Objective To study the apoptosis of pancreatic cancer cells induced by a retroviral vector containing antisense human telomerase RNA gene. Methods Sense and antisense telomerase RNA genes were transfected into Can-pan-2 cell line by electroporation,and selected with G418 to get stable transfection and retrovirus yielding cell lines. We transfected Can-pan-2 cell with concentrated retroviral supernatant,and selected with G418 to get stable transfection cell lines,The expression of the aim gene in Can-pan-2 cell was confirmed through PCR,while TRAP-PCR-ELISA was used to determine the telomerase activity,and immunofluorescence was adopted to determine reproductive activity of the infected cells,flow cytometry(FCM)was used to detect cell apoptosis.ResultsThe titer of sense and antisense recombinant retrovirus reached 0.95?106 CFU/mL and 1.1?106 CFU/mL respectively,PCR demonstrated that the telomerase RNA genes were successfully intergrated into the target cells’ genome,The cell growth and telomerase activity of Can-pan-2 were significantly inhibited and apoptosis occurred after antisense virus infection. Conclusion Antisense human telomerase RNA may inhibite telomerase activity of pancreatic cancer cells and promote apoptosis of the cells.

AIM: To detect the expression of microRNA (miRNA)-29a in pulmonary arteries of monocrotaline (MCT)-induced pulmonary hypertensive rats, and to investigate the effects of miRNA-29a on pulmonary hypertension.METHODS: MCT-induced pulmonary hypertensive model was established in Wistar rats.The expression of miRNA-29a in the lung tissue was determined by qPCR.miRNA-29a was overexpressed in the pulmonary hypertension rats by tail vein injection of miRNA-29a-mimic.Pulmonary artery pressure (PAP) and systemic arterial pressure (SAP) were measured.The morphological changes of the pulmonary arteries were observed by HE staining.The protein levels of p-Akt and p-eNOS were detected by Western blot.RESULTS: The mRNA expression of miRNA-29a was significantly decreased in the pulmonary arteries of MCT-induced pulmonary hypertensive rats.Furthermore, after overexpression of miRNA-29a, PAP was remarkably reduced, while SAP remained unchanged.In addition, the increased thickness of tunica media, the remodeling of pulmonary arteries and the decreased protein levels of p-Akt and p-eNOS in the pulmonary hypertensive rats were dramati-cally changed after miRNA-29a overexpression.CONCLUSION: Overexpression of miRNA-29a ameliorates pulmonary hypertension in rats.These effects may be associated with the activation of PI3K/Akt-eNOS signaling pathway.

Objective To study the clinical value of tamsulosin in prevention of urinary retention after radical hysterectomy.Methods Sixty-six patients after radical hysterectomy were enrolled in this study, and divided into two groups in random.Thirty-three cases in the treatment group were treated with tamsulosin 0.2 mg once every night,3 days before the catheter was drawn.While 33 cases in the control group only with placebo.Catheter keeping days, residual urine volume at first extraction,the rate of urinary retention and urinary tract infection,hospitalization time and drug adverse reaction were compared in two groups.Results Catheter keeping days and hospitalization time were shorter in the treatment group than those in the control group(P< 0.05).The residual urine volume at first extraction was(38.2 ± 5.6) ml in the treatment group and (168.5 ± 11.8) ml in the control group,there was significant difference between two groups (P< 0.05).The rate of urinary retention was 18.2%(6/33) in the treatment group and 36.4%(12/33) in the control group,there was significant difference between two groups (P <0.05).There was no adverse reaction resulted from tamsulosin in the treatment group.Conclusion Tamsulosin is an effective drug which could prevent and treat the urinary retention after radical hysterectomy.

BACKGROUND: Some studies have showed that after indirect co-culture, bone marrow mesenchymal stem cells can differentiate into myocardial cells and hepatocypte-like cells. OBJECTIVE: To investigate the possibility of human umbilical cord blood mesenchymal stem cells (UCB-MSCs) to differentiate into hepatocytes by co-culture with human hepatocyte line LO2 in vitro.METHODS: Full-term umbilical cord blood samples were obtained sterilely. The UCB-MSCs were isolated by density gradient centrifugation and directly adherence growth, then passaged with trypsin digestion at 80% cell fusion. By utilizing cell culture plate insets with microporous membrane combined with 6-well plate, the LO2-/UCB-MSCs co-culture system was established. UCB-MSCs were plated into the wells of 6-well plate at a density of 1×10~7/L. LO2 cells were plated into the cell culture plate insert at a density of 1×10~5/L. UCB-MSCs were plated in both layers in the control group. Surface markers of adhered cells were detected by flow cytometry. Morphological changes of UCB-MSCs were observed by inverted phase contrast microscopy. mRNA expression was detected by reverse transcriptase PCR. RESULTS AND CONCLUSION: HUCB MSCs expressed CD44 and CD29 strongly, but CD34 and CD45 were expressed negatively. After 5 days, fusiform-shaped cells were reduced in the co-culture group; while, the time passing by, cells shaped irregular round or polygonal were increased, which were similar to hepatocytes. At 4 weeks after culture, UCB-MSCs were still fusiform-shaped in the control group. At day 5 after culture, alpha fetoprotein mRNA expressed positively, but other expressed negatively in the co-culture group; at day 14 after culture, cytokeratin-19 mRNA and albumin mRNA expressions were observed; moreover, with the time passing by, the expression of albumin mRNA was increased, but the expression of alpha fetoprotein-19 mRNA was decreased. Antigenic expressions in the control group were negative. This suggested that UCB-MSCs could differentiate into hepatocypte-like cells by co-culture with human hepatocyte line LO2 in vitro.

OBJECTIVETo study the effect of ferment powder caterpillar fungus on cytochrome P450 isozymes CYP1A2, CYP3A4 and CYP2E1.

METHODThe methods of Cocktail probe drugs were used. The rats were randomly divided into two groups. One group were given ferment powder caterpillar fungus once daily orally for ten days. Another group received orally normal saline one daily as the blank control. After ten days of treatment, the rats were given probe drugs of coffine, dapsone and chlorzoxazone and the blood was taken out by femoral catheterization. The plasma concentration of probe drugs were determined by HPLC. Data of plasma drug level-time were disposed with DAS Ver 2.0.

RESULTThe metabolism of caffeine and dapsone speeded up after receiving ferment powder caterpillar fungus, but the metabolism of chlorzoxazone was hardly changed.

CONCLUSIONIt suggested that ferment powder caterpillar fungus tended to be the inducer of CYP1A2 and CYP3A4. But the CYP2E1 was hardly affected.

Animals ; Caffeine ; metabolism ; Chlorzoxazone ; metabolism ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Dapsone ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Fermentation ; Male ; Random Allocation ; Rats ; Rats, Wistar

Objective:To explore the effects of MiR-199a-3p on promoting the development of mycoplasma pneumonia via the silent information regulator 1(SIRT1)/nuclear factor-κB (NF-κB) pathway.Methods:Totally 80 SPF-grade BALB/c mice were randomly divided into a control group, a model group, a miR-199a-3p group, and a inhibitory group, with 20 rats in each group. Excep the control group, the model group was established as a mouse model of mycoplasma pneumoniae through a continuous nasal drip of a high-load mycoplasma pneumoniae bacterial solution for 3 days. After modeling, mice in each group had tail vein injections. The miR-199a-3p group and the inhibitory group mice were injected with agomir solution and antagonir solution, respectively. And the model group and control group mice were injected with the same amount of physiological saline through the tail vein. After the experiment, mice of all groups were killed and their blood was collected from the eyeballs, and interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) levels were detected by the enzyme-linked immunosorbent assay method. Subsequently, the lung tissues of the mice were taken for HE staining to observe pathological changes in the lung tissue. Real-time fluorescence quantitative PCR was used to detect miR-199a-3p gene expression levels in lung tissue, and Western Blot was used to detect SIRT1/NF-κB signaling pathway protein expression in lung tissue. Results:Compared with the model group, IL-4, IL-6, IL-1β, and TNF-α levels in the serum of the miR-199a-3p group were decreased, with a significant difference ( P < 0.05). The HE staining results showed that the lung tissue structure of the control group mice was nearly normal and there was no alveolar exudation or inflammatory response. The other three groups all had varying degrees of alveolar interstitial widening, alveolar exudation, and inflammatory cell infiltration. Compared with the model group, miR-199a-3p gene and SIRT1 protein expression levels in the miR-199a-3p group increased, with a significant difference (all P < 0.05). While NF-κB protein expression levels in the miR-199a-3p group decreased, there was a significant difference ( P < 0.05). Conclusions:The expression of miR-199a-3p gene is reduced in the lung tissue of mycoplasma pneumoniae mice. Increasing the level of miR-199a-3p gene has a protective effect on lung tissue damage in mycoplasma pneumoniae mice through anti-inflammatory effects, and its mechanism may be related to its regulatory effect on the SIRT1/NF-κB pathway.

Scutellaria baicalensis (S. baicalensis) and Scutellaria barbata (S. barbata) are common medicinal plants of the Lamiaceae family. Both produce specific flavonoid compounds, including baicalein, scutellarein, norwogonin, and wogonin, as well as their glycosides, which exhibit antioxidant and antitumor activities. Here, we report chromosome-level genome assemblies of S. baicalensis and S. barbata with quantitative chromosomal variation (2n = 18 and 2n = 26, respectively). The divergence of S. baicalensis and S. barbata occurred far earlier than previously reported, and a whole-genome duplication (WGD) event was identified. The insertion of long terminal repeat elements after speciation might be responsible for the observed chromosomal expansion and rearrangement. Comparative genome analysis of the congeneric species revealed the species-specific evolution of chrysin and apigenin biosynthetic genes, such as the S. baicalensis-specific tandem duplication of genes encoding phenylalanine ammonia lyase and chalcone synthase, and the S. barbata-specific duplication of genes encoding 4-CoA ligase. In addition, the paralogous duplication, colinearity, and expression diversity of CYP82D subfamily members revealed the functional divergence of genes encoding flavone hydroxylase between S. baicalensis and S. barbata. Analyzing these Scutellaria genomes reveals the common and species-specific evolution of flavone biosynthetic genes. Thus, these findings would facilitate the development of molecular breeding and studies of biosynthesis and regulation of bioactive compounds.
Evolution, Molecular ; Flavonoids/biosynthesis* ; Genome, Plant ; Plant Extracts/genetics* ; Scutellaria/metabolism* ; Whole Genome Sequencing