OBJECTIVETo explore the molecule mechanism of Salidroside inducing directional differentiation of mouse mesenchymal stem cells (MSCs) into neuronal cells.

METHODSThe mouse multipotent mesenchymal precursor cell line (D1) was taken as the objective. Cultured MSCs were divided into the negative control group (complete culture solution), the positive control group (containing 1 mmol/L β-mercaptoethanol), the Salidroside induced group (20 mg/L Salidroside), and the blocked group (20 ng/ ml DKK1, a special inhibitor of Wnt/β-catenin signal pathway). All cells were inoculated in a 6-well plate (1 x 10(4) cells/cm2) and grouped for 24 h. The expression of p-catenin was detected by fluorescence Immunochemistry in the negative control group, the positive control group, and the Salidroside induced group. The expression of neuron-specific enolase (NSE), beta 3 class III tubulin (β-tubulin III), nuclear receptor related factor 1 (Nurr1), glial fibrillary acidic protein (GFAP) mRNA, Wnt3a, β-catenin, low-density lipoprotein receptor-related protein6 (LRP6), Axin mRNA were detected using reverse transcrip- tion PCR (RT-PCR). The expression of β-catenin and NSE protein were analyzed by Western blot in the negative control group, the positive control group, and the Salidroside induced group. Ca2+ chelating agents (EGTA), L-type Ca2+ channel blocker (Nifedpine), and IP3Ks special inhibitor (LY294002) were used to block Ca2+ signal pathway respectively. The expression of Wnt3a, LRP-6, Axin, glycogen syn- thase kinase (GSK-3), and β-catenin mRNA were detected by RT-PCR. The β-catenin protein expression was analyzed using Western blot.

RESULTSCompared with the positive control group, β-catenin protein was strong positively expressed; the expression of Wnt3a, β-catenin, LRP6, Axin, NSE, β-tubulin III, Nurr1 mRNA, and NSE protein were obviously up-regulated in the Salidroside induced group (P < 0.01). Compared with the positive control group and the Salidroside induced group, β-catenin, NSE, Nurr1, and β-tubulin III mRNA expression decreased; β-catenin and NSE protein expression were also down-regulated in the blocked group (P < 0.01). Compared with the Salidroside induced group, the expression of Wnt3a, LRP-6, β-catenin, and Axin mRNA were down-regulated in the Ca2+ signal blocked group and the salidroside induced group (P < 0.01, P < 0.05).

CONCLUSIONSalidroside affected directional differentia- tion of MSCs into neuronal cells through Wnt/β-catenin and Ca2+ signal pathway.

Animals ; Cell Differentiation ; drug effects ; Glucosides ; pharmacology ; Glycogen Synthase Kinase 3 ; Lipoproteins, LDL ; Low Density Lipoprotein Receptor-Related Protein-6 ; Mesenchymal Stromal Cells ; physiology ; Mice ; Neurons ; Phenols ; pharmacology ; Phosphopyruvate Hydratase ; RNA, Messenger ; Signal Transduction ; Wnt Signaling Pathway ; physiology ; beta Catenin ; metabolism

Artificial intelligence technology is being widely applied in drug screening. This paper introduces the characteristics of artificial intelligence, and summarizes the application and progress of artificial intelligence technology especially deep learning in drug screening, from ligand-based and receptor structure-based aspects. This paper also introduces how to apply artificial intelligence to drug design from these two aspects. Finally, we discuss the main limitations, challenges, and prospects of artificial intelligence technology in the field of drug screening.

Proteasome inhibitors have shown remarkable success in the treatment of hematologic neoplasm. There has been a lot of attention to applying these drugs for solid tumor treatment. Recent preclinical study has signified the effectiveness on cell proliferation inhibition in lung adenocarcinoma treated by carfilzomib (CFZ), a second generation proteasome inhibitor. However, no insight has been gained regarding the mechanism. In this study, we have systematically investigated the CFZ functions in cell proliferation and growth, cell cycle arrest, and apoptosis in lung adenocarcinoma cells. Flow cytometry experiments showed that CFZ significantly induced G2/M cell cycle arrest and apoptosis in lung adenocarcinoma. MTS and colony formation assays revealed that CFZ substantially inhibited survival of lung adenocarcinoma cells. All results were consistently correlated to the upregulation expression of Gadd45a, which is an important gene in modulating cell cycle arrest and apoptosis in response to physiologic and environmental stresses. Here, upregulation of Gadd45a expression was observed after CFZ treatment. Knocking down Gadd45a expression suppressed G2/M arrest and apoptosis in CFZ-treated cells, and reduced cytotoxicity of this drug. The protein expression analysis has further identified that the AKT/FOXO3a pathway is involved in Gadd45a upregulation after CFZ treatment. These findings unveil a novel mechanism of proteasome inhibitor in anti-solid tumor activity, and shed light on novel preferable therapeutic strategy for lung adenocarcinoma. We believe that Gadd45a expression can be a highly promising candidate predictor in evaluating the efficacy of proteasome inhibitors in solid tumor therapy.
Adenocarcinoma of Lung/pathology* ; Apoptosis/drug effects* ; Cell Cycle Checkpoints/drug effects* ; Cell Cycle Proteins/genetics* ; Cell Line, Tumor ; Forkhead Box Protein O3/physiology* ; Gene Expression Regulation, Neoplastic/drug effects* ; Humans ; Lung Neoplasms/pathology* ; Oligopeptides/pharmacology* ; Proto-Oncogene Proteins c-akt/physiology* ; Up-Regulation

Alanine Transaminase ; metabolism ; Animals ; Liver ; drug effects ; Male ; Mice ; Plant Extracts ; pharmacology

Administration, Inhalation ; Animals ; Female ; Male ; Pesticides ; analysis ; toxicity ; Rats ; Rats, Wistar

Cerebral Infarction ; etiology ; Dextrocardia ; etiology ; Female ; Humans ; Infant ; Spleen ; abnormalities ; Syndrome

Objective To explore the relationship between detective time for serum cardiac troponin I(cTnI) and their positive rate in diagnosis of viral myocarditis(VM).Methods Twenty-one cases of VM were designed as the test group.The serum cTnI were dynamically detected and compared with the normal control group.Results The serum cTnI were all negative in the normal control group,of 6 cases(28.6%) in the test group were positive when admission,of 7 cases(33.3%),8 cases(38.1%),9 cases(42.9%),13 cases(61.9%) and 4 cases(19%) were positive 6,12,18,24,48,72 h and 10 to 14 days laters respectively.There were statistic significances,compared the accumulative total positive rate of 48 h and 72 h after hospitalization with of emission and of 10 to 14 days after hospitalization,respectively.Conclusion Monitoring serum cTnI dynamically may increase the positive rate of cTnI for the suspected patients.

OBJECTIVETo explore the relationship between microsatellite alterations of RASSF1A gene and the development of cervical carcinoma, and HPV16 infection.

METHODSTwo sites of microsatellite polymorphism of RASSF1A gene were selected, we used polymerase chain reaction (PCR) technique to detect the LOH and MSI of cervical tissues, and to detect the infection state of HPV16.

RESULTSThere were significant differences of LOH rates at the two sites between clinical stage and pathological grade (P < 0.05). Significant differences were noted between the cervical carcinomas with lymph node metastasis and those without lymph node metastasis in regard to their LOH and MSI at the two sites ( P < 0.05). The incidence of LOH of RASSF1A gene was higher in HPV16(+) than that in HPV16(-) ( P < 0.05).

CONCLUSIONThe change of RASSF1A gene is a relatively late event in cervical carcinomas. The detection of the LOH and MSI of RASSF1A gene might be helpful to the early diagnosis and the screening of cervical carcinoma. It might also be useful for predicting the prognosis of cervical carcinoma. Infection of HPV16 and LOH of RASSF1A gene had reacted together in the development of cervical carcinoma.

Cervical Intraepithelial Neoplasia ; diagnosis ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Loss of Heterozygosity ; genetics ; Microsatellite Repeats ; genetics ; NM23 Nucleoside Diphosphate Kinases ; genetics ; Tumor Suppressor Proteins ; genetics ; Uterine Cervical Neoplasms ; diagnosis ; genetics

OBJECTIVETo explore the value of the technique of multiplex fluorescence in sit hybridization (M-FISH) combined with interphase fluorescence in situ hybridization (FISH) in the identification of the chromosomal aberrations in multiple myeloma (MM) and to investigate the frequency of 13q14 deletion, IgH translocations and 17p13 deletion.

METHODSSeven MM patients with complex chromosomal abnormalities (CCAs) were analyzed by combining the technique of conventional cytogenetics (CC) with M-FISH and FISH.

RESULTSM-FISH identified the aberrations which were undetected by CC, including twelve kinds of numeral aberrations and twenty-nine kinds of structural aberrations, In addition, abnormalities of chromosome 1, chromosomes 13 deletion and IgH translocations were the most frequent aberrations. Using the LSI D13S319 probe specific for 13q14, we observed a deletion of 13q14 in 6 MM patients; using the LSI p53 probe specific for 17p13, we observed p53 deletion in 4 MM patients; using the LSI IGHC/IGHV probe specific for 14q32, we observed a translocation involving 14q32 in 5 MM patients (43.5%), two translocations in two cases (case 6 and 7).

CONCLUSIONM-FISH combined with FISH could refine the cytogenetics of MM patients and detect the missed abnormalities or correct the misidentified abnormalities analyzed by CC. It provides an ideal method for the research of chromosomal aberrations in MM.

Adult ; Aged ; Chromosome Aberrations ; Chromosome Deletion ; Chromosomes, Human, Pair 1 ; Chromosomes, Human, Pair 13 ; Female ; Humans ; Immunoglobulin Heavy Chains ; In Situ Hybridization, Fluorescence ; methods ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; Translocation, Genetic

AIMTo observe the effect of mesenteric lymph duct ligation on free radical and inflammatory mediator of myocardium with severe hemorrhagic shock in rats at different period, and explore the effect of intestinal lymphatic pathway on myocardium injury pathogenesis in shock rats.

METHODS78 male Wistar rats were divided into the sham group, shock group and ligation group. The model of serious hemorrhagic shock was established in shock group, ligation group, and mesenteric lymph was blocked by ligating mesenteric lymph duct in ligation group after resuscitate. All rats were executed and taken out heart making for homogenate of 10 percent to determine the MDA, SOD, tumor necrosis factor-alpha (TNFalpha), interleukin-6 (IL-6), myeloperoxidase (MPO), NO and NOS at after shock 90 min, after transfusion and resuscitate 0 h, 1 h, 3 h, 6 h, 12 h and 24 h etc. different times, and the expression of inducible nitric oxide synthase (iNOS) mRNA in myocardium was detected by RT-PCR.

RESULTSThe contents of MDA, TNFalpha, IL-6, MPO, NO, NOS and iNOS expression in myocardium of shock group were rising after transfusion and resuscitate, and that was higher level at 3 h to 12 h, and that was significantly higher than sham group, the activity of SOD was significantly lower than sham group. The contents of MDA, TNFalpha, IL-6, MPO, NO, NOS and iNOS expression in myocardium of ligation group were significantly lower than that of shock group at sameness points, and the SOD activity was higher.

CONCLUSIONThe mesenteric lymph duct ligation and blocking mesenteric lymph could reduce the PMN detaining, decrease the discharging of TNFa and IL-6, reduce the NO and expression of iNOS mRNA, and reduce the releasing of free radical and consuming of SOD.

Animals ; Free Radicals ; metabolism ; Inflammation Mediators ; metabolism ; Interleukin-6 ; metabolism ; Lymphatic Vessels ; metabolism ; Male ; Mesentery ; metabolism ; Myocardium ; metabolism ; pathology ; Neutrophils ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Peroxidase ; metabolism ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; metabolism ; pathology ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism