Objectives:This study aims to explore the clinical significance of lateral pelvic sentinel lymph node biopsy (SLNB) using indocyanine green (ICG) fluorescence navigation in laparoscopic lateral pelvic lymph node dissection (LLND) and evaluate the accuracy and feasibility of this technique to predict the status of lateral pelvic lymph nodes (LPLNs).Methods:The clinical and pathological characteristics, surgical outcomes, lymph node findings and perioperative complications of 16 rectal cancer patients who underwent SLNB using ICG fluorescence navigation in laparoscopic LLND in the Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College during April 2017 and October 2022 were retrospectively collected and analyzed. The patients did not receive preoperative neoadjuvant radiotherapy and presented with LPLNs but without LPLN enlargement (MRI showed the maximum short axes of the LPLNs were ≥5 mm and <10 mm at first visit).Results:All 16 patients were successfully performed SLNB using ICG fluorescence navigation in laparoscopic LLND. Three patients underwent bilateral LLND and 13 patients underwent unilateral LLND. The lateral pelvic sentinel lymph nodes (SLNs) were clearly fluorescent before dissection in 14 patients and the detection rate of SLNs for these patients was 87.5%. Lateral pelvic SLN metastasis was diagnosed in 2 patients and negative results were found in 12 patients by frozen pathological examinations. Among the 14 patients in whom lateral pelvic SLNs were detected, the dissected lateral pelvic non-SLNs were all negative. All dissected LPLNs were negative in two patients without fluorescent lateral pelvic SLNs. The specificity, sensitivity, negative predictive value, and accuracy was 85.7%, 100%, 100%, and 100%, respectively.Conclusions:This study indicates that lateral pelvic SLNB using ICG fluorescence navigation shows promise as a safe and feasible procedure with good accuracy. This technique may replace preventive LLND for locally advanced lower rectal cancer.

The aim of this study is to investigate the immune effect of H9 subtype avian influenza virus(AIV)NP protein on mice and lay the foundation for the development of avian influenza vi-rus(AIV)vaccine.The H9N2 virus NP gene amplification product was cloned into the pET-32a expression vector,and the protein expression was verified by SDS-PAGE and Western blot,and the immune effect was evaluated by measuring the secretion of supernatant multicytokines in mouse splenocytes culture.The results showed that the total length of the coding region sequence of NP gene was 1 497 bp,NP recombinant proteins exist in both soluble and insoluble protein forms,and the specific bands were visible in Western blot.After immunizing mice,serum produces IgG-bind-ing antibodies with antibody titers of 1∶40 000.Compared with the control group,IL-2,IL-5 and IL-13 were significantly increased(P＜0.001),and the secretion of IL-6 was significantly increased compared with the control group.IL-4 and IL-12 p70 secretions were elevated compared with con-trols,but there was no significant difference.Compared with the control group,the secretions of IL-1β,IL-18,GM-CMF,TNF-α and IFN-γ were inhibited,but the difference was not significant(P＞0.05).The results showed that NP recombinant protein is a good immunogen,laying a foundation for in-depth research on influenza vaccine.

Objectives:This study aims to explore the clinical significance of lateral pelvic sentinel lymph node biopsy (SLNB) using indocyanine green (ICG) fluorescence navigation in laparoscopic lateral pelvic lymph node dissection (LLND) and evaluate the accuracy and feasibility of this technique to predict the status of lateral pelvic lymph nodes (LPLNs).Methods:The clinical and pathological characteristics, surgical outcomes, lymph node findings and perioperative complications of 16 rectal cancer patients who underwent SLNB using ICG fluorescence navigation in laparoscopic LLND in the Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College during April 2017 and October 2022 were retrospectively collected and analyzed. The patients did not receive preoperative neoadjuvant radiotherapy and presented with LPLNs but without LPLN enlargement (MRI showed the maximum short axes of the LPLNs were ≥5 mm and <10 mm at first visit).Results:All 16 patients were successfully performed SLNB using ICG fluorescence navigation in laparoscopic LLND. Three patients underwent bilateral LLND and 13 patients underwent unilateral LLND. The lateral pelvic sentinel lymph nodes (SLNs) were clearly fluorescent before dissection in 14 patients and the detection rate of SLNs for these patients was 87.5%. Lateral pelvic SLN metastasis was diagnosed in 2 patients and negative results were found in 12 patients by frozen pathological examinations. Among the 14 patients in whom lateral pelvic SLNs were detected, the dissected lateral pelvic non-SLNs were all negative. All dissected LPLNs were negative in two patients without fluorescent lateral pelvic SLNs. The specificity, sensitivity, negative predictive value, and accuracy was 85.7%, 100%, 100%, and 100%, respectively.Conclusions:This study indicates that lateral pelvic SLNB using ICG fluorescence navigation shows promise as a safe and feasible procedure with good accuracy. This technique may replace preventive LLND for locally advanced lower rectal cancer.

The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β₂m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a Superdex^TM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.
Animals ; Capripoxvirus ; Epitopes, T-Lymphocyte/genetics* ; Peptides/genetics* ; Poxviridae Infections ; Sheep ; Sheep Diseases

Background: Chicken anemia virus (CAV) causes chicken infectious anemia, which results in immunosuppression; the virus has spread widely in chicken flocks in China. Objectives: The aim of this study was to understand recent CAV genetic evolution in chicken flocks in Guangxi Province, southern China. Methods: In total, 350 liver samples were collected from eight commercial broiler chicken farms in Guangxi Province in southern China from 2018 to 2020. CAV was detected by conventional PCR, and twenty CAV complete genomes were amplified and used for the phylogenetic analysis and recombination analysis. Results: The overall CAV-positive rate was 17.1%. The genetic analysis revealed that 84 CAVs were distributed in groups A, B, C (subgroups C1-C3) and D. In total, 30 of 47 Chinese CAV sequences from 2005-2020 belong to subgroup C3, including 15 CAVs from this study. There were some specific mutation sites among the intergenotypes in the VP1 protein. The amino acids at position 394Q in the VP1 protein of 20 CAV strains were consistent with the characteristics of a highly pathogenic strain. GX1904B was a putative recombinant. Conclusions Subgroup C3 was the dominant genotype in Guangxi Province from 2018–2020.The 20 CAV strains in this study might be virulent according to the amino acid residue analysis. These data help improve our understanding of the epidemiological trends of CAV in southern China.

Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes great economic losses. At the moment, no effective neutralizing antibody is available for scientific research and treatment. Therefore, developing a method for screening the neutralizing monoclonal antibodies is of great significance for the prevention and treatment of PRRSV and the screening of antigen sites. Monoclonal antibodies have been widely used in the treatment and diagnosis of many human and animal diseases. Therefore, screening effective neutralizing antibodies for different pathogens is an urgent task. Among the methods for monoclonal antibody screening, B cell immortalization is an effective method to obtain neutralizing monoclonal antibody. Specifically, in this study, the bcl-6 and bcl-xl genes were connected by f2a and then the yielded product was ligated to a vector for retrovirus packaging. The swine lymphocytes immunized with PRRSV were infected the yielded mature viruses and cultured in the complete medium containing CD40L and IL21 cytokines. Then, CD21 was used as the marker to screen B cells with the magnetic bead method. Finally, monoclonal B cells were obtained and the secretion of antibodies was tested. The results showed that the plasmid, either being transfected alone or with the packaged plasmids, could be expressed, and that the packaged retrovirus could infect the cells. Moreover, the infected lymphocytes secreted antibodies, so did the screened B cells. Therefore, the method for screening monoclonal antibody against PRRSV was successfully established.
Animals ; Antibodies, Monoclonal ; Antibodies, Neutralizing ; Antibodies, Viral ; Humans ; Porcine Reproductive and Respiratory Syndrome/prevention & control* ; Porcine respiratory and reproductive syndrome virus/genetics* ; Swine

Objective: To characterize the clinical epidemiological features of primary lung cancer patients based on massive clinical data. Methods: The demographic and histological information of 8 081 primary lung cancer patients who were initially identified from 1 January 2012 to 31 December 2013 in 16 hospitals from 6 provinces were retrospectively analyzed to determine the characteristics of different histological subtypes among different gender, age-group and birth cohort. Results: Among the 8 081 lung cancer patients, 5 422 (67.10%) were male and 2 659 (32.90%) were female, the average age was (60.96±10.14) years. The most common histological subtypes of lung cancer successively were adenocarcinoma (ADC), squamous cell carcinoma (SCC), small cell carcinoma and large cell carcinoma, and the proportions of these subtypes were 53.13%, 24.51%, 14.59% and 0.66%, respectively, which collectively accounted for 92.89%. The current highest smoking rate was observed in SCC patients, which was 62.45%, while lowest in ADC, which was 29.68%. The incidence of lung cancer in male was significantly higher that that of female, with a sex ratio of 2.04∶1. The highest sex ratio was observed in SCC, which was 9.14∶1, while lowest in ADC, which was 1.14∶1. The distribution of histological subtypes in male lung cancer patients was consistent with the general situation.While among the female lung cancer patients, the proportion of ADC was the highest (75.42%), followed by SCC (10.08%), squamous cell carcinoma (7.34%) and large cell carcinoma (0.39%). Analyzed by the birth cohort, the proportion of ADC gradually increased with the age, while the reduced tendency was observed in SCC (P<0.000 1). Conclusion Adenocarcinoma is the most important histological subtype of lung cancer, and the distribution characteristics of histological subtypes of lung cancer differs among genders, age and birth cohort.

Objective To characterize the clinical epidemiological features of primary lung cancer patients based on massive clinical data. Methods The demographic and histological information of 8 081 primary lung cancer patients who were initially identified from 1 January 2012 to 31 December 2013 in 16 hospitals from 6 provinces were retrospectively analyzed to determine the characteristics of different histological subtypes among different gender, age?group and birth cohort. Results Among the 8 081 lung cancer patients, 5 422 (67.10%) were male and 2 659 (32.90%) were female, the average age was (60.96± 10.14) years. The most common histological subtypes of lung cancer successively were adenocarcinoma ( ADC ), squamous cell carcinoma ( SCC ), small cell carcinoma and large cell carcinoma, and the proportions of these subtypes were 53.13%, 24.51%, 14.59% and 0.66%, respectively, which collectively accounted for 92.89%. The current highest smoking rate was observed in SCC patients, which was 62.45%, while lowest in ADC, which was 29.68%. The incidence of lung cancer in male was significantly higher that that of female, with a sex ratio of 2.04 ∶ 1. The highest sex ratio was observed in SCC, which was 9.14 ∶ 1, while lowest in ADC, which was 1.14 ∶ 1. The distribution of histological subtypes in male lung cancer patients was consistent with the general situation.While among the female lung cancer patients, the proportion of ADC was the highest (75.42%), followed by SCC (10.08%), squamous cell carcinoma ( 7.34%) and large cell carcinoma (0.39%). Analyzed by the birth cohort, the proportion of ADC gradually increased with the age, while the reduced tendency was observed in SCC ( P＜0.000 1). Conclusion Adenocarcinoma is the most important histological subtype of lung cancer, and the distribution characteristics of histological subtypes of lung cancer differs among genders, age and birth cohort.

Objective To characterize the clinical epidemiological features of primary lung cancer patients based on massive clinical data. Methods The demographic and histological information of 8 081 primary lung cancer patients who were initially identified from 1 January 2012 to 31 December 2013 in 16 hospitals from 6 provinces were retrospectively analyzed to determine the characteristics of different histological subtypes among different gender, age?group and birth cohort. Results Among the 8 081 lung cancer patients, 5 422 (67.10%) were male and 2 659 (32.90%) were female, the average age was (60.96± 10.14) years. The most common histological subtypes of lung cancer successively were adenocarcinoma ( ADC ), squamous cell carcinoma ( SCC ), small cell carcinoma and large cell carcinoma, and the proportions of these subtypes were 53.13%, 24.51%, 14.59% and 0.66%, respectively, which collectively accounted for 92.89%. The current highest smoking rate was observed in SCC patients, which was 62.45%, while lowest in ADC, which was 29.68%. The incidence of lung cancer in male was significantly higher that that of female, with a sex ratio of 2.04 ∶ 1. The highest sex ratio was observed in SCC, which was 9.14 ∶ 1, while lowest in ADC, which was 1.14 ∶ 1. The distribution of histological subtypes in male lung cancer patients was consistent with the general situation.While among the female lung cancer patients, the proportion of ADC was the highest (75.42%), followed by SCC (10.08%), squamous cell carcinoma ( 7.34%) and large cell carcinoma (0.39%). Analyzed by the birth cohort, the proportion of ADC gradually increased with the age, while the reduced tendency was observed in SCC ( P＜0.000 1). Conclusion Adenocarcinoma is the most important histological subtype of lung cancer, and the distribution characteristics of histological subtypes of lung cancer differs among genders, age and birth cohort.

Objective To investigate the serum levels of dickkopf-related protein 1 (DKK1) and sclerostin (SOST) in patients with axial spondyloarthritis treated with selective cyclo-oxygenase 2 inhibitor and its relation to clinical efficacy.Methods A randomized double-blind controlled trial with axial spondyloarthritis (ax-SpA) was carried out in our hospital.The data from patients in a single center was collected and analyzed.Serum DKK1 and SOST levels were measured by enzyme-linked immuno sorbent assay (ELISA)method before and after 12 weeks treatment,then correlation analysis were conducted for DKK1 and SOST levels with erythrocyte sedimentation rate (ESR),C reactive protein (CRP),Bath ankylosing spondylitis disease activity index (BASDAI),Bath ankylosing spondylitis functional index (BASFI) and SPARCC of the sacroiliac joint inflammation score.Chi-square tests were used for analyzing of categorical data.Fisher exact tests were performed when the expected frequencies were less than 5.Two independent samples t-test was used to compare the difference between groups.Single sample t-test was used to ompare the differences between data before and after treatment.Pearson or Spearman correlation was used for correlation analysis.Results After 12 weeks of treatment,a total of 116 patients completed the follow-up,including 57 cases of imrecoxib group and 59 cases of the celecoxib group.There were no statistically significant difference between the two groups (P＞0.05).The level of serum DKK1 was significantly increased after treatment [(393±137) pg/ml,vs (542±274)pg/ml,P＜0.05].The serum level of SOST increased significantly [(39±19) pg/ml vs (57±36) pg/ml,t=5.814,P＞0.05],too.The difference between the two groups was not statistically significant (P＞0.05).Spearman correlation analysis showed that serum DKK1 was positively correlated with serum SOST (r=0.226,P=0.015).A significantcorrelation was found between SOST level and ESR,CRP,finger to floor distance,left and fight lumbar side flexion and Schober's test (ESR:r=-0.379,P＜0.01;r=-0.309,P=0.001;r=-0.225,P=0.015;r=0.185,P=0.047;r=0.247,P=0.008;r=0.214,P=0.021).Conclusion Imrecoxib and celecoxib have similar efficacy on relieving the signs and symptoms of patients with ax-SpA.Short-term application of selective COX-2 inhibitors can increase DKK1 and SOST and possibly delay radiographic progression.