ObjectiveTo analyze the expression of molecular marker affecting the prognosis of acute myeloid leukemia （AML） patients from bioinformatics database， thus providing an experimental basis for further exploration of a novel molecular marker for the prognosis of AML. MethodsThe prognostic data of 179 AML patients from The Cancer Genome Atlas （TCGA） database were examined for differential gene analysis and survival analysis. The bone marrow samples of 74 healthy individuals （HI） and 542 de novo AML patients in the dataset GSE13159 downloaded from the Gene Expression Omnibus （GEO） database were analyzed to detect the difference in the expression levels of differential target genes. Peripheral blood and bone marrow samples were collected from 18 de novo AML patients and 20 age- and gender-matched healthy controls， and real-time fluorescent quantitative PCR was used to validate the expression levels of the differential genes in the AML patients. ResultsBioinformatics data analysis showed that the optimal cut-off value of Homo sapiens NK2 homeobox 3 （NKX2-3） calculated by R language was 0.051. Survival analysis revealed a statistically poorer overall survival in de novo AML patients with high NKX2-3 expression than in those with low NKX2-3 expression （P = 0.0036）. NKX2-3 was highly expressed in patients with de novo AML than in HI and the difference was statistically significant （P < 0.001）. Real-time fluorescence quantitative PCR verified the expression levels of the NKX2-3 gene in AML patients and confirmed that compared with those in HI， in the de novo AML patients， NKX2-3-1 and NKX2-3-2 were highly expressed and were significantly correlated （P = 0.000， P = 0.000）. ConclusionNKX2-3 is highly expressed in de novo AML patients， and the AML patients with high NKX2-3 expression have poor overal survival. NKX2-3 may be closely related to the clinical outcome and prognosis of AML.

Objective To identify the causative variants in 5 Chinese families with tuberous sclerosis complex(TSC)to provide genetic counseling and prenatal diagnosis.Methods Genetic counseling and clinical diagnosis were performed in 8 patients from five unrelated TSC families by teleconsultation.With informed consent obtained from the participants,3 to 5 mL peripheral blood samples were collected from the probands and their family mem-bers for the extraction of genomic DNA.Candidate pathogenic variants were screened by panel sequencing(PS).The candidate pathogenic variants found in TSC1 and TSC2 by PS were validated by PCR-Sanger sequencing and bioinformatics analysis.Results All the pathogenic mutations were identified in the probands and their available family members.Causative variants in TSC1 or TSC2 were detected in all patients,including three reported variants and two novel variants.The two novel variants,TSC2:c.245G＞A and TSC2:c.235delG,which were predicted to cause the nonsense variant p.(Trp82?)and the frameshift variant p.(Val79Lysfs27?)respectively was believed to introduce premature stop codons.The analysis of family co-segregation and bioinformatics were identified as very positive factors for pathogenicity.Conclusions This result provides more evidences for the genetic counseling and prenatal diagnosis in these families and expand the spectrum of TSC2 pathogenic variants.

Objective:To explore the genetic etiology of a fetus with cryptophthalmos detected by prenatal ultrasonography.Methods:A fetus undergoing induced labor at 32nd gestational week due to absence of bilateral eye fissures detected by prenatal ultrasonography in January 2017 was selected as the study subject. Umbilical cord blood sample from the fetus and peripheral blood samples from its parents were collected for the extraction of genomic DNA. Pathogenic variants were screened through whole exome sequencing (WES) and verified by Sanger sequencing. Pathogenicity of candidate variants was verified by bioinformatic analysis and protein structure simulation. Based on the results of genetic testing, prenatal diagnosis was provided to the couple upon their subsequent pregnancy.Results:The couple had four adverse pregnancies previously. The aborted fetus was the fifth, with fused bilateral upper and lower eyelids, poorly developed eyeballs, adhesion of the cornea with the upper eyelid, low-set ears, and abnormal plantar creases, and was diagnosed with cryptophthalmos. WES and Sanger sequencing revealed that the fetus has harbored compound heterozygous variants of the FREM2 gene, namely c. 4537G>A (p.D1513N) and c.7292C>T (p.T2431M). Both variants were unreported associated with cryptophthalmos previously. Protein structure simulation showed that they may lead to loss of hydrogen bonds in the protein product. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), both variants were predicted to be likely pathogenic (PM1_Supporting+ PM2_Supporting+ PM5+ PP3+ PP4; PM2_Supporting+ PM3+ PP3+ PP4). The mother was performed prenatal diagnosis in her sixth pregnancy based on the variants detected in this family, and delivered a daughter with normal phenotype. Conclusion:The FREM2: c. 4537G>A and c. 7292C>T compound heterozygous variants probably underlay the pathogenesis of cryptophthalmos in this fetus. Above finding has enriched the mutational spectrum of the FREM2 gene.

Hepatocellular carcinoma (HCC) is a common malignant tumor with poor prognosis and high mortality. In this study, we demonstrated a novel vaccine targeting HCC and tumor neovascular endothelial cells by fusing recombinant MHCC97H cells expressing porcine α-1,3-galactose epitopes (αGal) and endorphin extracellular domains (END) with dendritic cells (DCs) from healthy volunteers. END⁺/Gal⁺-MHCC97H/DC fusion cells induced cytotoxic T lymphocytes (CTLs) and secretion of interferon-gamma (IFN-γ). CTLs targeted cells expressing αGal and END and tumor angiogenesis. The fused cell vaccine can effectively inhibit tumor growth and prolong the survival time of human hepatoma mice, indicating the high clinical potential of this new cell based vaccine.

Objective:To study whether the "combination of medical care and pension" service model can effectively control the development of chronic disease in the elderly and reduce the direct economic burden caused by the disease.Methods:A total of 180 elderly participants who received the "combination of medical care and pension" service model for chronic disease management in Chongqing, China were investigated and analyzed statistically. Epidata 2.0 was used for data entry, SPSS 20.0 was used for data analysis, and paired sample t test was used for comparison between groups. Results:After 12 months of chronic disease management, hospitalization events and expenses of the elderly were reduced, among which the number of hospitalization was reduced by 0.24 timed on average, the length of hospitalization was reduced by 10.41 days on average, and the hospitalization expenses were reduced by 11 144 yuan on average. The direct economic burden due to chronic diseases was reduced by approximately 8 844.5 yuan, accounting for 33.8% of the average cost of hospitalization for the elderly without the application of the model.Conclusion:The chronic disease of the elderly is well controlled by chronic disease management through the "combination of medical care and pension" service model.

AIM: To evaluate the effect of TDF withdrawal time on changes of serum HBV-M, HBV DNA and ALT level in the mother-to-child blocking of the maternal population. METHODS: A prospective, randomized and controlled study was conducted. The 120 pregnant women with HBV who took TDF during 24 to 28 weeks of gestation were randomly divided into group A (withdrawal at delivery) and group B (withdrawal at 4 weeks postpartum), levels of HBV-M, HBV DNA, and ALT at different times were detected. The results were statistically analyzed by Wilcoxon Rank-sum test and χ

Objective:To identify pathogenicity of the potential splicing variants in two Chinese Han patients with osteogenesis imperfecta.Methods:Genomic DNA was extracted using the conventional phenol-chloroform method; whole exome sequencing (WES) was used to analysis the disease-related variants in the two probands; Minigene assay was used to identify pathogenicity of the variants found in the patients' genome that possibly affect RNA splicing.Results:Two potential splicing variants, c.858+1_858+5delGTAAG in intron 12 of COL1A1 and c.1405-7C>T in intron 24 of COL1A2, were found in proband 1 and proband 2, respectively. In addition, a missense mutation, c.2972G>T (p.G991V) in exon 45 of COL1A2, was detected in proband 2. Minigene assay revealed that the variant in proband 1 caused the skipping of exon 12, while the variant in proband 2 did not lead to aberrant splicing. G199 of the COL1A2 in proband 2 was a highly conserved amino acid site, and the results suggested that c.2972G>T (p.G991V) may be the real pathogenic variant by the means of bioinformatics analysis.Conclusion:The variant c.858+1_858+5delGTAAG in COL1A1 was a causative variant that led to OI in proband 1, while the missense variant c.2972G>T (p.G991V) in COL1A2 was the cause of OI in proband 2, instead of the variant c.1405-7C>T. Minigene assay for potential splicing variants detected by WES could not only validate the pathogenicity of the candidate variants and enrich the mutation spectrum of OI, but also lay the foundation for patients' prenatal diagnosis and subsequent mechanism research.

Background: High concentrations of particulate matter less than 2.5 μm in diameter (PM2.5) in poultry houses is an important cause of respiratory disease in animals and humans. Pseudomonas aeruginosa is an opportunistic pathogen that can induce severe respiratory disease in animals under stress or with abnormal immune functions. When excessively high concentrations of PM2.5 in poultry houses damage the respiratory system and impair host immunity, secondary infections with P. aeruginosa can occur and produce a more intense inflammatory response, resulting in more severe lung injury. Objectives: In this study, we focused on the synergistic induction of inflammatory injury in the respiratory system and the related molecular mechanisms induced by PM2.5 and P. aeruginosa in poultry houses. Methods: High-throughput 16S rDNA sequence analysis was used for characterizing the bacterial diversity and relative abundance of the PM2.5 samples, and the effects of PM2.5 and P. aeruginosa stimulation on inflammation were detected by in vitro and in vivo. Results: Sequencing results indicated that the PM2.5 in poultry houses contained a high abundance of potentially pathogenic genera, such as Pseudomonas (2.94%). The lung tissues of mice had more significant pathological damage when co-stimulated by PM2.5 and P. aeruginosa, and it can increase the expression levels of interleukin (IL)-6, IL-8, and tumor necrosis factor-α through nuclear factor (NF)-κB pathway in vivo and in vitro. Conclusions The results confirmed that poultry house PM2.5 in combination with P. aeruginosa could aggravate the inflammatory response and cause more severe respiratory system injuries through a process closely related to the activation of the NF-κB pathway.

OBJECTIVE: To investigate the changes in diversity, relative abundance and distribution of intestinal flora in patients with chronic rhinosinusitis and nasal polyps (CRSwNP) using high-throughput sequencing technology identify the intestinal flora significantly related to pathogenesis and progression of CRSwNP. METHODS: Ten patients with CRSwNP hospitalized in the Department of Otolaryngology-Head and Neck Surgery of Guangdong Provincial People's Hospital were selected as the case group with 10 healthy volunteers recruited in the same period as the control group. Fecal genomic DNA extraction kit was used to extract the DNA in the fecal samples, and the DNA fragment length was measured and quantified. The V3 and V4 highly variable regions of the 16S rDNA gene of prokaryotes were amplified followed by library construction, Illumina MiSeq sequencing, sequence alignment and species identification analysis. The relative abundance, diversity and distribution characteristics of the intestinal flora were analyzed, and the relevant metabolic pathways were predicted. RESULTS: Compared with the control group, the patients with CRSwNP had significant changes in the overall structure of the intestinal flora, highlighted by increased abundance of Saccharopolyspora and decreased contents of , , and . Among the metabolic pathways predicted to be associated with CRSwNP, 9 showed significant changes in patients with CRSwNP as compared with the control group ( < 0.05). CONCLUSIONS Patients with CRSwNP have significant changes in the structural characteristics of intestinal flora related with multiple metabolic pathways, and these changes may play an important role in the development of chronic rhinosinusitis.

OBJECTIVE：To provide reference for elucidating the anti-allergic asthma constituents in alkaloids-free part of Ephedrae Herba. METHODS ：Ephedrae Herba was extracted with 85％ ethanol and n-heptane，and then subjected to solid-phase extraction（filler AC 18）for pretreatment to enrich alkaloids-free part from the extract of Ephedrae Herba. HPLC method was adopted，and alkaloids-free fractions of Ephedrae Herba were performed on Unitary C 18 column and Eclipse XDB-C 18 column. Using high expression G protein coupled-receptor 35（GPR35 receptor）in HT- 29 cell as target ，GPR35 receptor agonist zaprinast （1 μmol/L）as positive control ，DMR response value as the detection index ，the agonistic and desensitizing activity of each fraction（100 μg/mL）on GPR 35 receptor was screened by label-free integrated pharmacological method ，so as to screen active anti-allergic asthma fraction. HPLC-Q-TOF-MS method was used to identify the chemical composition of the selected active fractions. RESULTS ：The alkaloids-free part of Ephedrae Herba was divided into two parts ，involving the precipitated part before solid phase extraction and the 95％ methanol elution part ；from them ，20 fractions were screened. Among them ，the precipitated fraction F 1.5-F1.10 and 95％ methanol eluted fraction F 2.5-F2.10 had a strong agonistic activity on GPR 35 receptor；at the same time，GPR35 receptor agonist zaprinast showed a relatively strong desensitization activity. The signal intensity of DMR induced by F1.5-F1.10 in the precipitated part of HT- 29 cells was even higher than that of reference drug zaprinast. By HPLC-Q-TOF-MS analysis，24 chemical components were identified from active fractions ，involving 14 flavonoids，2 volatile oils ，7 organic carboxylic acids ，1 anthraquinones. CONCLUSIONS ：The alkaloid-free part of Ephedrae Herba is mainly flavonoids and has anti-allergic asthma activity.