Rationale:Dysregulated phagocytic clearance within the synovial microenvironment contributes to persistent inflammation and impairs immune homeostasis in rheumatoid arthritis (RA). Because MerTK-positive macrophages are essential for efferocytosis and inflammation resolution, but are functionally impaired in RA, this study aimed to engineer macrophages capable of enhancing tumor necrosis factor-α (TNF-α) clearance and restoring inflammation-resolving macrophage function. Methods:We generated anti-TNFα chimeric antigen receptor macrophages (CAR-M) by replacing the MerTK antigen-binding domain with a TNF-R1 fragment. The phagocytic and degradative capacity of CAR-M toward soluble TNF-α was assessed, together with activation of Rho GTPases Rac1, Cdc42, and RhoA. Macrophage inflammatory mediator production, phenotypic polarization, SOCS1/3, NF-κB, and MAPK signaling were analyzed. The effects of CAR-M cells were further compared with adalimumab and control treatments under synovial fluid stimulation from patients with arthritis and in mice with collagen-induced arthritis (CIA). Results:Anti-TNFα CAR-M showed markedly enhanced phagocytosis and degradation of soluble TNF-α, accompanied by Rac1, Cdc42, and RhoA activation. CAR activation reduced inflammatory mediator production, including soluble TNF-α, interleukin-1 β (IL-1β), and interleukin-6 (IL-6), while increasing interleukin-10 (IL-10) secretion. This functional shift was associated with polarization toward an inflammation-resolving MerTK+CD206+ phenotype, SOCS1/3 upregulation, and NF-κB and MAPK pathway inhibition. Under stimulation with synovial fluid from arthritis patients, anti-TNFα CAR-M decreased TNF-α and IL-6 levels and drove macrophages toward an M2-like anti-inflammatory state. In CIA mice, local in situ CAR-M cell treatment produced stronger therapeutic effects than adalimumab or other controls, significantly alleviating inflammatory activation, clinical symptoms, and joint pathologies. Conclusions:Anti-TNFα CAR-M exert therapeutic effects via a dual mechanism: direct TNF-α clearance and synovial macrophage reprogramming toward a reparative phenotype. This synthetic biology strategy highlights the potential of precise cell engineering to correct phagocytic deficits and resolve chronic inflammation in RA.

Avian leukosis viruses (ALVs) include a group of avian retroviruses primarily associated with neoplastic diseases in poultry, commonly referred to as avian leukosis. Belonging to different subgroups based on their envelope properties, ALV subgroups A, B, and J (ALV-A, ALV-B, and ALV-J) are the most widespread in poultry populations. Early identification and removal of virus-shedding birds from infected flocks are essential for the ALVs’ eradication. Therefore, the development of rapid, accurate, simple-to-use, and cost effective on-site diagnostic methods for the detection of ALV subgroups is very important. Cas13a, an RNA-guided RNA endonuclease that cleaves target single-stranded RNA, also exhibits non-specific endonuclease activity on any bystander RNA in close proximity. The distinct trans-cleavage activity of Cas13 has been exploited in the molecular diagnosis of multiple pathogens including several viruses. Here, we describe the development and application of a highly sensitive Cas13a-based molecular test for the specific detection of proviral DNA of ALV-A, B, and J subgroups. Prokaryotically expressed LwaCas13a, purified through ion exchange and size-exclusion chromatography, was combined with recombinase polymerase amplification (RPA) and T7 transcription to establish the SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) molecular detection system for the detection of proviral DNA of ALV-A/B/J subgroups. This novel method that needs less sample input with a short turnaround time is based on isothermal detection at 37 °C with a color-based lateral flow readout. The detection limit of the assay for ALV-A/B/J subgroups was 50 copies with no cross reactivity with ALV-C/D/E subgroups and other avian oncogenic viruses such as reticuloendotheliosis virus (REV) and Marek’s disease virus (MDV). The development and evaluation of a highly sensitive and specific visual method of detection of ALV-A/B/J nucleic acids using CRISPR-Cas13a described here will help in ALV detection in eradication programs.

为研究B亚群禽白血病病毒(ALV-B)对新城疫疫苗免疫效力的影响,将50只SPF鸡随机分为2组,试验组鸡接种ALV-B,对照组鸡注射等量生理盐水,7 d后2组全部滴鼻免疫新城疫病毒(NDV)LaSota疫苗,试验期8周.结果显示,与对照组相比,当试验组的鸡感染ALV-B后,再免疫LaSota疫苗时,NDV抗体水平、IFN-γ和IL-2浓度以及外周血CD4+和CD8+的T淋巴细胞百分比均显著降低(P＜0.05).试验组鸡的肝脏、脾脏、法氏囊均发生肿瘤性组织学病变,其中肝脏超微组织学病变中可同时见到ALV-B和LaSota病毒.综合上述研究证明ALV-B可显著降低LaSota疫苗的免疫应答效果,两种病毒有共感染的现象.

GX0101 is the first field Marek's disease virus (MDV) recombinant with an REV LTR insert isolated in China. We speculated that there was a selective advantage of GX0101 becoming the more prevalent field strain from a very low percentage of recombinant virus. In the study, dual fluorescence quantitative real-time PCR (DF-qPCR) that detects GX0101 and GX0101ΔLTR simultaneously was established based on the genomic difference of GX0101 and its LTR deletion strain GX0101ΔLTR. MDV natural transmission was simulated in specific-pathogen-free (SPF) chicks, and continuous tracking of GX0101 and GX0101ΔLTR in chicks was carried out. The results showed that GX0101 possessed high horizontal transmission capacity, which could infect SPF chicks by contact in a short time and became the predominant strain following contact infections in chicken flocks. GX0101 still had a more significant advantage of horizontal transmission than GX0101ΔLTR after continuous passage even if the initially infectious dose was significantly lower. There were 72 differentially expressed MDV genes between GX0101 and GX0101ΔLTR, with the genes and gene products mainly involved in virus replication, tegument protein, glycoprotein, nucleocapsid protein, immune evasion, tumor development and/or pathogenesis, and hypothetical protein. Sixteen genes related to virus replication and transmission were significantly up-regulated. This is the first study to illuminate that increased horizontal transmission of recombinant MDV due to REV LTR was the competitive advantage of the virus being a prevalent strain and define the differential transcription profile of viral genes between GX0101 and GX0101ΔLTR. This will be helpful for in-depth study on the molecular mechanism of increased horizontal transmission of MDV by REV LTR.

Type 2 diabetes mellitus (T2DM) is a metabolic disorder characterized by β-cell loss, insulin resistance, islet inflammation and amyloid deposits derived from islet amyloid polypeptide (IAPP). Reducing toxic IAPP oligomers and inhibiting islet inflammation may provide therapeutic benefit in treating T2DM. Intravenous immunoglobulin (IVIg) is an efficient anti-inflammatory and immunomodulatory agent for the treatment of several autoimmune or inflammatory neurological diseases. However, whether IVIg has therapeutic potential on T2DM remains unclear. In present study, we showed that IVIg treatment significantly improved glucose control and insulin sensitivity, and prevented β-cell apoptosis by lowering toxic IAPP oligomer levels, attenuating islet inflammation and activating autophagy in human IAPP transgenic mouse model. These results suggest that IVIg is a promising therapeutic potential for T2DM treatment.

AIMS:Inflammation is strongly associated with the mechanism of β-cell failure in type 2 diabetes mellitus (T2DM). Blockade of the key proinflammatory cytokine IL-1β has been implicated as a promising therapeutic strategy for the prevention and treatment of type 2 diabetes. In this study, we developed an IL-1β-targeted therapeutic vaccine consisting of an IL-1β epitope peptide (A1β) and assessed its efficacy on a diabetic KK-Ay mouse model.MAIN METHODS:KK-Ay mice were immunized with A1β for three injections at a 2-week interval. The induced antibody titers, body weights and blood glucose levels were monitored every two weeks. Then the intraperitoneal glucose tolerance test and insulin tolerance test were performed. The β-cell mass, β-cell apoptosis and proliferation were evaluated by immunofluorescence. IL-1β gene expression in islets was also measured by quantitative RT-PCR.KEY FINDINGS:A1β immunization induced robust antibody responses, reduced body weight gain, improved glucose tolerance and insulin sensitivity in KK-Ay mice. Moreover, A1β restored β-cell mass, inhibited β-cell apoptosis, enhanced β-cell proliferation and downregulated IL-1β expression.SIGNIFICANCE:The novel IL-1β-targeted epitope vaccine has the therapeutic potential for T2DM.

A subgroup J avian leukosis virus (ALV-J), designated as SD1306, which was isolated and identified from inspection of suspected infection, a Salmonella pullorum (SP) strain was isolated at the same time. Sixty Hy-line brown chickens were used for the study. Detected the body weight, the immune organs index, histopathology, quantity change of the intestinal flora, the viremia and anti-ALV-J antibody after being inoculated the two pathogens. Results showed that the indicators had no significant change during the latent period, while at the 6th and 7th week, compared with other two groups inoculated separately, the body weight of co-infected group was significant lower; the thymus index was below the group infected with SD1306, while the spleen index was significant higher all the time. No evident tumour nodules and swelling in any tissues were seen at necropsy. Histopathological findings revealed large islands of leukomonocytes or myelocytes in tissues containing tumors. Salmonella in rectal feces of co-infected group was much higher than the group infected with Salmonella pullorum from the 3rd week, the number of E. coli significantly exceeded the group infected with Salmonella pullorum at the 7th week. The co-infected group positive rate of ALV-p27 antigen was 69.23%, while the group infected with SD1306 was 38.46% at the 5th week. The positive rate of both groups declined at the 6th week, rose again at the 7th week. Correspondingly the positive rate of anti-ALV-J antibody continued to decrease from the 6th week to the 7th week. It showed that ALV-p27 antigen and antibody level fluctuated strongly in some time and there was a negative correlation between them. The damage to the chicken was more serious in the co-infected chicken than the chicken infected separately. (C)2014 PVJ. All rights reserved

With the increasing use of genetically modified organisms (GMO) worldwide, the controversy of their safety would be unlikely to abate. Salt is a major stress limiting crop productivity and most alfalfa are salt-sensitive. In plants expression of the gene encoding for Betaine Aldehyde Dehydrogenase (BADH) confers tolerance to salinity and drought stresses. The safety of genetically modified alfalfa was assessed in rabbits consuming diets containing transgenic alfalfa (50%) and conventional non-genetically modified alfalfa (50%). Twenty four male rabbits were fed the two types of alfalfa for 22 weeks. Feed intake and weight gain were measured. Haematology and blood biochemistry of the two groups were compared at both 11 and the 22 week. All the animals were sacrificed at the end of the experiment and the parameters of the organ index, gross and microscopic appearance of tissues were compared between the two groups. No adverse effects were observed in rabbits consuming GM alfalfa and non-GM alfalfa. The results indicated that the feeding value of GM alfalfa and conventional non-GM alfalfa were equal in this study. No detrimental expected effects were observed in rabbits fed genetically modified alfalfa. We concluded that it is as safe and nutritious as existing alfalfa.

本研究用SPSS软件对生猪价格与猪肉、仔猪、玉米、鸡蛋价格及生猪和能繁母猪存栏量进行了相关性分析,结果为,生猪价格与猪肉、仔猪、玉米、鸡蛋价格的相关系数分别0.844、0.638、0.616、0.602。生猪价格的复回归方程为Y=-1.483+0.343X1+4.488X2+μ。生猪价格除受仔猪、玉米价格作用外,还受猪的品种、生产方式、规模以及供求关系、疫情等突发事件和国家宏观调控政策等多种因素的影响。

The study was designed to investigate the infection status and to finish the preliminary eradication of avian leukosis virus (ALV) and Salmonella pullorum (SP) in breeders of Chinese local "ShouGuang" chickens. ALV antigen and antibody was tested via ELISA, and SP antibody was detected by serum plate agglutination test (SPAT). The etiology and pathology was also studied. The ALV-P27 antigen, ALV-A/B and SP antibody positive chickens were eliminated in turn, and then the negative were retained as the breeder flocks. The results showed that the positive rate of antigen to ALV-P27, antibody to ALV-A/B, ALV-J and SP was 57.8, 6.7, 0 and 17.8% in this breeder farm, respectively. The co-infection of ALV and SP was confirmed and the positive rate of both SP and ALV-P27 or ALV-A/B was 10 and 1%, respectively. There were obvious tumor nodules and lymphoid tumor cells in the comb, liver and spleen of the co-infected chickens. The degenerative and atrophic ovarian follicles, inflammatory cell infiltration in muscle biopsies were also found. The elimination rate of ALV-p27, ALV-A/B and SP positive chickens was 55.4, 13 and 6.1%, respectively. The final amount of the breeder conservation was 309 chickens. In conclusion, the co-infection of ALV-B and SP was found and more emphasis should be given on its prevention; the preliminary eradication of "ShouGuang" breeder chickens was finished. (C) 2013 PVJ. All rights reserved

ALV antigen and antibody in "Bairi chickens" were tested with ELISA.Five flocks tested were negetive for anti-ALV-A/B antibody and 4 were positive for ALV-p27 antigen at high positive rates while only one was positive(57.14%) for anti-ALV-J antibody.One or more small tumour nodules were observed on the surface of livers in the early stage in "Bairi chickens" while the scattered tumour nodules were found on enlarged livers,spleens and kidneys in the late stage.Besides,tumour nodules were also detected in other viscera.Histologically,visceral tissues mainly contained myeloid cells with acidophilic granules and densely-arranged lymphoblast,and normal cells were squeezed and even destroyed by these tumour cells.Enveloped viral particles approximately 100 nm in diameter were observed in spleens and bursa Fabricii with transmission electron microscope.In some lymphocytes,nuclear membrane and plasma membrane were showed edema and even destroyed.The edema in endoplasmic reticulum and mitochondria with enlarged mitochodria cistermae was also observed.Numerous vacuoles emerged in cytoplasm.The increasing number of tumour cells and intra-and extra-cellular hydropic degeneration might be responsible for the enlargement of viscera organs.

Avain leukosis(AL) is infectious tumour diseases caused by avain leukosis viruses,which cause chickens to produce immunosuppression and atrophy of organs or tumors and break into great economical loss.The disease has been occuring reportedly over about 80% provinces in China between 1999-2009,especially most in Shandong and Beijing province et al.The disease occurs sustainedly and ascendly year by year and it occurred reportedly most in meat-type chickens before 2003 and most in egg-type chickens or both of them after 2003,meanwhile it is worth notifying that the disease is endangering to China land race chickens.Most AL in China were caused by J subgroup of ALV and few by A or B subgroups of ALV.It suggests that ALVs-J seperated from meat-type chickens variate less and more from egg-type chickens with analysis to more than 30 ALVs-J,and some ALVs can induce chicken to produce its antibodies and some can not,or some can produce antibodies without immunonetralization,and ALV can co-infect chickens with some viruses such as REV or MDV or CIAV et al and augument the immunosuppression and the pathological loss;and ALV can be detected positively from kidneys,livers,spleens,eggs,chicken embryos and claocas in chicken samples;and PCR,RFLP and ALV seperation from innoculated cells are used to identificate the subgroups of ALVs at present,but the last method is most effective.It can be concluded that the results of those researches will be helpful for understanding and controling to avain leukosis.

<正>2010年12月,某养殖基地饲养的100多只当地山羊发生了一种急性传染病,以体温升高、精神沉郁、呼吸困难、流浆液性或脓性鼻汁、阵发性咳嗽、全身皮肤均出现丘疹为基本特征。笔者对该病的病因进行了调查与分

<正>引起肉鸡猝死的原因很复杂,其中在冬季天气突变温差较大的季节,由于肉鸡自身抵抗力低,更容易暴发此病。笔者近期遇到一起多种病因造成的肉鸡猝死症病例,现将诊治情况报道如下。

DF1 cells were inoculated with three different doses(100,10,1 μL) of ALV-A,ALV-C,ALV-J-PY and ALV-J-WS separated from different chickens to evaluate three kinds of A,B,C ELISA kits.The samples of DF1 cells supernatant were obtained at different days post inoculation(dpi) and detected with three kinds of kits after freezing and thawing once.The results indicated that the samples inoculated with ALV-A or ALV-J-PY were both detected positively firstly at the 3rd dpi with A ELISA kit for 100 μL inoculation doses and at the 7th dpi for 1 μL dose;If with B ELISA kit,two ALVs of subgroup were detected positively within some days post inoculation,but the detectable time were longer than that of A ELISA kit;If with C ELISA kit,the detectable time of viruses were the longest.ALV-C and ALV-J-WS were detected positively with A ELISA kit at the 5th and 9th dpi for 100 μL doses,respectively;but for the other doses,no positive samples were detected at whole observed days.If with B and C ELISA kits,none of inoculated cells with three doses of inoculation was detected positively.It can be concluded that it is dramatically different between A,B and C ELISA kits in the detection of three exosomic subgroups of ALVs;The A ELISA kit is more sensitive than other two ELISA kits(B and C).Simultaneously,the positively detected days to ALVs are related to the doses of inoculation in DF1 cells.The results can be helpful for the application of three ELISA kits to the detection or isolation of exosomic ALVS.

Subgroup J avian leukosis virus(ALV-J) was isolated from both provetriculus suspensions and blood samples of 9 in 10 laying hens only with proventriculitis,and co-infection of ALV-J with reticuloendotheliosis virus was also confirmed in 4 chickens.The gp85 genes of ALV-J isolates were amplified by PCR,sequenced and compared to 20 reference ALV-J strains.Sequence comparisons indicated that the provetriculus isolate had 97.8% identity with the HPPS103 strain isolated from white meat-type chickens and 93.0% identity with SD07LK1 strain from egg-type chickens.The result demonstrated that ALV-J infection was common in some chickens with only proventriculus,further indicating the diversity of virus infections associated with proventriculitis.ALV-J may be take part in pathogenesis of provetriculitis in some chickens,but its pathogenic role needs to be further studied.

By the homology analysis for gp85 genes of 5 strains of ALV-J isolated from Hyline laying hens,the molecular evolution of ALV-J was studied in the present study.The 5 strains of ALV-J were obtained from Beijing,Shanxi and Shandong.The gp85 genes were cloned by PCR and the sequence was measured and compared with 14 published reference ALV-J strains.The homology analysis showed that the 5 isolated strains of ALV-J shared 96.6%(96.4%-96.8%) and 89.6%(89.3%-89.9%) homology with the HPRS-103 strain isolated from white meat-type broiler and SD07LK1 strain isolated from egg-type chickens.The homology of the 5 strains was exceeded 98.1%(98.1%-100%).The result indicated that ALV-J strains from Hyline egg-type chickens may have the same origin,the white meat-type broiler chickens imported abroad.