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OBJECTIVE@#To investigate the molecular mechanism in stable cell strains expressing Mini-hF9 gene with nonsense mutation.@*METHODS@#Mini-hF9 gene and its nonsense mutants were transfected into HeLa cells independently, and stable cell strains were obtained after G418 resistance screening and monoclonal transformation. The altered splicing and protein expression of mRNA in Mini-hF9 gene in stable cell strains were detected by using RT-PCR and Western blot.@*RESULTS@#The wild type and nonsense mutated human coagulation factor IX stable cell strains were constructed successfully, which were named HeLa-F9-WT, HeLa-F9-M1 and HeLa-F9-M2. Only normal splicing Norm was detected in the wild-type cell strain HeLa-F9-WT; Norm and Alt-S1 splicing were detected in HeLa-F9-M1; while Norm, Alt-S1 and Alt-S2 splicing were detected in HeLa-F9-M2.@*CONCLUSION@#The nonsense associated altered splicing (NAS) pathway, which generated alternately spliced transcripts, might be triggered in coagulation factor IX gene with nonsense mutation.
Subject(s)
Humans , Codon, Nonsense , Factor IX/metabolism , HeLa Cells , Mutation , RNA Splicing , RNA, Messenger/metabolismABSTRACT
OBJECTIVE@#To study the role of nucleotide-binding oligomerization domain-like receptor proteins 1 and 3 (NLRP1 and NLRP3) inflammasome signaling pathways in the immune mechanism of inflammatory bowel disease (IBD) in children.@*METHODS@#A total of 126 children with IBD were enrolled as the study group, including 32 children with Crohn's disease (CD) and 94 children with ulcerative colitis (UC). A total of 120 children who underwent colectomy were enrolled as the control group. The mRNA expression of NLRP1, NLRP3, Caspase-1, and interleukin-1β (IL-1β) was compared between groups.@*RESULTS@#The study group had significantly higher mRNA expression of NLRP1, NLRP3, Caspase-1, and IL-1β than the control group, and their mRNA expression levels tended to increase with the severity of CD or UC (P<0.05). In the children with UC or CD, the mRNA expression levels of NLRP1, NLRP3, Caspase-1, and IL-1β were positively correlated with serum IgM and IgG levels (P<0.05), and the mRNA expression levels of NLRP1 and NLRP3 were positively correlated with those of Caspase-1 and IL-1β (P<0.05).@*CONCLUSIONS@#The NLRP1 and NLRP3 inflammasome signaling pathways may regulate the immune mechanism of IBD in children by upregulating the expression of Caspase-1 and IL-1β.
Subject(s)
Child , Humans , Adaptor Proteins, Signal Transducing , Apoptosis Regulatory Proteins , Inflammasomes , Inflammatory Bowel Diseases , Interleukin-1beta , NLR Family, Pyrin Domain-Containing 3 Protein , Signal TransductionABSTRACT
Objective To solve the problems of easily being contaminated and being difficult to hold and poor sealing for traditional midstream urine collection cups,and to reduce the unqualified rate of midstream urine collection.Methods A total of 126 patients without indwelling urinary catheter were selected from July to December in 2016 as the experimental group.From January to May 2016,112 patients without urinary catheter were selected as the control group.On the basis of conventional therapy and nursing care,the experimental group was given the self-designed disposable closed midstream urine collector to collect the specimens,and the control group was given the traditional urine cup to collect specimens.The unqualified rate,patient satisfaction and compliance were compared between two groups.Results There were 126 specimens in the experimental group,ll5 were qualified specimens,11 were unqualified specimens,and the unqualified rate was 8.73％(11/126);there were 122 specimens in the control group,91 were qualified specimens,21 were unqualified specimens,and the unqualified rate was 18.75％ (21/122);the satisfaction scores of the experimental group and the control group were (4.3±0.2) and (2.8±0.6),respectively;the experimental group had 66 cases of full compliance,48 cases of partial compliance,12 cases of non-compliance,and the compliance rate was 90.47％ (114/126);the control group had 27 cases of full compliance,51 cases of partial compliance,34 cases of noncompliance,and the compliance rate was 39.28％(44/112).Conclusion The disposable closed midstream urine collector can reduce the unqualified rate of specimens and improve patient satisfaction rate and compliance.
ABSTRACT
Objective To solve the problems of easily being contaminated and being difficult to hold and poor sealing for traditional midstream urine collection cups,and to reduce the unqualified rate of midstream urine collection.Methods A total of 126 patients without indwelling urinary catheter were selected from July to December in 2016 as the experimental group.From January to May 2016,112 patients without urinary catheter were selected as the control group.On the basis of conventional therapy and nursing care,the experimental group was given the self-designed disposable closed midstream urine collector to collect the specimens,and the control group was given the traditional urine cup to collect specimens.The unqualified rate,patient satisfaction and compliance were compared between two groups.Results There were 126 specimens in the experimental group,ll5 were qualified specimens,11 were unqualified specimens,and the unqualified rate was 8.73％(11/126);there were 122 specimens in the control group,91 were qualified specimens,21 were unqualified specimens,and the unqualified rate was 18.75％ (21/122);the satisfaction scores of the experimental group and the control group were (4.3±0.2) and (2.8±0.6),respectively;the experimental group had 66 cases of full compliance,48 cases of partial compliance,12 cases of non-compliance,and the compliance rate was 90.47％ (114/126);the control group had 27 cases of full compliance,51 cases of partial compliance,34 cases of noncompliance,and the compliance rate was 39.28％(44/112).Conclusion The disposable closed midstream urine collector can reduce the unqualified rate of specimens and improve patient satisfaction rate and compliance.
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The chemical consitituents from cytotoxic fraction of the Callicarpa nudiflora extract were isolated and purified by a combination of HP-20 macroporous resin, silica gel and Sephadex LH-20 column chromatographies. The structures were elucidated on the basis of the spectroscopic data and comparison of their spectroscopic data with reported data. The cytotoxicity was evaluated by the MTT assay. The 50% and 70% EtOH elutions of EtOH-extract showed significant cytotoxic activities, leading to the isolation of twelve compounds, which were identified as luteoloside(1), lutedin-4'-O-β-D-glucoside(2), 6-hydroxyluteolin-7-O-β-glucoside(3), lutedin-7-O-neohesperidoside(4), rhoifolin (5), luteolin-7, 4'-di-O-glucoside (6), forsythoside B (7), acteoside (8), alyssonoside (9), catalpol(10), nudifloside(11), and leonuride(12). Compounds 3-6, 10 and 12 were isolated from this genus for the first time, and compound 9 was isolated from this plant for the first time. The cytotoxicity assay demonstrated that flavonoids 1-6, in various concentrations, showed monolithic proliferation inhibitory activities against Hela, A549 and MCF-7 cell lines. Compounds 3, 5 and iridoid glycoside 11 possessed higher cytotoxicacivities. In short, flavonoids are the main components of cytotoxic extract from C. nudiflora, while phenylethanoid glycosides are the predominant ingredient but inactive to cancer cell lines. In addition, the minor iridoid glycoside expressed weak cytotoxic activity.
Subject(s)
Humans , Callicarpa , Chemistry , Cell Proliferation , Cytotoxins , Chemistry , Pharmacology , Drugs, Chinese Herbal , Chemistry , Pharmacology , MCF-7 Cells , Molecular StructureABSTRACT
<p><b>OBJECTIVE</b>To study the effect of Triptolide (TL) on the growth of prostate carcinoma cell line, and analyze its function and mechanism in anti-prostate cancer.</p><p><b>METHODS</b>MTT experiments were performed to examine the inhibiting effect of TL on the proliferation of RM-1 cells, cell morphological changes observed by acridine staining, cellular cycles and apoptosis peak analyzed by flow cytometry, the apoptosis fracture zone investigated with DNA electrophoresis, and the expressions of caspase-3 and bcl-2 mRNA in RM-1 cells examined by RT-PCR.</p><p><b>RESULTS</b>The results of MTT experiments showed that after the treatment of TL (5, 10, 20, 40 and 80 ng/ml), the RM-1 cell proliferation inhibition rates were 9.8%, 25.1%, 39.2%, 48.8% and 53.2% respectively; 12, 24, 36 and 48 hours after the treatment of TL (10 and 20 ng/ml), the cell proliferation inhibition rates were 8.4%, 25.1%, 36.1%, 42.4% and 10.2%, 39.2%, 50.2% and 58.5% respectively. Acridine staining after the TL treatment revealed nucleus condensation, cell membrane invagination, irregular orange particles in the cells and apoptosis morphological changes; flow cytometry tests showed that 48 hours after the TL treatment (10, 20 ng/ml) of RM-1 cells, an obvious apoptosis peak appeared before the G1 stage; 24, 36 and 48 hours after it, DNA "trapezoid" strips could be seen; the caspase-3 mRNA expression in the TL treated cells was higher, and the bcl-2 mRNA expression was lower than in the controls.</p><p><b>CONCLUSION</b>TL can decrease bcl-2 expression, increase caspase-3 expression, induce apoptosis of prostate carcinoma cells, and consequently inhibit the proliferation of RM-1 cells in mice.</p>