1.Effects of specific small interfering RNA-mediated cyclinD1 gene silencing on the growth of human keloid fibroblasts
Daning LIANG ; Jianhua GAO ; Feng LU
Chinese Journal of Tissue Engineering Research 2007;0(02):-
AIM:Cyclin is a decisive factor of regulating cell cycle,and RNA interference as an effective and specific gene silencing technique,can induce cell express the phenotype of specific gene deficiency. This study is to apply cyclinD1 specific small interfering RNA(siRNA) on inhibiting cyclinD1 gene expression and investigate the effect of cyclinD1 specific siRNA on the cell cycle and multiplication in keloid fibroblasts. METHODS:The experiment was conducted at the Institute of Genetic Engineering(Grade BSL-2) in Southern Medical University from July 2006 to May 2007.①siRNA was designed with siRNA target finder of ambion Company,and synthesized chemically in Shanghai GeneChem,Co.,Ltd. Then double-strand siRNA was obtained following degenerative renaturation. Keloid fibroblasts were sampled from the patients in the Department of Plastic Surgery,Southern Medical University(informed consents were obtained from the patients or their relatives).②The keloid fibroblasts were divided into experimental group and control group. The cyclinD1 specific siRNA was transfected into the keloid fibroblast of the experimental group by the liposome-mediated gene transfection method. The untreated cells were set as controls.③At hours 24,48 and 72 of transfection,light microscope was used to observe cell morphologic change and flow cytometry was used to examine cell cycle. The viable cells were counted by MTT colorimetry and a cell growth curve was drawn. RESULTS:①Abnormal change of cell morphology that became into spherical shape or oval-shape from normal long fusiform after transfection may be the apoptosis or necrosis cells.②The G1 stage of cell cycle extended and the S stage decurtated. At 24,48 and 72 hours after transfection,the radio of G1 stage cell was 60.13%,66.22% and 67.53%,which were all significantly higher than that in the control group(54.53%);the radio of S stage cell(18.25%,17.11% and 11.15%) was also significantly lower than that in the control group(22.31%),indicating that the proportion of the cells blocked in G1 stage and those in S stage decreased in the keloid fibroblast.③siRNA-cyclinD1 inhibited the growth of keloid fibroblasts obviously by using MTT assay,and the cell growth curves indicated that the proliferation of cell transfected with cyclinD1 specific siRNA was inhibited significantly when compared with controls. CONCLUSION:CyclinD1 specific siRNA effectively inhibits the expression of cyclinD1 in keloid fibroblasts thus arresting the cell cycle at G1 stage and enhancing cell apoptosis.
2.THE EFFECT OF TAURINE ON nNOS POSITIVE NEURON IN HIPPOCAMPUS OF RAT INDUCED BY LEAD LESION
Feng YANG ; Jisheng LI ; Yahua LIU ; Daning WANG
Acta Nutrimenta Sinica 2004;0(06):-
Objective: To study the effect of taurine to resist the impact of lead on learning and memory. Methods: ABC immunohistochemistry method was used to study the quantity change of nNOS positive neuron in hippocampus of the rats which were fed with distinct dosage of lead acetate in drinking water (0.02, 0.2 g/L) and distinct dosage of taurine (5,10 g/kg) in feed. Rusults: Taurine could increase nNOS positive neuron quantity obviously in hippocampus of rats induced by lead lesion. Conclusion: Taurine could resist impact of lead on learning and memory obviously.
3.Treatment of Menopausal Syndrome Based on Phlegm,Stagnation,and Fire
Wanshi LIANG ; Jiajing WANG ; Yalin YOU ; Jingqi SHU ; Jian LIU ; Daning FENG ; Guangning NIE
Journal of Traditional Chinese Medicine 2023;64(20):2142-2145
Menopausal syndromes are mostly based on kidney deficiency, which could be expalined that kidney governing essence storage and controlling innateness, so when the kidney water was deficient and the liver fail to nourish, then led to liver depression and transform into fire; deficiency of the kidney, loss of warmth of the spleen, and inability to transport and transform the water and dampness will easily lead to phlegm and fire; the decline of the kidney yin and loss of water and fire will easily cause the exuberance of heart fire. Therefore, clinical symptoms of hot flashes, insomnia, and palpitations are common due to phlegm, depressions, and fire. Based on this, at the beginning of the treatment, we should treat the symptoms firstly by resolving phlegm, relieving depression and clearing fire, and commonly use Huanglian Wendan Decoction (黄连温胆汤), Yigan Powder (抑肝散), Chaihu plus Longgu Muli Decoction (柴胡加龙骨牡蛎汤), and Qingxin Zishen Decoction (清心滋肾饮), etc. After improving the symptoms of hot flashes and sweating, irritability, dreaming and frightening, then we should give the prescriptions to tonify kidney yang and nourish kidney yin, in order to eliminate the pathogens and reinforce healthy qi, and to treat both the manifestations and the root cause, so that the symptoms of the patients can be better alleviated.