Objective To analyze the virus subtypes, molecular evolutional and molecular transmission network features of the first confirmed mpox case in Huai’an City, Jiangsu Province, so as to provide insights into understanding of the transmission and evolution dynamics of mpox virus and formulation of the mpox control strategy in the city. Methods Genomic DNA was extracted from swabs of the first confirmed mpox case’s skin lesions in Huai’an City, and the amplicon sequencing library was constructed using the hypersensitive mpox virus whole-genome capture kit. High-throughput sequencing was performed using the GridION X5 nanopore sequencer on the Nanopore sequencing platform, and single nucleotide polymorphism (SNP) analysis of mpox virus genome sequences was performed following sequence assembly. In addition, phylogenetic analysis, genetic genealogy and molecular traceability analysis were performed. Results The virus whole genome sequence of the first confirmed mpox case was successfully obtained by high-throughput sequencing, with a full length of 197 182 bp, and was named hMpxV/China/JS-HA01/2023, which belonged to the clade IIb (West African clade) lineage B.1.3. Compared with the mpox virus reference sequence MPXV-M5312_HM12_Rivers-001 (GenBank accession number: NC_063383), the genome sequence of the Huai’an virus isolate carried 86 SNPs, including 40 SNPs in the coding region as non-synonymous mutations and 73 SNPs as nucleotide mutations caused by APOBEC3 (APOBEC3). Of the 97 mpox virus gene sequences, 79 sequences were included in the molecular network (81.44%), and the threshold of the genetic distance accessed to the network was 0.35/10⁵. There were two large molecular transmission clusters and one scattered cluster in the molecular transmission network of the mpox virus, andthehMpxV/China/JS-HA01/2023 sequence was located in the large cluster. The 97 gene sequences formed 92 haplotypes, including three shared haplotypes Hap_4, Hap_6 and Hap_38, and an exclusive haplotype Hap_1 of hMpxV/China/JS-HA01/2023 generated from mutation of the exclusive haplotype Hap_43, while the exclusive haplotype Hap_43 was generated from mutation of the shared haplotype Hap_38. Conclusions The whole genome sequence of the mpox virus isolated from the first confirmed mpox case in Huai’an City has been successfully obtained, and the molecular evolutionary and molecular transmission network characteristics of the virus have been preliminarily understood.

An innovative on-site real-time avian influenza virus(AIV)detection method was estab-lished by integratingrecombinase-aided amplification(RAA)with the clustered regularly inter-spaced short palindromic repeats(CRISPR)/CRISPR-associated protein(Cas)system.After analy-zing 120 sequences of the M gene of avian influenza viruses of different subtypes publicly available on NCBI,the RAA primers and crRNA were designed based on the identified highly conserved segment and used for RAA nucleic acid amplification.After the amplified products were transferred to a CRISPR/Cas13a detection system,the fluorescence values were monitored throughout the re-action process to indicate the results.The sensitivity and specificity of the RAA-CRISPR/Cas13a method were validated using gradient dilutions(106-100 copies/μL)of positive plasmids and sev-en other avian viruses.Fifty clinical samples were tested using this method and compared with the national standard fluorescence RT-PCR method.The results indicated that the detection limit for RAA-CRISPR/Cas13a method was 102 copies/μL,a two-fold improvement over the standard RAA.Specificity assay showed the established method only detected AIV with no cross-reactivity with other seven avian viruses.Compared to the national standard fluorescence RT-PCR method,this method exhibited 100％specificity,95.24％accuracy,and 98.00％consistency in detection of clinical samples.In conclusion,a universal and rapid RAA-CRISPR/Cas13a for detection of AIV was established with the capacity of achieving detection within 60 minutes at 37 ℃,which provides a rapid,sensitive,and specific on-site detection method for AIV.

Listeria monocytogenes is recognized as a significant foodborne pathogen, capable of causing listeriosis in humans, which is a global public health concern. This pathogen is particularly dangerous for pregnant women, as it can lead to invasive listeriosis in fetuses and neonates, posing a significant threat to both maternal and fetal health. Therefore, establishing suitable in vitro and in vivo models for L. monocytogenes placenta infection, as well as analyzing and exploring the infection process and its pathogenic mechanism, are important approaches to prevent and control L. monocytogenes infection in mothers and infants. In this study, we reviewed the in vitro and in vivo placental models used for studying the infection of L. monocytogenes in maternal and infant, summarized and discussed the advantages and limitations of each model, and explored the potential of in vitro cell models and organoids for the study of L. monocytogenes infection. This paper aims to support the study of the infection pathway and pathogenesis of listeriosis and provide scientific references for the prevention and control of L. monocytogenes infection.
Female ; Humans ; Pregnancy ; Listeria monocytogenes ; Listeriosis/prevention & control* ; Placenta/pathology* ; Public Health ; Infant, Newborn

Objective:To investigate the clinical characteristics of vascular neuro-ophthalmology in patients with central retinal artery occlusion (CRAO).Methods:A single-center, prospective clinical study. From January 2018 to December 2020, 49 eyes of 49 CRAO patients of The Neuro-ophthalmology Department of Xi'an First Hospital were included in the study. Data on patient demographic characteristics, vascular risk factors, disease characteristics, digital subtraction angiography (DSA) imaging characteristics of internal carotid arteries, treatment, treatment-related adverse events, and 1-month follow-up vascular events were collected. All patiens were examined by visual acuity, head CT and or magnetic resonance imaging. At the same time, 35 cases of internal carotid artery vascular DSA were examined; 14 cases of head and neck CT angiography were examined. The anatomical variation of the extracranial segment of the internal carotid artery was divided into tortuous, tortuous, and coiled; the aortic arch was divided into type Ⅰ , type Ⅱ , type Ⅲ, and bovine type. Intravenous thrombolysis, arterial thrombolysis, conservative treatment were performed. The follow-up time was1 month after treatment. Functional vision was defined as vision ≥20/100. Vascular events were strokes, cardiovascular events, deaths and neovascular glaucoma during follow-up.Results:Among 49 eyes of 49 cases, 40 eyes were male (81.6%, 40/49), and 9 eyes were female (18.4%, 9/49); the average age was 60.7±12.9 years. There were 33, 17, and 16 cases with hypertension, type 2 diabetes, and cerebrovascular disease, respectively; 27 and 34 cases had a history of smoking and tooth loss, respectively. Taking antihypertensive, hypoglycemic, antiplatelet aggregation/anticoagulation, and hypolipidemic drugs were 15, 5, 8, and 5 patients, respectively. There were 11 cases of transient amaurosis before the onset, and 17 cases of CRAO after waking up. There were 33 cases (67.3%, 33/49) with infarction of the affected side of the brain tissue. DSA was performed in 35 cases, and the stenosis rate of the internal carotid artery on the affected side was 70%-99% and 100% were 3 (8.6%, 3/35) and 4 (11.4%, 4/35) cases, respectively. The ophthalmic artery on the affected side originated from the external carotid artery in 5 cases (14.3%, 5/35). There were 17 (54.8%, 17/31) and 2 (6.5%, 2/31) cases of tortuousity and kinking in the extracranial segment of the internal carotid artery. There were 15 (42.9%, 15/35), 6(17.1%, 6/35), and 2 (5.7%, 2/35) cases of aortic arch type Ⅱ, type Ⅲ, and bovine type, respectively. Intravenous thrombolysis and arterial thrombolysis were performed in 13 and 29 cases, respectively. Complications occurred in 2 cases during treatment; 3 cases of symptoms fluctuated after treatment, and 10 cases of asymptomatic new infarcts occurred in imaging studies. Forty-eight cases were treated with antiplatelet aggregation/anticoagulation and hypolipidemic treatment. At discharge and 1 month after treatment, the recovery of functional vision was 7 and 17 cases, respectively. One month after treatment, 1 case died because myocardial infarction; 2 cases of neovascular glaucoma occurred.Conclusion:The proportion of CRAO patients with vascular risk factors and internal carotid artery abnormalities on the affected side is relatively high; the prognosis is relatively good after intravenous thrombolysis and/or arterial thrombolysis and secondary stroke prevention.

Objective:To investigate the risk factors for poor outcome and recurrence at 1 year after first-ever ischemic stroke in non-diabetic patients.Methods:Using Xi'an Stroke Registry Research Database, the clinical data of patients with non-diabetic first-ever ischemic stroke diagnosed in 4 tertiary A hospitals in Xi'an from January to December 2015 were collected. The National Institute of Health Stroke Scale (NIHSS) was used to evaluate the severity of stroke. Prognosis (functional outcome and recurrence) was followed up at 1 year after diagnosis. Functional outcome was assessed using the modified Rankin scale. 0-2 was defined as good outcome and >2 as poor outcome. Recurrence was defined as new focal neurological dysfunction caused by cerebral infarction or cerebral hemorrhage events during follow-up and confirmed by cranial CT or MRI. Multivariable logistic regression analysis was used to identify the independent influencing factors of clinical outcomes at 1 year. Multivariable Cox proportional hazard model was used to identify the independent influencing factors of recurrence within 1 year. Results:A total of 1 214 non-diabetic patients with first-ever ischemic stroke were included. One year follow-up showed that 210 patients (17.3%) had a poor outcome, 88 (7.2%) of them died, and 47 (3.9%) had recurrence. Multivariate logistic regression analysis showed that age (odds ratio [ OR] 1.065, 95% confidence interval [ CI] 1.042-1.090; P<0.001), atrial fibrillation ( OR 3.170, 95% CI 1.588-6.327; P=0.001), white blood cell count ( OR 1.106, 95% CI 1.006-1.216; P=0 037), baseline NIHSS score ( OR 1.210, 95% CI 1.147-1.277; P<0.001), and stroke associated-pneumonia (SAP; OR 3.677, 95% CI 1.451-9.316; P=0.006) were independently associated with poor outcomes. Multivariate Cox proportional hazards regression analysis showed that baseline NIHSS score (hazard ratio [ HR] 1.055, 95% CI 1.003-1.109; P=0.036) and SAP ( HR 7.067, 95% CI 3.154-15.836; P<0.001) were independently associated with recurrence. Kaplan-Meier survival curve analysis showed that the 1-year recurrence rate of patients with severe stroke was significantly higher than that of patients with mild to moderate stroke (log-rank test, P<0.001), and the 1-year recurrence rate of patients with SAP was significantly higher than that of patients without SAP (log-rank test, P<0.001). Conclusion:Age, atrial fibrillation, white blood cell count, baseline NIHSS score and SAP are the independent predictors of poor outcomes at 1 year after first-ever ischemic stroke in non-diabetic patients. Baseline NIHSS score and SAP are the independent predictors of recurrence within 1 year after first-ever ischemic stroke in non-diabetic patients.

Objective To evaluate the effect of hydrogen peroxide (H2O2) on autophagy in melanocytes,and to explore its possible regulatory mechanisms.Methods Normal human melanocytes at exponential growth phase were divided into several groups:blank control group receiving no treatment,positive control group treated with 100 nmol/L sirolimus solution,and experiment groups treated with H2O2 solution at different volume fractions of 10-7-10-3 respectively.After 4-hour treatment,cell counting kit-8 (CCK-8) assay and flow cytometry were performed to evaluate the cellular proliferative activity and detect apoptosis of melanocytes respectively.Acridine orange staining was performed to detect autophagosome formation,transmission electron microscopy to observe ultrastructural changes of autophagosomes,and Western blot analysis to measure the expression of autophagy-specific protein Beclin 1 and microtubuleassociated protein 1 light chain 3B (LC3B).A total of 84 autophagy-related genes were analyzed by RT2 Profiler PCR Array,so as to screen differentially expressed autophagy-related genes.Results After the treatment with H2O2 at different volume fractions of 10-3,5 × 10-4,10-4,5 × 10-5,10-5,5 × 10-6 and 10-6,experiment groups showed significantly decreased cellular proliferative activity,but significantly increased apoptosis rate compared with the blank control group (F =286.95,301.23,respectively,both P ＜ 0.05).With the increase in volume fractions of H2O2,the cellular proliferative activity was significantly gradually decreased (P ＜ 0.05),while the apoptosis rate showed an opposite trend (P ＜ 0.05),except that the 5 ×10-6 H2O2 group showed no significant differences in the apoptosis rate compared with the 10-5 H2O2 group and 10-6 H2O2 group.Acridine orange staining and electron microscopy showed autophagosome formation in the 10-5 H2O2 group,10-6 H2O2 group and positive control group.Western blot analysis revealed that Beclin1 expression and LC3B-Ⅱ/LC3B-Ⅰ ratio were significantly higher in the 10-5 H2O2 group,10-6 H2O2 group and positive control group than in the blank control group (all P ＜ 0.05).RT2 Profiler PCR Array showed significant up-regulation of ATG12,ATG3,ULK1,PIK3CG,PTEN and PIK3C3 genes and significant downregulation of EIF2AK3 gene in the 10-5 H2O2 group,10-6 H2O2 group and positive control group compared with the blank control group.In the 10-5 H2O2 group and positive control group,the mTOR gene was significantly up-regulated,and the ULK2 gene was significantly down-regulated.The 10-6 H2O2 group showed no obvious changes in the expression of mTOR gene,but significant up-regulation of AMPK and JNK1 genes.Conclusion H2O2 at volume fractions of 10-5 and 10-6 can induce autophagy in melanocytes,likely by influencing the expression of some related signaling molecules.

[Objectives]To investigate the protective effects of recombinant Trichinella spiralis excretory-secretory 53ku pro-tein(rTsP53)on acute lung injuries in mice.[Methods]Thirty Balb/c mice were randomly divided into normal group. ALI group and rTsP53 group(n=10,respectively). Macrophages were harvested by bronchoalveolar lavage. Mortality in 72 hours was counted and compared. Pathological damage of lung tissues was observed by HE staining and graded by Smith score. Wet/dry ratio was measured. The bronchoalveolar lavage fluid concertration of IL-6 and IL-4 was measured by ELISA. The mRNA expression of TNF-α,iNOS, IL-10 and Arg-1 in alveolar lavage macrophages was detected by RT-PCR.[Results]72 h mortality of ALI mice was 70%,which was reduced to 30% in mice received rTsP53 treatment. Compared with ALI mice,the pathological damage of in rTsP53 treated-mice was improved and Smith score was declined ,combined with descending W/D ratio. IL-6 level of alveolar lavage fluid was elevated in ALI mice compared with normal group. And alveolar lavage macrophage was polarized to M2 sub-type,appeared as higher mRNA expression of TNF-α and iNOS and lower level of IL-10 and Arg-1. Bronchoalveolar lavage fluid concentration of IL-6 was declined and IL-4 was elevated in rTsP53-treated mice compared with ALI group. The macrophages of alveolar wash had higher mRNA expression of IL-10 and Arg-1,while lower level of TNF-α and iNOS,manifesting M2 polarization characteristics.[Conclusion]Recombinant T.spiralis P53 protein could protect mice from acute lung injuries induced by LPS via modulating M2 macrophage polarization,which play a role in depression of inflammatory reaction and tissue repairment.

Objective: To understand the viral etiology of a clustered case of human infection outbreak in the middle school of Huai’an city. Methods: Nasopharyngeal swab samples from patients were collected and rapidly detected by Real-time RT-PCR and the target virus isolated in cells. Furthermore, HA1 segments of target virus were amplified by RT-PCR and sequenced. The genetic and phylogenetic analysis based on HA1 genes was computed. Results: Influenza A(H1N1)pdm09 viral nucleic acid in 11 nasopharyngeal swab samples from patients in the outbreak were positive. Compared to the vaccine strains A/California/07/2009, the Huai’an isolates, nucleotide identity was 97.7%-98.1%, and amino acid identity was 96.6%-97.4%. Phylogenetic analysis of HA1 segment sequences indicated that the Huai’an strains from the outbreak were related closely to the viruses isolated in the year of 2014. Sequence analysis indicated that the Huai’an isolates had no amino acid substitution in the receptor binding sites and glycosylation sites, while in the Ca1 of antigenic determinant of HA1 the Huai’an isolates had an amino acid substitution of S for T at 220. Conclusions The pathogen of the clustered case of human infection was Influenza A(H1N1)pdm09 virus. Though the Huai’an isolates had one animo acid substitution in the Ca1 of antigenic determinant, the antigenicity characteristic remained unchanged.

Objective To clone express and purify Schistosoma japonicum fructose?1 6?bisphosphate aldolase SjFBPA in E. coli and observe its expression in different developmental stages of S. japonicum. Methods FBPA gene was amplified from S. japonicum adult worm cDNA by using PCR. The amplified product was recombined into pET28a plasmid and inducibly expressed with IPTG in E. coli BL21. SDS?PAGE and Western blotting were employed to analyze and identify the recombinant protein SjFBPA rSjFBPA . Then rSjFBPA was purified by chromatographic purification and its purity was analyzed by SDS?PAGE. The protein concentration of rSjFBPA purified was measured by the BCA method. Furthermore SjFBPA mRNA was ana?lyzed in different developmental stages of S. japonicum by RT?PCR. Results SjFBPA was successfully amplified by using PCR and identified by restriction enzyme digestion and sequencing. The Western blotting analysis confirmed that the recombinant pro?tein could specifically reactive to the anti?His?tag monoclonal antibody. The concentration of the purified recombinant protein was about 4 mg/ml. The result of RT?PCR showed that SjFBPA mRNA was expressed in cercaria schistosomulum adult worm and egg of S. japonicum. Conclusion SjFBPA is successfully recombined and expressed in a prokaryotic system and SjFBPA mRNA is expressed in cercaria schistosomulum adult worm and egg of S. japonicum.

Objective To compare the accuracy of nucleic acid detection and antibody detection for diagnosing human immunode‐ficiency virus(HIV) infection .Methods Retrospectively analysed data of nucleic acid detection and antibody detection from 124 ca‐ses of patients diagnosed with HIV infection from 2005 to 2014 .The positive rates of the two methods were compared respectively in patients with early‐stage of HIV infection(76 cases) and patients with intermediate and advanced stage of HIV infection (48 ca‐ses) .Results In patients with early‐stage of HIV infectionn ,the positive rate of nucleic acid detection (94 .74% ) was higher than that of antibody detection (84 .21% );while in patients with intermediate and advanced stage of HIV infection ,the positive rate of antibody detection(97 .92% ) was higher than that of nucleic acid detection (81 .25% );both had statistically significant difference (P<0 .05) .Conclusion On the early stage of HIV infection ,the accuracy of nucleic acid detection is higher than that of antibody detection ;while on the intermediate and advanced stage of HIV infection ,antibody detection shows better accuracy .