Herpes simplex virus type 2 (HSV-2) is a common sexually transmitted pathogen known for causing genital herpes. Glycoprotein B (gB) of HSV-2 plays a crucial role in viral entry and infection. However, its post-translational modifications are not well understood. This study investigated the palmitoylation of HSV-2 gB, identifying cysteine residue 8 (C8) as a critical palmitoylation site. Using an acyl-biotin exchange assay, we confirmed that gB was indeed palmitoylated, and treatment with 2-bromopalmitate significantly reduced this modification. Further analysis revealed that ZDHHC14 interacted with gB and facilitated its palmitoylation. We also identified APT2 as a negative regulator of gB palmitoylation. Importantly, palmitoylation enhanced gB's localization to the plasma membrane, whereas the C8S mutation substantially impaired this localization. Functionally, palmitoylation of gB enhanced the infective efficiency of HSV-2 pseudotyped particles (pp) and live viruses, while both palmitoyltransferase and APT2 inhibition affected the entry efficiency of HSV-2pp. Our findings demonstrate that the palmitoylation of gB is crucial for its localization to the membrane and for facilitating efficient HSV-2 infection, highlighting palmitoylation as a promising target for antiviral interventions and providing new insights into the pathogenesis of HSV-2.

Nairobi Sheep Disease (NSD) is a typical tick-borne syndrome characterized by severe hemorrhagic gastroenteritis, spontaneous abortion, and a high case fatality rate in small ruminants. The pathogenic agent, Nairobi sheep disease virus (NSDV), has also been associated with human infections, indicating its possible zoonotic potential. Prior to this study, NSDV has been detected from ticks collected in Jilin, Hubei, and Liaoning provinces in China. In the present study, a total of 343 ticks (Haemaphysalis longicornis) were collected in Shandong province, China in 2020, and pooled into 16 libraries. Analysis of the meta-transcriptomic sequencing data identified NSDV strains SDWL07, SDWL08, and SDWL16 from three pools. The SDWL07 and SDWL16 strains were detected from unfed ticks, while SDWL08 was detected from cattle-feeding ticks. Phylogenetic analyses showed higher sequence identities between the three strains and other Chinese NSDV strains than those from India and Kenya. Phylogenetic analyses also revealed that they clustered together and fell within the China lineage, suggesting no potential genetic reassortment among them. In summary, this is the first report of the identification of NSDV in Shandong province, highlighting the continually expanding endemic regions of this pathogen. Surveillance of NSDV should be intensified in China, especially in areas where H. longicornis is endemic.

目的 评价以壳聚糖为佐剂的白喉类毒素(diphtheria toxoid,DT)疫苗对小鼠的免疫效果,为DT新型佐剂疫苗的制备提供实验依据.方法 将30只雄性C57BL/6小鼠分为黏膜免疫组和体液免疫组,每组再随机分为6组:阴性对照组(PBS)、阳性对照组(DT)、铝佐剂组、壳聚糖佐剂组(配成含壳聚糖0.5％的终溶液)、铝佐剂疫苗组和壳聚糖佐剂疫苗组,每组5只.黏膜免疫组和体液免疫组分别通过滴鼻和腹腔注射接种1次,PBS缓冲液500μL/只,DT 10 μg/只,铝佐剂500 μg/只,壳聚糖佐剂500μL/只.观察小鼠状态,并分别于接种前(0 d)、接种后7、21、35 d采集小鼠眼球血清,同时收集鼻洗液,采用ELISA法分别检测血清IgG抗体水平和鼻洗液sIgA抗体水平.结果 小鼠腹腔注射及鼻滴壳聚糖佐剂疫苗后,精神及行为状态正常.壳聚糖佐剂疫苗可有效诱导小鼠产生白喉IgG抗体,与经典铝佐剂疫苗组相比,免疫后35 d差异无统计学意义(F=0.742＜F0.05(1 8),P＞0.05);也可有效诱导小鼠产生sIgA,免疫后21 d明显高于经典铝佐剂疫苗组(F=127.926＞F0.05(1.8),P＜0.05).结论 壳聚糖佐剂DT疫苗安全性良好,可有效诱导小鼠黏膜免疫和体液免疫,具有替代铝佐剂制备疫苗的潜力,且可考虑制备鼻滴疫苗.

Estrogen has long been known to possess immune-modulatory effects in diseases, and multiple pathological conditions show great sex disparities. However, the impact of estrogen in Neisseria meningitidis infection has not been determined. The present study aimed to investigate the role of estrogen in N. meningitidis infection and the molecular mechanism. We selected 35 N. meningitidis isolates representing different clonal complexes (cc), serogroups, and isolation sources to infect the HBMEC cell line. Results showed that the expression of estrogen receptor (ER) β in N. meningitidis-infected cells was downregulated compared with that in normal cells. The expression of ERβ induced by invasive isolates was lower than that in carriers. Serogroup C isolates induced the lowest expression of ERβ compared with serogroup A and B isolates. We used four cc4821 N. meningitidis isolates to infect two kinds of host cells (human brain microvascular endothelial cells and meningeal epithelial cells). The results showed that 17 β-estradiol (E2) could inhibit the release of inflammatory factors interleukin (IL)-6, IL-8, and tumor necrosis factor-α after N. meningitidis infection via TLR4. E2 could inhibit the activation of the p38-MAPK signal pathway induced by N. meningitidis infection through binding to ERβ, and significantly inhibit the release of inflammatory factors in N. meningitidis-infected host cells. This study demonstrated that estrogen plays a protective role in N. meningitidis infection. ERβ is potentially associated with the release of inflammatory cytokines in N. meningitidis infection, which sheds light on a possible therapeutic strategy for the treatment of invasive diseases caused by N. meningitidis.

Ticks can transmit numerous pathogens and harbor diverse microbial communities. Considerable progress has been made in the characterization of the bacterial profiles of ticks, whereas other members of tick microbiota (such as fungi and viruses) and the functional characteristics of ticks warrant further exploration. To investigate the taxonomic and functional profiles and explore potential pathogens they were carrying, samples of different developmental stages and of both sexes of Haemaphysalis longicornis were collected in the present study and the metagenomic deep sequencing method was applied. Metagenomic deep sequencing results revealed that bacteria were predominant, followed by fungi, viruses, archaea and metazoans. Proteobacteria was the dominant phylum in the microbiota of H. longicornis. The abundance of microbial species varied significantly among groups, the bacteria of nymphs and female adults demonstrated unique characteristics, and the microbial community of males overlapped with those of nymphs and females. Functional annotation results demonstrated that the metagenomic sequences of the three groups were classified under metabolism, genetic information processing, environmental information processing and cellular processes. Differences in functional characteristics were observed in both the pathways composition and abundance of carbohydrate-active enzymes. Furthermore, whole metagenome sequencing helped to elucidate the diversity of pathogens carried by H. longicornis, which may facilitate further research attempting to prevent and control tick-borne diseases.

Shandong Province, China, has been implementing a malaria elimination program. In this study, we analyzed the epidemiological characteristics of malaria imported into Shandong Province between 2012 and 2017 to provide scientific data for the elimination of malaria. In this epidemiological study, we examined the status of malaria in 2012–2017 in Shandong Province, China. Data on all cases of malaria were collected from the online Infection Diseases Monitor Information System to describe and statistically analyze the sources of infection, species of parasite, populations affected, regional distributions, incidence, and temporal distributions of malaria. In total, 1053 cases of malaria were reported in 2012–2017, and all of them were imported. Plasmodium falciparum was the predominant species (77.6%) in Shandong Province; P. vivax malaria accounted for 10.9% of the total number of cases, P. ovale malaria for 2.9%, and P. malariae malaria for 8.2%. Most patients were male (96.8%), most were aged 21–50 years (87.2%), and migrant laborers (77.2%) and workers (6.6%) were at highest risk. The origin of the largest number of imported cases was Africa (93.4%), followed by Asia (5.9%) and Oceania (0.4%). Most cases of imported malaria occurred in June each year and 70% of cases were recorded in six cities during the period of 2012–2017. It is necessary to strengthen malaria surveillance among workers returning home from Africa and Southeast Asia, and to conduct timely blood tests to diagnose and treat imported infections.

Abstract The microbiota plays an important role in the growth of mosquitoes and the transmission of mosquito‐borne pathogens. The effects of changes in aquatic habitats in which mosquitoes live, as one of the major factors closely associated with the microbial communities of mosquitoes, on the microbiota of different developmental stages remain to be elucidated. Here, we compared the microbiota of larvae and pupae of Aedes albopictus exposed to different ampicillin concentrations and investigated the bacterial composition of adult females. The results demonstrate that the microbial community differed substantially between developmental stages and that samples of the same stages shared similarities, whereas differences were observed between adult females. Based on all observations, we hypothesize that the use of ampicillin caused dysbiosis rather than excluding bacteria from mosquitoes and that the disturbing effect of ampicillin was obvious in adults. Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed that most of the bacteria identified in this study were significantly associated with metabolism. Taken together, our results indicate that ampicillin can change the abundance of bacteria, while microbial communities of Ae. albopictus showed obvious stage‐specific characteristics. Further investigations are needed to characterize specific bacterial components that are affected by ampicillin exposure and to quantify their functions, thereby providing a better understanding of the influence of antibiotics on microbial communities at different life stages.

As one of the most important vectors, Haemaphysalis longicornis can transmit a variety of pathogens and is widely distributed in China. It has been reported that the bacterial community in ticks can impact tick fitness, development, and reproduction and even the transmission of tick-borne pathogens. In this study, bacterial diversity across all developmental stages (eggs, larvae, nymphs and adults) of H. longicornis was investigated using high-throughput sequencing technology. The results demonstrated that Proteobacteria was the dominant phylum and that Coxiella was the most abundant bacterial genus across all the samples. Alpha diversity analysis demonstrated that the eggs had the highest bacterial richness and diversity, and the bacterial community of the larvae was found to be similar to that of the eggs. However, there was a rapid increase in the relative abundance of Coxiella upon development of larvae to nymphs. Females exhibited the lowest bacterial diversity, and the proportion of Coxiella decreased from 85% in females to 45% in males. Our results suggest that H. longicornis lost most of the bacteria present in the early developmental stages and re-established the bacterial community after bloodmeals and molting.

Dermacentor silvarum is an important vector of tick-borne pathogens. The microbiota of ticks has been recognized to influence their development, fitness, and reproduction as well as the acquisition, establishment and transmission of pathogens. Eggs, larvae, nymphs and adults (females and males) of D. silvarum were used in this study to evaluate microbial community and diversity across different developmental stages. The results demonstrated that some developmental stages host different bacterial species, confirming that each stage of development could have a specific associated microbiota. Proteobacteria was the dominant phylum in most stages, while Actinobacteria was the most abundant in nymphs. The abundance of Brevibacterium significantly increased from the egg stage to the following stages, and there was a sharp increase in the abundance of this genus among nymphs. Eggs showed the highest microbial richness and diversity, and the microbial community of eggs exhibited high similarity to that of females. Moreover, the high abundance of Coxiella in eggs and females further indicated that members of this genus are vertically transmitted symbionts. Nymphs showed a microbial composition distinct from those of the other stages, and the microbial community associated with males possessed higher species richness and greater bacterial diversity than that of females. Further investigations are needed to determine the functions of the microbiota and provide information enabling a better understanding of the diversity of the microbial community at different life stages.

Haemaphysalis longicornis (Acari: Ixodidae) is an important vector tick that is widely distributed around the world. In many regions, this tick acts as vector of a wide range of pathogens to humans and animals, and its control is mainly based on the use of chemical pesticides. However, the occurrence of some adverse effects, such as tick resistance to pesticides and food and environmental contamination, are driving the need to develop more effective and environmentally sound approaches to control and prevent ticks. As an alternative control strategy, entomopathogenic fungi have been extensively used for the control of pests and cause high mortality in various ticks. In this study, we identified four isolates of Beauveria bassiana sensu lato from insects and investigated their pathogenicity against different developmental stages of H. longicornis (eggs, unfed larvae, unfed nymphs and engorged females). Phylogenetic analysis demonstrated that the four isolates of B. bassiana clustered into two clades. Four isolates showed different acaricidal qualities: the isolate from Cerambycidae (EF3) exhibited the highest pathogenicity to all developmental tick stages tested. High doses (1 × 107 conidia/ml) of the clade I fungi collected from Cryptotympana atrata fabricus (Cicadidae) (EF1), Cimicidae (EF2), and Boettcherisca peregrine (Sarcophagidae) (EF4) also showed virulence against H. longicornis, with high doses of the fungi application causing higher mortality than control group. Altogether, this study demonstrated that all four isolates of B. bassiana showed high virulence toward different developmental stages of H. longicornis, and therefore, they can be of potential use as biological control agents of ticks.

Haemaphysalis longicornis is a prominent tick species in China, and the major vector of an emerging tick‐borne disease: severe fever with thrombocytopenia syndrome (SFTS). Microbiome diversity of ticks is influenced by several factors. In this study, we investigated microbiome diversity in field‐collected female and male H . longicornis ticks and compared the microbial composition of fed and unfed ticks and of those feeding on different hosts using barcode sequencing of V3–V4 region of 16S RNA gene. Regardless of sex, host, and feeding status; the highest abundance among all samples was found for the genus Coxiella . The relative numbers of Coxiella sequences decreased with the length of the blood feeding, whereas the numbers of Staphylococcus and Corynebacterium increased gradually. The dominance of Coxiella across all samples indicates that it is an obligate symbiont of H . longicornis . Overall, higher microbiome richness was detected in male ticks than in female ticks. Fed ticks showed a more diverse microbe composition than unfed ticks, and ticks fed on goats exhibited the highest diversity. These findings of this study can serve as a basis for future studies of microbiota biology and interactions between the microbes and pathogens of H . longicornis .

The effects of deoxynivalenol in different dose including 100, 200, 500, 1000, and 1500 mu g/L on primary cranial osteoblasts from fetal mice were investigated. Fluorescence staining, flow cytometric measurement, 3-(4,5-dimethylthiazol 2-y1)-3,5-di-phenyltetrazolium bromide assay, quantitative PCR, and Western blot were used for the test. Mineralization and proliferation of osteoblasts decreased upon 100 mu g/L and higher deoxynivalenol treatment and apoptosis of osteoblasts was increased upon 500, 1000, and 1500 mu g/L deoxynivalenol treatment. Karyopyknosis, membrane breakage, and a decreased number of calcium nodes were also observed upon 500 mu g/L deoxynivalenol treatment. The mRNA and protein levels of B-cell lymphoma-2-associated X protein were upregulated, B-cell lymphoma-2 protein downregulated with increasing concentrations of deoxynivalenol treatment and their ratio increased. Deoxynivalenol induces apoptosis of osteoblasts, suggesting a mechanism by which deoxynivalenol can affect murine skeletal development.

Influenza A virus causes periodic outbreaks and seriously threatens human health. The drug-resistant mutants have shown an epidemic trend because of the abuse of chemical drugs. Aloe polysaccharides (APS) extracted from Aloe vera leaves have evident effects on the therapy of virus infection. However, the activity of APS in anti-influenza virus has yet to be investigated. Here, we refined polysaccharides from A. vera leaf. In vitro test revealed that APS could inhibit the replication of a H1N1 subtype influenza virus, and the most obvious inhibitory effect was observed in the viral adsorption period. Transmission electron microscopy indicated that APS directly interacted with influenza virus particles. Experiments on PR8 (H1N1) virus infection in mice demonstrated that APS considerably ameliorated the clinical symptoms and the lung damage of the infected mice, and significantly reduced the virus loads and mortality. Our findings provided a theoretical basis for the development of novel natural anti-influenza agents.

Objective To prepare Helicobacter pylori Omp18 protein and polyclonal antibodies against that protein Methods The Omp18 gene was amplified using PCR and cloned into the vector pMD-18T.It was then was cloned into the prokaryotic expression plasmid vector pTriEx(TM)-4.The vector was transformed into Escherichia coli JM109DE.Omp18 protein was produced and purified,and then rabbits were repeatedly administered the protein to produce polyclonal antibodies.Results The Omp18 gene was successfully cloned into the prokaryotic expression vector pTriEx(TM)-4 and the vector was successfully transformed into E.coli JM109DE.Omp18 protein was successfully produced,and purified Omp18 protein was administered to rabbits to yield polyclonal antibodies.Conclusion Omp18 protein was successfully expressed in prokaryotic cells,and polyclonal antibodies were successfully prepared in rabbits.This research will benefit study of the function and regulation of Omp18 and it will lay a foundation for detection of H.pylori and preparation of vaccines.

Notch1是参与调控细胞分化的重要分子,其表达异常与许多肿瘤的转移密切相关.在乳腺癌发展过程中,Notch1分子可能通过增强乳腺癌干细胞迁徙活性促进其转移,如果证实这一机制,对于研制靶向于乳腺癌干细胞Notch1分子的药物,阻断其转移有重要意义.

Objective To prepare Helicobacter pylori Omp18 protein and polyclonal antibodies against that protein Methods The Omp18 gene was amplified using PCR and cloned into the vector pMD-18T.It was then was cloned into the prokaryotic expression plasmid vector pTriEx(TM)-4.The vector was transformed into Escherichia coli JM109DE.Omp18 protein was produced and purified,and then rabbits were repeatedly administered the protein to produce polyclonal antibodies.Results The Omp18 gene was successfully cloned into the prokaryotic expression vector pTriEx(TM)-4 and the vector was successfully transformed into E.coli JM109DE.Omp18 protein was successfully produced,and purified Omp18 protein was administered to rabbits to yield polyclonal antibodies.Conclusion Omp18 protein was successfully expressed in prokaryotic cells,and polyclonal antibodies were successfully prepared in rabbits.This research will benefit study of the function and regulation of Omp18 and it will lay a foundation for detection of H.pylori and preparation of vaccines.

目的:评价PBL+ LBL双轨教学法在留学生医学微生物学教学中应用效果.方法:采用期末考试成绩和问卷调查综合评价PBL+ LBL双轨教学法的教学效果.结果:期末考试实验班学生成绩显著高于对照班学生成绩(P＜0.05).90％以上的学生认为PBL+ LBL双轨教学模式可激发学习兴趣,提高自身多重能力.结论:PBL+ LBL双轨教学法在留学生的教学效果明显优于传统LBL教学.

Objective The effects of Escherichia coli on the motility,ultrastructure,and apoptosis of human sperm in vitro were studied in order to identify the cause of male infertility and facilitate diagnosis and therapy for male infertility caused by pathogenic infections.Methods Semen samples were obtained from 30 healthy volunteers.Four groups of sperm suspensions were prepared: a normal sperm suspension serving as negative control(Group A),a sperm suspension mixed with suspended E.coli(Group B),a sperm suspension mixed with suspended E.coli treated with traditional Chinese medicine(Group C),and a sperm suspension mixed with suspended E.coli treated with furadantin(Group D).These samples were collected at 0,2,and 4 h.Motility parameters were analyzed using a computer-aided semen analysis(CASA) system.Changes in the ultrastructure of human sperm were observed with transmission electron microscopy.The rate of apoptosis of semen sperm was detected with flow cytometry combined with FITC-Annexin V/PI dual color fluorescence staining.Results Group B had significantly lower motility parameters,including straight line velocity(VSL),curve velocity(VCL),average path velocity(VAP),amplitude of lateral head displacement(ALH),and progressive motility,in comparison to the other groups at 4h(P0.01).There were no significant differences in the ultrastructure of sperm in the control group(A) and groups treated with medicine(C and D),while there were significant changes in the ultrastructure of sperm in Group B.In addition,the rate of sperm apoptosis increased significantly(P0.01) in Group B.Conclusion E.coli significantly inhibited sperm motility,changed the ultrastructure of sperm,and increased the rate of apoptosis of sperm in vitro.Therefore,E.coli infection may be one cause of male infertility.

IFN-γ plays an indirect anti-cancer role through the immune system but may have direct negative effects on cancer cells. It regulates the viability of gastric cancer cells, so we examined whether it affects their proliferation and how that might be brought about. We exposed AGS, HGC-27 and GES-1 gastric cancer cell lines to IFN-γ and found significantly reduced colony formation ability. Flow cytometry revealed no effect of IFN-γ on apoptosis of cell lines and no effect on cell aging as assessed by β-gal staining. Microarray assay revealed that IFN-γ changed the mRNA expression of genes related to the cell cycle and cell proliferation and migration, as well as chemokines and chemokine receptors, and immunity-related genes. Finally, flow cytometry revealed that IFN-γ arrested the cells in the G1/S phase. IFN-γ may slow proliferation of some gastric cancer cells by affecting the cell cycle to play a negative role in the development of gastric cancer.