Objective The study aims to investigate the immunological functions of the nucleocapsid (N) protein of the novel coronavirus Omicron (BA.1, BA.2) and evaluate the differences among different N proteins of mutant strains in immunogenicity. Methods By aligning sequences, the mutation sites of the Omicron (BA.1, BA.2) N protein relative to prototype strain of the novel coronavirus (Wuhan-Hu-1) were determined. The pET-28a-N-Wuhan-Hu-1 plasmid was used as template to construct pET-28a-BA.1/BA.2-N through single point mutation or homologous recombination. The three kinds of N protein were expressed in prokaryotic system, purified through Ni-NTA affinity chromatography, and then immunized into mice. The titer and reactivity of the polyclonal antibody, as well as the expression level of IL-1β and IFN-γ in mouse spleen cells, were detected using indirect ELISA and Western blot assay. Results The constructed prokaryotic expression plasmids were successfully used to express the Wuhan-Hu-1 N, BA.1 N, and BA.2 N proteins in E.coli BL21(DE3) at 37 DegreesCelsius for 4 hours. The indirect ELISA test showed that the titers of polyclonal antibody prepared by three N proteins were all 1:51 200. All three N proteins can increase the expression of IFN-γ and IL-1β cytokines, but the effect of Omicron N protein in activing two cytokines was more obvious than that of Wuhan-Hu-1 N protein. Conclusion The study obtained three new coronavirus N proteins and polyclonal antibodies, and confirmed that mutations in the amino acid sites of the N protein can affect its immunogenicity. This provides a basis for developing rapid diagnostic methods targeting N protein of different novel coronavirus variants.
Animals ; Mice ; SARS-CoV-2/genetics* ; Coronavirus Nucleocapsid Proteins/immunology* ; Nucleocapsid Proteins/isolation & purification* ; COVID-19/immunology* ; Antibodies, Viral/immunology* ; Mice, Inbred BALB C ; Interferon-gamma/metabolism* ; Interleukin-1beta/metabolism* ; Female ; Escherichia coli/metabolism* ; Mutation ; Humans

Objective:To investigate the mechanism of immunomodulatory effects of umbilical cord mesenchymal stem cells(UC-MSCs)modified by miR-125b-5p on systemic lupus erythematosus(SLE).Methods:The expression level of miR-125b-5p was detected by real-time fluorescence quantitative PCR in UC-MSCs and peripheral blood mononuclear cells(PBMCs)from SLE patients and health checkers.Annexin V-FITC/PI apoptosis detection kit was used to detect the effect of miR-125b-5p on apoptosis of UC-MSCs.MRL/lpr mice in each group were injected with UC-MSCs via tail vein,and T-lymphocyte subsets in the spleen of the MRL/lpr mice were detected by flow cytometry after 5 weeks.The expression levels of interleukin(IL)-4 and IL-17A in serum of MRL/lpr mice were detected by ELISA.Hematoxylin-eosin staining was used to observe the pathological manifestations of the lungs and kidneys of the MRL/lpr mice.Results:miR-125b-5p was significantly down-regulated in PBMCs of SLE patients compared with healthy controls(P＜0.01).Compared with the UC-MSCs group,the expression of miR-125b-5p in UC-MSCs modified by miR-125b-5p group was increased(P＜0.01).The survival rate of UC-MSCs was significantly increased by miR-125b-5p(P＜0.01).Compared with the untreated group of MRL/lpr mice,the expression level of IL-4 in serum was increased(P＜0.05);the expression level of IL-17A was decreased(P＜0.05);the proportion of Th17 cells in the spleen of MRL/lpr mice was decreased(P＜0.05);the inflammatory cells infiltration and micro-thrombosis of lungs and kidneys of MRL/lpr mice were significantly reduced in the UC-MSCs modified by miR-125b-5p treatment group.Conclusion:UC-MSCs modified by miR-125b-5p have immunomodulatory effects on systemic lupus erythematosus.

Mechanical ventilation(MV)provides life support for critically ill respiratory patients,but in the meantime can cause fatal ventilator-induced lung injury(VILI),and the latter remains a major challenge in respiratory and critical care medicine,because the pathological mechanism has not been fully elucidated.Recent studies show that on the one hand,in the lung with VILI,there exists airway collapse at multi-sites of an individual airway,which can not be explained by traditional airway collapse models.But on the other hand,under MV conditions,airway smooth muscle cells(ASMC)exhibit abnormal mechanical behaviors,accompanied by regulation of Piezo1 expression and endoplasmic reticulum stress.These phenomenons indicate that the MV-induced abnormal mechanical behavior of ASMC is closely related to multiple airway collapse and VILI.Therefore,by studying the MV-induced changes of ASMC mechanical behaviors and their relationship with airway collapse in lung injury,as well as the related mechanochemical signal coupling process,it is expected to reveal a novel mechanism of MV-associated airway collapse and lung injury from the perspective of cell mechanics.In this review,the recent research progress of airway collapse during MV,the regulation of ASMC mechanical behavior by MV-related high stretch,especially the related mechanochemical signal coupling mechanism is summarized.These advances may provide a novel insight for exploring the roles of ASMC abnormal mechanical behavior in the pathological mechanism of VILI,alternative targets of drug intervention for prevention and treatment of VILI,as well as for optimizing the ventilation mode in clinical practice.

Objectives:To investigate the association between social determinants of health(SDOH)and incident stroke and analyze the main risk factors for stroke among resident with different SDOH levels. Methods:From 2012 to 2015,30 036 residents(≥35 years old)from 30 districts in 14 provincial-level administrative divisions in China were enrolled this study based on stratified multi-stage-random-sampling method.The prevalence of cardiovascular diseases and related risk factors were investigated,and stroke events were followed up in 2018 to 2019.Principal component analysis was performed to establish SDOH scores based on 9 indicators related to socioeconomic and healthcare resources,participants were divided into low SDOH group(n=8 343)when it was≥-2.01 to<-1.14,middle SDOH group(n=7 257)when it was≥-1.14 to<0.10,and high SDOH group(n=8 457)when it was≥0.10 to≤5.79.Multivariate Cox regression was applied to estimate the association of SDOH levels with incident stroke.The random survival forest method was used to analyze the major risk factors in different SDOH levels. Results:A total of 24 057 participants were finally included,669(2.8%)participants developed stroke during a mean of(4.7±0.8)years follow-up.The incidence densities of stroke in the low,medium,and high SDOH groups were 468.39,628.85,and 700.39/100 000 person-years,respectively(Pdifference<0.05,Ptrend=0.01).Compared with individuals with low SDOH level group,fully HR for incident stroke among those with medium and high were 1.91(95%CI:1.54-2.36)and 1.59(95%CI:1.30-1.95),respectively(Ptrend<0.001).Advanced age is the primary risk factor for stroke in the population,especially in districts with high SDOH level.In districts with medium SDOH level,diabetes is an important risk factor for stroke.High blood pressure and alcohol consumption are important modifiable risk factors in low SDOH level districts. Conclusions:Present study shows that higher levels of SDOH are associated with increased risk of stroke.The main risk factors for stroke differ among participants with different SDOH level districts.Targeted interventions should be implemented to improve the prevention and treatment of stroke in populations with different levels of SDOH.

Objective:To explore the relationship among suicide risk,distress tolerance,and resilience in pa-tients with depression.Methods:A total of 130 patients with depression who met the Diagnostic and Statistical Manual of Mental Disorders,Fifth Edition(DSM-5)diagnostic criteria were included.The Chinese version of the Mini International Neuropsychiatric Interview(MINI)suicide module was used to assess suicide risk.According to the interview results,the patients with depression were divided into suicide risk group(98 cases)and non-suicide risk group(32 cases).The Hamilton Depression Rating Scale(HAMD),Distress Tolerance Scale(DTS),10-item Connor-Davidson Resilience Scale(CD-RISC-10)were used to assess severity of depressive symptoms,level of distress tolerance and resilience,respectively.Results:The DTS total scores were positively correlated with the CD-RISC-10 total scores(r=0.50,P＜0.01).The total scores of MINI suicide module were negatively correlated with the total scores of DTS and CD-RISC-10(r=-0.34,-0.34,Ps＜0.01).Distress tolerance had a direct effect on suicide risk(β=-0.26,P＜0.05),and resilience played a mediating effect on the relationship between distress tolerance and suicide risk(β=-0.13,P＜0.05),and the mediating effect accounted for 33.5％of the total effect.Conclusion:Patients with depression with lower levels of distress tolerance may have higher suicide risk,and resilience may play a partially mediating role in the relationship between distress tolerance and suicide risk.

Objective : To investigate the imunomodulatory effects of umbilical cord mesenchymal stem cells(UC⁃MSCs) modified by miR⁃125b⁃5p through JAK2/STAT3 pathway on systemic lupus erythematosus(SLE) . Methods: UC⁃MSCs were isolated and cultured under aseptic conditions ; Peripheral blood mononuclear cells ( PBMCs)were separated from SLE patients by density gradient centrifugation. The relative expressions of IL⁃17A , Foxp3 ,IFN⁃γ , IL⁃4 genes in PBMCs cultured with UC⁃MSCs for 48 h were detected by RT⁃qPCR; The relative expressions of JAK2 , p ⁃JAK2 , STAT3 , p ⁃STAT3 and IL⁃18 proteins in kidney tissues of MRL/lpr mice were detected by west⁃ern blot. Results : Compared with the PBMCs culture group , the expression of IFN⁃γ and IL⁃17A and the propor⁃tion of Th17/Treg cells were significantly down⁃regulated in UC⁃MSCs + miR⁃125b⁃5p group , UC⁃MSCs + miR⁃NC group and UC⁃MSCs group(P < 0. 01) , the expression of the proportion of Th1/Th2 cells (P < 0. 05) was down⁃regulated in UC⁃MSCs + miR⁃125b⁃5p group and UC⁃MSCs + miR⁃NC group ; Compared with the untreated group of MRL/lpr mice , the relative expressions of JAK2 , p ⁃JAK2 , STAT3 , p ⁃STAT3 and IL⁃18 proteins in kidney tissues of MRL/lpr mice were significantly down⁃regulated in UC⁃MSCs + miR⁃125b⁃5p group (P < 0. 01) , and the rela⁃tive expressions of IL⁃18 proteins was significantly down⁃regulated in UC⁃MSCs + miR⁃NC group and UC⁃MSCs group (P < 0. 01) . Conclusion miR⁃125b⁃5p plays a synergistic role in UC⁃MSCs ,which regulates the differenti⁃ation of Th2 cells in PBMCs of SLE patients and the relative expressions of p ⁃JAK2/JAK2 , p ⁃STAT3/STAT3 , and IL⁃18 proteins in kidney tissues of MRL/lpr mice. UC⁃MSCs modified by miR⁃125b⁃5p may play immunomodulato⁃ry effect on SLE by JAK2/STAT3 signaling pathway.

Objective:To explore the prevalence and risk factors of sarcopenia in patients with maintenance dialysis (MHD).Methods:It was a cross-sectional study. Patients who received MHD treatment in the Blood Purification Center of Hainan Provincial People's Hospital in October 2019 were included as study subjects. The patients were divided into sarcopenia group and non-sarcopenia group according to whether they had sarcopenia or not. Chest CT imaging and laboratory examination data were collected. Dual-energy X-ray absorptiometry was used to measure the skeletal muscle mass. Chi-square test or Mantel-Haenszel trend chi-square test was used to compare the clinical data of patients with and without sarcopenia. Multivariate logistic regression equation was used to analyze the risk factors of sarcopenia.Results:A total of 182 MHD patients were enrolled in the study, and the prevalence of sarcopenia was 33.5% (61/182). The proportions of age ≥60 years old, diabetic nephropathy, tunneled-cuffed catheter, body mass index <18 kg/m 2, serum albumin <40 g/L, low density lipoprotein cholesterol ≥3.37 mmol/L, left ventricular ejection fraction <50%, chest CT-suspected pulmonary tuberculosis (PTB) and PTB in sarcopenia group were higher than those in non-sarcopenia group (all P<0.05). Multivariate logistic regression analysis results showed that left ventricular ejection fraction <50% (≥50% as a reference, OR=3.250, 95% CI 1.035-10.206, P=0.044), low-density lipoprotein cholesterol ≥3.37 mmol/L (<3.37 mmol/L as a reference, OR=6.354 ,95% CI 1.675-24.108, P=0.007), chest CT-suspected PTB (normal as a reference, OR=7.433, 95% CI 1.531-36.083, P=0.013), and PTB (normal as a reference, OR=28.871, 95% CI 3.208-259.872, P=0.030) were independent influencing factors of sarcopenia in MHD patients. Conclusions:The prevalence of sarcopenia is higher in MHD patients. Blood low-density lipoprotein cholesterol ≥3.37 mmol/L, ejection fraction <50%, chest CT-PTB and suspected PTB are independent risk factors of sarcopenia in MHD patients. Correcting left ventricular systolic function, regulating blood lipids and preventing PTB as early as possible can reduce the prevalence of sarcopenia in MHD patients.

Objective To express the monkeypox virus (MPXV) A23R protein in Escherichia coli and purify by Ni-NTA affinity column, and to prepare mouse antiserum against MPXV A23R. Methods The recombinant plasmid pET-28a-MPXV-A23R was constructed and transformed into Escherichia coli BL21 to induce the expression of A23R protein. After optimizing the conditions of expression, A23R protein was highly expressed. Recombinant A23R protein was purified by Ni-NTA affinity column and identified by Western blot analysis. The purified protein was used to immunize mice for preparing the A23R polyclonal antibody, and the antibody titer was detected by ELISA. Results The expression of A23R recombinant protein reached the peak under the induced conditions of 0.6 mmol/L isopropyl-β-D-thiogalactoside (IPTG), 37 DegreesCelsius and 20 hours. The purity of the protein was about 96.07% and was identified by Western blot analysis. The mice were immunized with recombinant protein, and the titer of antibody reached 1:102 400 at the 6th week after immunization. Conclusion MPXV A23R is expressed highly and purified with a high purity and its antiserum from mouse is obtained with a high titre.
Animals ; Mice ; Monkeypox virus ; Antibodies ; Enzyme-Linked Immunosorbent Assay ; Blotting, Western ; Recombinant Proteins ; Escherichia coli/genetics*

OBJECTIVE: To compare the effectiveness of TiRobot-assisted and C-arm X-ray fluoroscopy assisted percutaneous kyphoplasty (PKP) via pedicle of vertebra in the treatment of osteoporotic vertebral compression fracture (OVCF) of thoracic vertebrae. METHODS: The clinical data of 85 patients with OVCF of thoracic vertebrae who were admitted between January 2020 and March 2023 and met the selection criteria was retrospectively analyzed including 40 patients (50 vertebrae) undergoing PKP assisted by TiRobot (group A) and 45 patients (50 vertebrae) undergoing PKP assisted by C-arm X-ray fluoroscopy (group B). There was no significant difference in the comparison of baseline data such as gender, age, body mass index, bone mineral density T-value, fracture segment, trauma history, and preoperative numerical rating scale (NRS) score, Oswestry disability index (ODI), and Cobb angle of injured vertebra between the two groups ( P>0.05). The effectiveness evaluation indexes of the two groups, including the operation time, the volume of injected cement, the times of fluoroscopies, the length of hospital stay, and the occurrence of postoperative complications were collected and compared. Anteroposterior and lateral X-ray films and CT of the injured vertebra were reviewed at 1 day after operation to observe whether there was cement leakage and to evaluate the distribution of cement in the injured vertebra. Before and after operation, pain was assessed using the NRS score, dysfunction was assessed using the ODI, and vertebral height recovery was assessed by measuring the Cobb angle of the injured vertebrae by X-ray films. RESULTS: Both groups of patients successfully completed the operation, the operation time, the volume of injected cement, the times of fluoroscopies, and the length of hospital stay in group A were significantly less than those in group B ( P<0.05). The patients in two groups were followed up 4-12 months (mean, 9.6 months). Bone cement leakage occurred in 5 vertebrae in group A and 15 vertebrae in group B after operation, all of which leaked to the intervertebral space and around the vertebral body, and the patients had no obvious clinical symptoms. The difference of bone cement leakage between the two groups was significant ( P<0.05). No severe complication such as intraspinal leakage, infection, or vascular embolism was found in the two groups. At 1 day after operation, the distribution index of bone cement in group A was mostly grade Ⅴ, which was well dispersed; while in group B, it was mostly grade Ⅱ and grade Ⅴ; the difference of bone cement distribution index between the two groups was significant ( P<0.05). The NRS score, ODI, and Cobb angle of injured vertebra in both groups were significantly improved at 1 day after operation when compared with preoperative ones ( P<0.05). There was no significant difference in the difference of the above indexes between the two groups before and after operation ( P>0.05). CONCLUSION TiRobot-assisted unilateral PKP in the treatment of OVCF of thoracic vertebrae is safe and effective, which can reduce the X-ray transmission times during operation, shorten the operation time, reduce the volume of bone cement injection, and thus decrease incidence of bone cement leakage.
Humans ; Thoracic Vertebrae/surgery* ; Fractures, Compression/surgery* ; Spinal Fractures/surgery* ; Kyphoplasty ; Bone Cements ; Retrospective Studies

Viruses are powerful tools for the study of modern neurosciences. Most of the research on the connection and function of neurons were done by using recombinant viruses, among which neurotropic herpesvirus is one of the most important tools. With the continuous development of genetic engineering and molecular biology techniques, several recombinant neurophilic herpesviruses have been engineered into different viral tools for neuroscience research. This review describes and discusses several common and widely used neurophilic herpesviruses as nerve conduction tracers, viral vectors for neurological diseases, and lytic viruses for neuro-oncology applications, which provides a reference for further exploring the function of neurophilic herpesviruses.
Herpesviridae/genetics* ; Neurosciences ; Genetic Vectors/genetics* ; Genetic Engineering ; Neurons