Hepatocellular cancer (HCC) is the sixth most common type of cancer worldwide. Guanosine monophosphate synthase (GMPS) participates in the regulation of chromatin and genes in various organisms, and is highly expressed in a number of human malignant tumors. However, the role of GMPS in HCC has not yet been fully studied and clarified. In this study, the differential fold changes in gene expression levels between HCC cancer tissues and correspondent adjacent normal tissue in The Cancer Genome Atlas Program and GEO datasets were analyzed using R language. GMPS expression levels in HCC cells were knocked down using specific siRNAs. In addition, CCK-8, EdU, TUNEL and immunofluorescence staining were conducted to explore the effects of GMPS siRNAs on HCC cell viability, proliferation, apoptosis and the STAT pathway level, respectively. The results indicated GMPS expression was significantly increased in HCC tumor tissues compared with the corresponding adjacent normal tissues. In addition, high expression of GMPS is negatively associated with the survival rate of patients with HCC. In vitro studies illustrated the knockdown of GMPS notably prevented HCC cell proliferation and induced HCC cell (Hep3B2.1-7 and MHCC97H) apoptosis by regulating the STAT3/c-Myc pathway. The apoptosis-specific marker cleaved caspase was significantly upregulated by GMPS knockdown in HCC cells. The findings of the present study revealed the association between GMPS and the prognosis of HCC. The results suggested that GMPS may serve as a promising marker for the prognosis of HCC, and it may also be a potential therapeutic target for HCC. These findings may lay the theoretical foundation for the clinical application of GMPS.

Outer membrane vesicles derived from Pseudomonas aeruginosa (PA-OMVs) play a crucial role in bacterial pathogenesis, mediating immune modulation and inflammation. Autophagy, a process that degrades damaged organelles, and pyroptosis, a form of programmed cell death, both regulate immune responses and contribute to infection defense. However, the relationship between PA-OMVs, autophagy, and pyroptosis remains insufficiently explored, particularly regarding their regulatory mechanisms. This study investigates how PA-OMVs influence cellular autophagy and pyroptosis, with the aim of identifying potential therapeutic strategies for infectious diseases. Bulk RNA sequencing and bioinformatics analysis were conducted on cells treated with PA-OMVs. Autophagy inhibitors, chloroquine (CQ) and 3-methyladenine (3-MA), were used to explore their effects on pyroptosis, with RT-PCR and ELISA applied to assess pyroptosis levels. The results revealed a complex interplay between autophagy and pyroptosis, with PA-OMVs modulating key immune and inflammatory pathways. Autophagy inhibition decreased the expression of pyroptosis markers, suggesting a regulatory role. These findings highlight the potential of targeting the autophagy-pyroptosis axis for new infection control strategies and vaccine development.

ABSTRACT The bacterium Pseudomonas aeruginosa is able to invade lung epithelial cells and survive intracellularly. During this process, it secretes outer membrane vesicles (OMVs), however, it is currently unclear how OMVs from P. aeruginosa (PA-OMVs) affect lung epithelial cells and their impact on oxidative stress, autophagy, and other physiological activities of lung epithelial cells. In this study, we found that PA-OMVs activated oxidative stress and autophagy in cells. We demonstrated that the NLRP3 (NLR family, pyrin domain containing 3) inhibitor MCC950 can enhance autophagy induced by PA-OMVs. The main function of NLRP3 is related to the body’s immune response and inflammation regulation. MCC950 is the most common inhibitor of NLRP3. Additionally, we showed that PA-OMVs not only enhanced the expression of AMP-activated protein kinase, a key regulator of cellular energy homeostasis, and reactive oxygen species, which play a crucial role in cellular signaling and oxidative stress, but also significantly enhanced the expression of NLRP3. Inhibiting the expression of NLRP3 further enhanced the process of PA-OMVs induced autophagy. These results demonstrate that PA-OMVs activate both autophagy and the NLRP3 inflammasome, with NLRP3 suppressing autophagy to a certain extent, hoping to provide broad ideas for the future applications of PA-OMVs. IMPORTANCE The discovery that lung epithelial cells exposed to outer membrane vesicles from Pseudomonas aeruginosa (PA-OMVs) activate cellular autophagy and induce protective immunity is significant. Specifically, the addition of an NLRP3 inhibitor, MCC950, has been found to decrease NLRP3 targets while simultaneously enhancing the autophagy activity induced by PA-OMVs. This finding unveils a novel theoretical framework for the development of PA-OMVs vaccines, highlighting new targets for enhancing the body’s anti-infective responses. By elucidating the mechanisms through which PA-OMVs trigger autophagy and bolster immune defenses, this research opens avenues for innovative vaccine design strategies aimed at combatting infections effectively.

Non-coding RNAs (ncRNAs) are genomic transcripts that do not encode proteins.They are also closely associated with tumor initiation and progression,as they play regulatory roles in gene transcription,post-transcription,and translation.Moreover,ncRNAs may enter circulatory system in the form of microvesicles or exosomes,or in combination with protein.The circulating ncRNAs are stable and widely existed in body fluids such as blood and urine.Certain circulating ncRNAs are differentially expressed in tumors,suggesting that they have potential value as novel tumor biomarkers.This article briefly reviews the progress of circulating microRNA (miRNA),long non-coding RNA (lncRNA) and circular RNA (circRNA) as breast cancer biomarkers,and discusses the clinical value of circulating ncRNA as a novel biomarker for breast cancer diagnosis and prognosis.

Long noncoding RNAs (lncRNAs) are dysregulated in cancer and involved in oncogenic or tumor inhibitory processes. The aim of the study was to investigate the expression pattern of lncRNA XLOC_013703 in multiple myeloma (MM) and to evaluate its biological role and potential significance. We found that XLOC_013703 was significantly decreased in CD138 positive plasma cells and serum of MM patients compared to normal controls, and the decreased XLOC_013703 expression was correlated with β2-MG, serum-free light chain (s-FLC) and revised international staging system. RNA-fluorescence in situ hybridization results revealed that XLOC_013703 was distributed both in the nucleus and in the cytoplasm of MM cells including H929, RPMI8226, and U266. Overexpression of XLOC_013703 inhibited the proliferation of U266 cells and blocked the cell cycle in G1 stage, thus contributing to MM cell apoptosis. By contrast, knockdown of XLOC_013703 promoted the growth of H929 cells. Western blot analysis confirmed that the expression of p-IκBα and nuclear P65 was substantially increased in shRNA transfection groups compared to control groups, whereas overexpression of XLOC_013703 reduced these expressions. In conclusion, we confirmed that the decreased expression of a novel lncRNA, XLOC_013703, in MM. XLOC_013703 was involved in MM cell survival and proliferation via nuclear factor-κB pathway which represents a potential therapeutic target for MM. © 2019 IUBMB Life, 71(9):1240-1251, 2019.

Objective To analyze the identification, drug resistance and clinical significance of a rare bacterium of Bordetella holmesii ( B. holmesii) to improve its detection and clinical diagnosis and treat-ment. Methods A strain isolated from a bacteremia case was identified by bacterial culture, biochemical tests and 16S rRNA gene sequencing. Mega 7. 0 software was used to conduct a similarity analysis of 16S rRNA gene sequences between the type strains of Bordetella spp. and the isolate, and then a phylogenetic tree was constructed. Antibiotic resistance of the isolate was determined by E-test. Changes in bacterial growth were measured after adding different concentrations of riboflavin or its inhibitor lumiflavin to the cul-ture medium. Results B. holmesii ABD2 was the pathogen causing bacteremia in the immunocompetent pa-tient. It was deposited under the number of CGMCC 1. 13721 in China General Microbiological Culture Col-lection Center (CGMCC), and the 16S rRNA gene sequences were deposited in National Center for Biotech-nology Information ( NCBI) with the accession number of KT828544. 1. Unrooted tree showed that the B. holmesii strain was highly homologous with B. pertussis. Antibiotic susceptibility test showed that the mini-mum inhibitory concentrations ( MIC) of piperacillin, ceftazidime, cefepime, imipenem, meropenem, cipro-floxacin, levofloxacin, gentamicin, amikacin, erythromycin, tetracycline and polymyxin B against the isolate were low, while the MIC values of cefazolin, cefuroxime, cefoxitin, cefotaxime, aztreonam and trime-thoprim-sulfamethoxazole were high. Riboflavin accelerated the growth of B. holmesii ABD2, while its inhibi-tor lumiflavin had an inhibitory effect. Conclusions As B. holmesii is hard to isolate and identify, limited clinical, microbiological and epidemiological data are available. It is an under-recognized pathogen with a considerable amount of information that remains to be studied.

Radiotherapy, chemotherapy, autologous/allogeneic stem cell transplantation, and targeted drug therapy are currently available therapeutic options for multiple myeloma (MM), but the clinical outcome remains unsatisfactory owing to frequent occurrence of drug resistance. Anti apoptosis is one of the main mechanisms to mediate drug resistance. Studies have shown that MCL-1 plays a key role in the growth of cancer cells “escaping” drug attacks, but the underlying mechanism remains unclear. Our previous study demonstrated that lncRNA H19 was highly expressed in the serum of MM patients. Bioinformatics predicts that miR-29b-3p is the downstream target gene, and MCL-1 is the downstream target protein of miR-29b-3p. Therefore, we speculated that MCL-1 may be involved in the occurrence of drug resistance through epigenetics. On the basis of these previous findings, the present study was intended to explore the biological function of H19, interactions between the downstream target genes, and the effect of H19 on BTZ resistance of myeloma cells. In addition, in vivo experiments we have also confirmed that H19 promoted tumor growth and may develop resistance to bortezomib partly. It was found that H19 reduced cell sensitivity to the chemotherapeutic drug BTZ by working as a miRNA sponge to inhibit the expression of miR-29b-3p, enhance MCL-1 transcriptional translation and inhibit apoptosis. These findings may help gain insights into the molecular mechanism of acquired BTZ resistance and develop new drug targets for the clinical treatment of MM.

目的 探讨2型糖尿病(T2DM)下肢血管病变患者血清腹腔脂肪型丝氨酸蛋白酶抑制剂(Vaspin)、抵抗素(RET)水平变化及意义.方法 选取T2DM患者110例,根据是否存在下肢血管病变分为NON-LVD组45例和LVD组35例,另选取同期健康体检者40例作为对照组.采集各组清晨空腹静脉血,应用自动生化仪检测NON-LVD组、LVD组治疗前后及对照组空腹血糖(FBG)、甘油三酯(TG)、总胆固醇(TC),采用放射免疫分析法测定空腹胰岛素(FINS),采用比色法测定糖化血红蛋白(HbA1c),采用ELISA法检测血清Vaspin、RET及游离脂肪酸(FFA)水平,计算胰岛素抵抗指数(HOMA-IR).采用Logistic多元逐步回归分析T2DM患者发生下肢血管病变的危险因素.结果 NON-LVD组、LVD组血清Vaspin水平均低于对照组,且LVD组低于NON-LVD组(P均<0.05).NON-LVD组、LVD组血清RET水平均高于对照组,且LVD组高于NON-LVD组(P均<0.05).血清Vaspin与RET水平呈负相关(r=-0.495,P<0.05).血清RET(β=0.194,OR=1.161)是T2DM患者发生LVD的独立危险因素,而血清Vaspin(β=-0.141,OR=1.159)是保护因素(P均<0.01).结论 发生下肢血管病变的T2DM患者血清Vaspin水平降低、RET水平升高,二者是导致T2DM患者发生下肢血管病变的危险因素.

目的:利用酶显色底物配制念珠菌显色平板,并评估其临床应用中的符合率、敏感性与特异性.方法:基础培养基沙保罗琼脂联合酶显色底物(H012、H026、H092),经过优化实验筛选最适宜配方,制备念珠菌显色平板,对112株临床分离株进行分离鉴定,并与API ID 32C鉴定结果比较.结果:白色念珠菌、光滑念珠菌、热带念珠菌和克柔念珠菌在配制的显色平板中分别呈现绿色、 紫色、 蓝灰色和粉红色,48h鉴定结果与API ID 32C总体符合率98.0%,敏感性与特异性分别达98.0%、99.4%.结论:本实验室配制的念珠菌显色平板灵敏度强、特异性高,更经济合理,能简便快捷地初步鉴别常见念珠菌,符合临床标本检测需求.

Objective To investigate the expression of B-cell activating factor ( BAFF ) and its specific receptor BAFF-R in patients with B-cell non-Hodgkin′s lymphoma ( B-NHL) and to analyze the cor-relations between BAFF and the development of B-NHL.Methods RTQ-PCR and Western blot assay were used to measure the expression of BAFF and its specific receptor BAFF-R in patients with B-NHL.Fluores-cence immunocytochemical staining was used to determine the localization of BAFF and BAFF-R in Raji cells, a B-NHL cell line.The expression of BAFF in tumor tissues from patients with B-NHL of different his-tologic subtypes was measured by immunohistochemistry.WST proliferation and TUNEL assays were used to evaluate the effects of BAFF and BAFF-R on the proliferation, survival rate and apoptosis of Raji cells.Lin-ear correlations between the concentrations of lactate dehydrogenase ( LDH) and the expression of BAFF and BAFF at mRNA and protein levels in patients with B-NHL were analyzed.Results BAFF and its specific receptor BAFF-R were expressed in Raji cells and played an important role in the survival and proliferation of B-NHL cell line.The expression of BAFF in tumor cells from patients with B-NHL varied with the different histologic subtypes of B-NHL.Patients with small B-cell malignant lymphoma, large B-cell lymphoma ( LBCL) , mucosa-associated lymphoid tissue lymphoma ( MALT lymphoma) and follicular lymphoma showed higher levels of BAFF, while those with mantle cell lymphoma showed lower levels of BAFF.Compared with the healthy subjects, patients with B-NHL showed significantly increased expression of BAFF at mRNA and protein levels.The levels of LDH were closely related to the expression of BAFF at mRNA and protein lev-els.Conclusion BAFF and its specific receptor BAFF-R might play an important role in the growth and survival of malignant B cells.

目的:探讨监护病区无菌体液标本中分离菌的分布及其耐药特点,以提高我院监护病区的感染防控与诊治水平.方法:回顾性分析2013年1月～2015年9月期间我院监护病区无菌体液标本中618株分离菌的分布及其耐药性,采用Whonet5.6统计软件进行数据分析.结果:618株分离菌主要来自血液、胸腹水和脑脊液等无菌体液标本;革兰阳性菌占53.1％、革兰阴性菌占35.9％和真菌占11.0％.革兰阳性菌对万古霉素、利奈唑胺和复方新诺明敏感率高,革兰阴性菌对碳青霉烯类和氨基糖苷类保持高度的敏感性,真菌对常用抗真菌药物耐药率均较低.结论:监护病区无菌体液中分离菌以革兰阳性菌为主,临床应结合药敏试验结果,选用合理的抗菌药物,减少耐药菌株产生.

Objective To verify the main analytical performance of AFP,CEA kits.Methods According to Health Industry Standards of People′s Republic of China WS/T xxx-2008,we performed the verification test of AFP,CEA commodities quantitative kits.Results The results of within-run CV and between-run CV in the precision verification experiments by using quality control serums were both lower than the corresponding CV values,which were manufacturers claimed.The average values of bias for trueness of verification experiments results were under the manufacturers claimed bias.The measuring interval of each testing items were close to the expected values.Coefficient of correlations(r)>0.975 and ideal linear(slope b was close to 1).Conclusion The results of verification test showed that the precision,trueness and measuring interval involved in this study were all met the requirements of clinical application.

Objective:To establish the examination method of BAFF mRNA expression in non-Hodgkin lymphoma peripheral blood mononuclear cells by real-time fluorescence quantitative PCR.Methods:The BAFF mRNA expression of 47 non-Hodgkin lymphoma(NHL) patients and 20 healthy peripheral blood mononuclear cells were examined by real-time fluorescence quantitative PCR(RFQ-PCR).Results:The sensitivity of RFQ-PCR was 10 pg /ml.The variation coefficients of intra-assay and inter-assay for low concentration sample were 8.56% and 11.32%,for high concentration sample were 0.76% and 4.58% respectively.The expression of BAFF mRNA in NHL patients were significantly higher compared to those in control groups(0.48±0.023,0.25±0.023).Conclusion:This assay has high sensitivity and reproducibility.It is suitable for measurement of the expression level of BAFF.The BAFF mRNA expression in NHL patients peripheral blood were higher compared to those in control groups,which may play important roles in the initiation and progression of NHL.

B-Lymphocyte stimulator (BLyS), a member of tumor necrosis factor superfamily, is a potent co-activator of B cells in vitro, and in vivo induces B cell proliferation and immunoglobulin secretion. Multiple myeloma (MM) is an incurable malignancy of terminally differentiated B cells (plasma cells). Previous studies have well ascertained that BLyS plays an important contributory role in the pathogenesis and propagation of multiple myeloma by virtue of its ability to promote B cell survival, expansion, and differentiation. However, the intracellular signaling of BLyS in human MM cells remains undefined. This study was designed to see whether there was interaction between MAPK signaling pathway and BLyS expression. It was found that the active protein p-JNK was expressed in KM3, U266 and PBMCs of MM patients, and that the expression of BLyS could be changed by JNK pathway activator and inhibitor. In addition, recombinant BLyS activated JNK pathway, while BLyS siRNA treatment inhibited the activation of JNK pathway. The level of BLyS expression and the activation of JNK pathway were positively correlated. These findings suggest that JNK activation and BLyS expression in MM cells may form a positive feedback loop that promotes the survival and proliferation of MM cells, and these may shed some light on the pathogenesis and treatment of MM.

目的 探讨B淋巴细胞刺激因子(BAFF)及其特异性受体BAFF-R对多发性骨髓瘤(MM)细胞增殖及凋亡的影响.方法 采用流式细胞术(FCM)、RT-PCR、Western Blot、荧光免疫细胞化学技术检测BAFF及其受体BAFF-R的表达与定位;WST细胞增殖实验及TUNEL法检测BAFF及其受体BAFF-R对MM肿瘤细胞生长、存活与凋亡的影响.结果 ①MM细胞能够表达BAFF及其特异性受体BAFF-R;②BAFF及其受体BAFF-R定位表达于KM3细胞的浆膜上;③BAFF及其受体BAFF-R促进MM细胞的增殖、存活,抗细胞凋亡.结论 BAFF及其受体BAFF-R对于骨髓瘤发生、发展可能起着重要的作用.

Objective To investigate the activation of MAPK signal pathway in multiple myeloma and the regulation of BLyS expression levels through MAPK signal pathway; preliminarily study the role of MAPK signal pathway in the up-regulation of BLyS expression levels induced by IFN-γ.Methods Activated MAPK pathway were detected by Western blot,while the expression of BLyS were detected with RT-PCR and Western blot,and Western blot investigated the effect of MAPK pathway on BLyS expression levels induced by IFN-γ.Results In addition to the expression of ERK,JNK,p38,p-JNK was also expressed in MM cell lines,the MAPK pathway inhibitor targeting JNK SP600125 can down-regulate the expression of BLyS,and its activator anisomycin can up-regulate the expression of BLyS.SP600125 restrained the proliferation and survival of MM cells.Conclusion JNK/SAPK pathway was activated in MM cells; The activated degree of JNK/SAPK pathway and the expression level of BLyS was positively correlated.JNK/SAPK pathway play an important role in the up-regulation of BLyS expression levels induced by IFN-γ.

Objective: To investigate the possible association between allelic polymorphism of tumor necrosis factor beta (TNF beta) in 1069 locus and severe post-trauma sepsis in the Han Chinese of Jiangsu Province, China.Methods: We studied the polymorphism of single-base mutation of the TNF beta gene in 58 patients with severe post-trauma sepsis and 88 healthy controls, using polymerase chain reaction (RCA) amplification and polyacrylamide gel electrophoresis (PAGE) analyses.Results: The frequency of the TNF beta*2 allele in patients complicated with post-trauma sepsis was significantly higher than that in healthy controls (68.9% vs 55.1%, P=0.015, RR=1.73).Conclusion: Our results implied an association between an allelic polymorphism of TNF beta in 1069 locus and severe post-trauma sepsis susceptibility in a Chinese population.

This paper firstly introduced the basic content of the comprehensive information emotional theory, which includes: basic model, basic principle, emotional changes produced by cognition fall, and then further discussed emotion space (feeling, emotion and affection space), emotional stimulus response threshold, migration mechanisms and rules of emotion space. The changes of emotional state (migration of emotion space) and cognition process and the relation between them were unveiled.

AIM:To investigate the effects of IFN-gamma and the inhibitor of NF-kappaB(BAY11-7082) on BAFF-R gene promoter activity.METHODS:The fragments with different lengths of the 5'-flanking region of BAFF-R gene were amplified by PCR.Then PCR products were cloned into Luciferase reporter vector (pGL3-Basic) to construct eight recombinant plasmids. These recombinant plasmids were transiently transfected into KM3 cells to locate the promoter region with the highest activity. Then cells transfected with recombinant plasmid with the highest promoter activity were cultured on media in the absence or presence of IFN-gamma and BAY11-7082 and relative luciferase activity were tested and compared. Meanwhile BAFF-R mRNA expression were also examined and compared before and after IFN-gamma and BAY11-7082 were added into cell medium.RESULTS:IFN-gamma can promote BAFF-R promoter activity and up-regulate BAFF-R mRNA expression. And BAY11-7082 can inhibit BAFF-R promoter activity and down-regulate BAFF-R mRNA expression.CONCLUSION:IFN-gamma and the NF-kappaB pathway could be involved in regulating the transcription and mRNA expression of BAFF-R gene.