Objective To explore the effect of the knockdown of transmembrane 9 superfamily protein member 2 (TM9SF2) on the replication of vesicular stomatitis virus (VSV), and investigate its role in the mechanism of antiviral innate immunity. Methods Small interfering RNA (siRNA) was used to knock down the TM9SF2 gene in human non-small cell lung cancer A549 cells. The CCK-8 method was used to assess cell proliferation. A VSV-green fluorescent protein (VSV-GFP) infected cell model was established. The plaque assay was used to measure the viral titer in the supernatant. RT-qPCR and Western blotting were employed to quantify the mRNA and protein levels of VSV genome replication in A549 cells following VSV infection, as well as the expression of interferon β (IFN-β) mRNA and interferon regulatory factor 3 (IRF3) protein phosphorylation following polyinosinic-polycytidylic acid (poly(I:C)) stimulation. Results Compared to the negative control, the knockdown of TM9SF2 exhibited a significant effect, with no observed impact on A549 cell proliferation. The VSV-GFP infected A549 cell model was successfully established. After viral stimulation, fluorescence intensity was reduced following TM9SF2 knockdown, and the mRNA and protein levels of VSV were significantly downregulated. The viral titer of VSV was decreased. After poly(I:C) stimulation, TM9SF2 knockdown significantly upregulated the mRNA level of IFN-β and the phosphorylation level of IRF3 protein. Conclusion The knockdown of TM9SF2 inhibits the replication of vesicular stomatitis virus, and positively regulates the type I interferon signaling pathway, thus enhancing the host's antiviral innate immune response.
Humans ; Virus Replication/genetics* ; Signal Transduction ; Membrane Proteins/metabolism* ; A549 Cells ; Vesiculovirus/physiology* ; Interferon-beta/metabolism* ; Interferon Regulatory Factor-3/genetics* ; Interferon Type I/metabolism* ; Vesicular Stomatitis/immunology* ; Gene Knockdown Techniques ; Vesicular stomatitis Indiana virus/physiology* ; RNA, Small Interfering/genetics*

The enterotoxigenic Escherichia coli (ETEC) infection is a major factor restricting the development of animal husbandry. However, the abuse of antibiotics will lead to the antibiotic residues and emergence of antibiotic-resistant bacteria. The existing vaccines face challenges in stimulating intestinal immunity, demonstrating limited prevention effects. Therefore, it is indispensable to develop a new vaccine that is safe and suitable as a feed additive to activate intestinal immunity. This study constructed yeast-cell microcapsules (YCM) carrying the DNA vaccine against ETEC by genetic engineering. Furthermore, animal experiments were carried out to explore the regulatory effects of feeding YCM on the intestinal immune system and intestinal microbiota. Saccharomyces cerevisiae was selected as the oral delivery vehicle (microcapsules) of the DNA vaccine. The codon-optimized nucleic acid sequence of K88, the main antigen of mammal-derived ETEC, was synthesized, and the yeast shuttle vector containing the corresponding DNA vaccine expression cassette was constructed by DNA recombination. The recombinant strain of YCM was prepared by transforming JMY1. Additionally, the characteristics of the YCM strain and its feasibility as an oral vaccine were comprehensively evaluated by the fluorescence reporter assay, gastrointestinal fluid tolerance assay, intestinal epithelial cell adhesion assay, intestinal retention assessment, antiserum detection, and intestinal microbiota detection. The experimental results showed that the DNA vaccine expression cassette was expressed in mammals, and the recombinant strain of YCM could tolerate up to 8 hours of gastrointestinal fluid digestion and had good adhesion to intestinal epithelial cells. The results of mouse feeding experiments indicated that the recombinant strain of YCM could stay in the intestinal tract for at least two weeks, and the DNA vaccine expression cassette carried by YCM entered the intestinal immune system and triggered an immune response to induce the production of specific antibodies. Moreover, feeding YCM recombinant bacteria also improved the abundance of gut microbiota in mice, demonstrating a positive effect in regulating intestinal flora. In summary, we prepared the recombinant strain of YCM carrying the DNA vaccine against ETEC and comprehensively evaluated its characteristics and feasibility as an oral vaccine. Feeding the recombinant YCM could induce specific immune responses and regulate intestinal microbiota. The findings provide a reference for the immunoprevention of ETEC-related animal diseases.
Animals ; Enterotoxigenic Escherichia coli/genetics* ; Saccharomyces cerevisiae/metabolism* ; Vaccines, DNA/genetics* ; Mice ; Escherichia coli Infections/immunology* ; Escherichia coli Vaccines/genetics* ; Capsules ; Mice, Inbred BALB C ; Female

OBJECTIVE To provide reference for the safe use of bevacizumab in cancer patients. METHODS The diagnosis and treatment of a 65-year-old female lung adenocarcinoma patient with diabetic ketoacidosis （DKA） induced by bevacizumab was retrospectively analyzed， and the possible mechanisms and causes were analyzed based on literature review. RESULTS & CONCLUSIONS The diagnosis and treatment process of patients were analyzed， and DKA caused by other drugs and disease factors were excluded. DKA was considered to be caused by the use of bevacizumab according to Naranjo’s ADR evaluation scale； the acidosis of the patient improved rapidly after one hemodialysis treatment. DKA caused by bevacizumab is rare in clinic， clinicians should be aware that bevacizumab may affect pancreatic function and induce DKA， and early detection and treatment should be achieved to improve the prognosis.

Objective: To analyze the relationship between microorganisms and endodontic disease by using 16 S rDNA sequencing to compare the composition of the microbial community in the root canals of teeth with pulpitis and apical periodontitis. Methods: Clinical samples were collected from teeth requiring root canal treatment.The total DNA of the bacteria in the samples and the gene fragments of the V3-V4 highly variable region on the 16S rDNA fragments were amplified through PCR.After sequencing by NovaSeq,statistical and bioinformatic analysis,including phylogenetic analysis,diversity analysis and analysis of group differences,were performed. Results: In total,6 teeth with pulpitis and 7 teeth with apical periodontitis were collected,and a total of 8 510 OTUs were obtained after next-generation sequencing,and the analysis of bacterial diversity showed that the difference between pulpitis and apical periodontitis in terms of the composition of the bacterial flora was statistically significant(P<0.05).In particular,the relative abundance of Proteobacteria,Acidobacteriota and Actinobacteriota phylum was significantly higher in the roots of teeth affected by pulpitis than apical periodontitis.The relative abundance of Bacteroidota phylum and Synergistota phylum was significantly higher in the root canals of teeth with apical periodontitis. Conclusion There is a complex diversity of infecting microorganisms in the root canals of teeth affected by endodontic diseases.The microbial communities in the infected root canals of pulpitis and apical periodontitis show some differences,and changes in the microbial composition of the root canals may be associated with the development of endodontic diseases.

OBJECTIVE To investigate the effect of eukaryotic translation elongation factor 1A1(eEF1A1)on the replication of vesicular stomatitis virus(VSV)and Herpes simplex virus 1(HSV-1)to identify a potential target for broad-spectrum regulation of viruses.METHODS Small interfering RNA(si-eEF1A)was transfected into human skin fibroblasts(BJ-5ta)to inhibit the expression of eEF1A1,and the negative control group was set up.The transfection efficiency was detected by real-time fluo-rescence quantitative PCR(RT-qPCR)and Western blotting,the cell model of eEF1A1 gene silencing was constructed.The cell model was infected with VSV,the gene copy number and protein expression of VSV in the cells were detected.The cell model was infected with HSV-1,the mRNA and protein expres-sion of HSV-1 in the cells were detected.The cell models were transfected with polyinosinic acid[Poly(I:C)]or sodium deoxyribonucleic acid(HT-DNA),respectively.The mRNA expression of interferon-β(IFN-β),C-X-C Motif Chemokine 10(CXCL10)and interferon-stimulated gene 56(ISG56)were detected by RT-qPCR.The phosphorylation expression of interferon regulatory factor 3(IRF3)and TANK binding kinase 1(TBK1)were detected by Western blotting.RESULT Compared with the negative control group,the mRNA and protein expression of eEF1A1 in the eEF1A1 gene silencing group were signifi-cantly decreased(P<0.01),the cell model of eEF1A1 gene silencing was successfully constructed.Compared with the negative control group,the VSV gene copy number of the eEF1A1 gene silencing group decreased by 70%-80%.The VSV protein expression decreased significantly(P<0.01).The mRNA expression of HSV-1 was decreased by 50%-60%,and the protein expression of HSV-1 was significantly decreased(P<0.01).After stimulation with Poly(I:C)or HT-DNA,compared with the negative control group.there was no significant difference in the mRNA expressions of IFN-β,ISG56 and CXCL10 and the protein phosphorylation expression of IRF3 and TBK1 in the eEF1A1 gene silencing group.CONCLUSION eEF1A1 silencing can inhibit VSV and HSV-1 virus replication,suggesting that eEF1A1 has a potential broad-spectrum regulatory effect on RNA viruses and DNA viruses,and may not recog-nize activated immune pathways through intracellular nucleic acid recognition.

Objective To analyze the relationship between microorganisms and endodontic disease by using 16S rDNA sequencing to compare the composition of the microbial community in the root canals of teeth with pulpitis and apical periodontitis.Methods Clinical samples were collected from teeth requiring root canal treatment.The to-tal DNA of the bacteria in the samples and the gene fragments of the V3-V4 highly variable region on the 16S rDNA fragments were amplified through PCR.After sequencing by NovaSeq,statistical and bioinformatic analysis,inclu-ding phylogenetic analysis,diversity analysis and analysis of group differences,were performed.Results In total,6 teeth with pulpitis and 7 teeth with apical periodontitis were collected,and a total of 8 510 OTUs were obtained after next-generation sequencing,and the analysis of bacterial diversity showed that the difference between pulpitis and apical periodontitis in terms of the composition of the bacterial flora was statistically significant(P＜0.05).In particular,the relative abundance of Proteobacteria,Acidobacteriota and Actinobacteriota phylum was significantly higher in the roots of teeth affected by pulpitis than apical periodontitis.The relative abundance of Bacteroidota phylum and Synergistota phylum was significantly higher in the root canals of teeth with apical periodontitis.Con-clusion There is a complex diversity of infecting microorganisms in the root canals of teeth affected by endodontic diseases.The microbial communities in the infected root canals of pulpitis and apical periodontitis show some differ-ences,and changes in the microbial composition of the root canals may be associated with the development of endo-dontic diseases.

Objective To systematically evaluate the efficacy and safety of electroacupuncture(EA)in the treatment of postoperative nausea and vomiting(PONV)after gynecological surgery.Methods PubMed,Cochrane Library,Web of Science,Embase,China national knowledge infrastructure(CNKI),Wanfang database,and China biomedical literature database(CBM)were systematically searched.The re-trieval period was from the establishment of the database to December 2022.Relevant randomized controlled trials on EA for the treatment of PONV in gynecological surgery were collected.RevMan 5.3 software was used for meta-analysis.Results Fourteen randomized controlled trials were accommodated,including 958 patients,477 patients in the EA group and 481 patients in the control group.Compared with the control group,the incidence of PONV was significantly lower in group EA at 0-48 hours postoperatively(RR=0.55,95％CI 0.47 to 0.65,P＜0.001),and the PONV scores were significantly lower in the postopera-tive period within 48 hours in group EA(MD=-0.40 scores,95％CI-0.65 to-0.16 scores,P=0.004),the incidence of postoperative remedial antiemetic were significantly lower(RR=0.28,95％CI 0.16 to 0.51,P＜0.001).Conclusion EA can reduce the incidence of PONV and the incidence of re-medial antiemetic after gynecologic surgery.

Objective To systematically evaluate the effect of transcutaneous electrical acupoint stimulation(TEAS)in the treatment of postoperative nausea and vomiting(PONV)after laparoscopic non-gastrointestinal surgery.Methods Databases such as PubMed,Cochrane library,Web of Science,Embase,CNKI,Wanfang,and Chinese biomedical database(CBM)were searched to find and screen ran-domized controlled trials(RCTs)of TEAS in the prevention and treatment of PONV after laparoscopic non-gastrointestinal surgery.The retrieval time was from the establishment of the database to July 2023.Meta-a-nalysis was performed using RevMan 5.3 software.Results Twenty-two RCTs involving 3 538 patients were included,including 1 799 in the TEAS group and 1 739 in the control group.The results of meta-analysis showed that the total incidence of PONV in the TEAS group was significantly lower than that in the control group 0-24 hours after operation(RR=0.54,95％CI 0.44-0.68,P＜0.001),and the incidence of postoperative remedial antiemetic was significantly reduced(RR=0.54,95％CI 0.38-0.77,P＜0.001).There was no significant difference in the incidence of postoperative acupoint stimulation-related adverse reactions between the two groups(RR=0.62,95％CI 0.15-2.51,P=0.500).Conclusion TEAS has good clinical efficacy and safety in the treatment of PONV after laparoscopic non-gastrointestinal surgery.

Objective:To investigate the early auditory discrimination of vowels, consonants and lexical tones in prelingually-deafened children with cochlear implants (CI) using auditory event-related potentials.Methods:Nineteen prelingually-deafened CI children and 19 normal hearing (NH) children were recruited in this study. A multi-deviant oddball paradigm was constructed using the monosyllable/ta1/as the standard stimulus and monosyllables/tu1/,/te1/, /da1/,/ra1/,/ta4/and/ta2/as the deviant stimuli. The event-related potentials evoked by vowel, consonant and lexical tone contrasts were recorded and analyzed in the two groups.Results:NH children showed robust mismatch negativities (MMNs) to vowel, consonant and lexical tone contrasts ( P<0.05), whereas CI children only showed positive mismatch responses (pMMRs) and P3a responses to the vowel ( P<0.05) and consonant contrasts ( P<0.05) and no significant event-related potential to the lexical tone contrasts ( P>0.05). The longer pMMR and P3a peak latencies ( P<0.01) but similar amplitudes ( P>0.05) were found in CI children than in NH children. CI children showed weaker phase synchronization of θ oscillations than NH children ( P<0.05). The duration of CI use was positively correlated with the scores of Categories of Auditory Performance (CAP) ( P=0.004), Speech Intelligibility Rate (SIR) ( P=0.044) and Meaningful Auditory Integration Scale (MAIS) ( P=0.001) in CI children. Conclusions:Prelingually-deafened CI children can process vowels and consonants at an early stage. However, their ability of processing speech, especially lexical tones, is still more immature compared with their NH peers. The event-related potentials could be objective electrophysiological indicators reflecting the maturity of CI children′s auditory speech functions. Long-term CI use is beneficial for prelingually-deafened children to improve auditory and speech performance.

Objective:To explore the feasibility of the maze design of a proton therapy system.Methods:According to the construction scheme of a medical proton therapy system, the ambient neutron dose equivalent rate outside the maze entrance was estimated by numerical analytical formula, and compared with the monitoring result.Results:The ambient neutron dose equivalent rate was estimated at between 3.85×10 -2 μSv/h and 7.57 μSv/h and measured to be ≤1.56 μSv/h. Conclusions:The maze entrance was designed to have 5 sections of tunnel for cyclotron room and 3 sections of tunnel for treatment room, respectively, each with designated length and width. The levels of ambient dose equivalent rate outside the maze entrance is acceptable.