Naringenin (4,5,7-trihydroxyflavonoid) is a naturally occurring bioflavonoid found in citrus fruits, which plays an important role in metabolic syndrome, neurological disorders, and cardiovascular diseases. However, the pharmacological mechanism and biological function of naringenin on anti-angiogenesis and anti-tumor immunity have not yet been elucidated. Our study firstly demonstrates that naringenin inhibits the growth of hepatocellular carcinoma (HCC) cells both in vivo and in vitro. Naringenin diminishes the ability of HCC cells to induce tube formation and migration of human umbilical vein endothelial cells (HUVECs) and suppresses neovascularization in chicken chorioallantoic membrane (CAM) assays. Meanwhile, in vivo results demonstrate that naringenin can significantly upregulate level of CD8⁺ T cells, subsequently increasing the level of immune-related cytokines in the tumor immune microenvironment. Mechanistically, we found that naringenin facilitate the K48-linked ubiquitination and subsequent protein degradation of vascular endothelial growth factor A (VEGFA) and mesenchymal-epithelial transition factor (c-Met), which reduces the expression of programmed death ligand 1 (PD-L1). Importantly, combination therapy naringenin with PD-L1 antibody or bevacizumab provided better therapeutic effects in liver cancer. Our study reveals that naringenin can effectively inhibit angiogenesis and anti-tumor immunity in liver cancer by degradation of VEGFA and c-Met in a K48-linked ubiquitination manner. This work enlightens the potential effect of naringenin as a promising therapeutic strategy against anti-angiogenesis and anti-tumor immunity in HCC.

OBJECTIVE: To explore the clinical characteristics and genetic basis of a child with autism spectrum disorder (ASD) in conjunct with congenital heart disease (CHD). METHODS: A child who was hospitalized at the Third People's Hospital of Chengdu on April 13, 2021 was selected as the study subject. Clinical data of the child were collected. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing (WES). A GTX genetic analysis system was used to analyze the WES data and screen candidate variants for ASD. Candidate variant was verified by Sanger sequencing and bioinformatics analysis. Real-time fluorescent quantitative PCR (qPCR) was carried out to compare the expression of mRNA of the NSD1 gene between this child and 3 healthy controls and 5 other children with ASD. RESULTS: The patient, an 8-year-old male, has manifested with ASD, mental retardation and CHD. WES analysis revealed that he has harbored a heterozygous c.3385+2T>C variant in the NSD1 gene, which may affect the function of its protein product. Sanger sequencing showed that neither of his parent has carried the same variant. By bioinformatic analysis, the variant has not been recorded in the ESP, 1000 Genomes and ExAC databases. Analysis with Mutation Taster online software indicated it to be disease causing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be pathogenic. By qPCR analysis, the expression level of mRNA of the NSD1 gene in this child and 5 other children with ASD was significantly lower than that of the healthy controls (P < 0.001). CONCLUSION The c.3385+2T>C variant of the NSD1 gene can significantly reduce its expression, which may predispose to ASD. Above finding has enriched the mutational spectrum the NSD1 gene.
Male ; Child ; Humans ; Autism Spectrum Disorder/genetics* ; Heart Defects, Congenital/genetics* ; Computational Biology ; Genomics ; Mutation ; RNA, Messenger/genetics* ; Histone-Lysine N-Methyltransferase/genetics*

Objective: To investigate the best amount of TPCK trypsin in Madin Darby canine kidney (MDCK) cell suspension for the culture of H7N9 avian influenza virus. Methods: Different concentrations of TPCK trypsin were added during the periods of cell growth and virus production. Their effects on cell growth, viability, glucose and lactate metabolism, and hemagglutination titer were monitored every 12 h. Inter-batch differences were analyzed. The amount of trypsin added in the cell growth phase was 0, 1 μg/ml, 2 μg/ml, 4 μg/ml, 6 μg/ml, 8 μg/ml, 10 μg/ml and 15 μg/ml. The amount of trypsin added during the virus production period was 0, 0.5 μg/ml, 1 μg/ml, 1.5 μg/ml, 2 μg/ml and 2.5 μg/ml. When the hemagglutination titers were same, the adding amount was further optimized at different multiplicity of infection (MOI) of 0.001, 0.005, 0.025 and 0.05. Results: No significant linear effects of TPCK trypsin concentration on cell number, viability, and glucose and lactate metabolism were observed. No toxicity to cell growth was observed when TPCK trypsin concentration reached 15 μg/ml. After the inoculation of H7N9 avian influenza virus, the hemagglutination titers in the 1 μg/ml, 1.5 μg/ml, 2 μg/ml and 2.5 μg/ml TPCK trypsin groups reached the peaks at 48 h, which were 1∶2^6.5. At 60 h, the hemagglutination titers of the latter two groups decreased faster than those of the former two groups. When the MOI was 0.005, the hemagglutination titer of the 1.5 μg/ml group at 48 h was 2^6.5 higher than 2⁶ in the 1 μg/ml group under the same condition. There were differences between different batches of TPCK trypsin. Conclusions Adding 1 μg/ml and 1.5 μg/ml of trypsin could better promote the proliferation of H7N9 avian influenza virus, and 1.5 μg/ml of trypsin had a wider range of MOI applicability.

Hepatocellular carcinoma is a malignant tumor with high morbidity and mortality globally.The therapeutic results on hepatocellular carcinoma with surgery and other classical treatments are not satisfactory.Tumor immunotherapies, such as dendritic cells and cytokine-induced killer cells (DC-CIK), tumor-infiltrating lymphocytes (TIL), chimeric antigen receptor T cells (CAR-T), immune checkpoint blockade (such as PD-1 blockade,CTL4 blockade) etc., are very promising new therapeutic approaches.In this paper, the progresses in the immunotherapy, mainly on PD-1, DC-CIK, TIL, are reviewed.Also pluripotent immune killer cells (PIK) which was developed in our lab was briefly introduced.

Objective To investigate the association of uncoupling protein 2 ( UCP-2 ) gene promoter -866G＞A polymorphism and ischemic stroke in diabetic patients.Methods A total of 844 type 2 diabetic patients including 404 cases with ischemic stroke and 440 cases without ischemic stroke were selected for the 4 year prospective study,Genomic DNA was extracted from the whole blood samples of subjects,UCP-2 gene promoter -866G ＞ A polymorphism was detected by TaqMan MGB probe method,and then the genotype and allele gene frequencies were compared.Results The risk of ischemic stroke in type 2 diabetic female patients with AA+GA genotypes of UCP-2 was higher than that with GG genotype (P＜0.05),but there was no difference among male patients with three genotypes.Conclusions UCP-2 gene promoter -866G ＞ A polymorphism increases the risk of ischemic stroke in Chinese diabetic women.

Objective To compare the accuracy of CT with other methods to measure the length of thoracic esophageal carcinoma. Methods 598 patients with thoracic esophageal squamous carcinoma were enrolled in this study.All the patients received three-field(cervical,thoracic:and abdominal)radical surgery without pre-operative radiotherapy or chemotherapy.The length of each Iesion was recorded and compared by measuring intraoperative specimen,formalin-fixed specimen,X-ray barium meal examination and CT,respectivelv. Results By the measurement of intraoperative specimen,formalin-fixed specimen,Xray barium meal examination and CT,the mean lengths of lesion were(5.22±1.94),(4.28±1.71),(5.12±1.92)and(6.71±2.52) cm,respectively.The measured length was significantly different between intraoperative specimen and formalin-fixed specimen or CT(t=16.01,P＜0.01;t=-15.54,P＜0.01),but not between intraoperative specimen and X-ray barium meal examination(t=1.62,P＞0.05).The measured lengths gradually decreased in the order of CT,intraoperative specimen,X-ray bailam meal examination and formalin-fixed specimen.For different pathological type(except intracavitary type)and different T staging,there was significant difference in lesion length between intraoperative specimen and CT(P＜0.05),but not between intraoperative specimen and X-ray barium meal examination(P＞0.05). Conclusions The length of esophageal carcinoma measured by intraoperative specimen is shorter than by CT,but longer than by X-ray barium meal examination.Specimen could shrink after foriBalin fixation.X-ray barium meal and other examinations should be referred when using CT to delineate tumor target volume of esophageal carcinoma for radiotherapy.