1.Mechanism of hsa-miRNA124-3p regulating the proliferation and migration of human lung cancer cell line NCI-H460
Ye WANG ; Wang XIE ; Jie ZHANG ; Jieying SHENG ; Zhongliang GUO
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(8):1051-1058
Objective · To study the regulation of hsa-miRNA124-3p on the proliferation and migration of human lung cancer NCI-H460 cells and its mechanism. Methods · Four pairs of lung cancer and para-carcinoma tissues were harvested in clinical and measured for hsa-miRNA124-3p and Krüppellike factor 4 (KLF4) levels. The theoretical binding site of hsa-miRNA124-3p in 3'-UTR of KLF4 was predicted by bioinformatics, and validated by luciferase report assay. NCI-H460 cells were transfected with pshRNA-Sponge-miRNA124 or pshRNA-KLF4, and 48 hours later, the proliferation of NCI-H460 cells after genetic intervention was assayed by the MTT method, and cell migration ability was observed by streak method. Results · For all four pairs of samples tested, hsa-miRNA124-3p was higher in the cancer tissues than in the adjacent tissue (P<0.01), and KLF4 protein was lower in the cancer tissues than in the adjacent tissue (P<0.01). The bioinformatic analysis showed there is a theoretical binding site (5'-UGCCUUAA-3') of hsa-miRNA124-3p in 3'-UTR of KLF4. Luciferase activity assay showed that hsa-miRNA124-3p could bind to the 3'-UTR region of KLF4 gene and negatively regulate the expression of protein. The proliferation of NC-H460 cells was suppressed by transfection with pshRNA-Sponge-miRNA12472 h after transfection (P<0.05 ). Compared with the control group, the proliferation activity of pshRNA-KLF4 transfection group was further enhanced (P<0.05) There was no significant difference in the proliferation of pshRNA-Sponge-miRNA124 and pshRNA-KLF4 cotransfection group and the control group (P>0.05). The data of cell migration assay showed that the changes of cell migration ability were the same as proliferation activity of the cells in groups 72 h after transfection. Conclusion · Hsa-miRNA124-3p increases the proliferation and migration in NCI-H460 cells via suppressing the expression of KLF4, and reducing the content of miRNA124-3p in NC-H460 cells can inhibit cell proliferation and migration via upregulating KLF4 expression.
2.Solubility peak and dielectric requirement of Buyang Huanwu docoction.
Huan ZOU ; Fuyuan HE ; Jieying LUO ; Sheng HUANG ; Yun QIU ; Feng CHEN
China Journal of Chinese Materia Medica 2009;34(13):1648-1653
OBJECTIVETo study the solubility peak and dielectric requirement of the Buyang Huanwu docoction materials, and provide theoretical and experimental foundation for selecting extraction solvent for extracting traditional Chinese drugs (TCD).
METHOD11 types of solvents were employed as single or complex solvent systems, whose solubility parameter and dielectric constant were from 14.11 to 47.86, dielectric requirement from 1 to 80 respectively, to lixiviate Buyang Huanwu decoction (5 g per samples) in nearly saturate volume as V0 for materials at 25 degrees C. The apparent solubilities of extracts were determined and calculated out according to the section of determination of extract in the appendix of 'Chinese Pharmacopoeia'.
RESULTThe saturate solvent V0 for materials powder were 0.21, 0.31, 0.49, 0.36, 0.77, 0.93, 0.86, 0.92, 1.08, 1.00, 1.14 mL x g(-1), respectively. The apparent solubility of Buyang Huanwu docoction for each solvent system were 114.0, 101.3, 73.40, 109.4, 210.7, 295.0, 501.4, 437.0, 355.6, 423.1, 210.6 g x mL(-1), respectively, among which the max apparent solubility, illustrated as solubility peak, was carried out by methanol-water (68: 32) with 47.5 corresponding to the Buyang Huanwu docoction dielectric requirement.
CONCLUSIONThe apparent solubilities of (TCD) and their formula are controlled by dielectric constant of extraction solvent, and are in accordance with stable dielectric requirement.
Drugs, Chinese Herbal ; chemistry ; Solubility ; Solvents ; chemistry
3.The Nomogram model in predicting prognosis of hepatocellular carcinoma patients based on four inflammatory markers
Zhiqiang FENG ; Zixuan YANG ; Shanshan HAN ; Yutao SHANG ; Junhui ZHAO ; Wanqing GU ; Qingmin YANG ; Jieying WU ; Jun SHENG ; Xiaodong GUO
Chinese Journal of Hepatobiliary Surgery 2020;26(6):443-448
Objective:To construct a Nomogram model in predicting recurrence-free survival (RFS) and overall survival (OS) at six months, one year and two years after hepatocellular carcinoma (HCC) resection by using inflammatory markers combined with other routine clinical indicators.Methods:The data of 314 patients with HCC who underwent first time hepatectomy at Beijing Chaoyang Emergency Rescue Center and Air Force Characteristic Medical Center from January 2013 to January 2018 were analyzed. HCC patients who underwent hepatectomy at the First Medical Center of PLA General Hospital from January 2011 to January 2016 ( n=106) were used as the external validation group. Univariate and multivariate Cox proportional risk model was used to analyze independent risk factors of recurrence and death in HCC patients. A Nomogram model was constructed based on independent risk factors. Validation of the efficacy of the Nomogram model was done based on external data. Results:In the experimental group, 174 patients relapsed. The median RFS was 26 months. The 6 months, 1 year and 2 years RFS were 26.8%, 43.9%, and 68.8%, respectively. A total of 142 patients had died. The median survival time was 30 months. The 6 months, 1 year and 2 years OS were 5.9%, 23.6% and 63.1%, respectively. In the external validation group, 63 patients had developed recurrence, with a median RFS time of 28 months. The 6 months, 1 year and 2 years RFS were 26.4%, 45.3%, 54.7%, respectively. The median survival time was 31 months. The 6 months, 1 year and 2 years OS were 7.5%, 25.5%, 46.6%, respectively. Tumor size (>6.0 cm, HR: 1.447), vascular invasion ( HR: 1.408), TBil (>0.94 mg/dl, HR: 1.949), NLR (>2.54, HR: 2.843), AGR (≤0.88, HR: 2.447) were independent risk factors of HCC recurrence ( P<0.05). Tumor size (>6.0 cm, HR: 2.207), vascular invasion ( HR: 1.529), and NLR (>2.54, HR: 2.708) were independent risk factors of death for HCC patients ( P<0.05). The C-indexes of half-year, one-year and two-year RFS were 0.764 (95% CI: 0.677-0.854), 0.710 (95% CI: 0.615-0.824) and 0.673 (95% CI: 0.601-0.786), respectively. The C-indexes of half-year OS, one-year OS and two-year OS were 0.729 (95% CI: 0.648-0.841), 0.708 (95% CI: 0.608-0.813) and 0.664 (95% CI: 0.618-0.771), respectively. Conclusion:In this study, the construction of a Nomogram model in predicting prognosis of HCC patients was helpful to guide clinicians in improving preoperative treatment plans and in providing ideas for individualized treatment of patients.
4.Calculation of retention and excretion fractions for uranium intake via wounds based on ICRP Report 137
Jieying LEI ; Yang ZHANG ; Bin ZHANG ; Ming XU ; Wei SHENG ; Gaofeng SUN
Chinese Journal of Radiological Health 2024;33(6):631-637
Objective To assess the retention and excretion fractions for uranium intake via wounds in the organs of adult reference computational phantom, and to improve the evaluation of health risks associated with radioactive isotopes intake via wounds. Methods A software for evaluation of the internal radiation dose was developed by combining the wound model in International Commission on Radiological Protection Report 156 and the systemic models in the International Commission on Radiological Protection Report 130 and the subsequent Occupational Intakes of Radionuclides series reports. This study was conducted to calculate the uranium retention fractions in adult reference individuals following ingestion through wounds of seven categories of uranium: weakly retained, moderately retained, strongly retained, very strongly retained, colloids, particles, and fragments. We assessed the retention fractions in the wound sites, bones, and liver, as well as the excretion fractions in urine and feces, and compared these results with authoritative results. Results The retention and excretion fractions of 238U and 235U in different forms of intake via wounds showed consistent variations. The retention fractions of weak and moderate uranium in wounds could be ignored after 1 000 d. The retention fractions of strong, avid, and colloid uranium were 10−4 to 10−5 after 10 000 d. A significant portion of the uranium present in the forms of particles and fragments exhibited long-term retention in wounds. The bone retention fractions were 1 to 2 orders of magnitude higher than the liver retention fractions. Following the intake of soluble and insoluble uranium, the retention fractions in these two organs decreased and increased, respectively, compared with those observed on the first day. The urinary excretion fractions were approximately 2 orders of magnitude higher than the fecal excretion fractions. Following the intake of soluble and particulate uranium, the excretion fractions decreased. At 1 000 d after the intake of uranium in the form of fragments, the urinary and fecal excretion fractions were 10−6 and 10−8, respectively. Conclusion The calculation results validated the accuracy of the established models, providing data support for the assessment of internal exposure doses in individuals following uranium wound contamination incidents.