Objective:To explore the genetic basis of a child with mental retardation and developmental delay.Methods:A child who had attended the genetic clinic of Linyi People′s Hospital from October 2023 to April 2024 was selected as the study subject. Intelligence and development were assessed with simplified Peabody scale. Electroencephalogram and imaging data were collected. Peripheral blood samples of the child and her parents were collected for the screening of genetic metabolic diseases, chromosomal karyotyping, and trio-whole genome sequencing (trio-WGS) analysis. Candidate variant was verified by Sanger sequencing, and RNAseq was carried out to verify the alternative splicing due to the candidate variant. This study has been approved by the Medical Ethics Committee of Linyi People′s Hospital (No. YX200083).Results:The patient was an 8-year-and-11-month-old girl. Both of her parents had normal phenotypes. The child was assessed by the simplified Peabody scale as having intellectual disability and developmental delay. MRI showed no definite abnormal signals within the brain parenchyma, and electroencephalogram was normal. Screening of genetic metabolic diseases showed no obvious abnormality. Chromosomal karyotype was normal. Trio-WGS has detected a c. 697+ 1G>A variant in the intron 4 of the NFIX gene, along with 9 other variants within eight genes. The c. 697+ 1G>A variant may cause abnormal splicing of the NFIX gene transcript. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c. 697+ 1G>A variant was predicted to be pathogenic (PVS1+ PS2+ PM2_Supporting), while the evidence for pathogenicity of the other 9 variants was insufficient. Conclusion:The novel de novo c. 697+ 1G>A variant of the NFIX gene probably underlay the pathogenesis of the child, which may have caused the disease by leading to abnormal splicing.

Objective:To analyze the treatment outcomes and prognoses of children with head and neck non-parameningeal rhabdomyosarcoma (HNnPM RMS).Methods:A retrospective analysis was performed on the clinical data of children with HNnPM RMS admitted to Beijing Children′s Hospital from September 2012 to September 2022. The clinical features, comprehensive treatment modes and prognoses of the patients were analyzed. The overall survival rate (OS) and event free survival rate (EFS) were calculated using the Kaplan-Meier method, and univariate analysis was performed using the Log-rank test.Results:A total of 70 children were included in this study, 38 males and 32 females, with a median age of 47 months (2-210 months). Pathological subtypes including the embryonal in 27 cases, the alveolar in 36 cases and the spindle cell and sclerosing in 7 cases. Thirty children (83.3%) with alveolar type were positive for FOXO1 gene fusion. All 70 children underwent chemotherapy, including 38 with neoadjuvant chemotherapy and 32 with adjuvant chemotherapy. Sixty of 70 children underwent surgery, of whom, 10 underwent two or more surgeries. There were 63 children underwent radiotherapy, including 54 with intensity-modulated radiation therapy, 4 with particle implantation and 5 with proton therapy. The median follow-up was 45 (5-113) months, the 5-year OS was 73.2%, and the 5-year EFS was 57.7%. Univariate analysis showed lymph node metastasis ( χ2=5.022, P=0.025), distant metastasis ( χ2=8.258, P=0.004), and high Intergroup Rhabdomyosarcoma Study (IRS) group ( χ2=9.859, P=0.029) as risk factors for poor prognosis. Before June 2016, the 5-year OS based on BCH-RMS-2006 scheme was 63.6%, and after 2016, the 5-year OS based on CCCG-RMS-2016 scheme was 79.6%. Conclusion:Multidisciplinary combined standardized treatment can offer good treatment outcome and prognosis for children with HNnPM RMS. Local control is a key to the efficacy of comprehensive treatment.

Objective:To develop the children′s postoperative health-related quality of life scale for thyroid cancer and to test its reliability and validity.Methods:The first draft of the scale was developed through literature search, focus group meetings, and a pre-survey, and 116 children (76 for testing and 40 for external validation) with thyroid cancer attending Beijing Children′s Hospital of Capital Medical University were selected to answer the scale, to screen and categorize the questions and to form the final scale with multiple dimensions.Results:The children′s postoperative health-related quality of life scale for thyroid cancer contained 5 dimensions and 29 questions. Exploratory factor analysis showed that the cumulative variance explained by the 5 factors was 64.343%. Confirmatory factor analysis showed correlations between the questions and dimensions of this scale, fair convergent validity for the scale, and good discriminant validity. The validity of the validity scale showed that there was a existing correlation between the questions and the validity scale of this scale. The item-dimension correlation coefficients showed that the questions in each dimension were well differentiated. The total Cronbach′s α coefficient of the scale was 0.930, the folded half reliability was 0.843, and retest reliabilities at 2 weeks, 1 month, and 3 months after the initial test were respectively 0.936, 0.922, and 0.910.Conclusion:The developed children′s postoperative health-related quality of life scale for thyroid cancer has good reliability and validity and can be used to assess the health-related quality of life of children after thyroid cancer surgery.

Objective:To determine the characteristics of cervical neuroblastoma and the effect of resection extent on survival and outcomes.Methods:We performed a retrospective review of 32 children with cervical neuroblastoma treated at Beijing Children′s Hospital between April 2013 and August 2020. Data were collected from the medical record. The individualized therapy was designed based on staging and risk group. Based on the extent of resection, patients were divided into incomplete and complete resection groups. Event free and overall survival rates were compared between two groups using the Kaplan-Meier method.Results:The ages of patients ranged from 1 month to 81 months, with a median age of 11 months, including 7 males and 15 females. Twenty-nine patients (90.6%) presented with cervical painless mass. The average diameter of the primary tumors was (5.12±1.43) cm. Tumors were located in the parapharyngeal space in 25 cases (78.1%) and in the root of the neck in 7 cases (21.9%). None had MYCN amplification. According to International Neuroblastoma Staging System (INSS), 15 patients (46.9%) were identified as stage 1, 11 patients (34.3%) as stage 2B, 3 patients (9.4%) as stage 3 and 3 patients (9.4%) as stage 4. There were 12 patients (37.5%) at low risk, 17 patients (53.1%) at intermediate risk and 3 patients at high risk according to Children′s Oncology Group (COG) risk classification system. All patients underwent tumor resection. Postoperatively Horner′s syndrome occurred in 13 patients (40.6%), pneumonia in 9 patients (28.1%), pharyngeal dysfunction in 8 patients (25.0%) and transient hoarseness in 4 patients (12.5%). At a median follow-up of 36.5 months, the overall survival rate was 96.4%, with no significant difference between incomplete and complete resection groups (100.0% vs. 96.3%, χ 2=0.19, P=0.667); the event free survival rate was 78.1%, with a significant difference between the two groups (40.0% vs. 85.2%, χ2=6.71, P=0.010). Conclusions:Primary cervical neuroblastoma has a young onset age, mostly in low and medium risk groups, and represents favorable lesions with good outcomes after multidisciplinary therapy. Less aggressive surgery with preservation of important structures is recommended. Complete resection should not be attempted if it would compromise vital structures.

Objective:To evaluate the efficacy and influencing factors of surgery combined with neoadjuvant chemoradiotherapy in the treatment of children with non-orbital head and neck rhabdomyosarcoma (HNRMS).Methods:Information from 45 children diagnosed as non-orbital HNRMS and subjected to surgery combined with neoadjuvant chemoradiotherapy in Beijing Children′s Hospital affiliated to Capital Medical University from August 2017 to July 2021 was analyzed. The patients included 25 males and 20 females, aged from 1 to 17 years old. The primary tumor site, pathological subtype, clinical stage, risk group, therapeutic regimen, resection range and outcome of all cases were also collected. The survival curves were made using the Kaplan-Meier method and the potential prognostic factors were investigated by Cox regression analysis.Results:Fifteen (33.3%) of 45 children achieved negative surgical margin under complete tumor resection. The postoperative pathological results showed that there were 20 cases of embryonic subtype, 19 cases of alveolar subtype and 6 cases of spindle sclerosis subtype. The postoperative follow-up time ranged from 4 to 71 months, with a median of 26 months. During the follow-up period, 13 children died, among whom brain metastasis was the most common cause of death, accounting for 7/13. The 3-year overall survival rate was 67.6%. Multivariate analysis showed that non-embryonic subtype ( HR=6.26, 95% CI: 1.52-25.87, P=0.011) and failure to reach R0 resection ( HR=9.37, 95% CI: 1.18-74.34, P=0.034) were independent risk factors affecting overall survival rate. Conclusion:Surgery combined with neoadjuvant chemoradiotherapy can offer a good efficacy for children with non-orbital HNRMS. Non-embryonic subtype and resection without negative operative microscopic margins are independent risk factors for poor prognosis, and brain metastasis is the main cause of death in these children.

Objective:To investigate the effect of LINC00261 on the radiosensitivity of nasopharyngeal carcinoma and tumor formation and its underlying mechanism in nude mice.Methods:qRT-PCR was used to detect the relative expression levels of miR-620 and LINC00261 in radiosensitive and radioresistant nasopharyngeal carcinoma tissues. After the 6-10B and HNE-3 cells were irradiated with 0, 2, 4, 6, and 8 Gy 60Coγ-ray, the relative expression levels of miR-620 and LINC00261 were measured by qRT-PCR. After over-expression or silencing of LINC00261 and inhibition of miR-620 expression, the cells were irradiated with 4 Gy 60Coγ-ray. Clone formation assay was performed to detect the radiosensitivity of nasopharyngeal carcinoma cells. Flow cytometry was used to detect cell apoptosis. Western blot was utilized to detect the expression levels of Cleaved caspase-3 and Cleaved caspase-9 proteins. Luciferase reporter assay was adopted to analyze the targeting relationship between LINC00261 and miR-620. The changes in tumor formation were observed in tumor-bearing nude mice. Results:Compared with the radiosensitive tissues, the expression of LINC00261 was significantly down-regulated, whereas that of miR-620 was significantly up-regulated in radioresistant tissues (both P<0.05). After different doses of irradiation, the expression of LINC00261 was significantly down-regulated, whereas that of miR-620 was significantly up-regulated in 6-10B and HNE-3 cells (both P<0.05). After overexpression of LINC00261 and interference with miR-620 expression, the expression levels of Cleaved caspase-3 and Cleaved caspase-9 were significantly up-regulated (both P<0.05), the cell apoptosis rate was remarkably increased ( P<0.05) and the cell survival fraction was significantly enhanced in 6-10B and HNE-3 cells ( P<0.05). LINC00261 targetedly regulated the expression of miR-620. Overexpression of miR-620 could attenuate the radiosensitization and pro-apoptotic effects of LINC00261 overexpression on nasopharyngeal carcinoma cells. LINC00261 overexpression could significantly reduce the tumor formation weight of nasopharyngeal carcinoma in nude mice ( P<0.05). Conclusion:Overexpression of LINC00261 can increase the radiosensitivity of nasopharyngeal carcinoma cells probably by targeted regulation of miR-620 expression.

Objective The purpose of this study was to explore common complications and their clinicopathological features in pediatric liver transplantation.Methods Clinical and pathological data of 240 liver biopsies from 168 children that conducted liver puncture from January 2015 to May 2018 in Tianjin First Central Hospital was retrospectively analyzed.We comprehensively analyzed incidence rate and pathological features of various complications,and correlations between acute rejection and C4d staining result or Banff score.Results A total of 86.67％ (208/240) liver biopsies could be definitely diagnosed with incidence rate of main complications in descending order as follows:T cell mediated rejection (TCMR) 60.57％ (126/208),drug-induced liver injury (DILI) 17.31％ (36/208),biliary complication 8.17％ (17/208),vascular complication 3.37％ (7/208),ischemia/reperfusion injury (IRI) 2.88％ (6/208),antibody mediated acute rejection (AMR) 1.92％ (4/208),HBV infection 1.92％ (4/208),non-alcoholic fatty liver disease (NAFLD) 1.44％ (3/208),chronic rejection (CR) 0.96 ％ (2/208) and HCV infection 0.48 ％ (1/208).TCMR and AMR in acute rejection (AR) accounted for 96.92％ (126/130) and 3.08％ (4/160),and into(portal-based,PB)type TCMR accounted for 96.03％(121/126) with the detectable rate of BP type subtype TCMR of 26.45％(32/121)within 30 d.There were 65.87％ (83/126)、25.40％ (32/126) 和4.76％ (6/126) of BP TCMR samples with "Banff ACR RAI" score within 3-5,6-7 and 8-9,and RAI score was negatively correlated with postoperative time (r =0.127,P =0.084).The incidence rate of central perivenulitis (CP) and portal eosinophils infiltration (PEI) in BP TCMR was 63.63％ (77/121) 和43.80％ (53/ 121),respectively,additionally,the PEI level was positively correlate with RAI score (P＜0.05).CP TCMR and AMR occurred within 30d-365 d and 8 d-180 d,respectively postoperative,while,the two CR occurred at 1095 d and 1335 d postoperative,and significant correlation was strikingly observed between rejection subtype and postoperative time (Z =9.231,P =0.026).C4d positive rate was 10％ (24/240),which was associated with Banff score and postoperative time,besides,C4d score was also correlated with rejection subtype and RAI score.The occurrence of DILI was mainly at time of ＜90 d or ＞180 d postoperative,and the detectable rate of biliary complication within 180 d postoperative was 82.35％ (14/17),IRI Appear in ＜30d.Hepatic artery complication account for nearly 57.14％ (4/7),occurrence time is ≤90 d.Occurrence of HBV infection,CMV infection and NAFLD were mainly at ＞365 d,＜90 d and ＜365 d,respectively.Conclusion There were lots of differences in clinical and pathological features among multi pediatric liver transplantation complications.Liver puncture plays an important role in rejection subtype classification and grading,as well as in non-rejection complications identification.

Objective To determine the levels of serum visfatin and HbA1c in patients with coronary heart disease and ex-plore the correlation with the severity of coronary atherosclerosis .Methods Totally 264 patients were enrolled who performed cor-onary angiography totally ,visfatin and HbA1c levels were detected respectively of 33 cases of control group ,51 cases of atheroscle-rosis group ,75 cases of single-vessel disease group ,72 cases of double-vessel disease group ,33 cases of triple-vessel disease group . Gensini score was used for evaluation of coronary artery lesion ,and to establish a multiple linear regression analysis of the relation-ship between each risk factor for coronary heart disease .According to the degree of coronary artery stenosis ,the patients also could be divided into the control group(33 cases) ,the non severe stenosis group (174 cases) and the severe stenosis group (57 cases) ,the changes of visfatin and HbA1c levels were analyzed in the three groups .Results In the groups by the coronary lesion count ,HbA1c levels increased with the degree of coronary artery lesions in the control group [(4 .98 ± 0 .21)% ] ,the atherosclerosis group [(5 .58 ± 0 .36)% ] ,the single-vessel disease group[(6 .17 ± 0 .48)% ] ,the double-vessel disease group[(6 .63 ± 0 .80)% ] ,the tri-ple-vessel disease group[(7 .97 ± 1 .49)% ] ,and comparisons had significant difference between any two groups(P< 0 .05) ;Visfatin level in the control group ,the atherosclerosis group ,the single-vessel disease group ,the double-vessel disease group ,the triple-ves-sel disease group were(0 .73 ± 0 .42)μg/L ,(1 .50 ± 0 .87)μg/L ,(3 .45 ± 2 .50)μg/L ,(5 .45 ± 2 .96)μg/L ,(9 .21 ± 6 .35)μg/L ,a-mong them coronary heart disease group (the single-vessel disease group ,the double-vessel disease group ,the triple-vessel disease group) is higher than the atherosclerosis group and the control group(P< 0 .05) ;the atherosclerosis group is higher than the con-trol group ,but there was no statistically significant difference(P> 0 .05) ;In according to the degree of coronary artery diameter ste-nosis ,the levels of visfatin ,HbA1c in severe stenosis group [(8 .25 ± 4 .86)μg/L ,(7 .35 ± 1 .43)% ] is significantly higher than the non severe stenosis group [(3 .22 ± 2 .74)μg/L ,(6 .14 ± 0 .70)% ] and the control group [(0 .73 ± 0 .42)μg/L ,(4 .98 ± 0 .21)% ] , P< 0 .01 ;The non severe stenosis group is significantly higher than the control group (P< 0 .01) .The levels of visfatin ,HbA1c ,hs-CRP ,LDL and TC had positive correlation with Gensini score(P< 0 .01) .The level of HDL was negatively correlated with Gensini score(r= - 0 .535 ,P < 0 .01) .The levels of visfatin ,HbA1c ,hs-CRP ,LDL and TC had positive correlation with visfatin ( P <0 .01) ,and the level of HDL and TG were negatively correlated with visfatin(P< 0 .01) .In multiple linear regression analysis ,the factors which finally entered the regression equation were HbA1c ,LDL ,hs-CRP ,visfatin and HDL .Conclusion The levels of visfa-tin and HbA1c is closely related to the severity of coronary atherosclerosis .Combined detection of visfatin and HbA1c can be used as important indicators for evaluating the severity of coronary atherosclerosis .

Objective To explore the inhibitory effects of endogenous hydrogen sulfide, a novel and important gas-eous transmitter generated in mammalian tissues mainly by cystathionine β-synthase (CBS) or cystathionineγ-lyase (CSE) on the apoptosis of the rat hepatic BRL cell line in physiological condition. Methods BRL cells were cultured, and divid-ed randomly into several groups in different phases of the experiment, including negative-siRNA (control) group, CBS siRNA (CBS 1~3) group and CSE siRNA (CSE 1~3) group, which were used to select the most efficient sequences of siRNAs at 48 or 24-hour transfection. Solution group and (CBS+CSE) siRNA group were added to detect the variation of apoptosis. The BRL cell line was observed and evaluated at 0, 4, 8, 12, and 24 hrs after siRNA transfection. When the mechanisms of the apoptosis were detected, CBS/CSE siRNAs were transfected individually or jointly into BRL cells, and compared with nega-tive-siRNA group to examine the variation. The genic and protein expression of CBS/CSE were detected by RT-PCR and Western blot assay. After transfection of CBS/CSE siRNA, the apoptosis of BRL cells was detected by Hoechst stain and flow cytometry (FCM). The mitochondrial membrane potential (MMP) changes were observed by fluorescent staining. Western blot assay was used to examine the protein expression of intracytoplasm cytochrome C (Cyt C) and cleaved-caspase 3. Re-sults CBS and CSE were observed in BRL cells. After transfection of CBS/CSE siRNA, endogenous H2S generation de-creased and the apoptosis of BRL cells increased. Accordingly, the expression of intracytoplasm-Cyt C and cleaved-caspase 3 increased. Conclusion The inhibition of endogenous H2S synthesis induced the apoptosis of BRL cells under physiologi-cal condition, which may be involved in mitochondrial pathway of apoptosis.