Long non-coding RNA(lncRNA)is a class of endogenous non-coding RNA with a length of more than 200 nt.Due to lack of open reading frame(ORF),they lack the ability to directly encode proteins,but they play a crucial role in gene regulation.They are widely involved in epigenetics,transcription,translation,modification and degradation,thereby affecting the life activities of the body,and their expression imbalance is closely related to the occurrence and development of diseases.Therefore,analyzing the inherent characteristics of lncRNA and revealing its intrinsic role can not only deepen our understanding of human physiological and pathological processes,but also provide new ideas and potential solutions for the diagnosis,prevention and treatment of diseases.So as to provide references for the related research of lncRNAs.

Objective:To explore the prenatal ultrasound phenotype and genetic basis of two fetuses with Wolf-Hirschhorn syndrome (WHS).Methods:A retrospective analysis was conducted on the ultrasound imaging data of two fetuses suspected for WHS at the Prenatal Diagnostic Center of Qingyuan People′s Hospital in July 2017 and August 2019, respectively. Amniotic fluid samples of the two fetuses were subjected to chromosomal karyotyping and chromosomal microarray analysis (CMA). This study was approved by Medical Ethics Committee of the Qingyuan People′s Hospital (Ethics No. IRB-2022-064).Results:Prenatal ultrasound examination of the two fetuses had consistently revealed WHS-associated traits including intrauterine growth restriction (IUGR), craniofacial abnormalities and cardiovascular anomalies. Karyotyping analysis suggested that both fetuses had harbored cryptic chromosomal translocations involving partial deletion of 4p. And parental verification revealed that it was de novo for fetus 1 and paternal for fetus 2. CMA has confirmed that fetus 1 had an approximately 8.7 Mb deletion at 4p16.3p16.1 and a 6.8 Mb duplication at 8p23.1p23.1, whilst fetus 2 had a 20.05 Mb deletion at 4p16.3p15.31 and a 7.66 Mb duplication at 9p24.3p24.1. The karyotype of fetus 1 was determined as 46, XN, der(4)t(4; 8)(p16.1; p23.1)dn.arr[hg19]4p16.3p16.1(68345_8721580)×1, 8p23.3p23.1(158048_6933745)×3, and that of fetus 2 was determined as 46, XN, der(4)t(4; 9)(p15.3; p24)dpat.arr[hg19]4p16.3p15.31(68345_20116061)×1, 9p24.3p24.1(208454_7868292)×3. Conclusion:The 4p deletion is probably the main cause for the WHS phenotype in both fetuses. WHS should be suspected when IUGR, renal anomalies, craniofacial and cardiovascular abnormalities are detected upon prenatal ultrasound screening.

Objective:To explore the diagnostic value of chromosome karyotype analysis, chromosomal microarray analysis (CMA) and whole exome sequencing (WES) in microcephaly.Methods:A total of 9 cases of microcephaly fetuses diagnosed by prenatal ultrasound or children with microcephaly diagnosed after birth were selected from the Sixth Affiliated Hospital of Guangzhou Medical University from January 2014 to August 2022.Karyotype analysis and/or CMA were used to detect. The cases with negative karyotype analysis and CMA results were further sequenced by trio-based WES (Trio-WES). Then the coding genes contained in the pathogenic copy number variation (CNV) fragments were analyzed by gene ontology (GO) enrichment. The genes related to the development of the central nervous system contained in the pathogenic CNV and the pathogenic genes found by Trio-WES were combined for gene interaction network analysis.Results:In this study, 9 cases of microcephaly were recruited, with the time of diagnosis ranged from 23 weeks of gestation to 7 years after birth, and the head circumference of fetus or children ranged from 18.3 to 42.5 cm (-7SD to -2SD). Karyotype analysis was detected in all 9 cases and no abnormality result was found. Eight cases were detected by CMA, and one abnormal was found. Five cases were detected by Trio-WES, and two cases were detected with likely pathogenic genes. The GO enrichment analysis of the coding gene in the 4p16.3 microdeletion (pathogenic CNV) region showed that: in biological process, it was mainly concentrated in phototransduction, visible light; in terms of molecular function, it was mainly concentrated in fibroblast growth factor binding; in terms of cell components, it was mainly concentrated in rough endoplasmic reticulum. Gene interaction network analysis suggested that CDC42 gene could interact with CTBP1, HTT and ASPM gene.Conclusions:CMA could be used as a first-line detection technique for microcephaly. When the results of chromosome karyotype analysis and/or CMA are negative, Trio-WES could improve the detection rate of pathogenicity of microcephaly.

Long non coding RNA(lncRNA)LINC01296 plays a carcinogenic role in different tumor growth,apoptosis,and metastasis,but its mechanism in nasopharyngeal carcinoma is still unclear.The aim of this study is to study the mechanism of knockdown of long non-coding RNA(lncRNA)LINC01296 in inhibiting immune escape of nasopharyngeal carcinoma cells by regulating programmed death ligand-1(PD-L1).In this study,human peripheral blood lymphocytes were isolated and cultured,and then co-cultured with human nasopharyngeal carcinoma cells CNE-2Z,meantime,C57BL/6 mice were injected subcutaneously with CNE-2Z cells to establish a nasopharyngeal carcinoma xenograft model(24 mice).All of them were randomly grouped into control group,lncRNA LINC01296 knockdown group(transfected with lncRNA LINC01296 small interfering RNA(siRNA)),negative control group(transfected with lncRNA LINC01296 siRNA negative control and empty plasmid),and lncRNA LINC01296 knockdown + PD-L1 overexpression group(transfected with lncRNA LINC01296 siRNA and PD-L1 overexpression plasmid).After grouping and transfection,the proportion of activated CD8+ T cells in human peripheral blood lymphocytes was detected by flow cytometry;the killing rate of human peripheral blood lymphocytes to CNE-2Z cells was detected by cell count kit-8 method;the tumor volume of tumor-bearing mice was measured;the CD8 and PD-L1 positive expression in tumor tissue of tumor-bearing mice was measured by immunofluorescence staining;the expression of lncRNA LINC01296 and PD-L1 messenger RNA(mRNA)in CNE-2Z cells and tumor tissues were detected by real-time PCR assay;the expression of PD-L1 protein in CNE-2Z cells and tumor tissues was detected by Western blotting.The results showed that compared with control group,the proportion of activated CD8+ T cells in human peripheral blood lymphocytes,the killing rate of CNE-2Z cells,and the positive proportion of CD8 in tumor tissue of the lncRNA LINC01296 knockdown group increased(P<0.05),the tumor volume and the expression of positive proportion of PD-L1,double positive proportion of CD8 and PD-L1,lncRNA LINC01296,PD-L1 mRNA and protein in CNE-2Z cells and tumor tissue decreased(P<0.05);there was no obvious change in each index of the mice in the negative control group(P>0.05);overexpression of PD-L1 can weaken the effects of lncRNA LINC01296 knocking down on various indicators in CNE-2Z cells and mice.In summary,knockdown of lncRNA LINC01296 can promote the activation and infiltration of CD8+ T cells by down-regulating PD-L1,attenuate the immune escape of nasopharyngeal carcinoma cells,and enhance the lethality of CD8+ T cells.

Objective:To explore the feasibility and clinical efficacy of watertight suture technique in skull base reconstruction after expanded endoscopic endonasal excision of tuberculum sellae meningioma.Methods:Fourteen patients with tuberculum sellae meningioma accepted expanded endoscopic endonasal excision of tuberculum sellae meningioma in Department of Neurosurgery, First Affiliated Hospital of Xinxiang Medical University from January 2018 to May 2022 were chosen. During reconstruction of skull base, femoral fascia was used to repair the dural defect of sellar base with watertight suture, and then the sellar base was covered with a larger layer of femoral fascia for reinforcement; no nasal septum mucosal flap was used. The clinical data and treatment efficacy of these patients were retrospectively analyzed.Results:Total resection showed by imaging was achieved in all 14 patients. During the surgery, Valsalva ventilation test confirmed that at least 12 stitches were needed to achieve watertight suture status; watertight suture status was achieved in 13 of the 14 patients, without cerebrospinal fluid (CSF) leakage; watertight suture status was not achieved in one patient due to tumor invasion of the sella floor dura and having an extensive excision, and CSF leakage appeared transiently after surgery but disappeared 2 weeks after surgery (bed rest). Among the 11 patients with visual damage and optic field defect, 9 patients improved obviously and 2 patients did not improve. Follow-up was performed for 5-53 months, with an average of (26.8±8.4) months; no tumor recurrence or CSF leakage were found in these patients; up to the last follow-up, the 2 patients with visual damage and optic field defect did not improve.Conclusion:Skull base reconstruction using watertight suture technique after expanded endoscopic endonasal excision of tuberculum sellae meningioma is reliable.

Objective:To explore the feasibility and clinical efficacy of resection of jugular foramen schwannomas via anterolateral approach. Methods:A retrospective analysis was conducted on clinical data of 15 patients with jugular foramen schwannomas admitted to Department of Neurosurgery, First Affiliated Hospital of Xinxiang Medical College from January 2018 to July 2022. Three patients had Samii type B, 7 had type C and 5 had type D. Resection of the schwannomas was performed via anterolateral approach. After surgery, regular follow-up was performed through outpatient review, telephone, or WeChat to evaluate tumor progression and neurological functions. Results:Adequate surgical exposure was obtained in all 15 patients. Total resection was obtained in 14 patients and subtotal resection in 1 patient. Posterior cranial nerve palsy was worsened in 1 patient and new-onset facial paralysis (House-Brackmann grading Ⅲ) was noted in 1 patient, without cerebrospinal fluid leakage, subcutaneous effusion or death. Choking and cough during drinking water, and dysphagia disappeared or relieved 3 months after surgery in patients with aggravated posterior cranial nerve palsy, but no significant recovery from hoarseness was noted 6 months after surgery. A patient with new-onset facial paralysis improved to House-Brackmann grading I 3 months after surgery. Up to the last follow-up, no tumor recurrence was observed in 15 patients.Conclusion:Resection via anterolateral approach is effective in jugular foramen schwannomas for its adequate surgical exposure, high total resection rate, and low postoperative complications.

Objective:To explore the application value of chromosome karyotype analysis, chromosomal microarray analysis (CMA) and whole exome sequencing (WES) in prenatal diagnosis of isolated corpus callosum abnormality (CCA) fetus.Methods:Fetuses diagnosed with isolated CCA by ultrasound and MRI and receiving invasive prenatal diagnosis in Guangzhou Women and Children′s Medical Center and Qingyuan People′s Hospital from January 2010 to April 2021 were selected. Karyotype analysis and/or CMA [or copy number variation sequencing (CNV-seq)] were performed on all fetal samples, and WES was performed on fetal samples and their parents whose karyotype analysis and/or CMA (or CNV-seq) results were not abnormal.Results:Among 65 fetuses with isolated CCA, 38 cases underwent karyotype analysis, and 3 cases were detected with abnormal karyotypes, with a detection rate of 8% (3/38). A total of 49 fetuses with isolated CCA underwent CMA (or CNV-seq) detection, and 6 cases of pathogenic CNV were detected, the detection rate was 12% (6/49). Among them, the karyotype analysis results were abnormal, and the detection rate of further CMA detection was 1/1. The karyotype results were normal, and the detection rate of further CMA (or CNV-seq) detection was 14% (3/21). The detection rate of CMA as the first-line detection technique was 7% (2/27). A total of 25 fetuses with isolated CCA with negative results of karyotyping and/or CMA were tested by WES, and 9 cases (36%, 9/25) were detected with pathogenic genes. The gradient genetic diagnosis of chromosomal karyotyping, CMA and WES resulted in a definite genetic diagnosis of 26% (17/65) of isolated CCA fetuses.Conclusions:Prenatal genetic diagnosis of isolated CCA fetuses is of great clinical significance. The detection rate of CMA is higher than that of traditional karyotyping. CMA detection could be used as a first-line detection technique for fetuses with isolated CCA. WES could increase the pathogenicity detection rate of fetuses with isolated CCA when karyotype analysis and/or CMA test results are negative.

Objective:To investigate the complement C3a receptor 1 (C3AR1) expression in glioma and its mechanism in progressing malignancy.Methods:(1) The C3AR1 mRNA expression data and clinical information were obtained in 607 glioma patients from The Cancer Genome Atlas (TCGA) database and 656 glioma patients from Chinese Glioma Genome Atlas (CGGA) database; the differences in C3AR1 mRNA expression were analyzed among gliomas with different World Health Organization (WHO) grading. The overall survival and disease-free survival were compared between high and low C3AR1 mRNA expression patients obtained from TCGA database by Gene expression profiling interactive analysis (GEPIA). Gene body (GO) function analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of C3AR1 related differentially expressed genes were performed by DAVID database. Correlation of C3AR1 mRNA expression with immune cell infiltration was analyzed using TIMER online website. (2) The brain tissues from 3 non-tumor patients and 9 glioma patients surgically resected in Department of Neurosurgery, First Affiliated Hospital of Xinxiang Medical University from January 2019 to September 2021 were collected; the C3AR1 protein expression was detected by Western blotting. (3) The in vitro cultured U87 and U251 cells were divided into negative control group and C3AR1 knockdown group ( C3AR1 being knocked down by lentivirus transfection); and CCK-8 assay, plate cloning assay and Transwell assay were used to detect the proliferation rate, number of colony formation and number of membrane penetrating cells. Western blotting was used to detect the nuclear factor-κB (NF-κB) signaling pathway protein expressions. Results:(1) In TCGA database, the C3AR1 mRNA expression in gliomas of WHO grading II, grading III and grading IV increased sequentially, with significant differences ( P<0.05). In CGGA database, the C3AR1 mRNA expression in glioma of WHO grading IV was statistically higher than that in gliomas of WHO grading II and grading III ( P<0.05). GEPIA showed that the overall survival and disease-free survival in the low C3AR1 mRNA expression group were statistically higher than those in the high C3AR1 mRNA expression group ( P<0.05). GO function analysis and KEGG pathway enrichment analysis revealed that C3AR1 related differentially expressed genes were more enriched in such biological processes and signaling pathways as calcium homeostasis, membrane structural valves, proton transmembrane transporter protein activity, chemokine signaling pathway and NF-κB signaling pathway. TIMER showed that C3AR1 mRNA expression in glioblastoma and low-grade glioma was positively correlated with infiltration degrees of B cells, CD4 + T cells, neutrophils, macrophages and dendritic cells, and C3AR1 mRNA expression in glioblastoma was negatively correlated with infiltration degree of CD8 + T cells ( P<0.05). (2) C3AR1 protein expression in glioma tissues was significantly higher than that in non-tumor tissues. (3) Compared with the negative control group, the C3AR1 knockdown group group had significantly lower proliferation rate, smaller numbers of colony formation and membrane penetrating cells, and lower expressions of NF-κB, phosphorylated (p)-NF-κB, p-NF-κB inhibitory protein (IκB)α, p-I-κB kinase (IKK)α and N-cadherin, and significantly higher E-cadherin expression. Conclusion:C3AR1 is highly expressed in glioma and progresses malignancy through NF-κB signaling pathway.

OBJECTIVE: To explore the phenotype and pathogenesis of a fetus with a rare chromosomal abnormality. METHODS: The fetus was analyzed by clinical prenatal ultrasonography, G-banding karyotyping and next generation sequencing (NGS). RESULTS: Prenatal ultrasonography of the fetus showed Dandy-Walker syndrome, growth restriction, and right-heart system dysplasia. The fetus had a chromosomal karyotype of 47,XY,t(11;22)(q23.3;q11.2),+der(22)t(11;22). Duplication of 11q23.3q25 and 22q11.1q21 were also detected by NGS. The chromosomal translocation carried by the fetus was derived from his father. CONCLUSION Duplications of chromosome 11q23.3q25 and 22q11.1q11.21 segments probably underlie the Dandy-Walker syndrome, growth restriction, and hypoplasia of the right heart system in the fetus.
Chromosome Disorders ; Chromosomes, Human ; Female ; Fetus ; Humans ; Karyotyping ; Pregnancy ; Prenatal Diagnosis ; Translocation, Genetic ; Trisomy

Objective: To investigate the correlation between fetal cranial nervous system malformation and chromosome abnormality. Methods: The pregnant women with fetal cerebral nervous system dysplasia were collected from January 2013 to August 2018 at the Prenatal Diagnostic Center of the Sixth Affiliated Hospital of Guangzhou Medical University.The fetus was diagnosed by ultrasonography and karyotype analysis. Results: A total of 18 cases of abnormal karyotypes were detected from 85 patient samples, and the abnormal rates were 21.18%.Single cranial nervous system malformation was found in 47 cases, abnormal karyotypes in 4 cases, multiple system malformation in 38 cases, and abnormal karyotypes in 14 cases, and the abnormal karyotype rate of multiple system malformation was higher than that of single cranial nervous malformation (36.84% vs.8.51%, χ²=10.101, P=0.001 5). And the 88.89%(16/18 cases)of abnormal karyotypes were founded in the early and middle pregnancy (≤28 weeks). The abnormal karyotype detection rates of cranial nervous system malformation associated with cardiovascular, skeletal and limb, facial neck abnormalities were 58.82% (10/17 cases), 50.00% (6/12 cases) and 50.00% (9/18 cases), respectively.In the fetal phenotypes, the abnormal karyotype detection rates of choroid plexus cysts were up to 64.29%, followed by arachnoid cysts (50.00%), craniocerebral abnormalities (45.45%) and holoprosencephaly (36.36%). Conclusions Chromosomal aneuploidy or structural abnormalities can lead to abnormal development of the fetal cranial nervous system, in which the rates of abnormal karyotypes on fetal cranial nervous with cardiovascular malformation and choroid plexus cysts are the highest.