Necrotizing enterocolitis (NEC) is an intestinal inflammatory and necrotic disease seen in premature infants, and remains the leading cause of death resulted from gastrointestinal diseases in premature infants. The specific pathogenesis of NEC is still unclear. In recent years, a lot of studies have reported that Toll-like receptor 4 (TLR4) plays a key role in the pathogenesis of NEC. TLR4, which is abundantly expressed in intestinal epithelial cells of premature infants, binds to bacterial lipopolysaccharide (LPS) to activate downstream signaling pathways, leading to disruption of intestinal epithelial integrity and bacterial translocation, resulting in intestinal ischemic necrosis and inflammatory responses, which may rapidly progress to severe sepsis, multiple organ dysfunction, and death. This paper reviews the mechanism of TLR4-related signaling pathways in intestinal epithelial injury and inflammatory responses in newborns with NEC, providing a reference to study new therapeutic targets for NEC.
Enterocolitis, Necrotizing/pathology* ; Toll-Like Receptor 4/metabolism* ; Humans ; Infant, Newborn ; Signal Transduction ; Inflammation/metabolism* ; Animals ; Intestines/immunology* ; Intestinal Mucosa/pathology* ; Infant, Premature

Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

Objective To verify the accuracy of Youssef's formula and evaluate whether fetal biacromial diameter(BA)and other fetal biological diameters estimated by ultrasound can be used to predict macrosomia and shoulder dystocia,so as to provide the possibility for clinical prediction of shoulder dystocia.Methods A total of 200 pregnant women with a gestational period of 37-42 weeks were examined with ultrasound within 3 days before delivery for collecting biparietal diameter(BPD),head circumference(HC),abdominal circumference(AC),humerus length(HL),femur length(FL),thoracic transverse diameter and midpoint diameter of upper arm;and the fetal BA was estimated by Youssef's formula.Neonatal BA,body mass and body length were measured within 1 day after delivery.The above data were analyzed for correlation.Newborns were grouped according to their body mass(macrosomia vs non-macrosomia)and whether they had shoulder dystocia or not(shoulder dystocia vs non-shoulder dystocia).Results(1)The fetal BA estimated by Youssef's formula was consistent with neonatal BA(P>0.05),and the estimated BA was positively correlated with BPD,HC,AC and neonatal body mass(P<0.001).(2)The BA,BA/AC and BA/HC in macrosomia group were different from those in non-macrosomia group(P<0.05).ROC curve showed that the sensitivity and specificity were 92.3%and 88.2%for macrosomia prediction when the estimated BA threshold was 16.05 cm,and those were 61.5%and 77.0%when BA/AC threshold was 0.455,and 76.9%and 72.7%when BA/HC threshold was 0.465.(3)Shoulder dystocia group had neonatal weight close to non-shoulder dystocia group(P>0.05),but higher BA/BPD,BA/HC and BA-BPD(P<0.05).ROC curve showed that the sensitivity and specificity were 100.0%and 66.8%for shoulder dystocia when BA threshold was 15.45 cm,100.0%and 80.6%when BA/BPD threshold was 1.695,100.0%and 81.6%when BA/HC threshold was 0.475,and 100.0%and 76.0%when the threshold difference between BA and BPD was 6.35 cm.Conclusion Fetal BA,BA/BPD,BA/HC,BA/AC and BA-BPD may be effective predictors of shoulder dystocia and macrosomia.

Objective To verify the accuracy of Youssef's formula and evaluate whether fetal biacromial diameter(BA)and other fetal biological diameters estimated by ultrasound can be used to predict macrosomia and shoulder dystocia,so as to provide the possibility for clinical prediction of shoulder dystocia.Methods A total of 200 pregnant women with a gestational period of 37-42 weeks were examined with ultrasound within 3 days before delivery for collecting biparietal diameter(BPD),head circumference(HC),abdominal circumference(AC),humerus length(HL),femur length(FL),thoracic transverse diameter and midpoint diameter of upper arm;and the fetal BA was estimated by Youssef's formula.Neonatal BA,body mass and body length were measured within 1 day after delivery.The above data were analyzed for correlation.Newborns were grouped according to their body mass(macrosomia vs non-macrosomia)and whether they had shoulder dystocia or not(shoulder dystocia vs non-shoulder dystocia).Results(1)The fetal BA estimated by Youssef's formula was consistent with neonatal BA(P>0.05),and the estimated BA was positively correlated with BPD,HC,AC and neonatal body mass(P<0.001).(2)The BA,BA/AC and BA/HC in macrosomia group were different from those in non-macrosomia group(P<0.05).ROC curve showed that the sensitivity and specificity were 92.3%and 88.2%for macrosomia prediction when the estimated BA threshold was 16.05 cm,and those were 61.5%and 77.0%when BA/AC threshold was 0.455,and 76.9%and 72.7%when BA/HC threshold was 0.465.(3)Shoulder dystocia group had neonatal weight close to non-shoulder dystocia group(P>0.05),but higher BA/BPD,BA/HC and BA-BPD(P<0.05).ROC curve showed that the sensitivity and specificity were 100.0%and 66.8%for shoulder dystocia when BA threshold was 15.45 cm,100.0%and 80.6%when BA/BPD threshold was 1.695,100.0%and 81.6%when BA/HC threshold was 0.475,and 100.0%and 76.0%when the threshold difference between BA and BPD was 6.35 cm.Conclusion Fetal BA,BA/BPD,BA/HC,BA/AC and BA-BPD may be effective predictors of shoulder dystocia and macrosomia.

Objective To investigate the efficacy and safety of Xihuang capsules as an adjuvant treatment for metastatic colorectal cancer and their impact on immune function. Methods A retrospective analysis was conducted on clinical data from 112 patients diagnosed with metastatic colorectal cancer. The patients were categorized into two groups: a control group (n=56) that did not take Xihuang capsules and an observation group (n=56) that did. The efficacy, improvement of quality of life, toxic and side effects and immune function of the two groups were analyzed and compared. Results After treatment, the disease control rate (DCR) and the rate of improvement in quality of life were significantly higher in the observation group compared to the control group. Additionally, levels of carcinoembryonic antigen (CEA) and the incidence of adverse reactions, including bone marrow suppression and liver and kidney function damage, were significantly lower in the observation group. Furthermore, the percentages of CD4⁺ and CD8⁺ T cells, the CD8⁺/CD4⁺ T cells ratio, as well as serum levels of high mobility group box-1 (HMGB1) and interleukin 2 (IL-2) in observation group were significantly elevated compared to pre-treatment levels. Subgroup analysis revealed that patients with a Karnofsky Performance Status (KPS) score ≤80, a high CD8⁺/CD4⁺ T cells ratio, and elevated HMGB1 levels experienced a significantly higher objective response rate (ORR) in the observation group. Conversely, patients with stage IVB disease, who had KPS score ≤80, a low CD8⁺/CD4⁺ T cells ratio and high CEA and IL-2 levels demonstrated a more pronounced DCR in the observation group. Conclusion Xihuang capsules exhibit promising clinical efficacy as an adjuvant treatment for advanced colorectal cancer. They not only enhance patients' quality of life and reduce the toxic and adverse effects of chemotherapy, but also improve immune function. These benefits are particularly significant in patients with a high tumor burden, indicating that Xihuang capsules are worthy of clinical application.
Humans ; Colorectal Neoplasms/immunology* ; Male ; Female ; Middle Aged ; Drugs, Chinese Herbal/adverse effects* ; Capsules ; Aged ; Carcinoembryonic Antigen/blood* ; Retrospective Studies ; Quality of Life ; Adult ; Neoplasm Metastasis ; Interleukin-2/blood* ; HMGB1 Protein/blood* ; Chemotherapy, Adjuvant

Objective To prepare mouse anti-human lymphocyte activation gene 3 (LAG3) monoclonal antibody (mAb) and perform immunological identification of the antibody. Methods BALB/c mice were immunized with LAG3-mLumin-3T3 cells, which stably express the extracellular and transmembrane regions of human LAG3 in mouse 3T3 cells. The secretion of anti-human LAG3 antibodies in mouse serum was assessed using flow cytometry and immunofluorescence. SP2/0 cells were injected subcutaneously into the mice to elicit solid myelomas, and mouse myeloma cells were subsequently isolated. Spleen cells from the immunized mice were fused with the myeloma cells to establish hybridomas, which were then separated using the limiting dilution method. Flow cytometry was used to detect LAG3 mAbs in the hybridoma culture medium. To map the epitopes recognized by these mAbs, 3T3 cells expressing individual extracellular domains of LAG3(LAG3 domains 1/-2/-3/-4-3T3) were used. Flow cytometry was also applied to analyze LAG3 expression on activated human peripheral blood mononuclear cells (PBMC) before and after co-culture with the LAG3 mAbs. Results Mice immunized with the recombinant eukaryotic cell antigen produced anti-LAG3 antibodies. The generated hybridomas secreted mouse anti-human LAG3 mAbs, with each hybridoma line recognizing different LAG3 antigenic domains. Conclusion Mouse anti-human LAG3 mAbs were successfully generated, with different hybridoma clones secreting antibodies that recognize distinct LAG3 epitopes. These findings lay the groundwork for further studies into the biological properties of LAG3 and the development of diagnostic reagents and therapeutic blocking antibodies for cancer treatment.
Animals ; Humans ; Mice ; Lymphocyte Activation Gene 3 Protein ; Antibodies, Monoclonal/immunology* ; Mice, Inbred BALB C ; Hybridomas/immunology* ; Antigens, CD/genetics* ; Immunization ; Recombinant Proteins/immunology* ; Female ; Eukaryotic Cells/immunology* ; Flow Cytometry ; Epitopes/immunology*

Objective To analyze the mortality and its changing trend, and composition and order of causes of death of residents in Mengzi City, and to provide a basis for further disease prevention and control. Methods The death causes surveillance data from 2018 to 2021 were derived from the all-cause-of-death surveillance system in Mengzi City. A retrospective analysis was performed on the mortality rate, life expectancy, life expectancy eliminating causes of death, and life loss. The annual percentage change (APC) was analyzed by the Joinpoint regression model to describe changes in mortality trends. Results The overall crude mortality rate was 633.88/100,000. The age-adjusted mortality was 866.87/100,000. There was a significant downward trend in the crude and standardized mortality (APC=-1.73% , APC=-5.96% , P<0.05). Deaths due to chronic non-communicable diseases (NCDs) accounted for 76.44% of the total deaths. The top 5 causes of death of the residents were cerebrovascular diseases (140.38/100 000), heart diseases (104.24/100 000), malignant neoplasms (92.75/100 000), injuries (79.37/100 000), and respiratory diseases (63.17/100 000) in order, accounting for 75.71% of all causes of death. Life expectancy was 75.67 years, 72.32 years and 79.49 years in the whole-population, males and females, respectively. The potential life expectancy loss due to injury, malignant tumor and cerebrovascular disease accounted for 65.45% of all causes of death. Conclusion Chronic non-communicable diseases are the focus of prevention and control work in Mengzi City. Particular attention should be paid to the damage to health and loss of life caused by injuries, malignancies and cerebrovascular diseases.

Tumor immune microenvironment is an important microecology for tumor development, where tumor-associated macrophages are the most abundant immune cells in the tumor immune microenvironment, with high plasticity and heterogeneity. Under the regulation of various environmental factors, tumor-associated macrophages can differentiate into different subgroups. Though complex and variable, all these environmental factors ultimately regulate tumor-associated macrophages by influencing the temporal and spatial heterogeneity of these cells’ internal components, structure, and functions. Mitochondrion are important organelles, responsible for energy production, metabolism, and centers of multiple signal transduction. More and more studies have found that mitochondria can regulate cell functions through various mechanisms such as morphological change, metabolic reprogramming, intermediate metabolites or mitochondrial genetic material. Mitochondrial disorders are involved in many diseases and pathological processes. Here, we review the mechanisms by which mitochondria regulate the polarization of macrophages and thus reshape the tumor immune microenvironment. Further, we discuss and prospect the current status of macrophage mitochondria-related tumor immunotherapy.

【Objective】 To analyze the association of overweight and obesity with elevated blood pressure(BP) among children and adolescents in Mengzi City, in order to provide reference for making effective interventions. 【Methods】 A total of 30 classes of students in 14 schools were selected into this study by a stratified random cluster sampling method from October 2020 to September 2021. The paticipants completed a questionnaire survey, and had their height, weight and BP measured. Logistic regression analysis was used to evaluate the association of overweight and obesity with elevated BP. 【Results】 A total of 4 015 children and adolescents aged 7 - 17 years were involved in the study. The detection rate of overweight, obesity and elevated BP was 12.53%, 10.01% and 12.38%, respectively. The detection rate of elevated BP was 9.96%, 19.48% and 25.62% in normal weight, overweight and obese groups, respectively. The risk of elevated BP in the overweight and obese groups was 2.190 times(95%CI: 1.696 - 2.926, P<0.001) and 3.243 times(95%CI: 2.501 - 4.204, P<0.001) as high as that in normal weight groups. The dietary and exercise behaviors of 1 604 adolescents were analyzed, it was found that the detection rate of obesity was significntly higher in non-boarding students(15.09%) and those who consumed fresh fruits at least once a day(13.49%)(χ²=18.012, 7.225, P<0.05), the detection rate of elevated BP was significantly higher in among those who did not consume sugary beverages(16.88%), and performed moderate-to-high intensity physical exercise of 60 minutes or more ≤ 2 days per week(16.88%)(χ²=9.403, 14.921,P<0.01). 【Conclusions】 The risk of developing hypertension increases with the high prevalence of overweight and obesity and is strongly associated with multiple factors. Therefore, it is essential to conduct effective weight control and behavioral lifestyle interventions among children and adolescents.

Objective To explore the value of vesical imaging reporting and data system(VI-RADS)combined with absolute tumor-wall contact length(ABTCL)and actual tumor-wall contact length(ACTCL)in diagnosing muscle invasive bladder cancer(MIBC).Methods The MRI data of 113 patients with pathologically confirmed bladder cancer(BCa)were analyzed retrospectively.All patients underwent conventional MRI,diffusion weighted imaging(DWI)and dynamic contrast enhanced(DCE)MRI before sur-gery.Two radiologists independently evaluated MRI images based on VI-RADS score,and measured quantitative parameters,inclu-ding ABTCL and ACTCL.The Chi-square test was used to compare the difference of VI-RADS scores between MIBC and non-mus-cle invasive bladder cancer(NMIBC).Quantitative parameters between MIBC and NMIBC were compared by Mann-Whitney U test.The receiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of VI-RADS,quantitative parameters and VI-RADS combined with quantitative parameters in the diagnosis of MIBC.Results VI-RADS,ABTCL and ACTCL had significant differences between MIBC and NMIBC(P＜0.05).The area under the curve(AUC)for VI-RADS,ABTCL and ACTCL in diagno-sing MIBC were 0.89,0.76 and 0.77,respectively.There was no significant difference between the AUC for ABTCL and ACTCL(P＞0.05).The AUC for VI-RADS combined with ABTCL or ACTCL in diagnosing MIBC was 0.93,higher than that of only VI-RADS(P＜0.05).Conclusion The combination of VI-RADS with either ABTCL or ACTCL can effectively improve the diagnostic performance of MIBC.ABTCL obtainedby linear measurement is easier to implement in clinical practice than ACTCL obtained by curved measurement.