Eukaryotic food sources analysis in situ of tropical common sea cucumber Holothuria leucospilota based on 18S rRNA gene high-throughput sequencing

Sea cucumber Holothuria leucospilota is one of the most widespread tropical holothurian species.In this study,eukaryotic organism composition in foregut and hindgut contents of H.leucospilota and surrounding sediments was assessed by 18S rRNA gene high-throughput sequencing.Eukaryon richness and diversity in the habitat sediments were significantly higher than those in foregut and hindgut contents of the sea cucumbers(P<0.05).The foregut content group,hindgut content group,and marine sediment group sequences were respectively assigned to 18.20±1.32,19.40±1.03,and 21.80±0.37 phyla.In the foregut contents,Nematoda(20.18%±9.59%),Mollusca(16.12%±10.49%),Chlorophyta(10.04%±4.85%),Annelida(8.72%±10.93%),Streptophyta(8.46%±4.65%),and Diatomea(5.99%±2.01%)were the predominant phyla,which showed the eukaryotic food sources of H.leucospilota were primarily belong to the above phyla.The predominant phyla in the hindgut contents were Streptophyta(45.55%±17.32%),Mollusca(4.93%±4.82%),Arthropoda(5.37%±3.08%),Diatomea(3.88%±2.34%),and Chlorophyta(3.79%±1.59%);and Annelida(37.80%±17.00%),Arthropoda(24.49%±12.53%),Platyhelminthes(7.14%±3.02%),Nematoda(4.14%±0.91%),and Diatomea(5.11%±1.35%)had large contents in the sediments.The comparatively high content of Paris genus in phylum Streptophyta in foregut contents indicated that land plants were one of the primary food sources of H.leucospilota,however the significantly higher contents of Streptophyta in hindgut contents than that in foregut contents might suggest a large part of the terrigenous detritus ingested might not be digested by H.leucospilota.UPGMA and PCoA analysis revealed that eukaryotic organism composition differed significantly between foregut contents of H.leucospilota and ambient sediments,indicating selective feeding feature of H.leucospilota.This study provided useful references for artificial feed of tropical sea cucumbers and enhanced understanding of the ecological roles of detritus-feeding macrobenthos.

一体化^(18)F-FET PET/MR术前评估成人胶质瘤患者MGMT基因启动子甲基化状态

目的 旨在研究一体化^(18)F-氟乙基酪氨酸(^(18)F-fluoroethyltyrosine, ^(18)F-FET)正电子发射断层成像(positron emission tomography, PET)/MR对成人胶质瘤的O6-甲基鸟嘌呤DNA甲基转移酶(O6-methylguanine DNA methyltransferase, MGMT)基因启动子甲基化状态的鉴别能力。材料与方法 回顾性分析未经活检或治疗的胶质瘤患者资料16例,均完成一体化PET/MR扫描,包括^(18)F-FET PET、常规MRI及体素内不相干运动(intravoxel incoherent motion, IVIM)成像。以靶本比(target-background ratio, TBR)=1.6为阈值对PET图像进行感兴趣体积(volume of interest, VOI)分割,通过刚性配准获得肿瘤VOI对应的IVIM图及其参数表观扩散系数(apparent diffusion coefficient, ADC)、真实扩散系数(true diffusion coefficient, D)、伪扩散系数(pseudo diffusion coefficient, D^(*))、灌注分数(perfusion fraction, f)、分布扩散系数(distributed diffusion coefficient, DDC)和异质性指数(heterogeneity index, α),使用pyradiomics对各参数进行特征提取,获得对应的一阶灰度直方图特征。计算每个IVIM参数图对应的特征值与^(18)F-FET PET参数的关系,并利用组间比较和受试者工作特征(receive operating characteristic, ROC)曲线分析探究PET和IVIM参数对MGMT启动子甲基化的区分能力。结果 IVIM-α的两个特征值90分位值(r=0.526,P<0.05)和最大值(r=0.520, P<0.05)与PET参数平均标准摄取值(meanstandarduptakevalue,SUVmean)呈正相关;IVIM-α和SUVmean在MGMT启动子甲基化状态阳性和阴性两组之间的差异有统计学意义(P<0.05),阳性组的IVIM-α均值和SUVmean显著高于阴性组;融合IVIM-α均值和SUVmean对MGMT启动子甲基化状态进行区分的曲线下面积(area under the curve, AUC)为0.77。结论 基于一体化PET/MR的^(18)F-FET PET和IVIM参数能够有效预测胶质瘤的MGMT启动子甲基化状态。

CDH18基因对宫颈癌细胞增殖和迁移的影响

目的 研究Ⅱ型钙粘蛋白18(Cadherin 18,CDH18)在宫颈癌中的表达及其对宫颈癌SiHa、HeLa和C33A细胞增殖和迁移能力的影响。方法 采用qRT-PCR、Western blot实验检测宫颈癌组织和细胞中CDH18的mRNA和蛋白质表达水平;分别构建过表达和敲低CDH18基因表达的质粒;在稳定转染质粒的宫颈癌细胞中,利用CCK-8、Transwell和划痕实验分析过表达及降低CDH18基因对SiHa、HeLa和C33A细胞增殖和迁移的影响。结果 高危型HPV阳性的宫颈组织和细胞中CDH18蛋白质和mRNA表达水平均显著低于正常宫颈组织和细胞(P<0.01),差异具有统计学意义;CCK-8、Transwell及划痕实验结果显示:过表达CDH18基因促进SiHa、HeLa和C33A细胞的增殖和迁移能力(P<0.01);敲低CDH18基因抑制了SiHa、HeLa和C33A细胞的增殖和迁移能力(P<0.05)。结论 CDH18基因在高危型HPV阳性宫颈癌组织和细胞中低表达,在高危型HPV阴性宫颈癌组织细胞中高表达;其过表达能够促进宫颈癌细胞的增殖和迁移。

lncRNA SNHG18在膀胱癌组织中的表达及其对细胞恶性生物学行为的影响

目的研究长链非编码RNA SNHG18基因在膀胱癌组织膀胱癌细胞系中的表达情况及其对细胞恶性生物学行为的影响。方法选取2021年8月至2022年10月行手术治疗的膀胱癌患者42例,应用RT-qPCR检测SNHG18基因在42例膀胱癌组织及3株膀胱癌细胞(5637、T24、SW780)中的表达情况;构建过表达载体pcDNA3.1-SNHG18与敲低载体SNHG18分别转染膀胱癌细胞,应用细胞增殖实验、细胞克隆形成实验、划痕实验、Transwell小室侵袭实验来观察膀胱癌细胞增殖、迁移、侵袭能力的变化。结果SNHG18在膀胱癌组织及细胞中的表达低于正常的膀胱上皮组织和膀胱上皮细胞,差异有统计学意义(P<0.05);SNHG18的表达量与性别、年龄、有无吸烟、有无高血压、有无糖尿病、有无淋巴结和远处转移及临床分期间无相关性(P>0.05);过表达SNHG18可抑制膀胱癌细胞的体外增殖、迁移和侵袭能力(P<0.05);敲低SNHG18可增强膀胱癌细胞的体外增殖、迁移和侵袭能力(P<0.05)。结论SNHG18在膀胱癌组织中表达量降低,与临床参数无关,与膀胱癌细胞增殖、迁移和侵袭能力有关。

^(18)F-FDG PET/CT代谢参数联合临床病理特征对晚期结直肠癌患者KRAS基因突变的评估价值

目的探讨^(18)F-氟代脱氧葡萄糖(^(18)F-FDG)PET/CT代谢参数联合临床病理特征对晚期结直肠癌患者鼠类肉瘤病毒癌基因(KRAS)基因突变的评估价值。方法回顾性分析64例晚期结直肠癌患者的临床资料。所有患者在治疗前均接受^(18)F-FDG PET/CT检查。分析最大标准化摄取值(SUV_(max))、不同阈值下的代谢肿瘤体积(MTV)和病变总糖酵解(TLG)、临床病理特征与患者KRAS基因突变状态之间的关系。通过多因素Logistic回归模型分析与患者KRAS基因突变相关的因素。采用受试者工作特征曲线分析^(18)F-FDG PET/CT代谢参数、临床病理特征及二者联合评估患者KRAS基因突变的效能。基于^(18)F-FDG PET/CT代谢参数和临床病理特征构建列线图模型。结果单因素和多因素Logistic回归分析结果显示,SUV_(max)≥19.55、MTV50%≥7.95、肿瘤组织中/高分化与患者KRAS基因突变有关。SUV_(max)、MTV50%和肿瘤组织分化程度评估患者KRAS基因突变的曲线下面积(AUC)分别为0.653、0.625和0.621,三者联合评估的AUC为0.800,均高于单个指标的AUC(P<0.05)。基于SUV_(max)、MTV50%和肿瘤组织分化程度构建的列线图模型的一致性指数为0.800,校准曲线与参考线基本拟合。结论^(18)F-FDG PET/CT的代谢参数SUV_(max)、MTV50%与晚期结直肠癌患者KRAS基因突变密切相关,联合肿瘤组织分化程度所构建的列线图模型对患者KRAS基因突变具有较高的评估价值。

Morphogenesis and Molecular Phylogeny of a Marine Urostylid Ciliate Apokeronopsis wrighti(Protista, Ciliophora, Hypotrichia)

Hypotrichs are one of the highly differentiated ciliated lineages which play important roles in ecological, environmental,evolutionary and basic biological studies. In the present study, we investigated the living characteristics, infraciliature, nuclear apparatus, ontogenesis and phylogenetic position of a marine hypotrichous ciliate, Apokeronopsis wrighti Long et al., 2008, which was isolated from coastal waters in Shenzhen, China. The new isolate resembles the type population in terms of morphological characteristics, morphometrics, and SSU rRNA gene sequence that is with a 99.7% similarity. Ontogenesis of A. wrighti is characterized by oral primordium for the proter as well as marginal and dorsal kineties anlagen in both filial products formed de novo, and the cirral row arranged along the paroral and endoral arises from several anterior frontoventral-transverse cirral streaks. Phylogenetic analyses based on SSU and concatenated gene data suggest that five species of Apokeronopsis form a monophyletic clade, and the genus Apokeronopsis is closely related to Thigmokeronopsis and Metaurostylopsis.

18号染色体多体在恶性周围神经鞘膜瘤的诊断价值及其与临床病理特征的相关性分析

目的探讨18号染色体多体在恶性周围神经鞘膜瘤(MPNST)的诊断价值及其与临床病理特征的相关性分析。方法收集2010—2024年西京医院病理科诊断的37例MPNST,采用荧光原位杂交(FISH)技术进行检测,并总结其临床病理学特征及免疫表型。结果37例患者中男性22例,女性15例,年龄4~78(平均37)岁。肿瘤平均大小为10.8 cm。MPNST最常见的原发部位为躯干(22/37)。FISH结果示37例患者中10例(27.0%)存在18号染色体多体。统计学结果示18号染色体多体与高核分裂象(P=0.013)和肿瘤分级(P=0.005)呈正相关,且多见于头颈部(P=0.003)。结论在本研究中,FISH显示MPNST中存在18号染色体多体,这可能与肿瘤组织学形态更具侵袭性和分级更高有关,并为其与其他梭形细胞肉瘤的鉴别诊断提供了线索。

肠易激综合征患者血清白细胞介素-18、降钙素基因相关肽与症状严重程度及肠屏障功能的相关性研究

目的:分析肠易激综合征(IBS)患者血清白细胞介素-18(IL-18)、降钙素基因相关肽(CGRP)与症状严重程度及肠屏障功能的相关性。方法:选取2021年2月—2022年10月北京市肛肠医院收治的101例IBS患者作为研究对象,根据IBS症状严重程度量表(IBS-SSS)评估结果分为轻度组(76~175分,n=31)、中度组(176~300分,n=38)、重度组(>300分,n=32)。检测患者血清白细胞介素-18(IL-18)、降钙素基因相关肽(CGRP)、血清二胺氧化酶(DAO)水平。采用Spearman相关系数分析血清IL-18、CGRP水平与症状严重程度的相关性,采用Pearson相关系数分析血清IL-18、CGRP水平与肠屏障功能的相关性。结果:中度组IL-18、CGRP水平高于轻度组,重度组IL-18、CGRP水平高于中度组与轻度组,差异有统计学意义(P<0.05)。Spearman相关性分析显示,IL-18、CGRP水平与症状严重程度呈正相关(r=0.723、0.823,P<0.05)。Pearson相关性分析显示,IL-18、CGRP水平P与DAO水平呈正相关(r=0.485、0.571,P<0.001)。结论:血清IL-18、CGRP水平异常偏高是IBS疾病检查与诊断的重要依据,在评估IBS患者症状严重程度和肠屏障功能方面具有一定价值。

Immunological Adjuvant Function of Aluminium Phosphate and Chicken IL-18 in NDV F Gene Vaccine

[Objective] This study aimed to investigate the immunological adjuvant function of aluminium phosphate and chicken IL-18 in NDV F gene vaccine. [Method] The vaccine （0.2 ml） containing aluminum phosphate adjuvant （90 μg）, pcDNA/F （200μg）, and pcDNA/chlL-18 （200 μg） was prepared. The 7 d old chick- ens to be tested were randomly divided into six groups （12 chickens in each group） and immunized through intramuscular injection with inactivated Newcastle disease vaccines, pcDNA/F＋pcDNA/chlL-18＋phosphate aluminum, pcDNA/F, pcDNA/F.＋pcDNA/ chlL-18, pcDNA/F＋aluminum phosphate, and physiological saline respectively; the secondary immunization was conducted with the same dose when the chickens were 21 d old. Their blood was sampled 0, 7, 14, 21, 28 d after first immunization. Anti- body titer was detected with ELISA and T cell transformation rate was measured with MIT. Experimental chicken will be challenged with 30 LD50 NDV virulence 28 d after first immunization. [Result] The survival rate of the chickens immunized with pcDNA/F＋aluminium phosphate＋pcDNA/chlL-18 achieved 8/12, higher than that of those immunized with pcDNA/F 4/12 and pcDNA/F＋pcDNA/chlL-18 （6/12）. The NDV antibody titer of the chickens immunized with pcDNA/F＋ aluminum phosphate, pcD- NA/F＋pcDNA/chlL-18 and pcDNA/F＋pcDNA/chlL-18＋aluminum phosphate is not differ- ent （P〉0.05）, but significantly lower than that of the chickens immunized with tradi- tional vaccine （P〈0.05）. The T cell transformation rate of the chickens immunized with pcDNA/F＋pcDNA/chlL-18＋aluminium phosphate was obviously higher than that of the chickens immunized with pcDNA/F （P〈0.05）. The T cell transformation rates of chickens immunized with pcDNA/F and the traditional vaccine showed no signifi- cant difference （P〉0.05）. [Conclusion] Combination of aluminium phosphate and pcD- NA/chlL-18 can significantly enhance the immune effect of NDV F gene vaccine.

Association of polymorphisms of interleukin-18 gene promoter region with chronic hepatitis B in Chinese Han population

AIM: To investigate the polymorphisms of interleukin-18 (IL-18) gene promoters, and to disclose whether such polymorphisms are associated with susceptibility to chronic hepatitis B in Chinese Han population. METHODS: Using polymerase chain reaction with sequence specific primers (PCR-SSP) method, the single nucleotide polymorphisms (SNPs) of the promoter region of IL-18 gene at position -607 and -137 were detected in 231 patients with chronic hepatitis B and 300 normal controls. RESULTS: Allele C at position -607 in the promoter of IL-18 gene was detected in 48.7% of normal controls and 51.9% of patients, while allele A at position -607 was detected in 51.3% of normal controls and 48.1% of patients. The frequencies of -607CC, -607 CA and -607AA genotypes in normal controls were 22.0%, 53.3% and 24.7% respectively and in chronic hepatitis B patients were 26.8%, 50.2% and 23.0% respectively. Allele G at position -137 in the promoter of IL-18 gene was detected in 82.3% of normal controls and 88.5% of chronic hepatitis B patients, while allele C at position -137 was detected in 17.7% of normal controls and 11.5% of patients. The frequencies of -137GG, GC and CC genotype were 67.3%, 30.0% and 2.7% in normal controls respectively, while in chronic hepatitis B patients were 78.8%, 19.5% and 1.7% respectively. The frequency of-137GG genotype in chronic hepatitis B groups was significantly higher than that in normal controls (x2=8.55, P=0.003 <0.05), whereas the frequencies of -607C/-137C and -607A/-137C haplotypes in chronic hepatitis B groups were significantly lower than that in normal controls. The association between genotypes of IL-18 promoter region polymorphisms and HBV copies showed that the frequency of -607AA genotype in high HBV-DNA copies groups was lower than that in low HBV-DNA copies groups (x2=6.03, P=0.014 <0.05). CONCLUSION: The polymorphisms of the promoter region of IL-18 gene at position -607 and -137 are closely associated with susceptibility to chronic hepatitis B. The people with allele C at position -137 in the promoter of IL-18 gene may be protected against HBV infection; moreover AA genotype at position -607 may be closely linked to inhibit HBV-DNA replication. These findings give some new clues to the study of pathogenesis of chronic hepatitis B.

Phylogenetic relationship of Podocopida (Ostracoda: Podocopa) based on 18S ribosomal DNA sequences

Nucleotide sequences from 18S rDNA of 11 ostracodes, which represent four suborders and six superfamilies of podocopidan, were determined. The phylogenetic relationships were analyzed based on three kinds of methods (maximum-likelihood, maximum-parsimony, and neighbor-joining), and the three topologies gained were basically similar. The results have showed that (1) a monophyletic Podocopida was supported strongly; (2) the phylogenetic relationships of four suborders were (Darwinulocopina plus (Bairdiocopina plus (Cytherocopina plus Cypridocopina))), which indicated that a close relationship between Cytherocopina and Cypridocopina, and Darwinulocopina had separated early from the main podocopinan; (3) Cypridocopinan formed a monophyletic group, among which the phylogenetic relationship of three superfamilies was (Cypridoidea plus (Macrocypridoidea plus Pontocypridoidea)).

THE BIOLOGICAL CHARACTERISTICS OF ADENOVIRUS-MEDIATED IL-18 GENE-MODIFIED MURINE COLORECTAL ADENOCARCINOMA CELL IN VIVO AND IN VITRO

Objective: Interleukin 18 (IL-18) is a strong activator of NK cells and promotes the generation of IL-2, IFN-γ and GM-CSF. In the present study, we constructed adenovirus encoding IL-18 gene (AdIL-18), and observed the biological characteristics of IL-18 gene-modified murine colorectal adenocarcinoma cell (CT26)in vivo andin vitro. Methods: Gene modification was mediated by adenovirus. The proliferation of the cells was determined by MTT and IL-18 was assayed by ELISA. The cytotoxicity of NK and CTL was detected by four-hour51Cr release assay. Results: IL-18 gene modification had no effect on the proliferation and morphology of CT-26 cellsin vitro, but the growth of IL-18-modified CT26 cells was obviously inhibitedin vivo. In addition, although IL-18-modified CT26 cells could form tumor nodulesin vivo as well as LacZ-modified CT26 cells or wild-type CT26 cells, the mean survival time of the mice inoculated with IL-18-modified CT26 cells was significantly prolonged as compared with that of control groups. Thus, the anti-tumor immune responses were induced in the group of mice inoculated with IL-18-modified CT26 cells, which might be related to the activation of NK cells and CTL. However, all the three groups ultimately died of tumor. Conclusion: IL-18-modified CT26 cells could induce the anti-tumor immune responses incompletely, which required other factors for effective anti-tumor responses.

Nematode Diversity of Qingdao Coast Inferred from the 18S Ribosomal RNA Gene Sequence Analysis

The 18S ribosomal DNA gene （18S rDNA） sequences （approxtmately 1300 bp in length） were amplified from the DNA extracted from the free-living marine nematodes collected from the inter-tidal sediment of Qingdao coast in bulk with nematode specific primers. The PCR products were cloned, re-amplified, digested with Rsa I and Hin61 restriction endonucleases and separated in agarose gel. Among 17 restriction fragment length types, types 1, 2 and 6 covered 61.2%, 14.4% and 9.3% of the clones analyzed, respectively, while the remaining 14 only covered 21 clones, which accounted for 15.1% of the total. Twenty-four representative clones were sequenced and phylogenetically analyzed by referring to those currently available in RDP and GenBank databases. Although it was hard to assign these sequences to known species or genera due to the lack of the 18S rDNA sequence data of known marine free-living nematodes, the obtained sequences were assigned to the nematodes of Adenophorea. Among them, twelve sequences were close to Pontonema vulgate and Adoncholaimus sp., four to Daptonema procerus and two （identical） to Enoplus brews. Our results showed that free-living marine nematode diversities could be determined by PCR retrieving and analysis of the 18S rDNA sequences and an 18S rDNA sequence could be assigned to a species or a genus only if the 18S rDNA sequences of the free-living marine nematodes were accumulated to some extent.

Comparative evaluation of microscopy,OptiMAL® and 18S rRNA gene based multiplex PCR for detection of Plasmodium falciparum & Plasmodium vivax from field isolates of Bikaner,India

Objective:To evaluate microscopy,OptiMAL? and multiplex PCR for the identification of Plasmodium falciparum（P.falciparum） and Plasmodium vivax（P.vivax） from the field isolates of Bikaner,Rajasthan（Northwest India）.Methods:In this study,a multiplex PCR（P.falciparum and P.vivax） was further developed with the incorporation of Plasmodium malariae（P.malariae） specific primer and also a positive control.The performance of microscopy,plasmodium lactate dehydrogenase（pLDH） based malaria rapid diagnostic test OptiMAL? and 18S rRNA gene based multiplex PCR for the diagnosis of P.falciparum and P.vivax was compared.Results:The three species multiplex PCR if.falciparum,P.vivax and P.malariae) with an inbuilt positive control was developed and evaluated.In comparison with multiplex PCR,which showed the sensitivity and specificity of 99.36%（95%CI,98.11%-100.00%） and 100.00%（95%CI,100.00%-100.00%）,the sensitivity and specificity of microscopy was 90.44%（95%CI,88.849-95.04%） and 99.22%（95% CI,97.71%-100.00%）,and OptiMAL? was 93.58%（95%CI,89.75%-97.42%） and 97.69%（95%CI, 95.10%-100.00%）.The efficiencies were 99.65%,95.10%and 95.45%for multiplex PCK.microscopy and OptiMAL?.respectively.Conclusions:Our results raise concerns over the overall sensitivities of microscopy and OptiMAL?,when compared to the multiplex PCR and thus stress the need for new molecular interventions in the accurate detection of the malarial parasites.This further highlights the fact that further developments are needed to improve the performance of rapid diagnostic tests at field level.

Synthesis of Codon-optimized Human Interleukin-18 Gene by Combination of Chemical and Enzymatic Method

According to the amino acid sequence and codon preference of E. coli, the human interleukin-18（IL-18） gene was optimized to avoid the rare codons. The total length of the synthesized gene is 571 bp; 18 oligonucleotides, DNA fragments were designed and synthesized by the phosphoramidite four-step chemical method. The whole DNA sequence was synthesized by a one-step total gene synthesis method, and then inserted in pUC18 vector. Five positive clones identified by blue-white colony screening were sent to Shanghai Sangon Biological Engineering Technology and Service Co., Ltd. for sequencing. The sequencing result shows that one clone contained the complete correct gene in all the five positive clones.

IL-18结合蛋白在创伤性颅脑损伤大鼠认知功能障碍中的作用及其机制

目的探究IL-18结合蛋白(IL-18BP)在创伤性脑损伤(TBI)大鼠认知功能障碍中的作用及其机制。方法120只健康雄性SD大鼠按随机数字表法分为假手术组、TBI组、TBI+IL-18BP组(经尾静脉注射IL-18BP 1.5 mg/kg)与TBI+IL-18BP+ADU-S100组(经尾静脉注射IL-18BP1.5 mg/kg,经腹腔注射ADU-S10020 mg/kg),每组30只。应用自由落体法建立TBI模型,造模后30 d行水迷宫实验检测大鼠认知功能,ELISA法检测大鼠血清IL-18、IL-18BP浓度,免疫荧光染色检测海马区星形胶质细胞GFAP、IL-18和cleaved-caspase-3荧光强度,Western blotting检测大鼠海马区干扰素基因刺激蛋白(STING)、磷酸化TANK结合激酶1(p-TBK1)、TBK1、磷酸化干扰素调节因子3(p-IRF3)、IRF3蛋白相对表达量。结果与假手术组比较,TBI组、TBI+IL-18BP组和TBI+IL-18BP+ADU-S100组大鼠逃逸潜伏期明显延长,穿越平台次数减少,目标象限活动时间百分比降低(P<0.0001),血清IL-18、IL-18BP浓度升高,海马区IL-18和cleaved-caspase-3荧光强度明显增强,STING、p-TBK1、p-IRF3表达明显上调(P<0.05);与TBI组比较,TBI+IL-18BP组大鼠逃逸潜伏期缩短,穿越平台次数增多,目标象限活动时间百分比增高(P<0.0001),血清IL-18浓度降低,IL-18BP浓度升高,海马区IL-18和cleaved-caspase-3荧光强度明显减弱,STING、p-TBK1、p-IRF3表达明显下调(P<0.05);与TBI+IL-18BP组比较,TBI+IL-18BP+ADU-S100组大鼠逃逸潜伏期延长,穿越平台次数减少,目标象限活动时间百分比降低(P<0.0001),血清IL-18浓度升高,IL-18BP浓度降低,海马区IL-18和cleaved-caspase-3荧光强度增强,STING、p-TBK1、p-IRF3表达明显上调(P<0.05)。结论IL-18BP可改善TBI大鼠的认知功能,其机制可能与抑制星形胶质细胞凋亡及抑制STING/TBK1/IRF3信号通路激活有关。

Construction and Identification of a Yeast Two-Hybrid Bait Vector and Its Effect on the Growth of Yeast Cells and the Self-Activating Function of Reporter Genes for Screening of HPV18 E6-Interacting Protein

By using a yeast two-hybrid system,a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins,and its effects on the growth of yeast cells and the activation of reporter genes were investigated.Total mRNA extracted from Hela cells was reversely transcribed into cDNA.Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector.The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing.Th...

Intratracheal administration of recombinant adenovirus containing IL-18 gene in treatment of experimental metastatic lung carcinoma

Objective: To study the treatment of experimental metastatic lung carcinoma by intratracheal injection of IL-l8 gene recombinant adenovirus. Methods: (1)The mouse IL-18 mRNA was detected by RT-PCR, and the concentration of IL-18 and associated cytokines in lung lavages and blood were determined by ELISA at different time points after intratracheal injection of IL-18 recombinant adenovirus. (2)The lung metastasis nodes, mouse survival periods and survival rates were evaluated. NK activity and CTL activity were determined by 51Cr 4 h release method. Results: (1) IL-18 mRNA was detectable in lung tissue 6 h after intratracheal use of IL-18 recombinant adenovirus. and the concentration of IL-18 in lung lavage was higher than that in peripheral blood. Neither IL-18 mRNA nor IL-18 was detectable in control group. (2) Intratracheal use of IL-18 recombinant adenovirus resulted in increased CTL and NK activity, longer survival time and higher survival rates compared with the control group, showing significant therapeutic effect on expermental lung metastasis. Conclusion: Intratracheal use of adenovirus vector containing IL- 18 gene has therapeutic effect on the lung metastasis, denoting that gene therapy of lung diseases could be applied through airway directly with recombinant adenovirus.

乳腺癌组织中泛素特异性蛋白酶18/干扰素刺激基因15表达水平及其临床意义

目的探究泛素特异性蛋白酶18(USP18)、干扰素刺激基因15(ISG15)在乳腺癌组织中的表达水平及临床意义。方法采用Ualcan数据库分析USP18、ISG15在正常乳腺组织和乳腺癌组织中的表达情况及二者与乳腺癌预后的关系;选取渭南市中心医院2012年5月至2015年7月诊治的99例乳腺癌病人癌组织、癌旁正常组织进行研究。分别检测USP18、ISG15mRNA及其蛋白表达情况;分析乳腺癌组织USP18、ISG15表达水平与病人临床病理特征、预后的关系;Cox回归分析乳腺癌病人预后的影响因素。结果Ualcan数据库中乳腺癌组织USP18为18.40±4.17、ISG15 mRNA表达水平为168.56±43.95高于正常乳腺组织(10.00±3.14、30.76±8.23)(P<0.05);乳腺癌组织USP18、ISG15 mRNA表达水平高低与乳腺癌预后无明显相关性。乳腺癌组织USP18为1.67±0.53、ISG15 mRNA为1.86±0.61及蛋白阳性表达率均高于癌旁正常组织(1.03±0.34、0.99±0.33)(P<0.05);乳腺癌组织USP18、ISG15表达水平均与淋巴结转移、乳腺癌分子亚型、肿瘤分化程度、TNM分期相关(P<0.05);乳腺癌病人癌组织中USP18表达水平与ISG15呈正相关(P<0.05);USP18阳性组、ISG15阳性组乳腺癌病人术后60个月生存率均低于USP18阴性组、ISG15阴性组(P<0.05);淋巴结转移、TNM分期、USP18、ISG15均是影响乳腺癌病人不良预后的独立危险因素(P<0.05)。结论乳腺癌病人癌组织USP18、ISG15表达水平均较高,两者均与乳腺癌病人预后关系密切,检测乳腺癌组织USP18、ISG15水平有助于评估乳腺癌病人预后。

Correlation of RCAS1 and Claudin-18 expression with proliferation and invasion gene expression in malignant pleural effusion

Objective: To investigate the correlation of RCAS1 and Claudin-18 expression with proliferation and invasion gene expression in malignant pleural effusion. Methods: A total of 187 patients with primary non-small cell lung cancer complicated by malignant pleural effusion were selected as malignant pleural effusion group and 56 patients with tuberculous pleural effusion were selected as tuberculous pleural effusion group. The expression of RCAS1 and Claudin-18 gene as well as proliferation and invasion-related genes in the pleural effusion were compared between the two groups, and Pearson test was used to evaluate the correlation of RCAS1 and Claudin-18 expression with proliferation and invasion gene expression in malignant pleural effusion. Results: RCAS1 and Claudin-18 mRNA expression in pleural effusion of malignant pleural effusion group were greatly higher than those of tuberculous pleural effusion group. Proliferation genes LRRC3B and TCF21 mRNA expression in pleural effusion of malignant pleural effusion group were lower than those of tuberculous pleural effusion group whereas SIRT1 and EZH2 mRNA expression were higher than those of tuberculous pleural effusion group;invasion genes DDX17, Nectin4, Vav3, NGAL and Snail mRNA expression were higher than those of tuberculous pleural effusion group whereas EFEMP1 and MCPH1 mRNA expression were lower than those of tuberculous pleural effusion group. The Pearson test showed that the RCAS1 and Claudin-18 expression in malignant pleural effusion were directly correlated with the expression of proliferation-related genes and invasion-related genes. Conclusion: RCAS1 and Claudin-18 expression increase abnormally in malignant pleural effusion, the specific expression is directly correlated with tumor cell proliferation and invasion activity, and they can be used as the reliable indicators for the identification of benign or malignant pleural effusion.