Objective:The whole genome of one porcine sapelovirus (PSV) strain from mosquitoes captured in Weishan county, Shandong province was sequenced and analyzed its genetic characteristics.Methods:Mosquitoes collected in pigpen in Weishan county of Shandong province in 2021 were homogenized and inoculated on C6/36 and BHK21 cell lines for virus isolation. Virus RNA was extracted from the positive culture and the whole genome was sequenced by next generation sequencing. Phylogenetic analysis, similarity comparison and recombination analysis were conducted via MEGA 7.0.26, BioEdit 7.0.9.0 and Simplot 3.5.1.Results:A PSV strain 265/SD was isolated from Culex tritaeniorhynchus. The clear cytopathic effects characterized by rounding, shrinkage and detachment of cells were observed on BHK21 cells. The whole genome of this study strain was 7 558 nt in length. Phylogenetic analysis based on VP1 sequences indicated that strain 265/SD belonged to traditional genotype. Different positions of 265/SD in the VP1 and 3D phylogenetic trees suggested the recombinant potential. Similarity plot revealed the existence of recombination but the exact origins remained unknown. The similarity analysis of VP1 regions showed that strain 265/SD had low nucleotide identity (77.3%) and amino acid identity (80.3%) with another strain SD/CHN/2016 from Shandong province. Conclusions:Our data enriched the whole genome information of PSV in Shandong province and provided scientific basis for prevention and control of PSV.

Objective:The whole genome of one porcine sapelovirus (PSV) strain from mosquitoes captured in Weishan county, Shandong province was sequenced and analyzed its genetic characteristics.Methods:Mosquitoes collected in pigpen in Weishan county of Shandong province in 2021 were homogenized and inoculated on C6/36 and BHK21 cell lines for virus isolation. Virus RNA was extracted from the positive culture and the whole genome was sequenced by next generation sequencing. Phylogenetic analysis, similarity comparison and recombination analysis were conducted via MEGA 7.0.26, BioEdit 7.0.9.0 and Simplot 3.5.1.Results:A PSV strain 265/SD was isolated from Culex tritaeniorhynchus. The clear cytopathic effects characterized by rounding, shrinkage and detachment of cells were observed on BHK21 cells. The whole genome of this study strain was 7 558 nt in length. Phylogenetic analysis based on VP1 sequences indicated that strain 265/SD belonged to traditional genotype. Different positions of 265/SD in the VP1 and 3D phylogenetic trees suggested the recombinant potential. Similarity plot revealed the existence of recombination but the exact origins remained unknown. The similarity analysis of VP1 regions showed that strain 265/SD had low nucleotide identity (77.3%) and amino acid identity (80.3%) with another strain SD/CHN/2016 from Shandong province. Conclusions:Our data enriched the whole genome information of PSV in Shandong province and provided scientific basis for prevention and control of PSV.

New threats posed by the emerging circulating variants of SARS-CoV-2 highlight the need to find conserved neutralizing epitopes for therapeutic antibodies and efficient vaccine design. Here, we identified a receptor-binding domain (RBD)-binding antibody, XG014, which potently neutralizes β-coronavirus lineage B (β-CoV-B), including SARS-CoV-2, its circulating variants, SARS-CoV and bat SARSr-CoV WIV1. Interestingly, antibody family members competing with XG014 binding show reduced levels of cross-reactivity and induce antibody-dependent SARS-CoV-2 spike (S) protein-mediated cell-cell fusion, suggesting a unique mode of recognition by XG014. Structural analyses reveal that XG014 recognizes a conserved epitope outside the ACE2 binding site and completely locks RBD in the non-functional "down" conformation, while its family member XG005 directly competes with ACE2 binding and position the RBD "up". Single administration of XG014 is effective in protection against and therapy of SARS-CoV-2 infection in vivo. Our findings suggest the potential to develop XG014 as pan-β-CoV-B therapeutics and the importance of the XG014 conserved antigenic epitope for designing broadly protective vaccines against β-CoV-B and newly emerging SARS-CoV-2 variants of concern.
Angiotensin-Converting Enzyme 2 ; Antibodies, Neutralizing ; Antibodies, Viral ; COVID-19 ; Epitopes ; Humans ; SARS-CoV-2/genetics* ; Spike Glycoprotein, Coronavirus/genetics*

Objective:To explore the efficacy and safety of anti-tumor necrosis factor alpha (TNFα) monoclonal antibodies (mAbs) for severe/refractory vasculo-Behcet′s disease (BD).Method:The clinical data of severe/refractory vasculo-BD patients treated with anti-TNFα mAbs were retrospectively analyzed. Response of anti TNFα mAbs was analyzed. The dosage changes of glucocorticoid, the level of erythrocyte sedimentation rate (ESR) and hypersensitive C-reactive protein (hsCRP) before and after treatment were recorded, as well as side effects.Result:Sixteen patients were enrolled. Arterial lesions were reported in 12 patients, including 9 with arterial aneurysm, 6 with arterial dilation, 2 with stenosis and 2 with occlusion. Seven patients presented venous thrombosis, including lower extremity veins ( n=6), cerebral venous sinus ( n=2) and inferior vena cava system ( n=2). Two cases had both arterial and venous involvement. Before the application of TNFα mAbs, all 16 patients failed to response to prednisone or its equivalent dose of 40 (7.5-90) mg/d in combination with cyclophosphamide, methotrexate, thalidomide or azathioprine for median 4 (0-156) months. After a mean duration of treatment for (17.1±6.5) months, 15 patients achieved complete remission and 1 patient achieved partial remission. Three patients received surgery without any postoperative complications. After using anti TNFα mAbs, the dosage of prednisone [5(0-12.5)mg/d vs. 40(7.5-90)mg/d, P<0.01], ESR [(7.3±4.6) mm/1h vs. (33.5±26.7) mm/1h, P<0.01] and hsCRP [1.9(0.2-11.4) mg/L vs. 24.3(0.4-113.9) mg/L, P<0.01] were significantly decreased. Side effects were observed in 2 patients. One developed pulmonary infection 12 months after adalimumab with conventional treatment. Another patient had allergy to infliximab then switched to adalimumab. Conclusion:In combination with corticosteroids and immunosuppressants, anti-TNF α mAbs are effective and well-tolerated in severe/refractory vasculo-BD, with a favorable steroid -sparing effect and rare postoperative complications.

Objective: To explore the mediating role of depression in the association between life events and Internet addiction, and to provide evidence for the intervention of Internet addiction. Methods: A total of 3 536 students randomly selected from 3 vocational colleges in Anhui Province completed the questionnaire survey, which included adolescents’ demographic characteristics, the Young Internet Addiction Inventory, the Adolescent Life Events Scale, and the Zung Self-Rating Depression Scale. Results: Of the 3 536 students surveyed, 427 were Internet addicts (12.08%), including 183 boys (14.89%) and 244 girls (10.58%). Negative life events were associated with depression and Internet addiction (r=-0.30，0.28, P<0.01); depression was mediated indirectly between negative life events and Internet addiction. There was statistical significance (a=0.30, b=0.13, P<0.01). Depression-mediated indirect effects accounted for 14.67% of the total effects. Conclusion Depression plays a mediating role in the relationship between negative life events and adolescents’ Internet addiction, suggesting that we can reduce the incidence of Internet addiction by reducing students’ depression through early psychological diagnosis and psychological quality training.

Objective To explore the effect of electroacupuncture at Quchi (LI11), Zusanli (ST36) on differentiation of neural stem cells after cerebral ischemia-reperfusion in rats. Methods Thirty-six male Sprague-Dawley rats were randomly divided into sham group (n=12), model group (n=12) and electroacupuncture group (n=12). The latter two groups were occluded the left middle cerebral arteries for 90 min-utes and reperfused. The electroacupuncture group received electroacupuncture at Quchi and Zusanli acupoints for 21 days. They were evalu-ated with modified Neurological Severity Scores 7, 14 and 21 days after electroacupuncture. Their infarct volumes were tested with MRI T2WI 21 days after electroacupuncture, while the differentiation of neural stem cells was observed with double-immunopositive BrdU/Dcx and BrdU/NeuN. Results Compared with the model group, the neurological deficits score improved in the electroacupuncture group in all the time points (P<0.05). The infarct volumes decreased in the electroacupuncture group (P<0.05), with less number of BrdU+/Dcx+cells in subventricular zone (P<0.001) and more number of BrdU+/NeuN+ cells in peri-infarct cortex (P<0.001) 21 days after electroacupuncture. Conclusion Electroacupuncture at Quchi and Zusanli acupoints can improve neurological function and decrease the infarct volumes in rats after cerebral ischemia-reperfusion, which may be associated with promoting differentiation of neural stem cells to neurons.

Objective To investigate the effect of electroacupuncture (EA) at Shenting (DU24) and Baihui (DU20) acupoints on hippocampal synaptic plasticity and learning and memory function, and its possible mechanism in rats with cerebral ischemia-reperfusion. Methods Thirty-two male adult Sprague-Dawley rats were randomly divided into sham group (n=8), model group (n=8), electroacupuncture (EA) group (n=8) and non-acupoint group (n=8). The model group, EA group and non-acupoint group were performed with left middle cerebral artery occlusion (MCAO). The EA group received electroacupuncture at Shenting and Baihui, while the non-acupoint group received electroacupuncture at two fixed non-acupoints from bilateral ribs, for fourteen days. Then they were tested with Morris Water Maze test, and their synaptic structure of hippocampal neurons was observed with transmission electron microscope. The level of total LIM-domain containing protein kinase 1 (T-LIMK1) and LIM-domain containing protein kinase 1 phosphorylation (p-LIMK1) were detected with Western blotting. Results Compared with the sham group, the latency increased (t>6.789, P<0.01) and the frequence crossing platform decreased (t= 8.695, P<0.001) in the model group, while the number of synapse in hippocampal neurons decreased, and the level of T-LIMK1 (t=7.568, P< 0.01) and p-LIMK1 (t=8.874, P<0.001) decreased. Compared with the model group, the latency decreased (t>4.938, P<0.01) and the frequence crossing platform increased (t=- 7.891, P<0.001) in the EA group, while the number of synapses increased, and the level of T-LIMK1 (t=-6.473, P<0.01) and p-LIMK1 (t=-6.579, P<0.01) increased. There was no significant difference between the the non-acupoint group and the model group in all the indices (P>0.05). Conclusion Electroacupuncture at Shenting and Baihui can improve the ability of learning and memory in rats with cerebral ischemia-reperfusion, which may relate with the increase of LIMK1 phosphorylation and hippo-campal synaptic plasticity.

Objective To study the clinical profile of patients with multicentric reticulohistiocytosis (MRH).Methods The clinical manifestations,image and histologic characteristics of ten patients with MRH were collected,and the relevant literatures were reviewed.Results Seven cases were women.The median (quartile) age was 46.5 (44.5-55.3) years.Clinical symptoms included papulonodular lesions on the skin (ten cases),erosive polyarthritis (ten cases),myositis (four cases),lung nodules (three cases),larynx nodules (one case) and serositis (one case).Biopsy demonstrated infiltration of multinucleated giant cells with CD68 positive and glassy cytoplasm.All of the ten patients were treated with corticosteroids and disease-modifying antirheumatic drugs,skin lesions of seven patients were ameliorated and arthritis was improved in three patients.Conclusion MRH is a systemic disease that has characteristic clinical manifestations and unique histological findings.Deep understanding of this disease can avcid.isdiagnose.Further studies are needed to explore effective treatment.

Objective To investigate the effects of downstream regulatory element antagonist modulator (DREAM) on the expression of glutamate transporter-1 (GLT-1) in spinal cord in rats with bone cancer pain and morphine tolerance.Methods Sixty female healthy Sprague-Dawley (SD) rats weighing 200 ～230 g were randomly divided into tow groups,group Ⅰ cancer pain (CP,n =48) and group Ⅱ Sham (S,n =12).Cancer pain in each group was produced by inoculation of syngenetic Walker 256 rat mammary gland carcinoma cells (5 × 105) to left tibia.Pain threshold to mechanical stimulus was measured before (baseline) and after the surgical procedure.From 14 d to 18 d after the inoculation of carcinoma cells,36 rats from group CP received subcutaneous injection of morphine at 3 times per day with doses increasingly from 10 mg/kg initially to 20 mg/kg,30 mg/kg,40 mg/kg,and 60 mg/kg.Equal volume of normal saline was applied to the 12 rats left in group CP.On 19th day after the carcinoma cells inoculation once subcutaneous injection of morphine at 3mg/kg was performed in all rats in group CP.From the next day,the rats in group CP ever receiving injections of morphine for 5 days were randomized into thre subgroups,including subgroups morphine tolerance (MT,n =12),vehicle (V,n =12),and RNAi (R,n =12).They were injected intrathecally with 20 μl of normal saline (NS),10 μl vehicle plus 10 μ1 NS,and 10 μ1 of DREAM-shRNA plus l0 μ1 NS,respectively,once a day for 5 days.Focusing on the affected limb,mechanical pain threshold was measured one day before surgery (T0),and at day7 (T1),day 14 (T2),day 18 (T3),day 19 (T4),day21 d (T5),day 25 (T6),and day 28 (T7) after surgery.The animals were sacrificed at day 28 after the procedure.The lumbar 4 segments in rats were removed for detection of DREAM and GLT-1.Results The mechanical threshold was significantly decreased at T1 compared to the baseline in all groups,returned to the baseline at T2 ～ T7 in group S,at T4 in group CP,and at T2 in group MT,V,and R,but remained low at T5 ～T7 in group CP,and at T3 ～T7 in group MT,V,and R.Compared to that at T1,it was decreased at T2 ～T3 and T5 ～ T7 in group CP,at T4 ～ T7 in group MT and V,and at T4 ～ T5 in group R,going back to the baseline at T4 in group CP and at T2 in group MT,V and R,and increased at T6 ～ T7 in group R.Compared to that in group S,the mechanical threshold in group CP,MT,V and R was decreased,and lower at T2 in group CP and at T4 in group MT,V and R.Compared to that in group CP,the mechanical threshold was significantly higher at T2 ～ T3 but lower at T4 in group MT,V,and R,decreased at T5 in group R and at T5 ～ T7 in group MT and V.The mechanical threshold was increased at T6 ～ T7 in group R and higher than that in groups MT and V.The expression of DREAM,compared to that in group S,was down-regulated in other groups.Compared to group CP,increment was shown in groups MT and V,and decrease was exhibited in group R.It was cut down in group R compared to that in groups MT and V.Compared to group S,GLT-1 was decreased in other groups.It was down-regulated in groups MT,V and R compared to group CP.Conclusions DREAM is involved in the development of allodynia after morphine tolerance in rats with bone cancer pain.No evidence in this study supports a link between DREAM and GLT-l in spinal cord.