BACKGROUND:Mesenchymal stem cells are susceptible to senescence during in vitro expansion,which greatly hinders their application in vivo and in vitro.How to improve the replicative senescence of mesenchymal stem cells is an urgent problem to be solved in tissue engineering. OBJECTIVE:To determine whether vascular endothelial growth factor combined with basic fibroblast growth factor can improve the aging of bone marrow mesenchymal stem cells caused by replicative passage. METHODS:Rat bone marrow mesenchymal stem cells were extracted by whole bone marrow adhesion method.Passage 2 cells were selected as normal control group.Passage 7 and later algebraic cells were selected as aging model group.Vascular endothelial growth factor(50 μg/L),basic fibroblast growth factor(10 μg/L),and their combination were administered.Cell proliferation was detected by CCK-8 assay.Cell senescence was observed by β-galactosidase activity staining.Cytoskeleton size and colony formation ability were observed by phalloidine staining and Giemsa staining,respectively,and the levels of senescence-related genes P16,P21,and P53 were detected by qRT-PCR.Gene expression levels of P16,P21,and P53 were tested by qRT-PCR. RESULTS AND CONCLUSION:(1)Vascular endothelial growth factor combined with basic fibroblast growth factor could promote the proliferation of aged bone marrow mesenchymal stem cells,which began to enter the plateau stage on day 9,and the absorbance value of the combined intervention group was significantly higher than that of the model group on day 9(P<0.05).(2)The phenotypic markers of the cells in the combined intervention group did not change,and the cell morphology changed from broad to slender.(3)Compared with the model group,the positive rate of β-galactosidase was significantly decreased(P<0.01);the number of nuclei increased(P<0.001);the total area of cytoskeleton increased(P<0.01);colony formation ability was enhanced(P<0.05);expression level of P16 was decreased(P<0.01)in the combined intervention group.These results indicate that vascular endothelial growth factor combined with basic fibroblast growth factor can improve the senescence of bone marrow mesenchymal stem cells caused by replicative passage without changing the cell phenotype.

Objective To investigate the efficacy of laparoscopic C-type radical hysterectomy through deep uterine vein approach for the treatment of cervical cancer.Methods From January 2021 to December 2022,58 cases of cervical cancer were treated with deep uterine vein approach laparoscopic C-type radical hysterectomy in our hospital.After establishing the operation channel,the ureter was identified in the posterior lobe of the broad ligament,and the uterine artery was exposed after separating the ureter.The connective tissue was separated along the dorsal side of the uterine artery to gradually expose the deep uterine vein.The parametrial lymph nodes surrounding the deep uterine vein were resected and sent to pathology alone.The deep uterine vein was continuously tracked towards the bladder,and its branches were freed.The deep uterine vein and its tributaries were double clipped by vascular clamp.Results The operation time was(307.2±54.1)min,the median bleeding volume was 50(20,100)ml,the lymph node dissection number was(26.3±6.9),the indwelling catheter time was(20.6±4.7)d,the time to removal of abdominal drainage tube was(9.4±4.1)d,the anal exhaust time was(36.7±4.1)h,the antibiotics use time was(9.2±4.2)d,and the hospital stay was(13.4±2.6)d.The postoperative complication rate was 3.4%(2/58),and the postoperative pathological staging upgrade rate was 22.4%(13/58).The postoperative European Organization of Research and Treatment of Cancer(EORTC)Quality of Life Questionnaire-Core 30(QLQ-C30)score was significantly higher than before surgery[(78.6±10.7)points vs.(47.1±7.6)points,t = 17.177,P = 0.000].All the 58 cases were followed up for 4-25 months(mean,13.5±6.2 months),with no recurrence.Conclusion Laparoscopic C-type radical hysterectomy through deep uterine vein approach is effective,safe,and reliable.

Objective:To investigate the safety and effectiveness of endoscopic retrograde cholangiopancreatography (ERCP) for the diagnosis and treatment of pediatric pancreaticobiliary maljunction (PBM).Methods:Data of 40 pediatric patients under 14 with PBM diagnosed and treated by ERCP at Department of Gastroenterology, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from November 2012 to September 2022 were collected. PBM types, ERCP-related diagnosis and treatment, adverse events and prognosis were retrospectively analyzed.Results:Nineteen cases were P-B type (joining of common bile duct with pancreatic duct), 17 were B-P type (joining of pancreatic duct with common bile duct), and 4 were complex type. Forty children with PBM underwent 50 ERCP-related operations, among which 48 procedures succeeded. One case failed during cannulation of ERCP, replaced by rendezvous-assisted endoscopic retrograde pancreatography (RV-ERP) afterwards. There were no serious postoperative adverse events such as bleeding, perforation or death. Thirty-four patients (85%) were followed up successfully, among which 14 underwent further surgery and 20 continued conservative treatment.Conclusion:ERCP is the golden standard to diagnose pediatric PBM, and it is also safe and effective treatment for PBM.

Objective:To understand the epidemiological characteristics and incidence trend of severe fever with thrombocytopenia syndrome (SFTS) in China.Methods:The incidence data of SFTS in China from 2018 to 2021 were collected from Chinese Disease Prevention and Control Information System for a statistical and descriptive epidemiological analysis by using software such as Excel 2016, Joinpoint 5.0.2, SPSS 26.0, and GraphPad Prism 8.0, especially, the SFTS cases reported monthly by key provinces were analyzed.Results:From 2018 to 2021, a total of 8 835 SFTS cases were reported in 25 provinces and the annual incidence showed an upward trend. The distribution of SFTS cases showed clustering, but the cases were mainly sporadic ones. The cases began to increase in March, mainly occurred during April to October (96.79%,8 551/8 835), and peaked during May to July. The cases were mainly distributed in middle-aged and old farmers, and slight more cases were women. The average case fatality rate was 5.38%, which varied greatly with areas. The case fatality rate tended to increase with age.Conclusion:From 2018 to 2021, the epidemiological characteristics of SFTS in China remained stable, but the number of reported cases gradually increased and the distribution showed an expanding trend, to which close attention should be paid.

Objective To investigate the effect of long non-coding RNA-LINC00342 in cervical cancer.Methods The bioinformatic analysis was performed by the cervical cancer data sets(GSE6791 and GSE63514)in GEO database.The potential oncogene LINC00342 was screened out.qRT-PCR was adopted to detect the expression level of LINC00342 in cervical cancer tissue and paracancerous normal cervical tissues of 50 cases of cervical cancer,and its relationship with the prognosis of the patients was analyzed.The correlation between LINC00342 with the other pathological features in the patients with cervical cancer was analyzed.In the cervical cancer cell lines Hela and SiHa cells,after transfecting si-LINC00342 and si-NCS,the LINC00342 expression level in the cells was detected by qRT-PCR.The cellular invasive and migration were detected by CCK-8/EdU proliferation test,Transwell invasion and migration test.Results The data sets of GSE6791 and GSE63514 and clinical tissue samples all showed that LINC00342 was highly expressed in cervical cancer tis-sues(P＜0.05).The clinical tissue samples related tests showed that the expression level of LINC00342 was correlated with the prognosis of the patients(HR=4.911,P=0.036 1);the LINC00342 level had no relation with the factors such as age of the patients and tumor histological type(P＞0.05),but was closely related to the tumor size and tumor stage(P＜0.05).The cellular experiments showed that knocking down LINC00342 expression could significantly inhibit the proliferation,invasion and migration abilities of cervical cancer cells(P＜0.05).Conclusion LINC00342 is highly expressed in cervical cancer tissues,which is a potential biomar-ker of the occurrence,development and poor prognosis of cervical cancer.

Objective:To explore the clinical phenotype and pathogenesis of a child with partial duplication in the long arm of chromosome 10 (10q), and conduct a review of relevant literature.Methods:A child presented at Lianyungang Maternal and Child Health Care Hospital in April 2018 for growth retardation, intellectual disability, and autism spectrum disorder (ASD) was selected as the study subject. Peripheral blood samples were collected from the child and his parents for G-banded chromosomal karyotyping analysis. Genomic DNA was also extracted for chromosomal microarray analysis (CMA). The clinical phenotype and relevant genes were searched in the Online Mendelian Inheritance in Man (OMIM) and the UK Database of Genomic Variation and Phenotype in Humans using Ensembl Resources (DECIPHER). The pathogenicity of chromosomal variation was analyzed based on guidelines from the American College of Medical Genetics and Genomics (ACMG). Relevant literature was searched from the CNKI, Wanfang Data, and PubMed databases by using keywords such as " 10q" " duplication" and " trisomy", with the time set as from the establishment of database to December 1, 2023. This study has been approved by the Medical Ethics Committee of the Lianyungang Maternal and Child Health Care Hospital (No. XM2023030).Results:The clinical phenotype of child had included growth retardation, intellectual disability, and ASD. G-banded chromosomal analysis suggested that the child has a karyotype of 46, XY, dup(10)(q23.31q24.33), whilst both of his parents were normal. CMA analysis of the child revealed that the child was arr[19]10q23.31q24.33(87603382_104948862)×3, with a 17.34 Mb duplication in the 10q23.31q24.33 region. Search of the OMIM database suggested that the duplicated segment has contained 171 genes associated with various diseases, and search of the DECIPHER database has identified cases with overlapping with the duplication. A search of the PubMed database has identified 2 publications involving 2 patients with chromosomal duplications overlapping the 10q23.31q24.33 region with a segment length of > 10 Mb. The 2 patients had mainly manifested growth retardation, intellectual disability, ASD, and facial and limb malformations. The main pathogenic genes had included PTEN, WNT8B, LZTS2, NFKB2, PAX2, KIF11, FRA10AC1, and CNNM2. No similar case was retrieved from the CNKI and Wanfang Data databases. Conclusion:The partial 10q duplication as a novel CNV involving genes such as PTEN and WNT8B probably underlay the growth retardation, intellectual disability and ASD in the child. This study has enriched the genotype-phenotype spectrum of patients with partial 10q23.31q24.33 duplications.

Objective:To explore the correlation between multidimensional nutritional evaluation indexes of folate and vitamin B 12 and mild cognitive impairment in community-dwelling older adults. Methods:In this study, cross-sectional study design was used, and the elderly people aged 60 years and above were recruited from three parallel communities in Shijiazhuang City.The survey was conducted including basic information questionnaire and semi-quantified food frequency questionnaires(FFQ)in the past year.The Montreal Cognitive Assessment Scale(MoCA)assessed the cognitive function.The subjects were divided into two groups of normal cognition and mild cognitive impairment according to the criteria categorized by education levels recommended by the 2018 China Dementia and Cognitive Impairment Diagnosis and Treatment Guidelines.Serum folate, vitamin B 12 and erythrocyte folate levels were measured by electrochemiluminescence immunoassay, serum homocysteine concentrations were measured by circulating enzyme assay, dietary folate and vitamin B 12 intake was calculated by FFQ survey and food composition tables.Logistic regression models were used to analyze the association between multidimensional nutritional indicators of folate and vitamin B 12 and mild cognitive impairment. Results:A total of 537 subjects were included in the study, involving 379(70.6%)with normal cognition and 158(29.4%)with mild cognitive impairment.Dietary vitamin B 12 intake and erythrocyte folate concentration was significantly lower in the elderly with mild cognitive impairment than in those with normal cognition[median(interguartile): 18.9(11.0, 32.9)μg/d vs.20.1(14.2, 34.8)μg/d, Z=2.010, P=0.044; median: 359.4(304.2, 413.4)μg/L vs.378.4(322.5, 433.1)μg/L, Z=2.168, P=0.031].The elderly with mild cognitive impairment had a higher prevalence of folate deficiency than those with normal cognition[17(10.8%) vs.22(5.8%), χ2=4.065, P=0.044].After adjusting for age, sex, and energy intake, the OR(95% CI)values for dietary vitamin B 12 intake and erythrocyte folate concentration in relation to mild cognitive impairment in the elderly were 0.99(0.987, 0.996)( P=0.039)and 0.98(0.974, 0.993)( P=0.021), respectively.The OR(95% CI)values for folate deficiency for mild cognitive impairment was 1.96(1.009, 3.795)( P=0.046). Conclusions:Long-term high dietary vitamin B 12 intake and better in vivo nutritional deposition of folate were negatively associated with mild cognitive impairment in community-dwelling elderly adults, whereas indicators reflecting the short-term nutritional status of folate and vitamin B 12 may be less associated with mild cognitive impairment.

Objective:To explore the clinicopathological and genetic characteristics of sporadic medullary thyroid carcinoma (MTC) and new therapeutic targets for sporadic MTC.Methods:Based on family and personal disease history, we identified 32 sporadic MTC who underwent surgical resection from Jan 2010 to Dec 2022. Clinicopathological and immunohistochemical features were analyzed in all patients, while 6 of them were subject to the whole exome sequencing (WES).Results:Compared with those of low-grade sporadic MTC, patients with high-grade tumors were more likely to have lymph node metastasis at presentation ( χ2=4.428, P=0.040); less likely to be cured by biochemical treatment ( χ2=4.072, P=0.044). Pathological grading scheme, biochemical cure, and TNM stage were independent risk factors of disease free survival. WES was performed on 6 pairs of normal tissues. We screened RET and RAS as driver mutations, and the mutation ratio was 3/6 respectively. Patients with RET or RAS mutations had no recurrence. In addition, we detected PDGFRA somatic mutation, with a mutation ratio of 1/6. Conclusions:For sporadic MTC cases, the pathological grading system has important prognostic value, and RET and RAS somatic mutations are the main driver mutations. PDGFRA are potential therapeutic targets for sporadic MTC.

Objective:To explore the effectiveness and safety of endoscopic ultrasound-guided pancreatic duct drainage (EUS-PD).Methods:A retrospective analysis was conducted on data of 16 patients who underwent EUS-PD because of endoscopic retrograde pancreatography (ERP) failure, poor effectiveness or anatomical changes and couldn't undergo the routine ERP in Nanjing Drum Tower Hospital from June 2018 to July 2022. The technical success of EUS-PD, clinical efficacy and post-procedure adverse events were analyzed.Results:In the 16 patients, there were 14 males and 2 females, with age of 50.69±12.95 years. A total of 19 times of EUS-PD operations were included, 3 of them were rendezvous-assisted endoscopic retrograde pancreatography (RV-ERP), 15 transgastric or transenteric EUS-guided stent placement and 1 was EUS-guided nasopancreatic duct placement. Technical success was achieved in 84.21% (16/19) patients, and among whom 93.75% (15/16) achieved clinical success. The overall incidence of postoperative adverse events was 52.63% (10/19) including 47.37% (9/19) abdominal pain, 15.79% (3/19) fever and 15.79% (3/19) postoperative pancreatitis. All adverse effects were relieved after general conservative treatment and no primary disease or surgery-related death occurred. The mean follow-up was 17.6 (8.2,22.3) months and 93.75% (15/16) of the patients were followed up. By the time of follow-up, 76.92% (10/13) of the patients who had successfully received EUS-PD had no recurrence of abdominal pain or distension.Conclusion:EUS-PD is a safe and effective alternative therapy for those with pancreatic diseases with ERP failure, poor efficacy or anatomical changes.

Objective:To investigate the risk factors for combined coronary microvascular dysfunction (CMD) in patients with ischemia and non-obstructive coronary artery disease (INOCA).Methods:From October 2020 to May 2022, 100 INOCA patients with myocardial ischemic symptoms who underwent coronary angiography (CAG) suggestive of <50% stenosis in all three coronary arteries at the Tenth People′s Hospital of Tongji University were prospectively recruited. Myocardial perfusion imaging (MPI), transthoracic echocardiography and cadmium-zinc-telluride (CZT) SPECT coronary flow quantification were performed in the same month, and 93 INOCA patients (36 males and 57 females, age (63.0±10.9) years) were finally included. CMD was defined as coronary flow reserve (CFR)<2.5. Independent-sample t test, Mann-Whitney U test and χ2 test were used to compare MPI results and left ventricular volume parameters between CMD and non-CMD groups. ROC curve analysis was used to analyze the efficacy of each index in predicting CMD, and independent risk factors for CMD were screened by multivariate logistic regression analysis. Results:Among 93 INOCA patients, 29 were in the CMD group and 64 were in the non-CMD group. The age, proportion of hypertension, left ventricular mass index (LVMI), summed stress score (SSS), summed difference score (SDS), left ventricular internal diameter systolic (LVIDS), interventricular septum thickness (IVST), and left ventricular posterior wall thickness (LVPWT) in the CMD group were higher than those in the non-CMD group ( t values: 2.42-3.76, χ2=8.94, z values: -3.31, -3.41, all P<0.05). ROC curve analysis showed that LVMI, SSS, SDS, LVPWT, IVST and age were significant in predicting CMD (AUCs: 0.67-0.72). Multivariate logistic regression analysis showed that LVMI (odds ratio ( OR)=1.08, 95% CI: 1.01-1.17), SDS ( OR=5.37, 95% CI: 1.95-14.78), hypertension ( OR=5.68, 95% CI: 1.34-24.18) and age ( OR=1.10, 95% CI: 1.03-1.18) were risk factors for CMD. Conclusion:LVMI, SDS, hypertension and age are strongly associated with combined CMD in INOCA patients, which can be used for early risk stratification of INOCA patients.