Background: Respiratory syncytial virus (RSV) RNA-dependent RNA polymerase (RdRp) complex is an essential molecular machine for viral genome replication. The L protein, the catalytic subunit of this complex (L-RdRp), is well-characterized structurally and represents a highly promising target for the development of novel small-molecule drugs against RSV. Methods: To address the limitations of current QSAR-based virtual screening strategies for RSV L-RdRp inhibitor development, we established a multi-dimensional computer-aided drug screening framework integrating activity, toxicity, drug-likeness, and stability. Results: Two OECD-compliant 2D-QSAR models were developed and rigorously validated to predict inhibitory activity and cytotoxicity, respectively. The optimal inhibitory activity model exhibited strong statistical performance, with R2 = 0.8281, QLOO2= 0.7653, Rtest2= 0.8713, QFn2= 0.8594 ∼ 0.8837, CCCtest = 0.9301, MAEtest = 0.1966. Similarly, the best cytotoxicity model achieved R2= 0.8263, QLOO2 = 0.7422, Rtest2 = 0.8951, QFn2 = 0.8108~0.8530, CCCtest = 0.9081, MAEtest = 0.1685. Based on these models, a four-step screening workflow—QSAR-based filtering and molecular docking (15,758 → 2446 → 162 → 19 compounds), ADMET evaluation (19 → 5), and molecular dynamics simulations (MDSs)—was implemented to identify promising L-RdRp inhibitors. Conclusions: Ultimately, five candidate compounds were selected, all of which demonstrated predicted higher inhibitory activity, lower predicted cytotoxicity, a stable predicted binding mode, and favorable oral bioavailability compared with the reference drug remdesivir. These findings provide valuable in silico-derived lead candidates and a reliable computational workflow for identifying experimental L-RdRp inhibitors targeting RSV.

Exosomes, as key mediators of intercellular communication, exhibit promising prospects in the biomedical field, particularly in stem cell-based replacement therapy. Current exosome isolation techniques face challenges in achieving batch enrichment and nondestructive release, which severely restrict their applications. Traditional specific enrichment strategies rely primarily on antibodies; however, aptamers can specifically recognize targets and serve as effective alternatives to antibodies, offering advantages such as small molecular size, high structural stability, and low cost, making them more suitable for scalable target isolation. Herein, we established the Graphene Oxide-anchored Releasable Aptamer Scaffold (GRAS) technique targeting CD63, a characteristic surface biomarker of exosomes. Leveraging the high affinity of single-stranded nucleic acids for graphene oxide (GO), along with the specific binding capacity of aptamers, we fabricated aptamer-displayed oligonucleotide scaffolds (Apt-Scaffold) and immobilized them onto GO to prepare Apt-Scaffold@GO. This configuration allows effective presentation of the CD63 aptamer on the GO interface for specific exosome recognition and capture. Different methods were further designed to achieve efficient and controllable exosome release. Our experimental results demonstrated that the GRAS technique outperformed the gold-standard ultracentrifugation (UC) in enriching exosomes from mesenchymal stem cell (MSC) culture medium. This method enables quantitative and specific enrichment of exosomes. With simple operation, low cost, and independence on large sophisticated instruments, it provides a robust technical foundation for advancing exosome-related biomedical applications.

IntroductionChimeric antigen receptor (CAR)-engineered T or natural killer (NK) cells are a promising approach for cancer immunotherapy. The leading region of the CAR structure is generally a single-chain antibody (scFv) fragment specific for a tumor cell surface molecule, and other structures are rarely reported.MethodsIn this study, we developed a novel anti-human epidermal growth factor receptor 2 (HER2) CAR-NK cell using an affibody molecule as the extracellular targeting domain instead of a conventional scFv. Affibody-based CAR-NK cells were generated from the NK-92 cell line. To enhance safety, CAR-NK cells were subjected to γ-irradiation, and their antitumor activity was further evaluated in combination with doxorubicin (DOX)-loaded nanoparticles.ResultsAffibody-based CAR-NK cells exhibited effective cytotoxicity against HER2-positive breast cancer cells, comparable to that of anti-HER2 scFv-based CAR-NK cells. γ-Irradiation at 10 Gy effectively inhibited malignant proliferation of CAR-NK cells but significantly reduced their cytotoxic activity. Notably, incorporation of DOX-loaded nanoparticles markedly enhanced the killing capacity of irradiated CAR-NK cells, restoring and even amplifying their antitumor efficacy.DiscussionThese findings demonstrate that affibody-based CAR-NK cells are a viable alternative to conventional scFv-based CAR constructs. Moreover, the combination of CAR-NK immunotherapy with chemotherapeutic nanomedicine effectively compensates for irradiation-induced cytotoxicity attenuation, offering a promising synergistic strategy for the treatment of HER2-positive breast cancer.

Berberine (BBR), a benzylisoquinoline alkaloid isolated from Chinese herb Coptis chinensis, has been widely used clinically to treat intestinal infectious diseases. Recently, it has been found to have multiple pharmacological effects, including anti-inflammatory activity and immune effects in inflammatory bowel disease (IBD). However, its exact targets remain to be elucidated. In this study, we used a mouse intestinal organoid-macrophage co-culture model to investigate the anti-inflammatory effects and immune effects of BBR. Our findings demonstrated that lipopolysaccharide (LPS) induced more robust inflammatory responses and epithelium damage in the co-culture system compared to the organoid alone. BBR effectively attenuated inflammation and restored epithelial barrier integrity by suppressing M1 macrophage polarisation and infiltration, alongside upregulating the expression and organisation of tight junction protein zonula occludens-1 (ZO-1). RNA sequencing and proteomic analysis revealed that BBR disrupted organoid-macrophage interaction by inhibiting chemokine (e.g., C-X-C motif chemokine ligand 1 (CXCL1) and macrophage migration inhibitory factor (MIF)) release from epithelial cells, thereby reducing macrophage recruitment. Collectively, our study establishes the organoid-macrophage co-culture system as a more physiologically relevant model for studying epithelial-immune interactions and elucidates the multi-target mechanism of BBR, which concurrently modulates epithelial cells, macrophages, and their crosstalk. These findings lay the foundation for further exploration of the therapeutic potential of BBR in inflammatory bowel disease and the development of targeted therapies that regulate cell interactions.

Nanovaccines represent a promising strategy for colon cancer immunotherapy, with the potential to elicit potent, tumor-specific immune responses. However, the efficacy of these vaccines is often compromised by the presence of cancer stem cells (CSCs) in the tumor microenvironment (TME). Nanoliposome (NLP) is a widely used delivery system in nucleic acid and drug delivery research. In this study, we enriched MC38-derived CSCs (MCSCs) and developed a manganese-CpG-nanoliposome (Mn@CpG@NLP) nanocomplex using MC38 colon carcinoma tumor lysates as antigens to induce immune responses against MCSCs-derived tumors. Manganese NLPs (Mn@NLP) are initially engineered by incorporating manganese ions to enhance their positive surface charge, thereby optimizing their interaction with cellular membranes and activating the STING signaling pathway to promote bone marrow-derived dendritic cells (BMDCs) maturation. Subsequently, these liposomes are co-assembled with the CpG oligonucleotides (CpG ODNs) 1826 adjuvant, a Toll-like receptor 9 agonist, through electrostatic interactions to form the Mn@CpG@NLP nano-adjuvant complex. The Mn@CpG@NLP complex is efficiently taken up by dendritic cells (DCs), leading to their maturation and activation. This nanocomplex also effectively stimulated cytotoxic T lymphocytes and promoted the secretion of cytokines. In vivo treatment with Mn@CpG@NLP significantly inhibited tumor growth and prolonged survival in a mouse model. This study highlights the potential of nano-adjuvant platforms in immunotherapy targeting cancers driven by colon CSCs.

Viral protein U (Vpu), an accessory protein of HIV-1, functions by antagonizing or hijacking various host factors to allow the virus to evade host immune surveillance and facilitate viral release. Nevertheless, there is limited understanding regarding the impact of Vpu on the biogenesis of HIV-1 RNAs. In this study, we utilized ascorbate peroxidase 2 (APEX2)-based proximity labeling techniques in combination with mass spectrometry and immunoprecipitation-mass spectrometry (IP-MS) to characterize interactions between HIV-1 Vpu and host proteins. We identified nine cellular targets of Vpu. Among these targets, our research demonstrated the interaction between Vpu and RNA-binding motif protein 10 (RBM10), which results in the degradation of RBM10 through the ubiquitin-proteasome pathway. The expression of RBM10 exerted an inhibitory effect on virus replication by binding to viral RNA and reducing the levels of incompletely spliced HIV-1 transcripts. Additionally, it promoted the transcription of various antiviral genes. The findings elucidate the role of HIV-1 Vpu in RNA replication and identify RBM10 as a novel regulator of HIV-1 transcription.IMPORTANCEA comprehensive analysis utilizing APEX2-MS and IP-MS techniques identified a total of 24 cellular targets of Vpu, three of which have been documented as restriction factors. Vpu-interacting proteins were found to be significantly enriched in pathways related to cell adhesion, RNA transport, and the spliceosome. The identification of RBM10 as a novel regulator of HIV-1 replication and infectivity, and RBM10 regulated transcription of both viral and host RNA transcripts. Vpu interacted with RBM10 and promoted its degradation through the ubiquitin-proteasome pathway.

Respiratory syncytial virus (RSV) is a leading cause of severe lower respiratory tract infections in infants, the elderly, and immunocompromised individuals worldwide. The pathogenic mechanism of RSV is closely linked to the membrane fusion process mediated by its fusion glycoprotein (F protein), which has consequently emerged as a critical target for developing anti-RSV therapeutics. At present, there is a lack of specific clinical treatments for RSV, and traditional drug discovery approaches are often time-consuming and expensive. In this context, quantitative structure–activity relationship (QSAR)–assisted drug design offers notable advantages. In this study, we collected a dataset consisting of 156 benzimidazole derivatives against F protein from publicly available sources. Transferable, reproducible, and interpretable 2D-QSAR inhibitory activity and cytotoxicity prediction models were constructed using Genetic Algorithm (GA) and Multiple Linear Regression (MLR). Following rigorous statistical validation, the best inhibitory activity model achieved R2 = 0.8740, Q_Loo^2 = 0.8272, R_test^2 = 0.8273, Q_Fn^2 = 0.8033–0.8492, CCCtest = 0.8782, MAEtest = 0.3014; the best cytotoxicity model was of R2 = 0.7573, Q_Loo^2 = 0.6926, R_test^2 = 0.7707, Q_Fn^2 = 0.7298–0.8656, CCCtest = 0.8639, MAEtest = 0.1342. The optimal inhibitory activity model was used to perform virtual screening on 912 benzimidazole derivatives retrieved from the PubChem, and identified 234 derivatives with better inhibitory activity than the reference JNJ-53718678. Among these, 152 derivatives were found to possess better docking binding energies than JNJ-53718678. Furthermore, we used the optimal toxicity model to assess their cytotoxicity, and identified 23 derivatives with predicted cytotoxicity lower than that of JNJ-53718678. Finally, through drug-likeness evaluation, ADMET analysis and molecular dynamics simulation, we obtained eight potential RSV inhibitors with higher inhibitory activity, lower cytotoxicity, and better pharmacokinetic properties compared to JNJ-53718678.

The outbreak of the novel coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has caused great harm to all countries worldwide. This disease can be prevented by vaccination and managed using various treatment methods, including injections, oral medications, or aerosol therapies. However, the selection of suitable compounds for the research and development of anti-SARS-CoV-2 drugs is a daunting task because of the vast databases of available compounds. The traditional process of drug research and development is time-consuming, labour-intensive, and costly. The application of chemometrics can significantly expedite drug R&D. This is particularly necessary and important for drug development against pandemic public emergency diseases, such as COVID-19. Through various chemometric techniques, such as quantitative structure-activity relationship (QSAR) modelling, molecular docking, and molecular dynamics (MD) simulations, compounds with inhibitory activity against SARS-CoV-2 can be quickly screened, allowing researchers to focus on the few prioritised candidates. In addition, the ADMET properties of the screened candidate compounds should be further explored to promote the successful discovery of anti-SARS-CoV-2 drugs. In this case, considerable time and economic costs can be saved while minimising the need for extensive animal experiments, in line with the 3R principles. This paper focuses on recent advances in chemometric modelling studies of COVID-19-related inhibitors, highlights current limitations, and outlines potential future directions for development.

Corona Virus Disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), poses a serious threat to human health and life safety. How to effectively prevent and treat COVID-19 is crucial. In this study, we used the inhibitors of nonstructural protein Nsp14 of SARS-CoV-2 to perform the quantitative structure activity relationship (QSAR) modelling for the first time. Based on different dataset division strategies, we selected partial least square (PLS) and multiple linear regression (MLR) methods to develop easily interpretable and reproducible QSAR models with 2D molecular descriptors. All models complied with the strict QSAR validation principles of OECD and internationally recognized validation metrics. The best model contained two molecular descriptors with the following statistical parameters: R2 = 0.7796, QLOO2= 0.7373, Rtest2 = 0.8539 and CCCtest = 0.9073. Obviously, the model exhibited good prediction performance and can be used for quickly predicting the inhibitory activity of unknown compounds against Nsp14. Mechanistic interpretation identified the detailed relationship between molecular structure information and inhibitory activity. The best QSAR model was used to predict the inhibitory activity of 263 true external compounds without experimental values against Nsp14, and the prediction reliability was analyzed and discussed. Molecular docking and ADMET analyses were conducted for compounds with higher similarity to the modelling compounds. Finally, two compounds were identified as potential candidate drugs of targeting Nsp14. The current work lays a solid theoretical foundation for the discovery of inhibitors targeting Nsp14, and has an important reference significance for the development of anti-COVID-19 drugs.

BACKGROUND:Abnormally regulated long non-coding RNAs (lncRNAs) functions in cancer emphasize their potential to serve as potential targets for cancer therapeutic intervention. LncRNA ASBEL has been identified as oncogene and an anti-sense transcript of tumor-suppressor gene of BTG3 in triple-negative breast cancer (TNBC).RESULTS:Herein, multicomponent self-assembled polyelectrolyte nanocomplexes (CANPs) based on the polyelectrolytes of bioactive hyaluronic acid (HA) and chitosan hydrochloride (CS) were designed and prepared for the collaborative modulation of oncogenic lncRNA ASBEL with antago3, an oligonucleotide antagonist targeting lncRNA ASBEL and hydrophobic curcumin (Cur) co-delivery for synergetic TNBC therapy. Antago3 and Cur co-incorporated CANPs were achieved via a one-step assembling strategy with the cooperation of noncovalent electrostatic interactions, hydrogen-bonding, and hydrophobic interactions. Moreover, the multicomponent assembled CANPs were ulteriorly decorated with a near-infrared fluorescence (NIRF) Cy-5.5 dye (FCANPs) for synchronous NIRF imaging and therapy monitoring performance. Resultantly, MDA-MB-231 cells proliferation, migration, and invasion were efficiently inhibited, and the highest apoptosis ratio was induced by FCANPs with coordination patterns. At the molecular level, effective regulation of lncRNA ASBEL/BTG3 and synchronous regulation of Bcl-2 and c-Met pathways could be observed.CONCLUSION:As expected, systemic administration of FCANPs resulted in targeted and preferential accumulation of near-infrared fluorescence signal and Cur in the tumor tissue. More attractively, systemic FCANPs-mediated collaborative modulating lncRNA ASBEL/BTG3 and Cur co-delivery significantly suppressed the MDA-MB-231 xenograft tumor growth, inhibited metastasis and extended survival rate with negligible systemic toxicity. Our present study represented an effective approach to developing a promising theranostic platform for combating TNBC in a combined therapy pattern.

In this study, we prepared a streptavidin magnetic bead based on graphene-coated iron nitride magnetic beads (G@FeN-MB) and tried to use it for the enrichment of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). The outer shell of our magnetic bead was wrapped with multiple graphene sheets, and there is no report on the application of graphene to the magnetic-bead-coating material. First, the graphene shell of G@FeN-MB was oxidized by a modified Hummer method so as to generate the carboxyl groups required for the coupling of streptavidin (SA) on the surface of the magnetic beads. X-ray photoelectron spectroscopy (XPS), Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM) were used to characterize the oxidized G@FeN-MB (GO@FeN-MB). Streptavidin was then linked to the surface of the GO@FeN-MB by coupling the amino of the streptavidin with the carboxyl on the magnetic beads by carbodiimide method; thus, the streptavidin magnetic beads (SAMBs) were successfully prepared. To prove the practicality of the SAMBs, biotinylated SARS-CoV-2 S1 antibody was linked with it to respectively capture SARS-CoV-2 Spike-protein-coupled polystyrene beads (S-PS) and pseudovirus with S-protein expressed. Microplate reader and fluorescence microscope results show that the SAMBs can effectively enrich viruses. In conclusion, the preparation of SAMBs with G@FeN-MB is feasible and has potential for application in the field of virus enrichment.

Pre-eclampsia (PE) is a type of hypertensive disorder during pregnancy, which is a serious threat to the life of mother and fetus. It is a placenta-derived disease that results in placental damage and necrosis due to systemic small vessel spasms that cause pathological changes such as ischemia and hypoxia and oxidative stress, which leads to fetal and maternal damage. In this study, four types of risk factors, namely, clinical epidemiology, hemodynamics, basic biochemistry, and biomarkers, were used for the initial selection of model parameters related to PE, and factors that were easily available and clinically recognized as being associated with a higher risk of PE were selected based on hospital medical record data. The model parameters were then further analyzed and screened in two subgroups: early-onset pre-eclampsia (EOPE) and late-onset pre-eclampsia (LOPE). Dynamic gestational week prediction model for PE using decision tree ID3 algorithm in machine learning. Performance of the model was: macro average (precision = 76%, recall = 73%, F1-score = 75%), weighted average (precision = 88%, recall = 89%, F1-score = 89%) and overall accuracy is 86%. In this study, the addition of the dynamic timeline parameter "gestational week" made the model more convenient for clinical application and achieved effective PE subgroup prediction.

目的 深入研究结肠癌肿瘤裂解物纳米疫苗的抗肿瘤作用.方法 制备纳米化胞嘧啶鸟嘌呤二核苷酸(Cytosine guanine dinucleotide,CPG)佐剂并与结肠癌细胞系(MC38)细胞裂解物(lysate)组成纳米疫苗.体外培养小鼠骨髓来源的树突状细胞(BMDC),分4组进行BMDC的激活实验,分别为PBS组、空纳米颗粒(NP)组、溶解态CPG组和负载CPG的纳米颗粒(CNP)组.建立接种MC38肿瘤细胞的C57BL/6结肠癌荷瘤小鼠模型,肿瘤体积增至50 mm3时分为4个免疫组,即PBS组、MC38细胞裂解物(lysate)组、CNP组和MC38细胞裂解物辅以CNP而成的疫苗(vaccine)组.每隔7 d进行皮下免疫,共免疫3次.最后一次免疫3 d后检测小鼠外周血、脾脏、腹股沟淋巴结T淋巴细胞比例及血清TNF-α、IFN-γ含量.结果 经CNP处理的BMDC成熟比例提高(P<0.01)且分泌大量细胞因子(P<0.0001),MC38肿瘤裂解物纳米疫苗免疫荷瘤小鼠后肿瘤体积明显小于其他组(P<0.0001),同时外周血T淋巴细胞、脾脏T淋巴细胞、淋巴结T淋巴细胞比例明显提高(P<0.05),血清中细胞因子含量亦明显增加(P<0.05).结论 MC38肿瘤裂解物纳米疫苗可以有效激活树突状细胞(DC),诱导肿瘤特异性细胞免疫应答,发挥显著的抗肿瘤作用.

教师的职责之一就是教书育人,从专业基础课生理学与其他医学基础课程的融合、与临床知识的融合、与日常生活常识的融合以及与学生专业知识的融合四个维度,探讨在培养人才的过程中,在讲授基础知识的同时如何更好地拓展学生的知识结构,与培养学生的能力相结合,达到学有所获、学有所用的目的,教师"真正把教学当作科学研究在做",从中领悟到教好书、育好人的真谛和内涵.

Abstract The restoration of suppressed miRNAs is a promising therapeutic approach for treating tumors; however, efficient delivery remains a challenge. Herein, we aimed to develop a polyethylenimine (PEI) functionalized graphene oxide nanosheets (GO‐PEI) as a suppressive miRNA delivery system to treat human intrahepatic cholangiocarcinoma (ICC). We found that the GO‐PEI complex possessed excellent transfection efficacy and acceptable toxicity. Furthermore, using our miRNA array analysis and The Cancer Genome Atlas (TCGA) dataset, we selected four miRNAs (miR‐194‐5p, miR‐125b‐5p, miR‐122‐5p, and let‐7c‐5p) that are remarkably downregulated in ICC samples compared to that in adjacent tissues. The GO‐PEI complexed with the four miRNAs (GO‐PEI‐4miR) achieved a higher miRNA transfection efficiency than Lipo2000, as verified by quantitative reverse transcription polymerase chain reaction (qRT‐PCR) assays, immunofluorescence, and flow cytometry analyses. In addition, in vitro experiments revealed that GO‐PEI‐4miR remarkably restored the repressed miRNAs, thereby significantly repressing tumor sphere formation, colony formation, drug resistance, and markedly inhibiting the target genes of these miRNAs and cancer stem cell‐related markers. Furthermore, both tumor weights and bioluminescent imaging analysis indicated that the GO‐PEI‐4miR significantly reduced tumor formation in vivo. Taken together, these results demonstrate that GO‐PEI could be a potential delivery system for multiple repressive miRNAs in ICC therapy.

Esophageal carcinoma (EC) is the sixth most deadly of all cancers. It is among the most malignant cancers due to its highly aggressive nature and low survival rate. The incidence of EC is high in Asia, particularly in Southern areas including China, Iran and Japan. There is a large body of evidence to suggest an association between the melanoma antigen gene (MAGE) family and the initiation of cancer; however, there is no clear evidence to suggest an association between EC and MAGE. Discovery of the chemical and physiological processes relevant to the occurrence of EC is vital for clinicians to diagnose and treat this highly aggressive cancer. The present study focused on the association of EC with the expression of MAGE family member A6 (MAGEA6) at the mRNA and protein levels using gene chip, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry. The expression of MAGEA6 in human esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC) tissue samples were compared with those in paracancerous tissue. The result of the gene chip assay revealed that as the generation grew, there was a significant increase in MAGEA6 transcription in the esophageal epithelial cell line, SHEE Different ESC cell lines also exhibited a significantly higher transcription of MAGEA6 compared with the HaCaT cell line, as determined via reverse transcription-quantitative PCR. An higher positive rate of MAGEA6 expression in ESCC and EAC tissues was also revealed when compared with paracancerous tissues, as determined via immunohistochemistry. The results indicated that MAGEA6 is highly transcribed and expressed in the development of EC and may therefore serve as a novel biomarker for the diagnosis or treatment of EC.

从"多元化教学方法融合,全面提升教学内容的有效性""教与学互动,助推教学内容的实用性""良好的教学方法是实现教学内容的优质载体"等三大方面,对教学方法的重要性、实际应用价值进行探索与研究,旨在为学生后续的学习、研究及工作奠定坚实的基石.

通过"我国公民的素养因素之一——多维健康观""专业教育与通识教育是相互促进的统一体"以及"'健康生活方式与健康传播'"是现代高等教育不可或缺的重要内容"几个方面,对"健康生活方式与健康传播"在高等教育中的价值进行初步分析和讨论,旨在依托通识教育载体使大学生摒弃陋习,养成健康生活习惯,倡导科学生活方式,形成知晓健康信息、认同健康信念、建立健康态度、自觉采纳健康行为的四个层次的健康新理念.

OBJECTIVE:The incidence of the upper gastrointestinal tumor has increased rapidly during recent decades. The relationship between local water pollution and the tumor is still not much clear, so this study was conducted to further investigate the local water pollution and its influence on the malignant cell transformation. Prevalence of human papillomavirus (HPV) in local esophageal cancer (EC) patients was also analyzed in Shenqiu County for the first time.METHODS:Two-step cell transformation was used to study different sources of water in the malignant cell transformation, and the existence of 3-methylcholanthrene (3-MC) in water was analyzed from the river and shallow and deep wells. HPV DNA in tissue samples of EC patients was detected by polymerase chain reaction (PCR) and HPV diagnostic kit.RESULTS:The river water has higher cytotoxicity than the shallow well water and induced significant cell malignant transformation, while deep well water has not shown the malignant cell transformation. In Huaihe River water, the 3-MC concentration was found higher than shallow and deep wells. An HPV infection rate was found high in patients with esophageal cancer.CONCLUSION:Long-term consumption of polluted water can induce malignant cell transformation, and the presence of HPV may be an important cause of cancer.

Objective To construct a recombinant double gene co-expressing plasmid of HIV-1 broadly neutralizing antibody and to detect its expression in 293T cell line.Methods HIV-1 neutralizing antibody 2G12 variable region of light chain (VL) and heavy chain (VH) was synthesized,and ligated with vectors containing human IgG constant regions of light and heavy chain to construct a complete 2G12 light and heavy Chain.The VL and VH of 2G12 and IRES were cloned into eukaryotic expression vector pVR by PCR amplification and then the recombinant plasmid was transfected into 293T cells.Expression of the antibody in cell supernatant was detected by ELISA.Binding and neutralizing activity of the cell supernatant were tested by ELISA and micro-neutralization assays.Results The recombinant double gene eukaryotic expression vector which can express human IgG was constructed successfully.The expression level of the supernatant was 6.43 μg/ml,and the antibody retained the binding and neutralizing activity.Conclusions The constructed vector can express the antibody with binding and neutralizing activity,this study provides a good platform for the expression of human HIV neutralizing antibody in the eukaryotic expression vector.