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

Pneumothorax during pediatric laparoscopic surgery is a potentially fatal complication that may not be promptly recognized. It can occur due to congenital anatomical abnormalities, pre-existing pulmonary disease, or operative factors during laparoscopy. Clinical presentation may range from asymptomatic to acute respiratory distress, pleuritic chest pain, and even life-threatening circulatory collapse. Here, we report a case of sudden intraoperative pneumothorax accompanied by extensive subcutaneous emphysema of the neck and chest wall during laparoscopic high ligation of the hernial sac in a child. The child presented with a reducible left lower abdominal mass and mild pain 3 days prior but did not seek medical attention. Symptoms worsened 1 day prior to admission, with difficulty reducing the mass. On April 8, 2021, the patient was admitted to the Department of Anesthesiology, Zhuzhou Hospital Affiliated to Xiangya School of Medicine of Central South University, with a diagnosis of "left inguinal hernia." On the second day of hospitalization, laparoscopic high ligation of the left inguinal hernia sac was performed under general anesthesia. During the procedure, the patient developed a sudden increase in airway pressure, marked hemodynamic fluctuations, crepitus in the neck and right anterior chest regions, and significantly diminished breath sounds in the right lung. Emergent bedside chest X-ray confirmed a right-sided pneumothorax. Immediate intervention including thoracic needle decompression, closed thoracic drainage, the lung re-expansion was performed. The patient was discharged on the 7th postoperative day with full recovery. This case highlights the need for clinicians to remain vigilant for iatrogenic pneumothorax during pediatric laparoscopic surgery. Close intraoperative monitoring of vital signs is crucial for early detection, recognition, and timely management of pneumothorax to ensure patient safety during minimally invasive procedures.
Humans ; Laparoscopy/methods* ; Pneumothorax/etiology* ; Ligation/methods* ; Hernia, Inguinal/surgery* ; Male ; Intraoperative Complications/etiology* ; Child ; Herniorrhaphy/methods* ; Female ; Subcutaneous Emphysema/etiology*

OBJECTIVE To investigate the effect of bs-5-YHEDA iron chelating peptide combined with semaglutide on the cognitive ability and pathological characteristics of D-Gal-induced Alzheimer's disease(AD) model mice. METHODS Forty mice were randomly divided into 5 groups, namely the healthy control group, PBS group, bs-5-YHEDA iron chelating peptide group, combined treatment group and positive control group, with 8 mice in each group, half of each sex. Except for the healthy control group, D-galactose was injected to induce the AD mice model for 6 weeks. For 3 consecutive weeks starting from the 4th week, the bs-5-YHEDA iron chelating peptide group was injected with bs-5-YHEDA(1 mg·mL–1) once every other day at 200 µL in the tail vein; the bs-5-YHEDA iron chelating peptide(1 mg·mL–1) and semaglutide(25 nmol·kg–1·d–1) were given alternately once a day in the combination treatment group; the positive control group was given memantine(3.3 mg·kg–1·d–1) by gavage every other day. The healthy control group and PBS group were injected with the equal dose of PBS. At the end of treatment, the learning memory ability of mice was detected by the Morris water maze method, whole brain and whole blood were dissected, and pathological changes in hippocampal region were observed by HE staining, and Aβ expression and Tau protein phosphorylation levels were detected by immunohistochemistry, enzyme-linked immunosorbent assay and immunoblotting. RESULTS In the Morris water maze spatial exploration experiment, the differences in the number of times the mice traversed the platform, the ratio of swimming distance to the target quadrant, and the time ratio were statistically significant in each group(P<0.05); compared with the PBS group, the ratio of swimming distance to the target quadrant increased in the combined treatment group, and the differences were statistically significant(P<0.05). The results of HE staining showed that compared with the healthy control mice, the hippocampal area in the PBS group showed reduced levels of pyramidal cells, disorganized arrangement, cell edema, and deep staining of nuclei consolidation. Cellular disorganization, deep staining of nuclei and apoptosis in the hippocampus were significantly improved in each treatment group after drug treatment. Immunohistochemistry and Western blotting results showed that the Aβ expression levels and Tau protein phosphorylation levels were significantly higher in the PBS-administered mice compared with the healthy control mice, and the Aβ expression levels and Tau protein phosphorylation levels were reduced in each group after drug treatment, with statistically significant differences(P<0.01 or P<0.001 ). CONCLUSION The combination of bs-5-YHEDA iron chelating peptide and semaglutide can effectively improve the learning and memory ability and pathological characteristics of AD mice, but from the results of immunohistochemistry and immunoblotting experiments, the improvement of pathological characteristics of AD mice in the combination treatment group is not obvious compared with the single bs-5-YHEDA iron chelating peptide group, suggesting that there may be a threshold effect of our designed dual-target combination treatment on the cognitive improvement of AD mice, and the optimization and validation of the effect of multi-target combination treatment need further study.

A family carrying a homozygous variant of DNAJB2 gene C.91C>T (p.His31Tyr) with distal hereditary motor neuropathy (dHMN) associated with early-onset Parkinson′s disease was reported. The patient presented with distal limb weakness and atrophy at the early stage of the disease, and developed Parkinson′s symptoms more than 10 years later. Neuroelectrophysiological examination suggested motor and sensory axonal involvement. This mutation site had not been reported and was considered to be a neogenic mutagenicity of dHMN, excluding other mutations that can cause early-onset Parkinson′s disease.

Objective:To investigate the clinical application value of commonly used preoperative indicators of sarcopenia in predicting postoperative pneumonia in patients aged 70 years and above with esophageal cancer.Methods:A retrospective analysis was conducted on the clinical data of 398 elderly patients(≥70 years old)with esophageal squamous cell carcinoma who underwent thoracic laparoscopic radical resection of esophageal cancer in our hospital from January 2020 to December 2021.The study aimed to investigate the correlation between clinical pathological indicators and commonly used measurement indicators of sarcopenia and postoperative pneumonia.Statistical analysis was performed to analyze the data.Results:The study found that the proportion of postoperative pneumonia in esophageal squamous cell carcinoma patients aged 70 years and above was 27.9%(111 out of 398). The pneumonia group had significantly lower preoperative BMI and peak expiratory flow(PEF)measurements compared to the non-pneumonia group, with statistically significant differences( t=2.799, 2.674, both P<0.05). Logistic multivariate analysis revealed that low PEF, low psoas major muscle index(PMI), and low psoas muscle density(PMD)were the primary risk factors for postoperative pneumonia in esophageal cancer patients aged 70 years and above(Wald χ2 values were 7.577, 6.091, 6.845, all P<0.05). The risk of postoperative pneumonia in esophageal cancer patients aged 70 years and above with low PEF, low PMI, and low PMD was found to be 1.969 times higher(95% CI: 1.215-3.185, P=0.006), 1.912 times higher(95% CI: 1.143-3.205, P=0.014), and 1.832 times higher(95% CI: 1.164-2.882, P=0.009)respectively, compared to patients with high PEF, high PMI, and high PMD. Conclusions:Low PEF, low PMI, and low PMD are significant risk factors for postoperative pneumonia in esophageal cancer patients aged 70 years and older.Preoperative PEF, PMI, and PMD, which are commonly utilized measurement indicators for sarcopenia, can be utilized as early screening indicators for postoperative pneumonia.

Objective:To investigate the efficacy and safety of separated R-CHOP in older patients with newly diagnosed diffuse large B-cell lymphoma(DLBCL).Methods:A total of 137 patients aged 65-80 years newly diagnosed with DLBCL between April 2013 and September 2022 at The First Affiliated Hospital of Zhengzhou University were enrolled.The patients were assigned into separated R-CHOP,full-dose R-CHOP,and reduced R-CHOP-like groups based on their different chemotherapy regimens.All individuals were treated in 21-day cycles for 4-8 cycles.The short-term and long-term efficacies and adverse reactions of the treatments were compared among the three groups,and factors influencing progression-free survival(PFS)and overall survival(OS)were analyzed.Results:The overall response rates(ORR)of patients in the separated R-CHOP,full-dose R-CHOP,and reduced R-CHOP-like groups were 89.7%,90.3%,and 86.1%,respectively,with no significant differences among them.The complete respond rate(CRR)of the separated R-CHOP group(64.1%)was significantly higher than that of the reduced R-CHOP-like group(33.3%)(P=0.008)but not significantly different from that of the full-dose R-CHOP group(66.1%).Survival curve analysis revealed no significant differences in PFS and OS between the separated and full-dose R-CHOP groups.Although the separated R-CHOP group showed improved PFS compared with the reduced R-CHOP-like group(P=0.036),there was no statistical difference in OS between these two groups.Multivariate analysis revealed that the international prognostic index(IPI)and separated R-CHOP had significant effects on PFS in patients with DLBCL(all P<0.05),whereas only IPI had a significant effect on OS(P<0.001).The incidence of leukopenia and grade 3-4 leukopenia in the separated R-CHOP group was significantly lower than that in the full-dose R-CHOP group(P=0.007,P=0.012),but there was no significant difference with the reduced R-CHOP-like group in this regard.Conclusions:In older patients with newly diagnosed DLBCL,separated R-CHOP showed good efficacy both in the short and long terms and had acceptable safety and tolerability profiles.

Objective:To investigate the association between serum lipid levels and inflammatory indicators in patients with primary open angle glaucoma (POAG).Methods:A case-control study was conducted.A total of 86 POAG subjects were collected as a POAG group at Joint Shantou International Eye Center of Shantou University and the Chinese University of Hong Kong from January 2016 to March 2022.Meanwhile, 86 with age-related cataract only, matched at age, sex and body mass index were collected as a control group.The intraocular pressure (IOP) and the mean defect of visual field (MD) were measured by Goldmann tonometer and Humphrey field analyzer, respectively.Human peripheral blood samples collected from subjects for serum lipid levels, including total cholesterol, total triglycerides, high density lipoprotein (HDL), and low density lipoprotein (LDL), were analyzed using an automated hematology analyzer and inflammatory markers including C-reactive protein, white blood cells (WBC), neutrophils, lymphocytes, monocytes, were analyzed using an automated biochemical analyzer.Indicators with statistically significant differences between the two groups were selected as independent variables, and multiple logistic regression analysis was used to determine the risk factors for POAG.Correlations between risk factors and ocular parameters (IOP and MD) were assessed using Pearson correlation analysis.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Joint Shantou International Eye Center of Shantou University and the Chinese University of Hong Kong (No.EC20210313[2]-P03). Written informed consent was obtained from each patient before any medical examination.Results:The HDL was (1.59±0.42)mmol/L in the POAG group, which was significantly lower than (1.76±0.50)mmol/L in the control group ( t=2.435, P=0.016). The LDL was (3.34±0.66)mmol/L in the POAG group, which was significantly higher than (3.08±0.71)mmol/L in the control group ( t=2.520, P=0.013). The WBC was (6.91±1.60)×10 9/L in the POAG group, which was significantly higher than (6.11±1.29)×10 9/L in the control group ( t=3.619, P<0.001). Multiple logistic regression analysis showed that high serum LDL level ( P=0.039, OR=2.354, 95% CI: 1.105-5.303) and high WBC level ( P=0.044, OR=1.310, 95% CI: 1.007-1.703) were risk factors for POAG.Pearson correlation analysis showed that the serum LDL and WBC levels of POAG patients were moderately positively associated with IOP ( r=0.610, P<0.001; r=0.358, P=0.001). LDL level was moderately negatively associated with MD ( r=-0.496, P<0.001). WBC level was weakly negatively associated with MD ( r=-0.235, P=0.030). Conclusions:The elevated peripheral blood LDL and WBC levels are risk factors for POAG onset.The elevated LDL and WBC levels are positively correlated with IOP and negatively correlated with MD in POAG 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*

Neurexin-3α, discovered in 2016, is a new type of autoimmune encephalitis antibody. Anti-neurexin-3α antibody-associated encephalitis is generally associated with prodromal symptoms or mood changes, having main clinical manifestations as seizures, memory disorders, confusion or loss of consciousness, central ventilation insufficiency, abnormal behavior, and speech disorders. This paper reviews the relevant research progress at home and abroad about pathogenesis, diagnosis and differential diagnosis, treatment and prognosis of anti-neurexin-3α antibody-associated encephalitis, so as to expand the understanding of clinicians for this disease.

Objective:To investigate the value of machine learning model based on MRI in predicting the abundance of tumor infiltrating CD 8+ T cell and prognosis of pancreatic cancer patients. Methods:The clinical data of 156 patients with pathological confirmed pancreatic cancer who underwent pre-operative MRI within 7 days before surgery in the First Affiliated Hospital of Naval Medical University from January 2017 to April 2018 was retrospectively analyzed. According to the international consensus on the predictive model, a total of 116 patients from January to December 2017 were included in the training set, and a total of 40 patients from January to April 2018 were included in the validation set. With the overall survival of patients as the outcome variable, X-Tile software was used to obtain cut-off values of the percentage of CD 8+ T cells, and all patients were divided into CD 8+ T-high and -low groups. The clinical, pathological and radiological features were compared between two groups. 3D slicer software was used to draw the region of interest in each layer of the primary MR T 1- and T 2-weighted imaging, arterial phase, portal venous phase, and delayed phase images for tumor segmentation. Python package was applied to extract the radiomics features of pancreatic tumors after segmentation and the extracted features were reduced and chosen using the least absolute shrinkage and selection operator (Lasso) logistic regression algorithm. Lasso logistic regression formula was applied to calculate the rad-score. The extreme gradient boosting (XGBoost) were used to construct the machine learning predicted model. The models′ performances were determined by area under the ROC curve (AUC), sensitivity, specificity, accuracy, positive predictive value, and negative predictive value. Results:The cut-off value of the CD 8+ T-cell level was 19.09% as determined by the X-tile program. Patients in the high CD 8+ T cell group had a longer median survival than those in the low CD 8+ T cell group (25.51 month vs 22.92 month, P=0.007). The T stage in the training set and tumor size in the validation set significantly differed between the groups (all P value <0.05). A total of 1 409 radiomics features were obtained, and 19-selected features associated with the level of CD 8+ T cell were determined after being reduced by the Lasso logistic regression algorithm. The rad-score was significantly lower in the CD 8- high group (median: -0.43; range: -1.55 to 0.65) than the CD 8- low group (median: 0.22; range: -0.68 to 2.54, P<0.001). The prediction model combined the radiomics features and tumor size. In the training set, the AUC, sensitivity, specificity, accuracy, and positive and negative predictive value were 0.90 (95% CI 0.85-0.95), 75.47%, 90.48%, 0.84, 0.87, and 0.81. In the validation set, the AUC, sensitivity, specificity, accuracy, and positive and negative predictive value were 0.79 (95% CI 0.63-0.96), 90.00%, 80.00%, 0.85, 0.82, and 0.89. The predictive model can accurately distinguish patients with high and low CD 8+ T cells in pancreatic cancer. Conclusions:The radiomics-based machine learning model is valuable in predicting the CD 8+ T cells infiltrating level in pancreatic cancer patients, which could be useful in identifying potential patients who can benefit from immunotherapies.