果将胃癌组进行分层分析。结果胃癌组的血清HSP-70为(2.33±0.76)pg/ml高于对照组(1.16±0.40)pg/ml(P<0.05);胃癌组的血清miR-204(0.462±0.117)、IL-2(2.26±0.78)ng/ml低于对照组miR-204(1.330±0.205)、IL-2(3.95±0.92)ng/ml(P<0.05);Ⅰ期和Ⅱ期胃癌患者的血清HSP-70水平低于Ⅲ期和Ⅳ期,血清miR-204、IL-2水平高于Ⅲ期和Ⅳ期(P<0.05);Hp感染阳性胃癌患者的血清HSP-70水平高于阴性患者,血清miR-204、IL-2水平低于阴性患者(P<0.05)。结论胃癌患者血清HSP-70、miR-204、IL-2水平异常表达与TNM分期、淋巴结转移及Hp感染有一定的关系。