江西省口岸区域白纹伊蚊特异性病毒Aedes albopictus anphevirus基因组序列研究

目的昆虫特异性病毒在白纹伊蚊中与致病性传染病病原体如登革病毒、基孔肯雅病毒和寨卡病毒等的传播和复制有着密切联系,本文对其基因组序列进行研究,为研发新型虫媒病毒防治策略提供理论基础。方法本研究利用二代测序技术,对江西省3个口岸区域(南昌昌北机场、九江城西港和赣州陆港)采集的野外白纹伊蚊种群携带的昆虫特异性病毒进行高通量测序。结果3个白纹伊蚊种群中广泛存在昆虫特异性病毒Aedes albopictus anphevirus(AealbAV)。利用MEGAHIT软件对这些病毒reads进行组装,得到2株AealbAV全基因组序列,与美国株MW147277.1高度同源(相似度均大于98%),基因登录号分别为九江株OR715784,赣州株OR729834,南昌株仅拼接出部分序列。结论AealbAV在江西白纹伊蚊种群中普遍存在,其全基因组研究为口岸区域利用昆虫特异性病毒进行白纹伊蚊综合防治提供了理论基础。

56 Years of the Marburg Virus—A Review of Therapeutics

Background: The Marburg virus (MARV) is the causative agent of Marburg virus disease (MVD). This filovirus first appeared in 1967 and has since caused several outbreaks with case fatality rates between 23% and 90%. The earliest cases of MVD are thought to be caused by exposure to an infected animal, either a reservoir host (some bat species, e.g., Rousettus aegyptiacus) or a spill-over host, such as non-human primates. The virus is spread between people by direct contact with blood or other bodily fluids (including saliva, sweat, faeces, urine, tears, and breast milk) from infected individuals. Despite the high fatality rate, the Marburg virus has no vaccine or drug treatment. Recent outbreaks of the virus in 2023 in Tanzania and Equatorial Guinea have reignited the need to develop effective therapeutics, especially in the wake of the COVID-19 pandemic. Purpose: This review seeks to highlight the drug discovery efforts aimed at developing vaccines or possible treatments as potential therapeutics. Several existing antiviral agents are being probed, and vaccines are in pre-clinical and clinical stages. Natural products are also an important source of possible drugs or lead compounds and when coupled with computational techniques, these strategies offer possible therapeutics for the Marburg virus, especially in Africa, which has a high disease burden. Methods: Using the search engines Google Scholar and PubMed;keywords e.g. Marburg virus, Marburg treatments, Marburg virus drug discovery were utilized. Several results were yielded, and articles published in recent years were accepted into the final list.Results and Conclusion: This study shows there is a growing interest in therapeutics for the Marburg virus, especially with the recent outbreaks and pandemic preparedness. Initiatives that to support vaccine development and access like the MARVAC consort time are critical to fighting this public health threat.

Virus Removal by Iron Coagulation Processes

Waterborne viruses account for 30% to 40% of infectious diarrhea, and some viruses could persevere for some months in nature and move up to 100 m in groundwater. Using filtration setups, coagulation could lessen virus charges as an efficient pre-treatment for reducing viruses. This work discusses the present-day studies on virus mitigation using coagulation in its three versions i.e., chemical coagulation (CC), enhanced coagulation, and electrocoagulation (EC), and debates the new results of virus demobilization. The complexity of viruses as bioparticles and the process of virus demobilization should be adopted, even if the contribution of permeability in virus sorption and aggregation needs to be clarified. The information about virion permeability has been evaluated by interpreting empirical electrophoretic mobility (EM). No practical measures of virion permeability exist, a clear link between permeability and virion composition and morphology has not been advanced, and the direct influence of inner virion structures on surface charge or sorption has yet to be conclusively demonstrated. CC setups utilizing zero-valent or ferrous iron could be killed by iron oxidation, possibly using EC and electrooxidation (EO) methods. The oxidants evolution in the iron oxidation method has depicted promising findings in demobilizing bacteriophage MS2, even if follow-up investigations employing an elution method are needed to secure that bacteriophage elimination is related to demobilization rather than sorption. As a perspective, we could be apt to anticipate virus conduct and determine new bacteriophage surrogates following subtle aspects such as protein structures or genome size and conformation. The present discussion’s advantages would extend far beyond an application in CC—from filtration setups to demobilization by nanoparticles to modeling virus fate and persistence in nature.

Effect of hepatitis C virus infection on expression of several cancer-associated gene products in hepatocellular carcinoma

AIM To study hepatocarcinogenesis of hepatitis C virus (HCV). METHODS Expression of HCV antigens (CP10, NS3 and NS5) and several cancer associated gene products (ras p21, c myc, c erbB 2, mutated p53 and p16 protein) in the tissues of hepatocellular carcinoma (HCC, n =46) and its surrounding liver tissue were studied by the ABC (avidin biotin complex) immunohistochemical method. The effect of HCV infection on expression of those gene products in HCC was analyzed by comparing HCV antigen positive group with HCV antigen negative group. RESULTS Positive immunostaining with one, two or three HCV antigens was found in 20 (43 5%) cases, with either of two or three HCV antigens in 16 (34 8%) cases, and with three HCV antigens in 9 (19 6%) cases. Deletion rate of p16 protein expression in HCC with positive HCV antigen (80%, 16/20) was significantly higher than that in HCC with negative HCV antigen. Whereas no significant difference of the other gene product expression was observed between the two groups. CONCLUSION HCV appears related to about one third of cases of HCC in Chongqing, the southwest of China, and it may be involved in hepatocarcinogenesis by inhibiting the function of p16 gene, which acts as a negative regulator of cell cycle.

Hepatitis C virus in human B lymphocytes transformed by Epstein-Barr virus in vitro by in situ reverse transcriptase-polymerase chain reaction

AIM: To study persistence and replication of hepatitis C virus (HCV) in patients' peripheral blood mononuclear cells (PBMC) cultured in vitro. METHODS: Epstein Barr virus (EBV) was used to transform the hepatitis C virus from a HCV positive patient to permanent lymphoblastoid cell lines (LCL). Positive and negative HCV RNA strands of the cultured cells and growth media were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) each month. Core and NS5 proteins of HCV were further tested using immunohistochemical SP method and in situ RT-PCR. RESULTS: HCV RNA positive strands were consistently detected the cultured cells for one year. The negative-strand RNA in LCL cells and the positive-strand RNA in supernatants were observed intermittently. Immunohistochemical results medicated expression of HCV NS3 and C proteins in LCL cytoplasm mostly. The positive signal of PCR product was dark blue and mainly localized to the LCL cytoplasm. The RT-PCR signal was eliminated by overnight RNase digestion but not DNase digestion. CONCLUSION: HCV may exist and remain functional in a cultured cell line for a long period.

Design,delivery and efficacy testing of therapeutic nucleic acids used to inhibit hepatitis C virus gene expression in vitro and in vivo

Despite major achievements in the treatment ofchronic hepatitis C with the combination ofinterferons and the nucleoside analog ribavirin themajority of patients with chronic hepatitis C virus(HCV) infection cannot be treated effectively.Toimprove this response rate we used antisensetechnologies to inhibit HCV translation as possibleadditional option for experimental treatment.Antisense oligodeoxynucleotides(ODN) are

Transfusion transmitted virus infection in general populations and patients with various liver diseases in south China

INTRODUCTIONAlthough several specific detecting methods hadbeen applied to determine the hepatitis virus,therewas a lot of cryptogenic hepatitis without anyknown hepatitis infectious marker.Theprevalence of hepatitis G virus (HGV) (also knownas GB-C virus) infection has been reported to be 5%-13% in patients with non-A-E hepatitis andcirrhosis,however,there is little evidencesuggesting that HGV causes hepatitis in human.

Could autophagy dysregulation link neurotropic viruses to Alzheimer’s disease?

Neurotropic herpesviruses have been associated with the onset and progression of Alzheimer’s disease,a common form of dementia that afflicts a large percentage of elderly individuals.Interestingly,among the neurotropic herpesviruses,herpes simplex virus-1,human herpesvirus-6A,and human herpesvirus-6B have been reported to infect several cell types present in the central nervous system and to dysregulate autophagy,a process required for homeostasis of cells,especially neurons.Indeed autophagosome accumulation,indicating an unbalance between autophagosome formation and autophagosome degradation,has been observed in neurons of Alzheimer’s disease patients and may play a role in the intracellular and extracellular accumulation of amyloidβand in the altered protein tau metabolism.Moreover,herpesvirus infection of central nervous system cells such as glia and microglia can increase the production of oxidant species through the alteration of mitochondrial dynamics and promote inflammation,another hallmark of Alzheimer’s disease.This evidence suggests that it is worth further investigating the role of neurotropic herpesviruses,particularly human herpesvirus-6A/B,in the etiopathogenesis of Alzheimer’s disease.

A duplex RT-PCR assay for detection of H9 subtype avian influenza viruses and infectious bronchitis viruses

H9 s ubtype avian influenza virus（AIV） and infectious bronchitis virus（IBV） are major pathogens circulating in poultry and have resulted in great economic losses due to respiratory disease and reduced egg production. As similar symptoms are elicited by the two pathogens, it is difficult for their differential diagnosis. So far, no reverse transcription-polymerase chain reaction（RT-PCR） assay has been found to differentiate between H9 AIV and IBV in one reaction. Therefore, developing a sensitive and specific method is of importance to simultaneously detect and differentiate H9 AIV and IBV. In this study, a duplex RT-PCR（d RT-PCR） was established. Two primer sets target the hemagglutinin（HA） gene of H9 AIV and the nucleocapsid（N） gene of IBV, respectively. Spec ific PCR products were obtained from all tested H9 AIVs and IBVs belonging to the major clades circulating in China, but not from AIVs of other subtypes or other infectious avian viruses. The sensitivity of the d RT-PCR assay corresponding to H9 AIV, IBV and mixture of H9 AIV and IBV were at a concentration of 1×10^1, 1.5×10^1 and 1.5×10^1 50% egg infective doses（EID_（50）） m L^–1, respectively. The concordance rates between the d RT-PCR and virus isolation were 99.1 and 98.2%, respectively, for detection of samples from H9N2 AIV or IBV infected chickens, while the concordance rate was 99.1% for detection of samples from H9N2 AIV and IBV co-infected chickens. Thus, the d RT-PCR assay reported herein is specific and sensitive, and suitable for the differential diagnosis of clinical infections and survei llance of H9 AIVs and IBVs.

Laboratory biosafety for handling emerging viruses

Emerging viruses are viruses whose occurrence has risen within the past twenty years,or whose presence is likely to increase in the near future.Diseases caused by emerging viruses are a major threat to global public health.In spite of greater awareness of safety and containment procedures,the handling of pathogenic viruses remains a likely source of infection,and mortality,among laboratory workers.There is a steady increase in both the number of laboratories and scientist handling emerging viruses for diagnostics and research.The potential for harm associated to work with these infectious agents can be minimized through the application of sound biosafety concepts and practices.The main factors to the prevention of laboratory-acquired infection are well-trained personnel who are knowledgable and biohazard aware,who are perceptive of the various ways of transmission,and who are professional in safe laboratory practice management.In addition,we should emphasize that appropriate facilities,practices and procedures are to be used by the laboratory workers for the handling of emerging viruses in a safe and secure manner.This review is aimed at providing researchers and laboratory personnel with basic biosafety principles to protect themselves from exposure to emerging viruses while working in the laboratory.This paper focuses on what emerging viruses are,why emerging viruses can cause laboratory-acquired infection,how to assess the risk of working with emerging viruses,and how laboratory-acquired infection can be prevented.Control measures used in the laboratory designed as such that they protect workers from emerging viruses and safeguard the public through the safe disposal of infectious wastes are also addressed.

Insertion site of FLAG on foot-and-mouth disease virus VP1 G-H loop affects immunogenicity of FLAG

The G-H loop of the foot-and-mouth disease virus（FMDV） virion contains certain dominant immunogenic epitopes, as well as an arginine-glycine-aspartic acid（RGD） motif that is recognized by cell surface integrin receptors. Previous experiments indicate that it is critical to maintain virus structural integrity when inserting an exogenous epitope into the surface of an FMDV structural protein. However, it remains to be determined how factors such as different insertion positions affect interactions among the virus, cells and host immune system. In this study, one infectious c DNA clone of the swine FMDV Cathay topotype strain O/CHA/90 was constructed. Then, a FLAG marker（DYKDDDDK） was inserted upstream（–4） or downstream（＋10） of the RGD motif to generate tagged viruses vFLAG-O/CHA/90 or vO/CHA/90-FLAG, investigating the possibility of expressing foreign antigen and effect on its immunogenicity. Compared to the parental virus, both tagged viruses exhibited similar plaque phenotypes, suckling mouse pathogenicity and antigenicity. Additionally, the FLAGtag insertion position did not change the use of integrin-mediated cell entry by the tagged viruses. Interestingly, both tagged vaccines protected pigs against challenge with the parental virus O/CHA/90 and induced immune responses against FMDV in BALB/c mice and pigs, but only vaccination with vFLAG-O/CHA/90 generated anti-FLAG antibodies. Our findings demonstrated that two sites（RGD–4 and RGD＋10） tolerated the insertion of an exogenous gene in the swine FMDV O/CHA/90 strain. However, only RGD–4 was a novel and appropriate inserting site which could tolerate exogenous FLAG. The resultant tagged virus is a promising candidate for FMD vaccine which can be differentiating infected from vaccinated animals（DIVA）.

Analysis of fig tree virus type and distribution in China

The common fig（Ficus carica L.）was one of the earliest horticultural crops to be domesticated.A number of different viruses can infect fig trees including Fig mosaic virus（FMV）that has been detected in several commercial fig trees in Xinjiang,China.However,the distribution of FMV and other fig-infecting viruses in China remains unknown.In the present study,a sample from an ancient fig tree growing in Xinjiang was investigated by electron microscopy（EM）followed by PCR/RT-PCR,and FMV,Fig badnavirus 1（FBV-1）and Fig leaf mottle-associated virus 1（FLMaV-1）were detected.Fig leaf samples（252）from commercial orchards across China were subjected to PCR/RT-PCR,and FMV,FBV-1 and Fig fleck-associated virus（FFka V）were relatively abundant（44.4,48.4 and 44%,respectively）,while FLMaV-1 and Fig mild mottle-associated virus（FMMa V）were much scarcer（5.6 and 0.4%,respectively）,and FLMaV-2,Fig cryptic virus（FCV）,and Fig latent virus（FLV）were not detected.The presence of disease-causing viruses in fig trees presents a significant challenge for fig producers in China.This study may help to promote actions aimed at controlling fig viruses,especially FMV.

Phylogenetic and Molecular Analysis of an H7N7 Avian Influenza Virus Isolated in East Dongting Lake in 2012

Objective In March 2012, an H7N7 subtype avian influenza virus （AIV） named A/wild goose/Dongting/PC0360/2022 （H7N7） （DT/PC0360） was recovered from a wild goose in East Dongting Lake. We performed whole-genome sequencing of the isolate, and analyzed the phylogenetic and molecular characterization. Methods RNA was extracted from environment samples （including fecal samples from wild bird or domestic ducks, and water samples） for detecting the presence of Influenza A Virus targeting Matrix gene, using realtime RT-PCR assay. The positive samples were performed virus isolation with embryonated eggs. The subtype of the isolates were identified by RT-PCR assay with the HI-HI6 and N1-N9 primer set. The whole-genome sequencing of isolates were performed. Phylogenetic and molecular characterizations of the eight genes of the isolates were analyzed. Results Our results suggested that all the eight gene segments of DT/PC0360 belonged to the Eurasian gene pool, and the HA gene were belonged to distinct sublineage with H7N9 AIV which caused outbreaks in China's Mainland in 2013. The hemagglutinin cleavage site of HA of DT/PC0360 showed characterization of low pathogenic avian influenza virus. Conclusion Strengthening the surveillance of AlVs of wild waterfowl and poultry in this region is vita for our knowledge of the ecology and mechanism of transmission to prevent an influenza pandemic.

Modification of the passive heamagglutination test for detection of <i>infectious bursal disease virus</i>

To modify the Passive Haemagglutination (PHA) test, a rapid test, used for qauntitative detection of viral antibodies, so that it can be used for determination of viral titres, dilutions of Infectious Bursal Disease Virus (IBDV) were used to sensitize the Red Blood Cells (RBCs) before reacting them with known IBD serum. Also, to improve sensitivity of the test, different RBC concentrations were used for the test. A standard IBDV gave positive PHA reaction upto its 1:2048 dilution. With different IBDV samples, positive PHA reactions occured upto dilutions, ranging from 1:16 to 1:4096. Different RBC concentrations gave different titres for same IBDV samples. With 0.6% and 0.2% RBC concentrations, mean PHA titres of IBDV samples increased from 454. 85 ± 315.32 to 2396.57 ± 489.55 (p < 0.05 ). It was concluded that PHA can be adopted for evaluation of viral titres. To improve sensitivity of the test, use of 0.2% RBC is recommended.

Physicochemical Properties of Musk Deer Pneumonia and Purulent Disease Viruses

[ Objective] To understand the physicochemical properties of musk deer pneumonia and purulent disease viruses. [ Method] The pneu- monia and purulent disease viruses were isolated from the abnormal and purulent lung tissues of musk deer. Then the isolated viruses were inocula- ted into the Vero cells. After culturing, the virus solution was collected and used to determine TCID50 and genoma types. The sensitivity to fat sol- vent, resistance to hydrochloric acid and trypsin as well as tolerance to heat of the musk deer pneumonia and purulent disease viruses were detec- ted, respectively. [ Result] The obvious cytopathic effects （CPE） were found in Veto cells infected by the isolated viruses. The virus was 2-1.43 TCID50/ml and its genome was RNA. The virus was not sensitive to chloroform, 1% trypsin and heats, and it had a certain tolerance to 0.1 mol/L hy- drochloric acid. [ Conclusion] The study on the physicochemical properties of musk deer pneumonia and purulent disease viruses lays a foundation for prevention and control of the musk deer pneumonia and purulent diseases.

Baculoviruses as Vectors in Mammalian Cells

The Baculoviridae are a large family of enveloped DNA viruses exclusively pathogenic to arthropods. Baculoviruses have been extensively used in insect cell-based recombinant protein expression system and as biological pesticides. They have been deomostrated to be safe to mammals, birds and fish. Recently, baculoviruses has been shown to transduce different mammalian cells in spite of the fact that they cannot replicate in mammalian cells (11, 73, 76). This has resulted in the development of baculoviruses as mammalian expression systems and even as vestors for gene therapy.

Non-Linear Mathematical Model of the Interaction between Tumor and Oncolytic Viruses

A mathematical modeling of tumor therapy with oncolytic viruses is discussed. The model consists of two coupled, deterministic differential equations allowing for cell reproduction and death, and cell infection. The model is one of the conceptual mathematical models of tumor growth that treat a tumor as a dynamic society of interacting cells. In this paper, we obtain an approximate analytical expression of uninfected and infected cell population by solving the non-linear equations using Homotopy analysis method (HAM). Furthermore, the results are compared with the numerical simulation of the problem using Matlab program. The obtained results are valid for the whole solution domain.

Solar Radiation, Perelman Entropy Mapping, DNA, Viruses etc.

A short note based on the homogeneous 5D space-time topological mappings is extended to cover DNAs of viruses and how the body’s immune system can be enhanced to recognize and remove it.

Synchronous Detection of DNA/RNA of Four Shrimp Viruses by Real-time Fluorescence Quantitative RT-PCR

[ Objective] This study aimed to establish a simultaneous detection method of shrimp viruses by real-time fluorescence quantitative RT-PCR, to improve the efficiency of inspection and quarantine. [ Method] A novel real-time fluorescence quantitative RT-PCR assay was established and optimized for simultaneously detecting DNA/RNA of four shrimp viruses （WSSV, IHHNV, TSV and YHV ）. [ Result] The optimized real-time fluorescence quantitative RT-PCR system gener- ated typical amplification curves with high amplification efficiencies （E = 1.06, 1.07, 0.92 and 0.92, respectively）, good hnear relationship （ r = 1 ）, uniform repeatability （ standard deviation = 0.05 - 0.46 ; variation coefficient = 0.26% - 1.62% ） and high sensitivity, exhibiting no significant differences compared with re- al-time fluorescence quantitative PCR （average error of Ct value = 0.04 -0.40; T = 0.53 -2.50; P 〉 0.05 ）. The total detection time was about 1 h. [ Conclusion] The optimized real-time fluorescence quantitative RT-PCR system can be used for rapid detection of WSSV, IHHNV, TSV and YHV.

Stealth Adapted Viruses: A Bridge between Molecular Virology and Clinical Psychiatry

Cytopathic “stealth-adapted” viruses bypass the cellular immune defense mechanisms because of molecular deletion or mutation of critical antigen coding genes. They, therefore, do not provoke the inflammatory reaction typical of infections with the conventional viruses from which stealth adapted viruses are derived. Stealth adapted viruses establish persistent, systemic virus infections, which commonly involve the brain. The brain damage can cause major mood and cognitive disorders, fatigue, seizures and various manifestations of an impaired autonomic nervous system. Symptoms can also result from: 1) induced autoimmunity, 2) antibody formation against virus antigens, 3) virus-induced cellular damage to non-brain tissues and 4) induced heightened overall immune reactivity, such that normally unrecognized components of the virus begin to become targeted by the cellular immune system. This last mechanism is relevant to the reported neurological and psychiatric adverse effects of vaccination in certain individuals. It is also appropriate to consider the infectious component of stealth adapted virus infections since family members and others may be at risk for becoming infected.