1.Effect of IGF-1 on expressions of VEGF under suppression of HIF-1α by shRNA in cultured human RPE cells
Chinese Journal of Experimental Ophthalmology 2012;30(4):316-319
BackgroundProliferative vitreo-retinal disease (PVD)is one group of ocular complications marked by the enhanced proliferation of various cells included retinal pigment epithelial (RPE) cells.Insulin-like growth factor-1 (IGF-1)and vascular endothelial growth factor(VEGF) are implicated in the aberrant cell proliferation and pathological neovascularization that characterizes PVD,but the signaling mechanism is unclear now. Objective This study was to explore the effect of IGF-1 on VEGF in cultured human RPE cells under the small hairpin loop RNA (shRNA) keeping hypoxia-inducible factor-1 α ( HIF-1 α) silencing. Methods Human retinas were isolated from 4 healthy male donors,and the RPE cells were harvested and cultured.The ceils were identified using anti-human keratin antibody.The third to fifth generation of human RPE cells were used in the experiment.One target site of HIF-1α mRNA was chosen by certain design principle,and shRNA was designed and synthesized by the target site and transferred into the cells in vitro,and then the cells were cultivated with 50 μg/L IGF-1 for 24 hours.The mRNA and protein expressions of HIF-1α and VEGF were detected by RT-PCR and Western blot respectively. Results Cultured human RPE cells showed the flat irregularly multangular shape,and 97%cells appeared the positive response for keratin.HIF-1α mRNA expression in human RPE cells was significantly lower in 50 μg/L IGF-1 group than the 0 pg/L IGF-1 group ( 1.49±0.18 vs 1.46±0.17 ) ( t =0.335,P =0.743 ),however,the expressing levels of HIF-1α protein( 1049.86±172.54 vs 0.00±0.00) and VEGF mRNA(0.95±0.15 vs 0.35±0.07) and VEGF protein (391.98±56.77 vs 214.36±37.15)were raised in the 50 μg/L IGF-I group compared with 0 μg/L IGF-1 group (t=16.098,9.935,6.928,P<0.05).After the HIF-1α-specific shRNA was transferred into cultured RPE cells,the expressions of both HIF-1α mRNA and its protein significantly decreased in RPE cells under 50 μg/L IGF-1 concentration condition( F=68.679,89.904,P=0.000),moreover,the expression of VEGF mRNA and its protein were significantly lowed(F=21.770,6.205,P<0.05). ConclusionsIGF-1 promotes the accumulation of HIF-1α protein and induce the expression of VEGF in human RPE cells,which probably play a pivotal role in the development of PVD.
2.Electrogastrographic parameters from 40 healthy subjects 6 to 14 years of age.
Yan YANG ; Hui-min YAN ; Ming LI
Chinese Journal of Pediatrics 2005;43(9):697-698
Adolescent
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Electromyography
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Female
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Humans
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Male
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Stomach
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physiology
3.Mode and developing tendency of diagnostic and treated system of TCM
Jiansheng LI ; Min ZHAO ; Ming CHEN
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(06):-
Based on the formation and developing course of the system of diagnosis and treatment of Traditional Chinese Medicine(TCM),to explain three modes of diagnosis and treatment,including differential diagnosis of diseases therapy,determination of treatment based in pathogenesis obtained through differentiation of symptoms and signs,illness-syndrome coalescent.The latter one consist of the mode of illness-syndrome coalescent of TCM,which is based on diagnosing disease in TCM,to carry out determination of treatment based in pathogenesis obtained through differentiation of symptoms and signs and that based on diagnosing disease in modern medicine,to carry out determination of treatment based in pathogenesis obtained through differentiation of symptoms and signs of TCM.Meanwhile,to approach the advantages and flaws among different modes of diagnosis and treatment,accordingly we could offer theory evidences for elevating the level of curing disease of TCM.
4.Detection of anti-HLA antibodies and its clinical implication after kidney transplantation
Liuyang LI ; Lijuan HU ; Min LI ; Ming ZHAO
Chinese Journal of Laboratory Medicine 2001;0(01):-
Objective To evaluate the level of specific anti human leukocyte antigen IgG antibodies and its effect on renal transplantation Methods Specific anti HLA IgG antibodies of 685 serum samples were measured by the micro enzyme linked immunosorbent assay with Lambda Antigen Tray and Lambda Antigen Tray Mixed Results There were 12% positive recipients of anti HLA IgG antibodies among 685 renal transplantation patients The rates of allografts rejection after transplantation were 50% and obviously higher in the sensitized recipients than in non sensitized ones ( P
5.Investigation and analysis on current situation of clinical teaching
ming-min, GU ; ming-de, ZHU ; ai-li, NUER ; xiao-bo, LI ; hui, XIA ; gang, HUANG
Journal of Shanghai Jiaotong University(Medical Science) 2008;0(S1):-
Objective To investigate the clinical teaching situation by using developmental inspection of School of Medicine,Shanghai Jiaotong University(SJTU-SM),and to put forward some suggestions. Methods By checking questionnaires and informal discussions,the relevant information was collected and analyzed by using SPSS statistics sofware. Results The clinical teaching quality of SJTU-SM was basically satisfied.The satisfaction from internship of grade 2004 was better than that of grade 2003.However,some problems in clinical teaching must be improved.Conclusion The investigation showed that the clinical teaching quality of SJTU-SM is being improving.However,in order to achieve the international accreditation standards,the quality guarantee system of clinical teaching need to be further perfected.
6.Ghrelin attenuates renal fibrosis and apoptosis of obstructive nephropathy
Guangxi SUN ; Lipei FAN ; Xianghua SHI ; Min LI ; Ming ZHAO
The Journal of Practical Medicine 2015;(7):1102-1106
Objective To investigate the effect and underlying mechanisms of ghrelin in a rat model of renal fibrosis. Methods Male Sprague-Dawley rats were divided into 4 groups , including sham operation +saline or brain gut peptide treatment group , model + saline or brain gut peptide treatment group. Unilateral ureteral obstruction (UUO) was established by left ureteral ligation. 7 days and 14 days after operation, the rats were sacrificed , while the kidney tissue of obstruction side was harvested for pathlogical changes through Masson coloration. Expression of α-smooth muscle actin (α-SMA), transforming growth factor beta1 (TGF-β1) and phosphorylated Smad3 (p-Smad3) in renal tissues were analyzed through immunohistochemistry. Expression of α-SMA and TGF-β1 mRNA was detected by real-time-PCR. Apoptosis kidneys cells were marked with TUNEL. Results Ghrelin inhibited renal fibrosis by reducing the production of collagen , restraining extracellular matrix (ECM) deposition and decreasing the expression of α-SMA. Meanwhile, ghrelin inhibited the accumulation of myofibroblasts by blocking the transforming growth factor-β1/Smad3 (TGF-β1/Smad3) signaling pathway. Moreover, ghrelin could attenuate renal tubular cell apoptosis induced by UUO injury. Conclusion Ghrelin can reduce renal fibrosis and renal cell apoptosis induced by UUO , demonstrating that ghrelin is a potent antifibrotic agent that may have therapeutic potential for patients with obstructive nephropathy.
7.Inhibiting cyclooxygenase and 5-lipoxygenase activities is an anti-inflammatory mechanism of Huzhang Gout Granule.
Yifei WANG ; Wenjing WU ; Ming ZHANG ; Min ZHOU ; Bin LI
Journal of Integrative Medicine 2009;7(10):963-8
Objective: To study the role of estrogen receptor (ER) and progesterone receptor (PR) in the formation of cholesterol calculus and investigate the effects of Shengqing CapsuObjective: To observe the effects of Huzhang Gout Granule (HZGG), a compound traditional Chinese herbal medicine, on cyclooxygenase (COX) and 5-lipoxygenase (5-LOX) activities, the two important oxidases in the course of inflammation, so as to investigate the possible anti-inflammatory mechanism of HZGG. Methods: After stimulating the blood sample of healthy volunteer with calcium ionophore A23187, concentration of thromboxane B(2) (TXB(2)) in the healthy volunteer's blood was detected by enzyme-linked immunosorbent assay (ELISA) to observe the effects of HZGG at low- and high-dose on the activity of COX-1, with aspirin as control drug. The concentration of prostaglandin I(2) (PGI(2)) in the healthy volunteer's blood sample, in which aspirin was added to destroy activity of COX-1 beforehand and which was stimulated with lipopolysaccharide, was detected by ELISA method to observe the effects of HZGG on the activity of COX-2, with celecoxib as control drug. In the animal experiment, 40 rats were implanted with sponges soaking in 0.5% arachidonic acid solution in the back to induce inflammatory effusion. Content of leukotriene B4 (LTB4) in the polymorphonuclear leukocytes (PMNs) from the inflammatory effusions was detected with reversed-phase high-performance liquid chromatography (RP-HPLC) to observe the impacts of different doses of HZGG on the activity of 5-LOX, with dexamethasone as control drug. Results: The concentration of TXB(2) in the low-dose HZGG group was higher than those in the high-dose HZGG group and the aspirin group (P<0.05). The concentrations of PGI2 in the low- and high-dose HZGG groups were higher than that in the celecoxib group (P<0.05), but there was no significant difference between the low-dose HZGG group and the high-dose HZGG group (P>0.05). The content of LTB4 in the blank control group was higher than those in the low-dose HZGG group, the high-dose HZGG group or the dexamethasone group (P<0.05) Conclusion:HZGG can reduce the releasing of inflammatory mediators, such as TXB2, PGI2 and LTB4, by inhibiting the activities of COX and 5-LOX.
8.5-Fluorouracil-loaded polylactic acid nanoparticles have a killing effect on gastric cancer cell lines in vitro
Xiaoli LI ; Min NIU ; Ming ZHANG ; Nana ZHANG ; Yao SHI
Chinese Journal of Tissue Engineering Research 2015;(38):6179-6183
BACKGROUND:5-Fluorouracil occupies an important position in the treatment of gastric cancer, but its long-term use can easily induce adverse reactions such as myelosuppression and leukopenia. Polylactic acid and its copolymers have a higher biocompatibility, and their decomposer cannot gather in the body.
OBJECTIVE:To investigate the in vitro cytotoxicity mechanism of 5-fluorouracil-loaded polylactic acid nanoparticles on gastric cancer cel lines.
METHODS:Ten mice were selected in this study. 5-fluorouracil-loaded polylactic acid nanoparticles (1×10-7, 1×10-6, 1×10-5, 1×10-4 mol/L) were prepared using ultrasonic emulsification method. Kiling effect of polylactic acid nanoparticles on gastric cancer cel lines in vitrowere detected. Then, the inhibition rate was calculated at different concentrations.
RESULTS AND CONCLUSIONS: Under the transmission electron microscope, 5-fluorouracil-loaded polylactic acid nanoparticles had good shape and relatively evenly distributed with no adhesions. After drug administration, the drug concentration was 50% at 24 hours and 62.9% at 72 hours. After 48 hours co-culture with single 5-fluorouracil or 5-fluorouracil-loaded polylactic acid nanoparticles, the viability of gastric cancer cels showed a decrease trend with the increase of drug concentrations, and moreover, 5-fluorouracil-loaded polylactic acid nanoparticles had a better cel inhibition ability than the single 5-fluorouracil (P < 0.05). The IC50value of 5-fluorouracil-loaded polylactic acid nanoparticles was significantly lower than that of 5-fluorouracil (P < 0.05). These findings indicate that polylactic acid nanoparticles as good drug carriers have a strong drug loading capacity and increase drug concentration in the body, but cannot reduce the biological activity of 5-fluorouracil, which provide new ideas for the treatment of gastric cancer.
9.Significance of CD20-positive lymphocytes infiltrating in renal allograft biopsies with chronic allograft nephropathy
Jianmin HU ; Ming ZHAO ; Ying GUO ; Hua CHEN ; Min LI
Chinese Journal of Organ Transplantation 2012;33(1):9-13
ObjectiveTo investigate the action mechanism of CD20 lymphocyte infiltration in the renal allograft biopsy with chronic allograft nephropathy (CAN).MethodsCAN cases confirmed by renal biopsy within 2 years after renal transplantation served as study subjects. By using immunohistochemistry,the deposition of C4d and the CD20-positive lymphocytes infiltration in the renal grafts were examined.The clinical follow-up data were analyzed.ResultsForty-four cases of CAN were enrolled in the study, including 13 cases (29.5% ) of CD20-positive lymphocytes infiltration,and 31cases (70.5% )of CD20-negative lymphocytes infiltration. CD20-positive lymphocytes in biopsy showed nodular and scattered lymphocytes infiltration.There were 5 (26.3%)cases of CAN Ⅰ,4 cases (25.0%) of CAN Ⅱ,and 4 (44.4%) of CAN Ⅲ in CD20-positive group.There was no statistically significant difference between the only CAN group and CAN with AR group in CD20-positive rate.Immunohistochemical staining showed there were 12 cases (27.3%) with C4d linear deposition in peritubular capillary endothelial cells (PTC).C4d positive rate had no significant difference among the CAN classifications. There was no significant relationship between C4d deposition and CD20-positive lymphocytic infiltration.The average serum creatinine in CD20-negtive group and CD20-posigtive group was 140.8 ± 22.0 and 183.5 ± 25.5μmol/L before biopsy,and 165.6 ± 37.6 and 242.2 ± 59.1 μmol/L one year after biopsy.The average serum creatinine level in CD20-positive group was higher than in CD20-negtive group before and after biopsy.ConclusionProgressive chronic humoral immunity is high risk in the process of CAN. The CD20-positive lymphocyte infiltration has no relevance with CAN grade and C4d deposition in PTC,but is associated with circulating antibody PRA and allograft long-term outcome. Pathogenetic mechanism may not contribute to chronic humoral immune,but B cells presenting donor antigens,are recognized and activated by T cells as antigen-presenting cells.
10.Effect of Wuzhi capsules on whole blood concentration of Tacrolimus in renal transplantation recipients
Jianmin HU ; Hua CHEN ; Min LI ; Ming ZHAO
Chinese Journal of Tissue Engineering Research 2010;14(5):887-890
BACKGROUND: It has reported that the combination of Tacrolimus and Wuzhi capsules can increase the peak concentration and area under curve of Tacrolimus recipients, however, the effects of this combination on recipients is poorly understood. OBJECTIVE: To investigate the effects of Wuzhicapsules on the blood concentration of Tacrolimus recipients. METHODS: A total of 38 renal transplantation recipients receiving triple therapy regime (Tacrolimus+mycophenolate +Prednisolone) were involved in the study and were divided into the experimental group (n=21) and control group (n=17). Recipients in the experimental group were taking Wuzhi capsules simultaneously, and those in the control group were not taking Wuzhicapsules. The Tacrolimus dosage to therapeutic concentration window and renal function were compared between the two groups at months 1,3 and 6 after transplantation. The incidence rates of acute rejection and diabetes mellitus, and renal and liver functions were observed at 6 months after transplantation. RESULTS AND CONCLUSION: The Tacrolimus dosage to therapeutic concentration window of the experimental group was significantly smaller than that of the control group (P < 0.05). However, incidence rate of acute rejection, diabetes mellitus, and renal and liver functions had no dramatically difference in 2 groups (P > 0.05). Results suggest that Wuzhi capsules may improve the immunosuppressive efficacy of Tacrolimus in renal recipients, therefore, can decrease the toxic effect of Tacrolimus with reduction dosage.