OBJECTIVE: To explore the genetic basis of a patient with clinically suspected Loeys-Dietz syndrome (LDS). METHODS: A child who had presented at Beijing Anzhen Hospital in September 2018 was selected as the study subject. Clinical data and family history of the patient were collected, along with peripheral blood samples of the proband and his parents. Whole exome sequencing (WES) was carried out through next-generation sequencing. RESULTS: Candidate variants were searched through bioinformatic analysis focusing on genes associated with hereditary aortic aneurysms. Candidate variant was verified by Sanger sequencing. The patient was found to have cardiovascular abnormalities including early-onset aortic dilatation and coarctation, and LDS syndrome was suspected. WES revealed that he has harbored a heterozygous c.1526G>T missense variant of the TGFBR2 gene. The same variant was not found in either parent and was predicted as likely pathogenic (PM1+PM2_Supporting+ PM6+PP3+PP4) based on the guidelines from the American College for Medical Genetics and Genomics (ACMG). CONCLUSION The TGFBR2 c.1526G>T variant probably underlay the LDS in this patient and was unreported previously in China. Above finding has enriched the mutational spectrum of the TGFBR2 gene associated with the LDS and provided a basis for the genetic counseling for the patient.
Child ; Humans ; Male ; China ; Computational Biology ; Family ; Loeys-Dietz Syndrome/genetics* ; Mutation ; Receptor, Transforming Growth Factor-beta Type II/genetics*

Objective:To investigate the influencing factors of textbook outcomes in liver surgery (TOLS) after radical resection of gallbladder carcinoma.Methods:The retrospective case-control study was conducted. The clinicopathological data of 530 patients who underwent radical resection of gallbladder carcinoma in 15 medical centers, including the First Affiliated Hospital of Army Medical University et al, from January 2014 to January 2020 were collected. There were 209 males and 321 females, aged (61±10)years. Patients underwent radical resection of gallbladder carcinoma, including cholecystectomy, hepatectomy, invasive bile duct resection, and lymph node dissection. Observation indicators: (1) situations of TOLS; (2) influencing factors of TOLS. Measure-ment data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data between groups was conducted using the Mann-Whitney U test. The univariate analysis was conducted using the corresponding statistical methods based on data type, and variables with P<0.10 were included in multivariate analysis. Multivariate analysis was conducted using the Logistic stepwise regression model. Results:(1) Situations of TOLS. All 530 patients underwent radical resection of gallbladder carcinoma, and there were 498 cases achieving R 0 resection, 508 cases without ≥grade 2 intra-operative adverse events, 456 cases without postoperative grade B and grade C biliary leakage, 513 cases without postoperative grade B and grade C liver failure, 395 cases without severe com-plications within postoperative 90 days, 501 cases did not being re-admission caused by severe com-plications within postoperative 90 days. Of the 530 patients, 54.53%(289/530) of patients achieved postoperative TOLS, while 45.47%(241/530) of patients did not achieve postoperative TOLS. (2) Influencing factors of TOLS. Results of multivariate analysis showed that American Society of Anesthesiologists classification >grade Ⅱ, preoperative jaundice, T staging as T3?T4 stage, N staging as N2 stage, liver resection as right hemi-hepatectomy, and neoadjuvant therapy were independent factors influencing TOLS in patients undergoing radical resection of gallbladder carcinoma ( odds ratio=2.65, 1.87, 5.67, 5.65, 2.55, 3.34, 95% confidence interval as 1.22?5.72, 1.18?2.95, 2.51?12.82, 2.83?11.27, 1.41?4.63, 1.88?5.92, P<0.05). Conclusion:American Society of Anesthesiologists classification >grade Ⅱ, preoperative jaundice, T staging as T3?T4 stage, N staging as N2 stage, liver resection as right hemi-hepatectomy, and neoadjuvant therapy are independent factors influencing TOLS in patients undergoing radical resection of gallbladder carcinoma.

Objective:To explore the short-term and long-term prognostic outcomes of anatomical liver resection(AR)for patients with perihilar cholangio-carcinoma. Methods:This is a retrospective study.All data were obtained from 4 centers,including The First Affiliated Hospital of Army Medical University,Eastern Hepatobiliary Hospital of Naval Medical University,Sichuan Provincial People's Hospital and Affiliated Hospital of Qinghai University,of a multi-center database.A total of 305 consecutive perihilar cholangiocarcinoma patients receiving radical resection between January 2013 and June 2021 were included in this study.According to the method of liver resection,all patients were divided into the AR group(n=205)and the non-anatomical liver resection(NAR)group(n=100).The baseline characteristics,short-term prognosis and long-term prognosis of the 2 groups were compared. Results:The perioperative transfusion rate and the 30-day complication rate were significantly lower in the AR group than those in the NAR group(P＜0.05).There was no statistically significant difference in the survival rates between the AR and the NAR groups(P＞0.05). Conclusion:The 2 hepatic resection modalities had no obvious effect on the long-term prognosis of perihilar cholangiocarcinoma patients after radical resection,but choosing AR tends to achieve a better short-term prognosis and is worth promoting in clinical practice.

Objective:To investigate the expression level of CD70 and receptor CD27 in hepatocellular carcinoma (HCC) and their relationship with clinicopathological features and prognosis, and to explore the possible role of CD70 in the tumor microenvironment.Methods:Download the RNA sequencing data related to HCC from The Cancer Genome Atlas (TCGA), and analyze the expression differences of CD70 and CD27 between HCC and normal tissues, and their relationship with clinicopathological features. Kaplan-Meier survival curve was used to compare the survival rate of patients with different expressions of CD70 and CD27. The correlation between CD70 and other genes that affect the expression of tumor microenvironment was analyzed.Results:The data of 371 HCC tumor samples and 50 normal tissue samples were obtained by TCGA database. The expression of CD70 in HCC tumor tissues was significantly higher than that in normal tissues. There was no significant correlation between CD70 expression and age, gender, tumor stage, distant metastasis and survival ( P>0.05). The expression of CD27 in HCC tumor tissues was not significantly different from that in normal tissues, but the overall survival rate of patients with high CD27 expression was significantly higher than that of patients with low CD27 expression. The expression of CD70 in HCC tumor tissue was significantly positively correlated with the expressions of IL-12, FOXP3, CD25, CD39, CD27, PD-1, PD-L1, LAG3, CTLA4, Tim3, VEGF, IDO, CXCR4 and CCL2 ( P<0.05, | r|>0.3). Conclusion:CD70 is abnormally expressed in HCC. Although CD70 expression is not significantly associated with pathological staging and survival in HCC patients, it is related to the immunosuppressive tumor microenvironment, which may be one of the mechanisms of immune escape in HCC.

Objective:To investigate the inhibitory effect of esomeprazole on proliferation and chemosensitizing effect of breast cancer cells.Methods:Human MBA-MD-231, MCF-7 breast cancer cell line and human Huh7 liver cancer cell line were cultured by conventional methods; cells were treated with different concentrations of esomeprazole, and CCK8 kit was used to detect the proliferation of different tumor cells after stimulation. Cells were treated with different concentrations of esomeprazole, and the effects of esomeprazole on cell cycle of different cells were analyzed by flow cytometry. Cells were treated with different concentrations of paclitaxel and epirubicin combined with esomeprazole, and CCK8 kit was used to detect the proliferation of different tumor cells after stimulation. Measurement data were expressed as mean ± standard deviation ( ± s), and analysis of variance was used for comparison among multiple groups. Results:CCK8 results showed that esomeprazole could inhibit the proliferation of MBA-MD-231 cells, MCF-7 cells and Huh7 cells in a dose-dependent manner. Flow cytometry results showed that cells in G 0/G 1 phase were significantly increased by esomeprazole treatment. Esomeprazole can enhance the inhibitory effect of paclitaxel and epirubicin on the proliferation of MBA-MD-231 cells and MCF-7 cells, and improve the chemosensitivity. Conclusion:Esomeprazole blocks breast cancer cell MBA-MD-231, MCF-7 and liver cancer cell Huh7 in G 0/G 1 phase, thereby inhibiting cell proliferation. Esomeprazole can enhance the inhibitory effect of chemotherapeutic drugs on the proliferation of MBA-MD-231 and MCF-7 cells.

Objective: To analyze the expression of tight junction protein 3(claudin-3) in colorectal cancer and its relationship with the development, metastasis and prognosis of colorectal cancer. Methods: From February 2013 to February 2015, 78 patients with colorectal cancer operated in the People's Hospital of Sanmen County were selected in this study.The tissues of colorectal cancer and adjacent tissues were collected and claudin-3 expression was detected.The relationship between claudin-3 and clinicopathological features of colorectal cancer was analyzed. Results: The positive rate of claudin-3 in cancer tissues(83.33%) was higher than that in paracancerous tissues(48.72%), and the difference was statistically significant(χ²=20.832, P<0.01). The positive rate of claudin-3 in the poorly differentiated group(100.00%) was significantly higher than that in the well-differentiated group(85.71%) and the well-differentiated group(52.63%)(χ²=19.209, P<0.01). The claudin-3 positive rate with lymphatic metastasis(97.30%) was higher than that without lymphatic metastasis(70.73%), and the difference was statistically significant(χ²=9.882, P<0.01). The claudin-3 positive rate in patients with less than 3 years of survival (91.11%) was higher than that in patients with more than 3 years of survival(72.73%)(χ²=4.633, P<0.05). Conclusion The expression of claudin-3 is related to the occurrence, development and lymphatic metastasis of colorectal cancer, and can predict the prognosis of the patients.

[Abstract] Objective: To investigate the effect of 18H12, a functional monoclonal antibody that can target gastric cancer stem cells, on the self-renewal and invasion ability of gastric cancer cells. Methods: The gastric cancer cell line PAMC-82 was used as cell model, the expression of ENO1 (enolase-1) on the membrane surface of its parental cells and enriched stem cells by sphere culture was detected by Flow cytometry. Flow cytometry was used to separate ENO1+ cells and ENO1- cells to detect their self-renewal ability and invasion ability. With the commercial ENO1 antigen and antibody as the samples, CoIP (co-immunoprecipitation) was used to verify whether 18H12 antibody targeting ENO1 could able to accurately recognize ENO1. After being treated with 18H12 for 12 h, 24 h and 48 h, the selfrenewal and invasion ability of PAMC-82 cells were detected by methylcellulose pelletization experiment and Transwell chamber assay, respectively. Results: Flow cytometry showed that the expression of ENO1 on the membrane surface of PAMC-82 sphere cells was significantly higher than that of its parental cells (P<0.01), so ENO1 could be a potential target for targeting gastric cancer stem cells. The self-renewal ability and invasion ability of the sorted ENO1+ cells were significantly stronger than those of the ENO1- cells and the parental cells (P<0.05 or P<0.01). 18H12 antibody could accurately recognize ENO1, which was consistent with the commercial antibody recognition band. 18H12 could significantly inhibit self-renewal ability and invasion ability of PAMC-82 cells (P<0.01). Conclusion: Monoclonal antibody 18H12 can significantly inhibit the self-renewal and invasion of gastric cancer stem cells and is expected to be a candidate antibody drug targeting gastric cancer stem cells.

Objective: To explore the effects of Oxcarbazepine combined with Levetiracetam treatment on electroencephalography (EEG) and serum levels of S-100β, glial fibrillary acidic protein (GFAP) in children with epilepsy. Methods: One hundred and ten children with epilepsy who were admitted to Zhumadian Central Hospital from March 2016 to May 2017 were selected as research subjects.The random number table method was used to divide 55 patients into combined group (treated with Oxcarbazepine plus Levetiracetam) and 55 patients in control group (only treated with Oxcarbazepine). The clinical efficacy, EEG background activity, serum S-100β and GFAP levels were compared between the 2 groups after treatment. Results: The control rate of the combined group was 69.09%, the markedly effective was 18.18%, the effective was 9.09%, and ineffective was 3.64%; the control group had a control rate of 52.73%, markedly effective 20.00%, effective 20.00%, and ineffective 7.27%, respectively.The differences between the 2 groups were statistically significant (Z=-2.012, P=0.044). Before and after treatment, the background activity of EEG in both groups was dominated by alpha wave activity.There was no significant difference in the activity rate of alpha wave, theta wave and delta wave between the 2 groups (all P>0.05). Before treatment, there was no significant difference in serum S-100β and GFAP levels between the 2 groups (all P>0.05). After treatment, serum S-100β [(0.415±0.086) μg/L, (0.473±0.091) μg/L], GFAP [(2.60±0.44) ng/L, (2.93±0.40) ng/L] in the combined group and the control group were significantly lower than before treatment, and the differences were statistically significant (t=6.339, 6.703, 3.001, 3.364, all P<0.05). After treatment, serum S-100β and GFAP levels in the combined group were significantly lower than those in the control group, and the differences were statistically significant (t=3.435, 4.116, all P<0.05). Conclusions Oxcarbazepine combined with Levetiracetam is superior to Oxcarbazepine alone in the treatment of children with epilepsy, as it can reduce the levels of S-100β and GFAP and is worthy of clinical application.

Objective To provide new evidences for understanding the mechanisms of promo-tive role of orexins in anesthetic emergence and the effect of microinjection of orexin-A/orexin-B into cerebral ventricle on the release of histamine.Methods Male SD rats were randomly divided into sa-line (control ), orexin-A and orexin-B groups. The microdialysis probe was inserted into hypothalamus under stereotaxic apparatus.The perfused fluid from the area of hypothalamic tube-romammillary nucleus was collected using in vivo microdialysis at 1 h,2 h and 3 h after 1 nmol or 5 nmol orexin-A or orexin-B microinjection into the cerebral ventricle (n =5 each).The concentrations of histamine at each time point in dialysates of perfused fluid were detected by high performance liquid chromatography (HPLC)to analyze its dynamic changes.After one week,each group was microin-jected with 10 nmol,15 nmol and 20 nmol orexin-A and orexin-B (n =5)into the cerebral ventricle respectively,dialysates was collect and histamine was detected at 1 h to analyze its dosage response. After one week,each group was microinjected 0.3 μl saline orexin-A and orexin-B (n =6)into the tu-beromammillary nucleus.Results Compared with the control group,microinjection of 1 nmol orexin-A significantly increased histamine release at 1 h,but the same dose of orexin-B had no such effect,5 nmol of orexin-A or orexin-B injections significantly facilitated histamine release at 2 h and 3 h (P <0.01).Microinjection of 10 nmol,15 nmol and 20 nmol orexin-A and orexin-B into ventricle caused an significant increase of histamine release at 1 h while the effect was the strongest in 20 nmol (P <0.05).Compared with the control group,microinjection of orexin-A significantly decreased time of the righting reflex (P <0.01),but the same dose of orexin-B had no such effect.Conclusion Micro-injection of both orexin-A or orexin-B into cerebral ventricle could promote the release of histamine, while the effect of orexin-A was stronger.Microinjection orexin-A into tuberomammillary nucleus sig-nificant facilitated recovery from isoflurane.

Objective:To discuss the screening results and clinical characteristics of children of Miao and Dong nationalities with mediterranean anemia in ethnic minority areas of Qiandongnan State of Guizhou Province,and to clarify the differences of the mediterranean anemia among different minorities.Methods:A total of 1 623 children of Miao and Dong nationalities with mediterranean anemia in minority areas of Qiandongnan State were selected by multistage stratified random sampling method;quantitative analysis of HbA2 and HbF was used to screen the selected children with mediterranean anemia initially;phenol chloroform extraction method was applied to extract the DNA from the children with mediterranean anemia;ASO/RDB-PCR reverse dot blot hybridization method was used to analyze the gene characteristics of the children with mediterranean anemia.Results:A total of 1 623 children of Miao and Dong nationalities were selected as the subjects.Among 938 children with Miao nationality,there were 18 children with positive α-mediterranean anemia and 36 children with positive β-mediterranean anemia,and the positive detection rate was 1.92%.Among 685 children with Tong nationality,there were 13 children with positive α-mediterranean anemia and 24 children with positive β-mediterranean anemia,and the positive detection rate was 3.50%.The detection rates of composite of α-and β-mediterranean anemia in the children of Miao nationality and Tong nationality were 1.49% and 4.61%.There was no significant difference in the detection rates of different kinds of mealiterranean anemia between two nationalities (P<0.05).The major gene mutations in α-mediterranean anemia were——SEA/-αα and-α3.7,and the major gene mutations in β-mediterranean anemia were CD17/N and CD14-15/N,while the major gene types of the composite of α-and β-mediterranean anemia were——SEA/β41-42 and——SEA/β17.There was no difference in the positive rates of major gene types of different kinds of mediter ranean anemia between two nationalities(P<0.05).Conclusion:There is no difference in the positive rate of children of Miao and Dong nationalities with mediterranean anemia in minority areas of Qiandongnan State.CD17/N,——SEA/-αα and ——SEA/β41-42 are the major gene types of α-,β-,and αβ-mediterranean anemia,respectively.