Objective:To investigate the effect of microwave on human adenovirus type 2 (HAdV-2) capsid protein in simulated infectious wastes.Methods:Droplets of HAdV-2 virus suspension were added to medical disposable gloves to simulate infectious waste, irradiated under different microwave conditions, the temperature change was recorded, and the irradiated viral supernatant was treated with Dnase I and detected by PCR and qPCR to determine the effect of microwave on the integrity of the viral capsid. ELISA was used to detect the effect of microwave irradiation on the structure of viral hexon protein. The virus was treated alone at the highest temperature during microwave irradiation to investigate whether there were non-thermal effects during microwave disinfection.Results:The maximum temperature during microwave irradiation was 76 ℃, and the PCR and qPCR result showed that Dnase I could significantly damage the viral nucleic acid after microwave irradiation, while the virus control group and heat treatment group were not significantly affected, indicating that microwave irradiation could destroy the integrity of the viral capsid. The result of ELISA showed that microwave irradiation could significantly weaken the binding ability of Hexon protein and antibody, and the result of heat treatment group were similar.Conclusions:Microwave irradiation can destroy the integrity of the HAdV-2 capsid and the structure of Hexon protein, in which the damage to the integrity of the capsid is mainly due to non-thermal effects, and the structural changes of hexon protein are mainly due to thermal effects.

Humans ; RNA, Long Noncoding/genetics* ; MicroRNAs/genetics* ; Cell Line, Tumor ; Inflammation/genetics* ; Apolipoproteins E ; Apoptosis

Purpose To investigate the feasibility of CT-based feature tracking technology to quantify left ventricular myocardial strain(MS)and its significance in hypertrophic obstructive cardiomyopathy(HOCM).Materials and Methods A total of 35 HOCM patients who underwent cardiac coronary angiography from March 2019 to December 2021 in the First Affiliated Hospital of the Air Force Military Medical University were retrospectively included,and a total of 60 cases who were negative for cardiac coronary angiography among those who visited the hospital with suspected coronary artery disease were randomly enrolled.Conventional cardiac functional parameters and MS parameters were quantified via post-processing software,and differences of parameters between the groups were analyzed.The diagnostic efficacy of MS parameters for HOCM was further evaluated.Results Compared to the control group,the HOCM group exhibited significant increases in various conventional left ventricular functional parameters,including left ventricular wall thickness,mass,mass index,end-diastolic volume and stroke volume(t=2.119 to 24.861,all P<0.05).However,there were no statistically significant differences in end-systolic volume and cardiac output between the two groups(P>0.05).The global longitudinal and radial strain values of HOCM group were significantly lower than those of control group(t=12.857,-6.427,P<0.01),while the endocardial global circumferential strain of HOCM group was significantly higher than that of control group(t=-2.369,P<0.05).Among MS parameters,global longitudinal strain exhibited the best diagnostic efficacy for HOCM,with an area under the curve of 0.997.A cutoff value of≤20.78%for global longitudinal strain showed that the sensitivity and specificity was 100%and 95%,respectively.Conclusion The MS parameters quantified by the CT-based feature tracking technique are superior to left ventricular ejection fraction in quantifying left ventricular function,with the highest sensitivity and specificity for early myocardial function impairment of longitudinal strain.In addition,the technique has good repeatability and is expected to become a new indicator for the assessment of myocardial function in HOCM.

Objective:To investigate the form of programmed cell death (PCD) induced by severe fever with thrombocytopenia syndrome virus (SFTSV) infection in Vero cells and further explore the existence of pyroptosis, so as to provide new ideas for studying the pathogenic mechanism of SFTSV.Methods:Vero cells were infected with SFTSV at different multiplicity of infection (MOI), cytopathic effect (CPE) was observed daily, cell viability was detected by CCK-8 method, and cell membrane damage was detected by LDH release test to determine the optimal amount of virus infection and cell death time. Vero cells were pretreated with different PCD inhibitors and infected with SFTSV. CCK-8 kit was used to detect the cell viability and determine the death form of PCD caused by SFTSV. The expression of pyroptosis related proteins was detected by Western blotting to further explore the existence of pyroptosis.Results:At 48 h and 72 h after SFTSV infected Vero cells with MOI=10, the optimal infection amount and time of subsequent experiments were observed. At this time, the CPE of cells was obvious, the cell viability decreased to 51% and 41% of the control group ( P<0.001, P<0.001), and the LDH release amount reached 24% and 37% of the maximum enzyme activity release wells, were 3.8, 3.4 times of LDH release in the control group ( P<0.001, P<0.001). The inhibition of SFTSV-induced cell death by different PCD inhibitors showed that pan-caspase inhibitor and receptor-interacting serine/threonine-protein kinase 3 (RIP3) inhibitor had inhibitory effects at 48 h and 72 h. The cell viability was 2.1 and 1.6 times of the viral control group at 48 h, 2.3 and 1.7 times of the viral control group at 72 h, and the effect of pan-caspase inhibitor was significantly higher than that of the other inhibitor groups. Caspase-1 and caspase-3 inhibitors only had inhibitory effect at 48 h, and the cell viability was 1.5 and 1.3 times of the viral control group. After SFTSV infection of Vero cells, the expressions of caspase-1 and IL-1β increased gradually with the prolongation of time, and reached 3.4 and 9.5 times of the control group at 48 h, respectively ( P<0.001, P<0.001). Conclusions:SFTSV infection of Vero cells can lead to various forms of PCD, including apoptosis, pyroptosis and programmed necrosis, and pyroptosis related protein activation can be detected in the process of PCD, which further explored the existence of pyroptosis.

Objective:To study the inactivation effects of microwave on human adenovirus 2 (HAdV-2) in simulated infectied wastes, and to explore its molecular mechanism.Methods:25 μl of HAdV-2 virus suspension was dripped with medical disposable gloves, masks and cotton swabs to simulate infectied wastes, and irradiated under different microwave conditions: gloves and masks were irradiated for 30 s, 60 s, and 90 s at 300 W, 500 W, and 700 W, respectively. Cotton swabs irradiate 60 s, 90 s, 120 s at 500 W and 700 W respectively. Temperature changes were recorded, and the inactivation logarithmic values were calculated by the 50% endpoint method to evaluate the microwave inactivation effects, and the proliferation ability of the virus was detected by qPCR. The damage of Penton, Fiber, Hexon and E2 B genes was detected by PCR. The virus was treated with the highest temperature of 76 ℃ during microwave irradiation to study whether there was non-thermal effect during microwave disinfection. Results:After microwave irradiation of infectied waste, the temperature of masks and gloves carriers rises rapidly, with the highest temperature of 76 ℃. The temperature of the cotton swab carriers rose slowly, and the highest temperature is 65 ℃. The inactivation effect of microwave on HAdV-2 was positively correlated with microwave power and irradiation time. In the mask and glove group, microwave power of 700 W irradiated for 60 seconds, and the inactivation logarithm value could reach 3.0, In the cotton swab group, microwave power of 700 W irradiated for 120 seconds, and the inactivation logarithm value was still less than 3.0. This indicated that there were differences in the conditions for microwave inactivation of the virus in different carriers. The qPCR result showed that microwave irradiation could weak the proliferation ability of the virus. Microwave irradiation had no effect on the virus's Penton and Fiber genes, but caused some damage to the Hexon and E2 B genes. The inactivation effect of individual heat treatment on HAdV-2 was weaker than that of microwave irradiation, and there was no damage to the Hexon, Penton, Fiber, and E2 B genes. This indicated the presence of non thermal effects during the microwave inactivation process. Conclusions:Microwave irradiation can inactivate HAdV-2 in simulated infectied wastes through thermal and non-thermal effects, and its damage to viral DNA is one of the mechanisms of virus inactivation.

Objective:To explore the clinical efficiency evaluation and prognostic factors of aspiration guided by neuronavigation in the treatment of pediatric brain abscess (PBA).Methods:A total of 47 patients with PBA were treated with aspiration guided by neuronavigation between January 2013 and January 2019 at the First Affiliated Hospital of Zhengzhou University.All clinical data were retrospectively analyzed.According to Glasgow Outcome Scale on discharge, all children were divided into 2 groups, namely good prognosis group and poor prognosis group.Prognostic factors were analyzed by using univariate analysis and binary Logistic regression multivariate analysis. Results:Among the 47 children, 38 children (80.9%) were assigned to the good prognosis group, and 9 children (19.1%) were assigned to the poor prognosis group.Univariate analysis proved that abscess volume>4 cm( χ2=5.650, P=0.017), multiple or multilocular abscess ( χ2=3.258, P=0.027), and abscess located in functional areas ( χ2=6.187, P=0.013) were correlated with poor prognosis.Multivariate analysis revealed that abscess volume>4 cm( OR=5.913, 95% CI: 2.241-25.917, P=0.023) and abscess located in functional areas ( OR=10.519, 95% CI: 3.918-62.513, P<0.001) were independent risk factors for poor prognosis. Conclusion:The treatment of PBA with aspiration guided by neuronavigation is safe, effective and minimal invasive, and the clinical efficiency is satisfactory.Abscess volume>4 cm and abscess located in deepbrain/functional areas are independent risk factors for poor prognosis.

Objective:To explore the diagnosis, treatment and prognostic of pediatric intracranial atypical teratoid/rhabdoid tumor(AT/RT).Methods:A total of 15 pediatric patients with intracranial AT/RT were treated between January 2012 and June 2019 at the First Affiliated Hospital of Zhengzhou University.The clinical data were retrospectively analyzed.Overall survival (OS) rate and progression free survival (PFS) rate were calculated by adopting Kaplan- Meier method.The differences between the 2 groups were tested by performing Log- rank method, and the prognostic factors were analyzed by COX regression. Results:There were 12 males and 3 females, with the median age of 5.5 years (ranging from 8 months to 17.1 years). All patients underwent surgical resection.Gross-total resection (GTR) was achieved in 10 cases and subtotal resection (STR) was carried out in 5 patients.The conducted treatments were as follows: surgery+ radiotherapy+ chemotherapy+ intrathecal injection in 6 cases, surgery+ chemotherapy+ intrathecal injection in 4 cases, surgery+ radiotherapy in 2 cases, and surgery alone in 3 cases.Until January 2020, the median survival time of all the 15 patients was 18 months (ranged 1-27 months), and the survival rate was 33.3%.The 1-year OS rate and PFS rate for all 15 cases were 71.5% and 49.7%, respectively.The 2-year OS rate and PFS rate were 17.9% and 0, respectively. Log- rank analyses revealed that the 1-year OS rates of children less than 3 years old and those older than 3 years were 87.5% and 57.1%, respectively ( χ2=6.057, P=0.014). The 1-year OS rates of children with GTR and those with STR were 90.0% and 40.0%, respectively ( χ2=6.057, P=0.014). The 1-year OS rates of children with tumor dissemination and those without tumor dissemination were 100.0% and 33.3%, respectively( χ2=9.865, P=0.002). The 1-year OS rates of children in the standard-risk group and those in the high-risk group were 88.9% and 41.7%, respectively ( χ2=5.111, P=0.024). COX regression analyses proved that age, the extent of tumor resection, tumor dissemination and risk stratification are independent risk factors for prognosis [hazard radio( HR)=3.411, 3.795, 5.245, 3.397; P=0.025, 0.011, 0.001, 0.017]. Conclusions:Pediatric intracranial AT/RT is rare.The preliminary diagnosis and prognosis are difficult and poor, respectively.The complete resection of tumors with maximal safety remains the primary treatment.Age, the extent of tumor resection, tumor dissemination and risk stratification are independent prognostic factors for AT/RT children.

Objective:To study the amino acid site variation of HIV-1 Tat protein from different parts of AIDS patients with HAD and non HAD and its effect on oxidative stress of U87 cells.Methods:HIV-1 Tat amino acid sequences were analyzed by BLAST and MEGA6 software to study the variation of amino acid sites in four parts of central nervous tissue basal ganglia(BG) and peripheral spleen (SPL) of an HIV-associated dementia (HAD) patient(H) and a non-HAD patient(N), The HIV-1 tat genes were transfected into U87 cell. The green fluorescent protein was observed under microscope to determine the Tat protein expression. The expression of Tat protein in U87 cells was detected by Western blotting. CK-8 method , Western blotting and malondialdehyde (MDA) detection kit were used to study the effect of Tat protein on cell activity, oxidative stress index glutathione peroxidase (GPX), MDA level. Results:Amino acid sequence analysis showed that the key amino acid sites of HIV-1 Tat protein from N-BG, N-SPL, H-BG and H-SPL were different; Tat protein could inhibit the activity of U87 cells, which could be reversed by antioxidant N-acetyl-L-cysteine (NAC). Compared with the control group, the levels of MDA were increased and the expression of GPX protein was decreased in the four experimental groups ( P<0.05). And different sources of Tat protein had different ability to induce oxidative stress, the level of MDA in H-BG group was higher than that in N-BG group( P<0.05). The expression of GPX protein in BG group of both HAD and non-HAD patients was lower than that of SPL group( P<0.05). Conclusions:There are differences in the key amino acid sites of Tat protein in peripheral and central nervous system between HAD and non-HAD patients, and their effects on oxidative stress were also different.

Objective:To investigate the role of HIV-1 negative regulator (Nef) in HIV-1 neuropathogenicity.Methods:Five different HIV-1 nef genes were obtained from the central nervous system (CNS) and peripheral regions (basal ganglia, frontal cortex, meninges, temporal cortex and spleen) of a patient with HIV-1-associated dementia (HAD). The recombinant pcDNA3.1- nef eukaryotic expression vectors were constructed by connecting them with pcDNA3.1 vector and transfected into human glioma cell line U87 respectively. The expression of Nef protein was detected by immunohistochemistry and Western blotting at 22ndhour, 27 th hour, 32nd hour, 37th and 42nd hour after transfection. The result were analyzed quantitatively by JEDA801D and JD-801 software. The supernatant of U87 cells was collected every 5 hours from 27th hour to 62nd hour after transfection. The levels of TNF-α and IL-1 β in the supernatant were determined by ELISA, and the dynamic expression of the two cytokines was analyzed. Results:Five recombinant pcDNA3.1- nef eukaryotic expression vectors of nef genes from different tissues of an HAD patient were successfully constructed and transfected into U87 cells. The result of immunohistochemistry showed that Nef protein began to express at 42nd hour after transfection, which was further confirmed by Western blot. The result of ELISA showed that the levels of cytokines in the supernatant of each group increased gradually with time from 22ed hour to 37th hour after transfection, but there was no significant difference among the groups (TNF-α: F=0.445, P=0.837; F=0.579, P=0.742; F=0.617, P=0.714; F=2.728, P=0.057. IL-1β: F=2.656, P=0.062; F=0.485, P=0.809; F=0.165, P=0.982; F=2.463, P=0.093); The levels of TNF-α and IL-1 β in the experimental group were significantly higher than those in the control group from the 42nd hour ( P<0.05); after 42nd hour, the levels of cytokines in each group gradually decreased, and the levels of TNF-α and IL-1 β remained stable from the 57th hour to the 62nd hour, while the levels of TNF-α and IL-1 β in the experimental group were higher than those in the control group from the 42nd hour to the 62nd hour, the difference was still statistically significant (TNF-α: F=241.310, P<0.001; F=242.638, P<0.001; F=250.114, P<0.001; F=143.877, P<0.001; F=146.172, P<0.001. IL-1β: F=251.578, P<0.001; F=188.816, P<0.001; F=276.240, P<0.001; F=238.136, P<0.001; F=163.361, P<0.001), and there was no significant difference among the experimental groups ( P>0.05). Conclusions:HIV-1 Nef protein can induce and enhance the secretion of TNF-α and IL-1 β in U87 cells. However, the amino acid variation of HIV-1 Nef protein from different sources in an HAD patient had no effect on the secretion of TNF-α and IL-1 β.

Objective: To investigate the toxic effect of HIV-1 Vpr protein on neurons. Methods: HIV-1 vpr gene was amplified by nested PCR in four parts of peripheral spleen (SPL) and central nervous tissue meninges (MG) of HIV-associated dementia (HAD) patients and non-HAD patients. Eukaryotic expression vector pEGFP-N1-vpr was constructed. The gene sequence and key amino acid sites were analyzed by BLAST and MEGA6. The expression of Vpr protein in N2a cells was detected by Western-blotting. The effects of Vpr proteins from different sources on the activity and cell cycle of N2a cells were studied by flow cytometry. Results: HIV-1 vpr gene was successfully amplified by PCR. Sequence analysis showed that the vpr gene sequence belonged to HIV-1B subtype. There were amino acid mutations at C-terminal 84, 86 and 87 sites of central Vpr protein from HAD and non-HAD patients. Vpr protein could inhibit the activity of nerve cells, leading to G2 phase arrest. Different sources of Vpr had different intensity of action. Compared with other groups, Vpr protein from the meninges of HAD patients showed stronger inhibition of cell activity and G2 phase arrest ability. Conclusions Variations in key amino acid sites of Vpr protein could cause significant changes in its biological functions, and its significance in the pathogenesis of HAD remains to be further studied.