Objective To investigate the effect of a disintegrin and metalloprotease domain(ADAM8)gene on colon cancer HCT8 cell proliferation and proliferation signal transduction pathways PI3K-Akt-mTOR. Methods Colon cancer HCT8 cells were cultured in vitro,and transfected with ADAM8 overexpression plasmid and RNA interfering plasmid. Cell proliferation were detected by EdU and MTS method assays. PI3K activity was measured by PI3K activity detection kit,Western Blot method was performed to detect the ratio of Akt and p-Akt and expression of mTOR. Results Compared with control group(1.00 ±0.12),the cell proliferation in ADAM8 overexpression group(1.22 ±0.13)was significantly higher (P<0.05)and in RNA interfering group(0.78 ±0.11)was significantly lower while PI3K activity had no significant changes in three groups. After ADAM8 overexpression,and the ratio of p-Akt/Akt and mTOR expression were increased significantly,while reduced significantly after RNA interferered. Conclusion ADAM8 can promote HCT8 cell proliferation through enhancing the phosphorylation of Akt and promoting the expression of mTOR.

Objective: To obtain the optimum prescription of the rhubarb lactobacillin dispersible tablets.Methods: The kind and amount of the excipient were investigated by using the disintegrating time to establish the optimum prescription and preparation process.Results: The tablets were prepared by compressing moist granulation with the PVPP and CMS-Na as the disintegrant,MCC as diluent,sodium alginate as sweller,magnesium stearate as lubricant and 80% alcohol as adhesive agent.The preparation complied with the requirement of Pharmacopeia of the People's Republic of China(2005edition).Conclusion: This prescription of the tablet is reasonable and the preparation process is feasible.

Objective To develop a specific trans-splicing intron ribozyme type Ⅰ-mediated dual reporter gene system (Rib53-Fluc-tk) for targeting CEA.Methods The novel CEA-targeting trans-splicing ribozyme with the downstream reporter system (Rib53-Fluc-tk) was constructed by genetic engineering technology.The trans-splicing reaction product was evaluated using the 131I-5-iodo-2'-fluro-l-beta-D-arabinofuranosy-luracil (FIAU) cellular uptake rates and the bioluminescence.Two-sample t test,the analysis of variance and the least significant difference (LSD) t test was performed for data analysis.Results The sequence of Rib53-Fluc-tk was proved by gene-sequencing test.Human MCF-7 breast cancer cells showed a high ratio of firefly luciferase/renilla luciferase (0.64±0.10,n =4).A 520 bp band of product existed,which matched with the predicted size using RNA from cells transfected with Rib53-Fluc-tk in MCF-7.Signals were detected by bioluminescence in human embryonic kidney 293T cells co-transfected with Rib53-Fluc-tk and pCDNA3.1-CEA.The labelling rate of 131I-FIAU was (64.02±4.79)％ (n =3).The radiochemical purity was (95.96± 1.07)％ (n=3),and the stability of the radiocompound remained high in human serum at least for 24 h.The uptake of 131I-FIAU in 293T cells transfected with Rib53-Fluc-tk was (0.31±0.01)％ (n=4),while it increased with the incubation time in 293T cells co-transfected with pCDNA3.1-CEA and Rib53-Fluc-tk and reached (1.40±0.06)％ at 4.5 h (F=1 007.29,t=136.34,both P＜0.01).Conelusions A novel and specific reporter gene in the cellular level was established.Taking advantage of trans-splicing reaction of the ribozyme,it could improve the specificity of the reporter gene imaging.

Objective: To observe the role of Qudu medicinal granules (祛毒冲剂) on enterogenous endotoxemia . Methods: Sixty-one cases with enterogenous endotoxemia were randomly divided into two groups:Qudu medicinal granule group (n=30) that was treated with Qudu medicinal granules combined with western medicine, and smecta group which was treated with smecta and western medicine (n=31). Changes of symptoms and signs were observed before treatment and 1, 3, 7 days after treatment. Blood samples were collected in the morning to measure the white blood cell (WBC), plasma lipopolysaccharide (LPS) and tumor necrosis factor (TNF) levels. Results: Recovery speed of WBC count in Qudu medicinal granule group was faster than that of the smecta group, there was significant difference on the third day after treatment (P

OBJECTIVETo identify the casual mutation of a Chinese pedigree with hypertrophic cardiomyopathy (HCM), and to analyze the genotype-phenotype relationship.

METHODSThe coding exons of 26 reported disease genes were sequenced by targeted resequencing in the proband and the identified mutation were detected with bi-directional Sanger sequencing in all family members and 307 healthy controls. The genotype-phenotype correlation was analyzed in the family.

RESULTSA missense mutation (c.2191C > T, p. Pro731Ser) in the 20th exon of MYH7 gene was identified. This mutation was absent in 307 healthy controls and predicted to be pathogenic by PolyPhen-HCM. Totally 13 family members carried this mutation, including 10 patients with HCM and 3 asymptomatic mutation carriers. The proband manifested severe congestive heart failure and 8 patients expressed various clinical manifestations of heart failure, including dyspnea, palpitations, chest pain, amaurosis or syncope. Five patients were diagnosed as HCM at the age of 16 or younger. One family member suffered sudden cardiac death.

CONCLUSIONSThe Pro731Ser of MYH7 gene mutation is a causal and malignant mutation linked with familiar HCM.

Adolescent ; Asian Continental Ancestry Group ; Base Sequence ; Cardiomyopathy, Hypertrophic ; ethnology ; genetics ; Death, Sudden, Cardiac ; Exons ; Humans ; Mutation, Missense ; Myosin Heavy Chains ; genetics ; Pedigree ; Phenotype ; Research Design ; Ventricular Myosins

Objective To investigate the relationship between prognostic nutritional index (PNI) and neutropenia after adjuvant chemotherapy in patients with colorectal cancer. Methods The clinical data of 44 patients with colorectal cancer performed adjuvant chemotherapy in Shunyi District Hospital from December 2014 to January 2018 were retrospectively analyzed, and the patients were divided into group A (grade 0-2 neutropenia) and group B (grade3-4 neutropenia) according to the degree of neutropenia. The serum albumin, peripheral lymphocyte counts, and neutrophil counts within 1 week before chemotherapy were collected, and the PNI was calculated. The chi-square test and rank sum test were used to compare the clinical data, body mass index (BMI), baseline neutrophil count, and PNI between the two groups. Logistic regression analysis was used to analyze the risk factors for neutropenia after chemotherapy. Results The baseline median neutrophil counts and median PNI in group A were 3.17×109/L [(1.38-7.79)×109/L] and 50.40 (37.40-57.05), and in group B were 2.54 ×109/L [(1.22-3.87) ×109/L] and 45.50 (37.95-50.95). The baseline neutrophil counts and PNI in group A were significantly higher than those in group B, the differences between the two groups were statistically significant (Z= -2.085, P= 0.037; Z= -2.615, P= 0.009). Logistic regression analysis showed that PNI was an independent risk factor for neutropenia after chemotherapy (HR=0.803, 95％CI 0.646-0.998, P= 0.048). Conclusion PNI has a certain role in predicting neutropenia after adjuvant chemotherapy in patients with colorectal cancer.

Objective To identify the casual mutation of a Chinese pedigree with hypertrophic cardiomyopathy (HCM),and to analyze the genotype-phenotype relationship.Methods The coding exons of 26 reported disease genes were sequenced by targeted resequencing in the proband and the identified mutation were detected with bi-directional Sanger sequencing in all family members and 307 healthy controls.The genotype-phenotype correlation was analyzed in the family.Results A missense mutation (c.2191C>T, p.Pro731Ser) in the 20th exon of MYH7 gene was identified.This mutation was absent in 307 healthy controls and predicted to be pathogenic by PolyPhen-HCM.Totally 13 family members carried this mutation , including 10 patients with HCM and 3 asymptomatic mutation carriers.The proband manifested severe congestive heart failure and 8 patients expressed various clinical manifestations of heart failure , including dyspnea , palpitations , chest pain , amaurosis or syncope.Five patients were diagnosed as HCM at the age of 16 or younger.One family member suffered sudden cardiac death.Conclusions The Pro731Ser of MYH7 gene mutation is a causal and malignant mutation linked with familiar HCM.

Objective To observe the efficacy of gray-scale ultrasound-based radiomics for differentiating peripheral pulmonary adenocarcinoma and squamous cell carcinoma.Methods Data of 88 patients with single peripheral lung adenocarcinoma and 58 patients with single peripheral lung squamous cell carcinoma proved pathologically with puncture biopsy and clearly visualized with lung ultrasound were retrospectively analyzed.The patients were divided into training set(n=103)and test set(n=43)at the ratio of 7:3.Based on gray-scale ultrasound of training set,radiomics features associated with differential diagnosis of peripheral lung adenocarcinoma and lung squamous cell carcinoma were extracted and screened.Using 4 different classifiers,including support vector machine(SVM),linear discriminant analysis(LDA),logistic regression(LR)and the least absolute shrinkage and selection operator combined with logistic regression(LASSO-LR),4 corresponding radiomics models were obtained,and the relative best models were selected according to their performances under 10-fold cross validation.The receiver operating characteristic curves were drawn,the areas under the curve(AUC)were calculated to evaluate the differentiating efficacy of each model,and DeLong test was used for the comparison.The differentiating accuracy of models were obtained under the best cutoff value with the maximum Youden index.Results The AUC of SVM,LDA,LR and LASSO-LR radiomics models for differentiating peripheral lung adenocarcinoma and lung squamous carcinoma in test set was 0.864,0.867,0.880 and 0.844,respectively,and no significant difference was found among 4 models(all P＞0.05).Under the best cutoff value of each model,the corresponding accuracy of SVM,LDA,LR and LASSO-LR radiomics models for differentiating peripheral lung adenocarcinoma and lung squamous cell carcinoma was 86.05％,83.72％,88.37％and 86.05％,respectively.Conclusion Radiomics models based on gray-scale ultrasound could be used to differentiate peripheral lung adenocarcinoma and lung squamous cell carcinoma.

Hepatitis C virus (HCV) is a leading cause of liver disease worldwide. Although several HCV protease/polymerase inhibitors were recently approved by U.S. FDA, the combination of antivirals targeting multiple processes of HCV lifecycle would optimize anti-HCV therapy and against potential drug-resistance. Viral entry is an essential target step for antiviral development, but FDA-approved HCV entry inhibitor remains exclusive. Here we identify serotonin 2A receptor (5-HTR) is a HCV entry factor amendable to therapeutic intervention by a chemical biology strategy. The silencing of 5-HTR and clinically available 5-HTR antagonist suppress cell culture-derived HCV (HCVcc) in different liver cells and primary human hepatocytes at late endocytosis process. The mechanism is related to regulate the correct plasma membrane localization of claudin 1 (CLDN1). Moreover, phenoxybenzamine (PBZ), an FDA-approved 5-HTR antagonist, inhibits all major HCV genotypes in vitro and displays synergy in combination with clinical used anti-HCV drugs. The impact of PBZ on HCV genotype 2a is documented in immune-competent humanized transgenic mice. Our results not only expand the understanding of HCV entry, but also present a promising target for the invention of HCV entry inhibitor.

The secondary structures of hepatitis C virus (HCV) RNA and the cellular proteins that bind to them are important for modulating both translation and RNA replication. However, the sets of RNA-binding proteins involved in the regulation of HCV translation, replication and encapsidation remain unknown. Here, we identified RNA binding motif protein 24 (RBM24) as a host factor participated in HCV translation and replication. Knockdown of RBM24 reduced HCV propagation in Huh7.5.1 cells. An enhanced translation and delayed RNA synthesis during the early phase of infection was observed in RBM24 silencing cells. However, both overexpression of RBM24 and recombinant human RBM24 protein suppressed HCV IRES-mediated translation. Further analysis revealed that the assembly of the 80S ribosome on the HCV IRES was interrupted by RBM24 protein through binding to the 5'-UTR. RBM24 could also interact with HCV Core and enhance the interaction of Core and 5'-UTR, which suppresses the expression of HCV. Moreover, RBM24 enhanced the interaction between the 5'- and 3'-UTRs in the HCV genome, which probably explained its requirement in HCV genome replication. Therefore, RBM24 is a novel host factor involved in HCV replication and may function at the switch from translation to replication.
Cells, Cultured ; Hepacivirus ; genetics ; growth & development ; metabolism ; Humans ; Protein Biosynthesis ; RNA-Binding Proteins ; metabolism ; Virus Replication ; genetics