ObjectiveTo investigate the mutation status of the MET gene in tumor patients， with a focus on its expression in lung cancer tissues and its impact on patient prognosis， providing theoretical basis and potential targets for precise diagnosis and treatment of the tumors. MethodsA total of 1，627 tumor patient samples were collected from January 2021 to September 2025. High-throughput sequencing technology was used to conduct DNA-level gene detection on 137 tumor patients with MET mutations. The distribution of mutation sites in exons and introns， as well as the occurrence frequencies of MET gene amplification and gene fusion were analyzed， with particular attention paid to the mutation frequencies of each exon and the high-frequency variant types. Immunohistochemical staining was performed to analyze the expression levels of c-MET protein in tumor tissues of the 137 patients. Immunohistochemical analysis was conducted on tumor tissues of 9 patients with MET gene exon 14 skipping mutations to study the association between c-MET protein expression and this specific mutation type. RNA sequencing results， immunohistochemical staining data， and follow-up prognosis of lung cancer from the GEPIA2 database and TCGA database were comprehensively analyzed to explore the potential relationship between MET gene expression in tumor tissues and patient prognosis. ResultsAmong 1，627 tumor patients， 137 MET mutation patients were detected， including 93 males with an average age of （66.85±9.61） years and 44 females with an average age of （59.43±11.08） years. Lung cancer was the most common tumor type （117 cases）. MET gene expression was lower in breast cancer and glioblastoma multiforme， but higher in lung cancer and colorectal cancer as compared with the expression in normal tissues. Survival analysis showed that patients with high MET gene expression in lung adenocarcinoma had a shorter overall survival time than those with low expression. A total of 961 mutations were detected by next-generation sequencing， including 547 exon mutations and 374 intron mutations. MET gene amplification was found in 13 cases and gene fusion in 27 cases. The highest mutation frequency was in exon 2， followed by exon 5， exon 4， and exon 19. High-frequency variant types included c.2890C>A （p.L964M） on exon 4 and c.3028G>T （p.D1010Y） on exon 5. The c-MET protein expression was closely related to the type of gene mutation. Missense mutations often led to positive or strongly positive c-MET protein expression， while nonsense mutations often resulted in weak or absent c-MET protein expression. Among the 9 patients with MET gene exon 14 skipping mutations， the c-MET protein expression score was mostly 0 - 1， with only 1 case scoring 2. Among the 117 lung cancer patients， the incidence rate was higher in males than in females， and more common among stage Ⅲ-Ⅳ patients than among stage I-Ⅱ patients. Out of the 41 patients treated with surgery + targeted drugs （gutemitinib， savolitinib）， 8 cases of recurrence or metastasis were found during follow-up. ConclusionMET gene mutations exhibit specific mutation characteristics in different tumor types， and its expression levels show significant differences between various tumor tissues and normal tissues. It is closely related to the poor prognosis of lung adenocarcinoma patients. We found that c-MET protein expression is closely related to the type of MET gene mutation， especially in patients with MET gene exon 14 skipping mutations， which presents a characteristic expression pattern. This provides an important basis for the preliminary screening using immunohistochemistry in clinical practice.

Antibody (Ab) humanization is critical to reduce immunogenicity and enhance efficacy in the preclinical phase of the development of therapeutic Abs originated from animal models. Computational suggestions have long been desired, but available tools focused on immunogenicity calculation of whole Ab sequences and sequence segments, missing the individual residue sites. This study introduces Site-specific Immunogenicity for Therapeutic Antibody (SITA), a novel computational framework that predicts B-cell immunogenicity score for not only the overall antibody, but also individual residues, based on a comprehensive set of amino acid descriptors characterizing physicochemical and spatial features for antibody structures. A transfer-learning-inspired framework was purposely adopted to overcome the scarcity of Ab-Ab structural complexes. On an independent testing dataset derived from 13 Ab-Ab structural complexes, SITA successfully predicted the epitope sites for Ab-Ab structures with a receiver operating characteristic (ROC)-area unver the ROC curve (AUC) of 0.85 and a precision-recall (PR)-AUC of 0.305 at the residue level. Furthermore, the SITA score can significantly distinguish immunogenicity levels of whole human Abs, therapeutic Abs and non-human-derived Abs. More importantly, analysis of an additional 25 therapeutic Abs revealed that over 70% of them were detected with decreased immunogenicity after modification compared to their parent variants. Among these, nearly 66% Abs successfully identified actual modification sites from the top five sites with the highest SITA scores, suggesting the ability of SITA scores for guide the humanization of antibody. Overall, these findings highlight the potential of SITA in optimizing immunogenicity assessments during the process of therapeutic antibody design.

Antibody humanization is critical to reduce immunogenicity and enhance efficacy in the preclinical phase of the development of therapeutic antibodies originated from animal models.Computational suggestions have long been desired,but available tools focused on immunogenicity calculation of whole antibody sequences and sequence segments,missing the individual residue sites.This study introduces Site-specific Immunogenicity for Therapeutic Antibody(SITA),a novel computational framework that predicts B-cell immunogenicity score for not only the overall antibody,but also individual residues,based on a comprehensive set of amino acid descriptors characterizing physicochemical and spatial features for antibody structures.A transfer-learning-inspired framework was purposely adopted to overcome the scarcity of Antibody-Antibody structural complexes.On an independent testing dataset derived from 13 Antibody-Antibody structural complexes,SITA successfully predicted the epitope sites for Antibody-Antibody structures with a receiver operating characteristic(ROC)-area unver the ROC curve(AUC)of 0.85 and a precision-recall(PR)-AUC of 0.305 at the residue level.Furthermore,the SITA score can significantly distinguish immunogenicity levels of whole human antibodies,therapeutic antibodies and non-human-derived antibodies.More importantly,analysis of an additional 25 thera-peutic antibodies revealed that over 70％of them were detected with decreased immunogenicity after modification compared to their parent variants.Among these,nearly 66％antibodies successfully iden-tified actual modification sites from the top five sites with the highest SITA scores,suggesting the ability of SITA scores for guide the humanization of antibody.Overall,these findings highlight the potential of SITA in optimizing immunogenicity assessments during the process of therapeutic antibody design.

Objective:To analyze vaginal microecological changes in patients with vaginal infectious diseases and their correlation with human papillomavirus (HPV) infection.Methods:A case-control study was conducted involving 416 patients who visited the Gynecology Outpatient Department and the Cervical Disease Clinic at Yongkang Maternity and Child Care Hospital between December 2022 and December 2023. All patients underwent testing for vaginal microecology and HPV to evaluate the prevalence of vaginal infectious diseases and HPV infection. Vaginal microecological indicators were compared between patients with vaginal infections and those without identifiable pathogenic bacterial dysbiosis. The indicators included microbial density, lactobacilli levels, pH, hydrogen peroxide levels, abnormal leukocyte esterase, and the grading of vaginal lactobacilli. Additionally, the HPV infection status was compared among patients with different vaginal microecological environments, and the correlation between vaginal infections and HPV infections was analyzed.Results:In a study involving 416 participants, 216 were diagnosed with vaginal infections, with an incidence rate of 51.92% (216/416). Among these cases, 118 were classified as a single infection, accounting for 54.63% (118/216), while 98 were identified as mixed infections, accounting for 45.37% (98/216). The rate of HPV infection among the 216 patients with vaginal infectious diseases was 37.04% (80/216). This rate was significantly higher than the 15.31% (30/196) observed in patients without identifiable pathogenic bacterial dysbiosis ( χ2 = 24.79, P < 0.001). Patients with vaginal infectious diseases also displayed elevated rates of abnormal lactobacilli levels, abnormal pH, abnormal leukocyte esterase, and abnormal lactobacilli grading, with rates of 59.26%, 86.57%, 72.69%, and 57.41%, respectively. In comparison, patients without identifiable pathogenic dysbiosis had rates of 31.12%, 18.88%, 51.53%, and 34.18%, respectively. All differences were statistically significant ( χ2 = 32.76, 86.83, 19.64, 22.28, all P < 0.001). Additionally, significant differences in HPV infection rates were observed among patients who tested positive for bacterial vaginosis, vulvovaginal candidiasis, and trichomoniasis when compared to those who tested negative ( χ2 = 12.46, 4.04, 6.14, P < 0.001, 0.044, 0.013). Both bacterial vaginosis and vulvovaginal candidiasis were recognized as high-risk factors for HPV infection ( OR = 4.039, 2.902, both P < 0.05). Conclusions:Vaginal infectious diseases are significantly linked to HPV infection, particularly bacterial vaginosis and vulvovaginal candidiasis. Analyzing the characteristics of the vaginal microbiota can enhance the clinical management of HPV infection.

Objective:To analyze vaginal microecological changes in patients with vaginal infectious diseases and their correlation with human papillomavirus (HPV) infection.Methods:A case-control study was conducted involving 416 patients who visited the Gynecology Outpatient Department and the Cervical Disease Clinic at Yongkang Maternity and Child Care Hospital between December 2022 and December 2023. All patients underwent testing for vaginal microecology and HPV to evaluate the prevalence of vaginal infectious diseases and HPV infection. Vaginal microecological indicators were compared between patients with vaginal infections and those without identifiable pathogenic bacterial dysbiosis. The indicators included microbial density, lactobacilli levels, pH, hydrogen peroxide levels, abnormal leukocyte esterase, and the grading of vaginal lactobacilli. Additionally, the HPV infection status was compared among patients with different vaginal microecological environments, and the correlation between vaginal infections and HPV infections was analyzed.Results:In a study involving 416 participants, 216 were diagnosed with vaginal infections, with an incidence rate of 51.92% (216/416). Among these cases, 118 were classified as a single infection, accounting for 54.63% (118/216), while 98 were identified as mixed infections, accounting for 45.37% (98/216). The rate of HPV infection among the 216 patients with vaginal infectious diseases was 37.04% (80/216). This rate was significantly higher than the 15.31% (30/196) observed in patients without identifiable pathogenic bacterial dysbiosis ( χ2 = 24.79, P < 0.001). Patients with vaginal infectious diseases also displayed elevated rates of abnormal lactobacilli levels, abnormal pH, abnormal leukocyte esterase, and abnormal lactobacilli grading, with rates of 59.26%, 86.57%, 72.69%, and 57.41%, respectively. In comparison, patients without identifiable pathogenic dysbiosis had rates of 31.12%, 18.88%, 51.53%, and 34.18%, respectively. All differences were statistically significant ( χ2 = 32.76, 86.83, 19.64, 22.28, all P < 0.001). Additionally, significant differences in HPV infection rates were observed among patients who tested positive for bacterial vaginosis, vulvovaginal candidiasis, and trichomoniasis when compared to those who tested negative ( χ2 = 12.46, 4.04, 6.14, P < 0.001, 0.044, 0.013). Both bacterial vaginosis and vulvovaginal candidiasis were recognized as high-risk factors for HPV infection ( OR = 4.039, 2.902, both P < 0.05). Conclusions:Vaginal infectious diseases are significantly linked to HPV infection, particularly bacterial vaginosis and vulvovaginal candidiasis. Analyzing the characteristics of the vaginal microbiota can enhance the clinical management of HPV infection.

Objective:To improve the accuracy of cone-beam CT (CBCT) image registration by using a dose-guided registration algorithm based on multi-objective optimization.Methods:A total of 28 sets of CBCT images of 6 patients with lung cancer and 5 patients with cervical cancer admitted to Harbin Medical University Cancer Hospital in 2022 were retrospectively analyzed. Using the results of bone-based registration as the starting points for dose registration algorithm, the dose fluence weighted mean square errors of each displacement point in surrounding three-dimensional space were calculated, and the candidate displacement points were selected by unsupervised k-means clustering method. The three-dimensional dose distribution of each candidate displacement point was calculated by using the limited size pencil beam algorithm, and the dose histogram indexes were extracted as the optimization variables of the multi-objective optimization algorithm. After the Pareto optimal solution set obtained using multi-objective genetic algorithm, the optimal unique solution was then determined based on the predetermined objective weight scheme.Results:After dose optimization registration, the values of D 90%, D 95%, D 98%, D mean and conformity index (CI) of planning target volume (PTV) of lung cancer patients were increased by 0.23 Gy, 0.49 Gy, 1.05 Gy, 0.15 Gy, 0.03 compared with conventional registration, respectively, and no significant difference was found for the organs at risk (OAR). For cervical cancer cases, the values of D 90%, D 95%, D 98%, D mean and CI of PTV were increased by 0.72 Gy, 1.15 Gy, 2.53 Gy, 0.24 Gy, 0.05 compared with conventional registration, respectively, whereas the evaluation indexes of partial OAR were decreased by 1.06-1.81 Gy. Conclusion:The proposed dose-guided registration algorithm can improve the dose coverage for the target area, decrease the dose for OAR and reduce residual error of rigid registration algorithm, which can be implemented as part of online adaptive radiotherapy.

Objective:To analyze and clarify the causes, types, risk factors and treatment principles of early postoperative tachycardia in children with congenital heart disease.Methods:A retrospective analysis of the clinical data of 2 126 children with primary radical congenital heart surgical procedure in Shanghai Children′s Hospital from January 2014 to December 2020, including 1 322 cases of ventricular septal defect or ventricular septal defect combined with atrial septal defect, 421 cases of atrial septal defect, 194 cases of tetralogy of Fallot, D-transposition of the great artery or double outlets of right ventricle combined with pulmonary stenosis, and 189 cases of other complex congenital heart disease. The surgical method is a median sternal skin incision or a small right axillary skin incision, and cardiopulmonary bypass is established routinely. The age, body mass, disease type, cardiopulmonary bypass and aortic occlusion time, vasoactive drug use, ECG monitoring and other indexes were observed and monitored.Results:There are 425 cases of early postoperative tachycardia in 2 126 children with congenital heart disease, with an incidence of 20.0%. The incidences of sinus tachycardia, borderline ectopic tachycardia, atrial tachycardia and ventricular tachycardia were 14.8%(314/2 126), 4.5%(96/2 126), 0.8%(17/2 126) and 0.5%(10/2 126), respectively. Logistic regression analysis revealed that the low age ( OR=1.98, 95% CI: 1.25-2.65, P<0.01), low weight ( OR=2.35, 95% CI:1.86-2.75, P<0.01), large ventricular septal defect ( OR=1.56, 95% CI:1.09-2.06, P=0.02), complex congenital heart disease ( OR=2.03, 95% CI: 1.57-2.52, P<0.01), long duration of cardiopulmonary bypass ( OR=1.77, 95% CI: 1.23-2.28, P<0.01), long aortic cross-clamp time ( OR=1.89, 95% CI:1.20-2.55, P<0.01), acidosis ( OR=1.63, 95% CI:1.11-2.14, P<0.01), and the combination usage of vasoactive drugs ( OR=1.86, 95% CI:1.23-2.48, P<0.01) were significantly associated with the occurrence of early postoperative tachycardias. Conclusion:This study has important clinical guiding value for predicting early postoperative tachycardia in children with congenital heart disease, clarifying its causes and types, and timely handling, so as to improve the postoperative survival rate of children.

Objective    To study the safety of right vertical infra-axillary thoracotomy (RVIAT) in the repair of ventricular septal defect (VSD) and the optimal age for RVIAT. Methods    Between June 2014 and June 2018, 441 children underwent VSD repair via RVIAT in our hospital. According to the age, they were divided into four groups: a 4 months to 1 year old group (R1 group, n=123), a 1-2 years old group (R2 group, n=106), a 2-5 years old group (R3 group, n=166), a >5 years old group (R4 group, n=46). The clinical effects of the patients were compared. Results    All the operations were successfully performed and no serious complication was found in all groups. No statistical difference was observed in the operation time, blood loss during operation, thoracic drainage 24 h after operation among groups (P>0.05). The cardiopulmonary bypass time, aortic cross-blocking time and ICU stay time in the R1 and R2 groups were longer than those in the R3 and R4 groups (P<0.05). In the R1 group, the postoperative ventilating time and postoperative hospital stay time were longer, and the blood transfusion volume was more than those in the R3 and R4 groups (P<0.05). The incidence of postoperative complications was higher in the R4 group than that in the R1 and R3 groups (P<0.05). Conclusion    VSD repair via RVIAT may be more effective in children >2 years old, and 2-5 years old may be the optimal age.

Objective: To investigate the association between genetic variation of kinase insert domain receptor (KDR) and the prognosis in colorectal cancer (CRC) patients received 5-FU based adjuvant chemotherapy. Methods: The clinical data of 176 CRC patients, who underwent surgical treatment at the Department of Anus and Intestine Surgery, People’s Hospital of Zhengzhou during January 2012 and December 2017, were retrospectively analyzed, and 93 cases of tumor tissues were collected for this study. The genotype of KDR polymorphism locus was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). qPCR was used to detect the expression of KDR mRNAin colorectal cancer tissues. The correlation between the polymorphism genotypes and other variables was analyzed by logistic regression model. The expression of different genotypes of KDR was analyzed by nonparametric test. The relationship between KDR genotype and prognosis of patients was analyzed by Kaplan-Meier survival analysis, and the other variables were adjusted by Cox risk scale model. Results: Of the polymorphisms analyzed, only rs2071559 was of clinical significance. The distribution frequency of KDR rs2071559 in 176 CRC patients was as follows: TT genotype in 95 cases (53.98%), TC genotype in 70 cases (39.77%) and CC genotype in 11 cases (6.25%); the minor allele frequency was 0.26; and the distribution of three genotypes was in accordance with Hardy-Weinberg's Equilibrium (P=0.690). The median disease free survival (mDFS) of patients carrying C allele and wild type TT genotype was 4.4 and 3.2 years, respectively (P<0.05); The median overall survival (mOS) of patients with TC/CC genotype and TT genotype was 5.2 and 4.0 years, respectively (P<0.05). After COX model modification, the effect of TC/CC genotype on mOS was still statistically significant (OR=0.55, P<0.05). The mRNA expression of KDR in cancer tissues of the patients with TC/CC genotypes were significantly lower than those of the wild type TT genotype (P<0.01). Conclusion: The polymorphism of KDR rs2071559 is associated with clinical outcomes in patients with colorectal cancer. KDR rs2071559 may affect the prognosis of colorectal cancer patients by affecting the mRNAexpression of KDR.

OBJECTIVETo investigate the expression of miR-29b in cholangiocarcinoma and explore its effects on cell proliferation and apoptosis of cholangiocarcinoma cells.

METHODSReal-time PCR was used to detect the expression of miR-29b in cholangiocarcinoma cells line QBC939 and cholangiocarcinoma tissues. The lentiviral vector LV-hsa-miR-29b and blank vector were constructed to infect QBC939 cells. MTT assay and cell clone formation assay were performed to assess the changes in the cell proliferation and clone formation, respectively; flow cytometry was employed to evaluate the effect of miR-29b overexpression on cell cycle and apoptosis.

RESULTSThe expression of miR-29b was significantly down-regulated in QBC939 cells and cholangiocarcinoma tissues as compared with H-69 cells and normal tissues ( < 0.01). Compared with the blank vector, the lentiviral vector LV-hsa-miR-29b caused significantly increased expression of miR-29b in QBC939 cells ( < 0.01), which exhibited suppressed cell proliferation and clone formation ( < 0.01 or 0.05), cell cycle arrest at the S phase ( < 0.05), and significantly increased cell apoptosis ( < 0.01).

CONCLUSIONSAs a tumor-suppressing miRNA, miR-29b is down-regulated in cholangiocarcinoma, and its overexpression can suppress the proliferation and induce apoptosis of cholangiocarcinoma cells.