Objective:To investigate the association between Cyclin D1 and metastasis and determine the relationship between Cyclin D1 proteins expression and P21 WAF1?P53 proteins in ovarian cancer.Methods:Cyclin D1,P21 WAF1 and P53 proteins were detected from 61 ovarian carcinoma and 7 ovarian tissues without cancer by using Western blot method.Results:The positive rate of Cyclin D1 and P53 proteins expression in cases with ovarian cancer were significantly higher than those in without cancer tissues(P0.05).However, the expression of P21 WAF1 protein was correlated with clinical stages(P0.05).The expression of Cyclin D1,P53 and P21 WAF1 protein in ovarian carcinoma were related to metastasis(P

Objective To study the effects of 5-azacytidine’s demethylation for P16 gene on hemangioma cell’s proliferation and apoptosis.Methods Bisulfite sequencing PCR was applied to detect P16 gene′s promoter methylation status in 5-azacytidine treated and untreated EOMA cell line.RT-PCR and western blot were used to detect the P16 gene mRNA and protein expressions.Flow cytometry was used to detect cell proliferation, cell cycle and cell apoptosis.The differences of P16 gene′s promoter methylation status,mRNA and protein expressions,cell proliferation,cell cycle and apoptosis in two groups were compared. Results The methylation rates in 1st and 13th CGs were 0%after 5-azacytidine treatment in EOMA hemangioma cell line,which were lower than in untreated cells.The mRNA and protein expressions increased after 5-azacytidine treatment,which were significantly higher than in untreated cells.The absorbance,S phase and G2/M phase and PI after 5-azacytidine treatment were lower than untreated cells,while the G0/G1 phase and apoptosis rates were higher.Conclusion The P16 gene promoter is hypermethylated in hemangioma cells with silent gene expressions.5-azacytidine could reverse P16 gene′s promoter methylation and silent gene expressions,which inhibit hemangioma cell’s proliferation and promote apoptosis.

Objective To study the effects of Lentinan on bleomycin A5’s inhibition effects in hemangioma-derived endothelial cell. Methods Hemangioma-derived endothelial cell line EOMA were divided into LTN group (only lentinan treatment), BLE group (only bleomycin A 5 treatment), L+B group (Lentinan and bleomycin A5 treatment) and CON group (no lentinan and bleomycin A5 treatment). Cell proliferation, cell cycle, PI and apoptosis were detected and compared among four groups. Results The differences of absorbance in LBE group was significantly higher than that in L+B group after treatment for d 1-d 7 (P<0.05), but both of them were significantly lower than in CON group and LTN group (P<0.05). The G 0/G 1 stage and apoptosis rate in L+B group was significantly higher than in BLE group(P<0.05), while it was significantly lower in S stage, G 2/M stage and PI(P<0.05). Conclusion Lentinan could significantly promote bleomycin A5’s inhibition effects on hemangioma-derived endothelial cell.

Objective:To investigate the predictive value of gadobenate dimeglumine (GD-BOPTA) multi-phase enhanced MRI for the expression of cytokeratin19 (CK19) in hepatocellular carcinoma (HCC).Methods:A total of 153 patients of HCC confirmed by pathology from June 2016 to February 2020 in First Affiliated Hospital of Fujian Medical University were enrolled retrospectively. According to the post-operative pathology, the patients were divided into CK19-negative group ( n=122) and CK19-positive group ( n=31). All the patients underwent MRI scan and Gd-BOPTA multi-phase enhanced scan before operation. MRI features on Gd-BOPTA MRI were compared between two groups. The qualitative indicators included tumor morphology, mosaic signs, intratumoral hemorrhage, intratumoral fat, non-rim arterial phase hyper-enhancement (APHE), non-peripheral washout, targetoid manifestation, enhanced capsule, corona enhancement, DWI signal, vascular invasion and hepatobiliary phase (HBP) enhancement. The quantitative indicator of tumor-to-liver signal ratio (SR) on HBP was recorded. The χ 2 test or Fisher exact probability method was used to compare the qualitative parameters between two groups, and student′s t test or Mann -Whitney U test was used for quantitative data. Predictive parameters were identified by univariate and multivariate logistic regression analysis to predict the value of the expression of CK19. The ROC curve was used to analyze the diagnostic efficacy of MRI parameters. Results:There were statistically significant differences between CK19-positive and CK19-negative groups ( P<0.05) in alpha fetoprotein, tumor morphology, non-rim APHE, non-peripheral washout, targetoid manifestation, corona enhancement, HBP enhancement and SR. Multivariate logistic regression analysis showed tumor morphology, corona enhancement, HBP enhancement and SR were independent predictors of CK19 expression in HCC. The area under the ROC curve of the combined four indicators for predicting CK19 expression in HCC was 0.823, and the sensitivity and specificity were 80.7% and 75.4%, respectively. Conclusions:Gd-BOPTA multi-phase enhanced MRI has an important value in the prediction of the expression of CK19 in HCC. The combination of signs of HBP can improve the prediction efficiency of CK19.

OBJECTIVETo investigate the insulin-like growth factor-I (IGF-I) gene and polypeptide expression in cultured rat osteoblast (ROB) and the role of IGF-I in mediating the cell-to-cell communication by mimicking the pharmacokinetics of parathyroid hormone (PTH).

METHODSThe ROB was cultured with three kinds of treatment: (1) Control (Ctr), the cells were cultured without PTH during the first 6 hours and the subsequent 42 hours in a 48-hour cycle; (2) Intermittent exposure to PTH (Itm), the cells were cultured with PTH during the first 6 hours, but without PTH in the subsequent 42 hours; and (3) Continuous exposure to PTH (Ctu), the cells were cultured with PTH during the first 6 hours and the subsequent 42 hours.

RESULTSThe bone-forming activities of ROB were increased in Itm and inhibited in Ctu. The IGF-I mRNA content in Itm cells was elevated only during the first 6 hours and that in Ctu cells was elevated at any time during an incubation cycle. The free IGF-I concentration in the medium of Itm cells was generally higher and that of the Ctu cells was generally lower compared with those of the Ctr cells. The IGF-I antibody significantly reduced the alkaline phosphatase activity within the cells of Ctr and Itm.

CONCLUSIONSPTH rapidly and constantly stimulates the IGF-I gene transcription of osteoblast. There was an obvious discrepancy between the IGF-I mRNA content within the osteoblast and the free IGF-I level around the osteoblast in either mode of PTH action. The IGF-I might be important for osteoblast-osteoblast communication and bone-forming activity, not only in intermittent PTH administration, but also in the physiological functioning of osteoblasts.

Animals ; Cells, Cultured ; Gene Expression ; drug effects ; Insulin-Like Growth Factor I ; genetics ; physiology ; Osteoblasts ; Parathyroid Hormone ; pharmacology ; Peptides ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transcription, Genetic ; drug effects

Objective: To establish a method for simultaneous and rapid detecting of the polymorphisms in Cytochrome P450 2C9 (CYP2C9), CYP2C19, CYP4F2, Vitamin K epoxide reductase (VKORC1) and ATP-binding cassette subfamily B member1 (ABCB1) gene, which were associated with warfarin and clopidogrel, based on liquid phase chip technology. Methods: Method establishment. The eight gene sequences near targeted sites related to warfarin and clopidogrel were found in Genbank, and the specific primers and probes were designed. Through multiple PCR amplification, followed by allele specific primer extension (ASPE), and MagPlex-Tag microspheres hybridization, the suspension array Luminex 200 system step-by-step, the genotypes were determined by fluorescence signal. The reaction system was optimized and its methodological evaluation was performed. 260 patients with antithrombotic therapy from Dongguan houjie hospital were recruited in this study form June 2017 to December 2018. The eight genotypes of the 260 patients were detected by the established method, and the results were compared with the sequencing results. Results: The results of 260 samples showed that allelic median fluorescence intensity (MFI) ratios of homozygotes (mutant/wild-type) were all greater than 0.9 or less than 0.1, and all the allelic MFI ratios of heterozygotes were between 0.3 and 0.6. The within run and between run coefficients of variance for allelic MFI ratios were lower than 6.4% and 10.9%, respectively. The minimum DNA template requirements was 0.75ng. The genotypes of 260 patients determined by the established method were completely concordant with the sequencing results. Conclusion A method was established successfully for rapid detecting the genotypes which associated with warfarin and clopidogrel based on liquid phase chip technology.

ObjectiveTo investigate the influence of different diagnostic criteria on the short-term prognosis of patients with acute-on-chronic liver failure （ACLF）. MethodsA total of 115 ACLF patients who were hospitalized in Department of Gastroenterology， The Second Affiliated Hospital of Kunming Medical University， from January 2018 to January 2022 were enrolled， and all patients received internal medical treatment combined with artificial liver therapy. According to the guidelines， the patients were divided into CMA guideline group （Diagnostic and treatment guidelines for liver failure by Chinese Medical Association）（n=100）， APASL guideline group （Consensus statements of Asian Pacific Association for the Study of the Liver）（n=94）， and EASL guideline group （Criteria proposed by European Association for the Study of the Liver）（n=36）. The above three guidelines were compared in terms of 90-day mortality rate. A one-way analysis of variance was used for comprision of continuous date between groups； the chi-square test was used for comprision of categorical date between groups. The receiver operating characteristic （ROC） curve of related variables. ResultsThe 90-day mortality rate was 50.0% in the CMA guideline group， 51.1% in the APASL guideline group， and 77.8% in the EASL guideline group， and the EASL guideline group had a significantly higher 90-day mortality rate than the CMA guideline group （χ²=8.351， P=0.004） and the APASL guideline group （χ²=7.650， P=0.006）. EASL guideline had a sensitivity of 22.2% and a specificity of 92.3% in predicting the risk of short-term mortality， with an area under the ROC curve was 0.576. ConclusionACLF patients who meet EASL guideline tend to have a worse short-term prognosis， and this guideline may help to identify patients at a relatively high risk of short-term death.