Inosine:A broad-spectrum anti-inflammatory against SARS-CoV-2 infection-induced acute lung injury via suppressing TBK1 phosphorylation / 药物分析学报

Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)-induced cytokine storms constitute the primary cause of coronavirus disease 19(COVID-19)progression,severity,criticality,and death.Gluco-corticoid and anti-cytokine therapies are frequently administered to treat COVID-19,but have limited clinical efficacy in severe and critical cases.Nevertheless,the weaknesses of these treatment modalities have prompted the development of anti-inflammatory therapy against this infection.We found that the broad-spectrum anti-inflammatory agent inosine downregulated proinflammatory interleukin(IL)-6,upregulated anti-inflammatory IL-10,and ameliorated acute inflammatory lung injury caused by mul-tiple infectious agents.Inosine significantly improved survival in mice infected with SARS-CoV-2.It indirectly impeded TANK-binding kinase 1(TBK1)phosphorylation by binding stimulator of interferon genes(STING)and glycogen synthase kinase-3β(GSK3β),inhibited the activation and nuclear trans-location of the downstream transcription factors interferon regulatory factor(IRF3)and nuclear factor kappa B(NF-κB),and downregulated IL-6 in the sera and lung tissues of mice infected with lipopoly-saccharide(LPS),H1N1,or SARS-CoV-2.Thus,inosine administration is feasible for clinical anti-inflammatory therapy against severe and critical COVID-19.Moreover,targeting TBK1 is a promising strategy for inhibiting cytokine storms and mitigating acute inflammatory lung injury induced by SARS-CoV-2 and other infectious agents.

Packaging of Rift Valley fever virus pseudoviruses and establishment of a neutralization assay method

Rift Valley fever (RVF) is an acute, febrile zoonotic disease that is caused by the RVF virus (RVFV). RVF is mainly prevalent on the Arabian Peninsula, the African continent, and several islands in the Indian Ocean near southeast Africa. RVFV has been classified by the World Organisation for Animal Health (OIE) as a category A pathogen. To avoid biological safety concerns associated with use of the pathogen in RVFV neutralization assays, the present study investigated and established an RVFV pseudovirus-based neutralization assay. This study used the human immunodeficiency virus (HIV) lentiviral packaging system and RVFV structural proteins to successfully construct RVFV pseudoviruses. Electron microscopy observation and western blotting indicated that the size, structure, and shape of the packaged pseudoviruses were notably similar to those of HIV lentiviral vectors. Infection inhibition assay results showed that an antibody against RVFV inhibited the infective ability of the RVFV pseudoviruses, and an antibody neutralization assay for RVFV detection was then established. This study has successfully established a neutralization assay based on RVFV pseudoviruses and demonstrated that this method can be used to effectively evaluate antibody neutralization.

Application of 23G minimally vitrectomy trocar in acute angle closure glaucoma / 眼科新进展

Objective To evaluate the outcome of 23G minimally vitrectomy trocar on eyes with acute angle closure glaucoma.Methods A retrospective review was performed of patients with acute angle closure glaucoma who underwent combined compound trabeculectomy from September,2014 to June,2015 in Beijing MEM care system.Intraoperative 23G minimally vitrectomy trocar was used in all of the patients.30 eyes of 30 patients were enrolled in the study.All patients were followed up for 12 months.Visual acuity,intraocular pressure and complications were observed.Results No serious complications such as explosive choroid hemorrhage and retinal hemorrhage occurred during operation.All of 30 eyes maintained adequate pressure control.The average postoperative IOP was (15.93 ± 1.35) mmHg (1 kPa =7.5 mmHg),was less than the preoperative (56.34 ± 6.96) mmHg (P =0.00).And the visual acuity in 23 eyes were improved.Conclusion Combined trabeculectomy with 23G minimally vitrectomy trocar in acute angle closure glaucoma patients is a kind of safe and effective operation method,can obviously reduce the intraoperative explosive choroid hemorrhage.

Effect of Deletion of the Carboxyl Terminal of the NS1 Protein on Pathogenicity of the Influenza B Virus / 病毒学报

To analyze the molecular basis of the variation of the pathogenicity of the influenza B virus, we rescued a recombinant virus with a deletion in the carboxyl terminal of the NS1 protein using reverse genetics based on the parental virus B-S9 of B/Yamagata/16/88. A mutant strain with a deletion of 171 amino acids in the carboxyl terminal of the NS1 protein was named "B-L5". BALB/c mice were inoculated with 3 X 105 EID50 of B-L5 and the parental virus B-S9, respectively. Then, weight changes, survival, and viral titers were documented. During 3 days post-inoculation (dpi) to 7 dpi, the weight of mice infected with B-S9 decreased. However, the weight of mice infected with B-L5 showed weight decreases only at 2 dpi, and quickly recovered at 3 dpi. B-S9 and B-L5 could replicate in the lungs of BALB/c mice. However, viral titers in the lungs of mice infected with B-L5 were 7900-times lower than those of mice infected with B-S9 at 3 dpi. Viral titers in the lungs of mice infected with B-L5 were not detected at 6 dpi. These results showed that, compared with the parent virus B-S9, the mutant virus B-L5 showed lower pathogenicity in BALB/c mice. Our study suggests that deletion of the carboxyl terminal of the NS1 protein decreases the pathogenicity of the influenza B virus. Establishment of a reverse-genetics system for the B influenza virus will provide a platform for studying its pathogenesis, and mechanism of transmission, and for developing live-attenuated influenza B virus vaccines.

An epidemiological study of H1N1 influenza A / 中国免疫学杂志

H1N1 influenza A spread around the world in 2009.There were lots of patients in China too.We did this research to know the epidemiological feature and transmission route and strengthen the prevention and control measure of the influenza.Methods:We collected the nasopharyngeal swab and serum samples of 56 patients who had flu symptom from the infectious disease department of 1st hospital of Jilin University from October in 2009 to December in 2009.The specific antibody of the serum samples were detected by the blood clots suppression method and the H 1N1 RNA of the nasopharyngeal swab was detected by the Nest-RT-PCR assay.Results:The results of nucleic acid test showed that 21(37.5%) and 16(27.8%) samples were found NP and M of influenza A positive respectively and only 2 ( 3.6%) were found H1N1 of influenza A positive.The results of the blood clots suppression method showed that the serum samples of 27 patients (48.2%) could suppress the red blood cells blot of H 1N1 influenza A specifically and all the antibody titer was more than 1∶320.The antibody titer was more than 1∶5 120 in 8 of them.There′s significant difference of the serum antibody titer between the recovery phase and the acute phase.The specific H1N1 influenza A antibody of 27 (48.2%) serum samples in the recovery phase were positive and it was much higher than the result of nucleic acid test .Conclusion:The nucleic acid could be detected in the acute phase and the serum antibody detection could be done in the later stage .Using both the assays could increase the positive rate of H 1N1 influenza.

Preparation and application of a colloidal gold strip to detect the rabies antibody / 生物工程学报

To develop a specific, rapid, and convenient immunochromatography assay (ICA) to detect the rabies antibody in clinical sample from immuned dogs by rabies vaccine. Colloidal gold particles labeled with purified rabies virus (CVS11) were used as the detector reagent. The staphylococcal protein A (SPA) and pured rabbit anti-rabies virus IgG were blotted on the test and control regions of nitrocellulose membrane. Then the strip was assembled with sample pad, absorbing pad, and dorsal shield. The assay samples (261 dog's serum) were collected from Wildlife Rabies Disease Diagnostic Laboratories of Ministry of Agriculture in China, Institute of Military Veterinary, Academy of Military Medical Sciences and other six provinces, including rabies virus positive and negative serum. The performance of the strip was compared to fluorescent antibody virus neutralization test. The neutralizing antibody titer could be detected above 0.5 IU. The strip did not change of performance when stored at room temperature for 12 months. It may offer reference of neutralizing antibody titer level after dogs immuned rabies vaccine and determin whether the dogs need to be immuned again.