1.Prevention effect of 5-fluorouracil natural delayed-release microballoons on proliferative vitreoretinopathy
Xiao, CHEN ; Shuai, QIN ; Xu-dong, FU ; Qin, DING
Chinese Journal of Experimental Ophthalmology 2011;29(6):501-504
Background Proliferative vitreoretinopathy(PVR) is a tissue repair prevention and treatment of PVR in clinic.Natural delayed release microballoons are therefore becoming a hot spot for its easy manipulation,large lading dose and long acting duration.Objective This study was to evaluate the effect of 5-fluorouracil natural delayed release microballoons on the prevention of PVR.Methods The lymphocytes were collected from clean pigment rabbit to prepare the 8×107/ml cell suspension with complete culture fluid.PVR models were established in 45 healthy pigment rabbits by intravitreal injection of lymphocyte suspension.The animals were randomly divided into 3 groups and 15 rabbits for each.0.1ml normal saline,10g/L or 20g/L 5-fluorouracil natural delayed release microballoons were injected into vitreous cavity respectively.PVR was graded on Fastenberg's method under the slit lamp in 1,2,4,8 weeks.The animals were sacrificed and retinas were obtained for the histopathological and ultrastructural examination in the eighth week after administration of drug.Results The numbers of eyes with different grades of PVR were significantly different among 3 groups in 1 week,2,4,8 weeks(P<0.05).The eye numbers with PVR was significant less in 20g/L Fu group than those of 10g/L Fu group and normal saline group(P<0.05).There was statistical difference in PVR ranking among these 3 groups in 8 weeks after injection of drug(H=46.795,P<0.05).The morphology and ultrastructure of retinas under the light microscope and transmission electron microscope were near normal in all of the three groups.Conclusion Implantation of 5-fluorouracil natural delayed release microballoons into vitreous cavity is effective and safe in preventing PVR in experimental model,and the therapeutic effect of microballoons with 20g/L 5-Fu is better.
2.The role of eNOS on the regulatory effects of EPO on mitochondrial biogenesis in cardiomyocytes exposed to chronic hypoxia
Chuan QIN ; Lin CHEN ; Yinbing XIAO
Chongqing Medicine 2015;(16):2167-2169,2173
Objective To explore the role of endothelial nitric oxide synthase(eNOS) in the regulatory effects of erythropoie‐tin (EPO) on mitochondiral biogenesis in cardiomyocytes exposed to chronic hypoxia .Methods H9c2 cardiomyocytes were cultured in the environment of hypoxia for 1 week(94% N2 ,5% O2 ) ,establishing the chronic hypoxic cardiomyocyte model .All the cells were divided into 3 groups :HC(chronic hypoxic control) ,HE[treated with chronic hypoxia and 20 U/mL recombinant human EPO (rhEPO) ]and HR(cells transfected with eNOS shRNA plasmid and treated with 20 U/mL rhEPO and chronic hypoxia) .Fluores‐cent probe was used to detect the changes of mitochondial number .Mitochondial DNA (mtDNA) relative express level was assayed by RT‐PCR .The expression and phosphorylation of eNOS protein were analyzed with Western blot .Results rhEPO significantly increased the phosphorylation of eNOS and elavated the mitochondialt number and mtDNA (P< 0 .05) .shRNA interference on eNOS significantly blocked all the above changes induced by rhEPO (P<0 .05) .Conclusion Phosphory lation of eNOS is the im‐portant signalling pathway for the enhanced mitochondrial biogenesis in cardiomyocytes exposed to chronic hypoxia by EPO .
3.Preparation of recombinant firefly luciferase by a simple and rapid expression and purification method and its application in bacterial detection
Qin XIAO ; Hui CHEN ; Jinming LIN
Journal of Pharmaceutical Analysis 2010;22(2):97-101
A simple and rapid expression and purification method of recombinant firefly luciferase was developed for bacteria detection. A modified luciferase gene from North American firefly Photinus pyralis was cloned into pET28a expression vector and the recombinant protein was produced in Escherichia coli BL21. The recombinant luciferase, equipped with a polyhistidine affinity tag, was purified by immobilized metal ion affinity chromatography (IMAC). The approach generated an abundant expression and an efficient purification of a recombinant luciferase with final yield 1.995mg/L of cell culture. Experiments on the recombinant luciferase also showed that the relative light units (RUL) of the enzyme were 5.8×108, and the specific activity was 2.9×1010 RLU/mg. By applying adenosine triphosphate (ATP) bioluminescence to detection of the coin bacteria using the recombinant protein, the ATP content of bacteria was 9.48×10-16mol/mL, and was identical to the bacteria counts (4500CFU/mL) in order of magnitude. Taken together, our results provided a simple and efficacious method of the preparation of recombinant luciferase, which could be applied in the determination of bacteria via ATP bioluminescence.
4.Toxicity of levodopa and dopamine on PC12 Cells and neuroprotective effect of several anti-Parkinson drugs on the toxicity
Qin XIAO ; Shengdi CHEN ; Jian FEI
Chinese Journal of Geriatrics 2003;0(07):-
Objective To investigate the toxicity of levodopa and dopamine on PC12 cells and neuroprotection of several anti-Parkinson drugs i.e. amantadine,pergolide and selegiline. Methods The possible cytotoxicity of levodopa and dopamine at different dosage on rat pheochromocytoma PC12 cells and the effects of some anti-Parkinson drugs (amantadine,pergolide and selegiline) on levodopa- or dopamine-induced cytotoxicity were determined by MTT assay and flow cytometry. Results There was a concentration- and time-dependent decrease in cell viability and a concentration-dependent increase in apoptotic cells induced by levodopa and dopamine ( P
5.Effects of morphine on PTEN expression and NF-κB activity in human gastric carcinoma cell line MGC-803
Yi QIN ; Jing CHEN ; Zhiling XIAO ; Yubo XIE ; Qiang XIAO
Chinese Journal of Anesthesiology 2010;30(12):1446-1448
Objective To investigate the effects of morphine on PTEN expression and NF-κB activity in human gastric carcinoma cell line MGC-803.Methods The human gastric cancer cell line MGC-803 was purchased from Cell Biology Research Institute,Chinese Academy of Sciences,and cultured in DMEM liquid culture medium.The cells were randomly divided into 2 groups(n = 6 each): control group and morphine group.The cells was exposed to 0.1 μmol/L morphine in morphine group.The apoptosis was assessed by flow cytometry after being incubated with morphine for 24 h.PTEN expression and NF-κB activity were detected using RT-PCR and Western blot.Results The apoptotic rate was significantly increased,PTEN expression was up-regulated and NF-κB activity was significantly decreased in morphine group compared with control group(P < 0.05).Conclusion Morphine can promote the apoptosis in human gastric cancer cells by up-regulating PTEN expression and decreasing NF-κB activity.
6.Consistency analysis of optical coherence tomography angiography and fundus fluorescein angiography in the diagnosis of central retinal vein occlusion
Miao ZENG ; Xiao CHEN ; Yanping SONG ; Qin DING ; Zhongshan CHEN
Chinese Journal of Ocular Fundus Diseases 2016;32(4):362-366
Objective To assess the consistency of diagnostic results using optical coherence tomography angiography (OCTA) and fundus fluorescein angiography (FFA) in the central retinal vein occlusion (CRVO).Methods This is a retrospective case series of 26 eyes of 26 patients with CRVO.There were 10 females (10 eyes) and 16 males (16 eyes).The mean age was (49.19±10.50) years.The mean course of the disease was (27.81± 21.60) days.Simultaneous OCTA and FFA were performed in all patients using 7-standard field of Early Treatment Diabetic Retinopathy Study (ETDRS) to evaluate the microaneurysms,nonperfused areas,optical disc/retinal neovascularization and maeular edema.The consistency was evaluated using weighted Kappa statistic values.Kappa≥0.76,consistency is excellent;0.60≤Kappa<0.75,consistency is good;0.40≤Kappa<0.60,consistency is general;Kappa<0.40,consistency is poor.Results Based on OCTA,microaneurysms were found in 23 eyes,nonperfused areas in 16 eyes,optical disc/retinal neovascularization in 8 eyes and macular edema in 21 eyes.Based on FFA,23 eyes were diagnosed to have microaneurysms,14 eyes have nonperfused area,8 eyes have optical disc/retinal neovascularization,22 eyes have macular edema.The consistency was excellent for microaneurysms and optical disc/retinal neovascularization (Kappa=0.772,0.766;P<0.01),good for nonperfused areas and macular edema (Kappa =0.703,0.600,P< 0v01).Conclusion There is high consistency between OCTA and FFA in the diagnosis of microaneurysms,macular edema,nonperfused areas and optical disc/retinal neovascularization in CRVO patients.
7.Effects of chronic hypoxia on left and right ventricular function and the expression of cardiac TRPC channels in rats.
Hui-Qin CHEN ; Mo-Jun LIN ; Xiao-Ru LIU
Chinese Journal of Applied Physiology 2014;30(3):274-278
OBJECTIVETo explore the effects of chronic hypoxia on left and right ventricular function and the expression of cardiac transient receptor potential canonical (TRPC) channels in rats.
METHODSForty eight SD male rats were randomly divided into control group (CON) and chronic hypoxic pulmonary hypertension model group (CH) (n = 24). In CH group, rats were exposed in chronic hypoxia environment (10% +/- 0.2% O2) to induce myocardial hypertrophy. After 3 weeks, mean systemic arterial pressure (mSAP), right ventricular systolic pressure (RVSP), left ventricular systolic pressure (LVSP), left or right ventricular pressure maximum rate of rise (LV/RV + dp/dt(max)), left or right ventricular pressure maximum rate of descent (LV/RV-dp/dt(max)), right ventricular hypertrophy index (RVMI) and left ventricular hypertrophy index (LVMI) were measured. Left and right ventricular myocardium tissue sections were stained by HE and observed under light microscope. Real-time polymerase chain reaction (real-time-PCR) and Western blot were performed to detect the expression of TRPC subfamily.
RESULTSRVSP, RVMI, RV + dp/dt(max) and RV-dp/dt(max) were markedly elevated in CH group (P < 0.01) in comparison to CON group. LVMI was markedly reduced in CH group in comparison to CON group (P < 0.01). LVSP, LV + dp/dt(max) and LV- dp/dt(max) had no significant changes in CH group in comparison to CON group. Right ventricular myocardial cells of CH group became thick, the nuclei stained deeply, the shape of nuclei became not regularity. Left ventricular myocardial fibers did not change significantly. There was significant difference in the levels of mRNA and protein of TRPC1 between CON and CH groups.
CONCLUSIONFor three weeks exposed to chronic hypoxia induced right ventricular hypertrophy specifically, raised the mRNA and protein expression of TRPC1 on right ventricular myocardial cells . TRPC1 might be involved in the development of cardiac hypertrophy.
Animals ; Disease Models, Animal ; Hypertension, Pulmonary ; metabolism ; physiopathology ; Hypoxia ; metabolism ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley ; Transient Receptor Potential Channels ; metabolism ; Ventricular Function, Left ; physiology ; Ventricular Function, Right ; physiology
8.How to improve psychological quality of medical students in histology and embryology teaching
Xingshu CHEN ; Yunlai LIU ; Maolin QIN ; Hongli LI ; Lan XIAO
Chinese Journal of Medical Education Research 2011;10(10):1208-1210
It is important and necessary for the teaching process in histology and embryology integrated by psychological quality.The psychological quality of teachers can be improved by professional training and by themselves.Teachers should teach everyone differently according to the different psychological character of medical students who are born after 1990.Teachers can improve psychological quality of medical students in teaching process including the discussion,visiting the embryo sample and the second class.
9.Clinical study of acute promyelocytic leukemia with expression of BCR 3 isoform of PML-RARa fusion gene
Xin WANG ; Lin LIU ; Jianbin CHEN ; Jianyu WANG ; Qin XIAO
Chongqing Medicine 2013;(28):3382-3384,3387
Objective To assess the efficacy of combination therapy with all-trans-retinoic acid (ATRA ) and arsenic trioxide (ATO) in treating patients with acute promyelocytic leukemia (APL) .Methods A retrospective study was conducted to evaluate the efficacy of combining ATO with ATRA based induction therapy ,followed by 3 courses of consolidation chemotherapy and 2-year sequential ATO/ATRA maintenance therapy in newly diagnosed APL ,and the efficacy between high risk group and low/inter-mediate risk group ,also between different PML-RARa isoform sub-group were compared .Results In high risk group and low/in-termediate risk group ,the complete remission(CR)rates were 70 .0% and 96 .9% (P=0 .04) ,respectively ;the 3 years overall sur-vival rates(OS) and disease free survival rates (DFS) were(70 .0 ± 14 .5)% ,(96 .9 ± 3 .1)% ,P= 0 .01 and(66 .7% ± 19 .2)% , (93 .8 ± 6 .1)% ,P=0 .08 ,respectively .In BCR1 group and BCR3 group ,the CR were 78 .6% and 95 .6% (P=0 .14) ,respectively ;the rates of 3 years OS and DFS were(95 .7 ± 4 .3)% ,(78 .6 ± 11 .0)% ,P=0 .18 ,and(92 .9 ± 6 .9)% ,(87 .5 ± 11 .7)% ,P=0 .24 , respectively .Conclusion The results indicate that ATO based first-line protocol is highly effective for treatment of newly diagnosed APL ,especially for the PML-RARa BCR3 isoform APL .
10.Ginsenosides-induced bone marrow mesenchymal stem cells promote nerve regeneration in traumatic brain injury
Jun QIN ; Jiakang CHEN ; Xuedong LI ; Yongjun MAI ; Zhenyong XIAO
Chinese Journal of Tissue Engineering Research 2015;(45):7292-7297
BACKGROUND:Previous studies have shown that bone marrow mesenchymal stem cels in the treatment of neurological diseases have achieved some success, which can promote neurological alterations; however, there is no breakthrough on gene and drug regulation. OBJECTIVE:To investigate the influence of ginsenosides-induced differentiation of bone marrow mesenchymal stem cels on nerve regeneration after traumatic brain injury. METHODS: A traumatic brain injury model was built in rats using hydraulic shock method, and then rat models were randomly divided into model group (traumatic brain injury group), bone marrow mesenchymal stem cel group, ginsenosides group (ginsenosides induced differentiation of bone marrow mesenchymal stem cels). At 2 weeks after transplantation, western blot assay was used to detect protein expression levels of nerve growth factor and brain-derived neurotrophic factor, immunohistochemistry assay used to detect the number of BrdU-positive cels. At 1, 3 days and 1, 2 weeks after transplantation, modified neurological severity scores were recorded. RESULTS AND CONCLUSION: The expression levels of nerve growth factor and brain-derived neurotrophic factor protein were significantly higher in the ginsenosides group than the bone marrow mesenchymal stem cel group and model group (P < 0.05). The number of BrdU positive nerve cels was also higher in the ginsenosides group than the bone marrow mesenchymal stem cel group and model group (P < 0.05). At 3 days and 1, 2 weeks after transplantation, the modified neurological severity scores in the ginsenosides group were lower than those in the bone marrow mesenchymal stem cel group and model group (P< 0.05). These findings indicate that ginsenoside-induced bone marrow mesenchymal stem cel transplantation can promote nerve regeneration in rats with traumatic brain injury, which has better outcomes than bone marrow mesenchymal stem cel transplantation alone.