1.Construction of p16 Gene Recombinant Expression Vector and Its Antitumorigenic Effects on Human Lung Adenocarcinoma Cells
Xiaoying FU ; Shilong CAO ; Ruiqiong RAN
Chinese Journal of Cancer Biotherapy 1994;0(01):-
p16 gene was a tumor supressor gene found recently. The p16 protein is a negative regulator of cell proliferation . Loss of normal p16 function is associated with the development of neoplasms. To detect antitumorigenic effect of p16 on human lung adenocarcinoma cells, we cloned a p16 cDNA into the pcDNA3 vector at the sites of BamHl and Xho I to gain a p16 gene recombinant expression vector plasmid. We then transferred the p16 gene recombinant plasmid into human lung adenocarcinoma cell line (NCI-H460) by electroporation method. After G418 selection we assessed cell growth properties and cell cycle pattern by flow cytometry to G418-resistant clones of NCI-H460-pl6 and NCI-H460-vect respectively. The results show that human p16 gene could suppress the phenotype of NCI-H460 cell line and p16 gene therapy plays a positive role in human lung adenocarcinoma treatment.
2.Fugene 6——a new approach to induce the gene transfection in high efficiency for eukaryotic cells in vitro
Yide HU ; Nan GAO ; Xiaoyun CAO ; Jue ZHOU ; Shilong CAO
Immunological Journal 2001;(2):138-140
Objective To develop a new method to induce the gene transfection in high efficiency for eukaryotic cells in vitro. Methods Four kinds of p14ARF gene primarily deleted human carcinoma cell line including H460,A549,U251,and PC-3 were transfected with the human p14ARF expression vector (pCI-neo-p14ARF) by using the new nonliposomal transfection reagent Fugene 6. The efficiency of gene transfer was determined by screening the cells in G418. Results After 21 days' selection, G418-resistant clones were shown in all the transfected plate. PCR product of p14ARF gene was positive in all the G418-resistant clones. Cytotoxicity of Fugene 6 was detected. The cell proliferation activity was not affected when it was cultured in a high dose of Fugene 6. Conclusion These results demonstrate that Fugene 6 is a rapid, feasible, reproducible, and noncytotoxic gene transfection approach for eukaryotic expression vector in vitro.
3.Percutaneous puncturing drainage for the treatment of liver abscess: analysis of its curative effect and influence factors
Meiling ZHANG ; Chuanwu CAO ; Shilong HAN ; Xiaoyun XIE ; Maoquan LI
Journal of Interventional Radiology 2017;26(5):458-461
Objective To evaluate the curative effect of percutaneous puncturing drainage in treating liver abscess,to analyze the factors affecting curative effect,and to discuss the methods ior reducing mortality and complication rate as well as for shortening hospitalization time.Methods Clinical data of 121 patients with liver abscess,who were admitted to authors' hospital during the period from January 2011 to January 2016,were retrospectively analyzed.For the patients with confirmed liver abscess,adequate antiinfective therapy was adopted,at the same time CT scan was performed to evaluate the liquefaction of lesion,and under CT guidance percutaneous puncturing drainage was carried out.The mortality,complication rate,hospitalization time and the factors affecting curative effect were analyzed.Results A total of 121 patients with liver abscess were enrolled in this study.Two patients died after percutaneous puncturing drainage,the mortality was 1.6%.The factors affecting mortality included old age,underlying disease,the diameter and solid components of abscess.Two patients developed peripheral hepatic abscess and abdominal wall abscess,the complication rate was 1.6%,and clinical cure was achieved after active treatment in these two patients.The main factor affecting complication rate was inappropriate surgical manipulation.Clinical cure was achieved in all 119 patients,with a cure rate of 98.3%,and the average hospitalization time was (15.1±6.0)days.The risk factors that affected hospitalization time included the number of abscess X6 (r=0.232,P=0.021),abscess size X7 (r=0.26,P=0.005) and white blood cell count X8 (r=0.238,P=0.009).Multiple linear regression equation analysis indicated that statistically significant correlation existed between the above influence factors and hospitalization time (P<0.05).The multiple regression equation was as follows:Y=-3.438+3.055X6+0.527X7+0.297X8,F=5.819,R2=0.416.No statistically significant correlation existed between the hospitalization time and other factors,including gender,age,diabetes mellitus,pathogenic bacteria and location of abscess (P>0.05).Conclusion Percutaneous puncturing drainage is an effective treatment for liver abscess,it carries lower mortality and lower complication rate,and its hospitalization time is short.(J Intervent Radiol,2017,
4.Transfection of wild-type p14ARF gene leads to growth inhibition of human lung cancer cell lines
Nan GAO ; Yide HU ; Kangmei HUANG ; Xiaoyun CAO ; Jue ZHOU ; Shilong CAO
Chinese Journal of Lung Cancer 2001;4(1):15-19
Objective To examine whether wild-type p14ARF gene is a candidate suppressor gene for lung cancer. Methods Human lung cancer cell lines having various endogenous backgrounds in INK4a, p53 and Rb genes were used as the recipients of the wild-type p14ARF gene. The expression of p14ARF mRNA and protein was detected with RT-PCR, immunohistochemistry and Western blot after G418 selection. Clones which expressed both p14ARF mRNA and protein were identified and selected for further experiements. By comparing with the parental and negative control cells treated with empty vectors, the effects of exogenously transfected p14ARF on cell division rate, cell cycle distribution and morphologic alteration were analyzed. In vivo evaluation of the growth rate was also made with the experiment of nude mice tumor formation. Results Upon transfection with p14ARF gene, cells were arrested at G1 or G1/G2 phase of cell cycle in three wtp53 lung cancer cell lines and their proliferation rates were also inhibited. Conclusion Human wild-type p14ARF gene has suppressive effect on abnormal proliferation of lung cancer cells, especially in some wtp53 lung cancer cells, and it might be an ideal candidate for gene therapy of human lung cancer.
5.Research progress on the involvement of macrophages in myocardial ischemia reperfusion injury mediated by NF-κB signaling pathway
Shilong LI ; He WANG ; Yingjie CAO
Chinese Journal of Comparative Medicine 2024;34(4):139-146
Myocardial ischemia reperfusion injury(MIRI)has become one of the most serious complications affecting the clinical outcome of patients with cardiovascular diseases.The immune inflammatory response of macrophages is closely related to the occurrence and development of MIRI.Many studies have shown that the NF-κB signaling pathway can participate in MIRI regulation by influencing the polarization and inflammatory state of macrophages,pyrodeath,infiltration and other functions,and is a potential target for MIRI therapy.Therefore,this article will review the research progress of NF-κB signaling pathway between macrophage function and MIRI regulation.
6.Anti-osteoporosis Effect of Isorhamnetin: A Review
Shilong MENG ; Xu ZHANG ; Yawei XU ; Yang YU ; Wei LI ; Yanguang CAO ; Xiaolin SHI ; Wei ZHANG ; Kang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):347-352
Osteoporosis is a common senile bone metabolism disease, clinically characterized by decreased bone mass, destruction of bone microstructure, increased bone fragility, and easy fracture. It tends to occur in the elderly and postmenopausal women, seriously threatening the quality of life and physical and mental health of the elderly. At present, the treatment of osteoporosis is mainly based on oral western medicines, such as calcium, Vitamin D, and bisphosphonates. Still, there are drawbacks such as a long medication cycle and many adverse reactions. In recent years, due to the advantages of multi-component, multi-pathway, and multi-target, some traditional Chinese medicines and effective ingredients can regulate the osteogenic and osteoclastic differentiation process in both directions and are widely used in the prevention and treatment of osteoporosis. Hippophae rhamnoides is a commonly used herbal medicine, and its fruits are rich in flavonoids, polyphenols, fatty acids, vitamins, and trace elements, which have been proven to have a good anti-osteoporosis effect. Isorhamnetin is the main effective ingredient of Hippophae rhamnoides fruits, which has many pharmacological effects such as anti-inflammation, anti-oxidative stress, anti-aging, and anti-tumor. Studies have shown that isorhamnetin can participate in the regulation of bone metabolism and has a good anti-osteoporosis effect. However, the pharmacological effects and related mechanisms of isorhamnetin against osteoporosis have not been systematically summarized. Therefore, this paper reviewed the pharmacological effects and related mechanisms of isorhamnetin against osteoporosis by referring to relevant literature to provide more basis for the development and application of isorhamnetin.