1.Effect of suppression of platelet-derived growth factor-α receptor expression with antisense oligonucleotide on proliferation and apoptosis of retinal pigment epithelium cell
Yan-yi, PENG ; Mei-yuan, QIU ; Zhi-xiang, DING ; Miao-yun, LIAO ; Cai-wen, FAN
Chinese Journal of Experimental Ophthalmology 2012;30(4):341-345
BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10% fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.
2.Mechanism of Alisma plantago-aquatica Linn. improving chronic glomerulonephritis based on network pharmacology and experimental verification
Zhi-miao QIU ; Bin LU ; Meng-juan WEI ; Li-li JI
Acta Pharmaceutica Sinica 2023;58(6):1430-1440
This study aims to explore the improvement and the mechanism of the
3.Foscarnet sodium for treatment in patients with severe chronic hepatitis B.
Yan-yan YU ; Da-zhi ZHANG ; Xiao-hui MIAO ; Chuan-lin ZHU ; Xia-qiu ZHOU ; Hao YU ; Chong-wen SI
Chinese Journal of Hepatology 2006;14(11):814-816
OBJECTIVETo investigate the effectiveness of foscarnet sodium in the treatment of severe chronic hepatitis B.
METHODSTwo hundred and eight patients were enrolled in a multicenter, double-blind, controlled study. The patients received foscarnet sodium (foscarnet group) or saline (control group) injections for 4 weeks, and were then followed for 24 weeks.
RESULTSHBV DNA negative rate was 12.8% in the foscarnet group and 7.1% in the control group at the end of treatment; and it was 5.5% and 3.0% at the end of the follow-up period respectively (P > 0.05). The rate of HBV DNA decrease of more than 2 log copies/ml was 53.2% in the foscarnet group and 16.2% in the control group at the end of treatment, and 23.9% and 8.1% (P < 0.01) respectively at the end of the follow-up period. The rate of HBV DNA < 10(5) copies/ml was 64.2% and 30.3% at week 4 in the two groups respectively, and 40.4% and 22.2% (P < 0.01) at the end of the follow-up period. HBeAg negative rate was 17.3% and 5.8% at the end of the treatment, and 22% and 5.4% at the end of the follow-up period (P < 0.01). The rate of HBeAg seroconversion was 12.7% and 3.7% at week 4, and 16.7% and 1.5% at the end of the follow-up period. Response rate was 60.6% and 21.2% at the end of week 4 (P < 0.05).
CONCLUSIONFoscarnet sodium injection has a good effect on severe chronic hepatitis B patients and it is safe to use on them.
Adolescent ; Adult ; Antiviral Agents ; adverse effects ; therapeutic use ; Double-Blind Method ; Female ; Foscarnet ; adverse effects ; therapeutic use ; Hepatitis B, Chronic ; drug therapy ; Humans ; Male ; Middle Aged ; Young Adult
4.Effect of kaolin combined with propranolol on paraquat concentration in lungs of poisoned mice
Xin-Jun MIAO ; Xiu-Hua ZHU ; Yu-Xi CHEN ; Xian-Ke QIU ; Yong LI ; Zhi-Li CHEN ; Ruo-Si ZHANG
Shanghai Journal of Preventive Medicine 2015;27(9):536-539
[ Objective ] To observe the effect of kaolin and propranolol on paraquat ( PQ ) concentration in the lungs of poisoned mice. [ Methods] A group of 144 ICR mice were randomly divided into 3 groups:PQ, treatment, and control.Then 100 mg/kg PQ were intragastrically administrated ( ig) in PQ group and treatment group, while only the same volume normal saline was given in control group.And then 48 g/kg kaolin combined with 3.2 mg/kg propranolol were administered in treatment group immediately after poisoning while only the same volume of normal saline was given in the other two groups. Pathological examination was done and PQ concentration in lungs of the mice detected 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, 24 h after poisoning. [ Results] In the lungs of the mice in PQ group occurred alveolar capillary expansion, endothelial cell swelling, small or large sheet-shaped inflammation cell infiltration and mainly neutrophils while in treatment group the above lesions were apparently alleviated.In PQ and treatment groups, PQ concentration in lungs of both groups rose significantly 0.5 h after poisoning and up to peak at 4 h.But PQ concentration in lungs decreased significantly in treatment group from 4 h to 24 h after poisoning ( P<0.05) , as compared with that in PQ group. [ Conclusion] PQ concentration in lungs of the poisoned mice was decreased and the injury alleviated when they were treated with kaolin combined with propranolol.It is held that further research is worth doing in clinical practice.
5.A phase I dose-finding trial of hyperthermic intraperitoneal docetaxel combined with cisplatin in patients with advanced-stage ovarian cancer
Zhi-yao YOU ; Miao-fang WU ; Hui LI ; Yan-fang YE ; Li-juan WANG ; Zhong-qiu LIN ; Jing LI
Journal of Gynecologic Oncology 2024;35(1):e1-
Objective:
To identify the maximum tolerated dose (MTD) of docetaxel combined with a fixed dose of cisplatin (75 mg/m 2 ) delivered as hyperthermic intraperitoneal chemotherapy (HIPEC) in patients with ovarian cancer.
Methods:
In this phase I trial, a time-to-event Bayesian optimal interval design was used.Docetaxel was given at a starting dose of 60 mg/m2 and was increased in 5 mg/m2 increments until the MTD was determined or the maximum dose level of 75 mg/m2 was reached. The doselimiting toxicity (DLT) rate was set at 25%, with a total sample size of 30 patients. HIPEC was delivered immediately following debulking surgery at a target temperature of 43°C for 90 minutes.
Results:
From August 2022 to November 2022, 30 patients were enrolled. Among the patients who received a dose of docetaxel ≤65 mg/m2 , no DLT was reported. DLTs were observed in one patient who received 70 mg/m2 docetaxel (grade 3 anaemia) and in three patients who received 75 mg/m2 docetaxel (one case of grade 3 anaemia, one case of grade 3 hepatic impairment and one case of grade 4 thrombocytopenia). Patients treated with docetaxel 75 mg/m2 in combination with cisplatin 75 mg/m2 had an estimated DLT rate of 25%, which was the closest to the target DLT rate and was therefore chosen as the MTD.
Conclusion
Docetaxel, in combination with a fixed dose of cisplatin (75 mg/m2), can be used safely at intraperitoneal doses of 75 mg/m2 in ovarian cancer patients who received HIPEC (43°C, 90 minutes) following debulking surgery.
6.A phase I dose-finding trial of hyperthermic intraperitoneal docetaxel combined with cisplatin in patients with advanced-stage ovarian cancer
Zhi-yao YOU ; Miao-fang WU ; Hui LI ; Yan-fang YE ; Li-juan WANG ; Zhong-qiu LIN ; Jing LI
Journal of Gynecologic Oncology 2024;35(1):e1-
Objective:
To identify the maximum tolerated dose (MTD) of docetaxel combined with a fixed dose of cisplatin (75 mg/m 2 ) delivered as hyperthermic intraperitoneal chemotherapy (HIPEC) in patients with ovarian cancer.
Methods:
In this phase I trial, a time-to-event Bayesian optimal interval design was used.Docetaxel was given at a starting dose of 60 mg/m2 and was increased in 5 mg/m2 increments until the MTD was determined or the maximum dose level of 75 mg/m2 was reached. The doselimiting toxicity (DLT) rate was set at 25%, with a total sample size of 30 patients. HIPEC was delivered immediately following debulking surgery at a target temperature of 43°C for 90 minutes.
Results:
From August 2022 to November 2022, 30 patients were enrolled. Among the patients who received a dose of docetaxel ≤65 mg/m2 , no DLT was reported. DLTs were observed in one patient who received 70 mg/m2 docetaxel (grade 3 anaemia) and in three patients who received 75 mg/m2 docetaxel (one case of grade 3 anaemia, one case of grade 3 hepatic impairment and one case of grade 4 thrombocytopenia). Patients treated with docetaxel 75 mg/m2 in combination with cisplatin 75 mg/m2 had an estimated DLT rate of 25%, which was the closest to the target DLT rate and was therefore chosen as the MTD.
Conclusion
Docetaxel, in combination with a fixed dose of cisplatin (75 mg/m2), can be used safely at intraperitoneal doses of 75 mg/m2 in ovarian cancer patients who received HIPEC (43°C, 90 minutes) following debulking surgery.
7.A phase I dose-finding trial of hyperthermic intraperitoneal docetaxel combined with cisplatin in patients with advanced-stage ovarian cancer
Zhi-yao YOU ; Miao-fang WU ; Hui LI ; Yan-fang YE ; Li-juan WANG ; Zhong-qiu LIN ; Jing LI
Journal of Gynecologic Oncology 2024;35(1):e1-
Objective:
To identify the maximum tolerated dose (MTD) of docetaxel combined with a fixed dose of cisplatin (75 mg/m 2 ) delivered as hyperthermic intraperitoneal chemotherapy (HIPEC) in patients with ovarian cancer.
Methods:
In this phase I trial, a time-to-event Bayesian optimal interval design was used.Docetaxel was given at a starting dose of 60 mg/m2 and was increased in 5 mg/m2 increments until the MTD was determined or the maximum dose level of 75 mg/m2 was reached. The doselimiting toxicity (DLT) rate was set at 25%, with a total sample size of 30 patients. HIPEC was delivered immediately following debulking surgery at a target temperature of 43°C for 90 minutes.
Results:
From August 2022 to November 2022, 30 patients were enrolled. Among the patients who received a dose of docetaxel ≤65 mg/m2 , no DLT was reported. DLTs were observed in one patient who received 70 mg/m2 docetaxel (grade 3 anaemia) and in three patients who received 75 mg/m2 docetaxel (one case of grade 3 anaemia, one case of grade 3 hepatic impairment and one case of grade 4 thrombocytopenia). Patients treated with docetaxel 75 mg/m2 in combination with cisplatin 75 mg/m2 had an estimated DLT rate of 25%, which was the closest to the target DLT rate and was therefore chosen as the MTD.
Conclusion
Docetaxel, in combination with a fixed dose of cisplatin (75 mg/m2), can be used safely at intraperitoneal doses of 75 mg/m2 in ovarian cancer patients who received HIPEC (43°C, 90 minutes) following debulking surgery.
8.Clinical study of umbilical cord-derived mesenchymal stem cells for treatment of nineteen patients with steroid-resistant severe acute graft-versus-host disease.
Guang-hua CHEN ; Ting YANG ; Hong TIAN ; Man QIAO ; Hui-wen LIU ; Cheng-cheng FU ; Miao MIAO ; Zheng-min JIN ; Xiao-wen TANG ; Yue HAN ; Guang-sheng HE ; Xu-hui ZHANG ; Xiao MA ; Feng CHEN ; Xiao-hui HU ; Sheng-li XUE ; Ying WANG ; Hui-ying QIU ; Ai-ning SUN ; Zhi-zhe CHEN ; De-pei WU
Chinese Journal of Hematology 2012;33(4):303-306
OBJECTIVETo evaluate the safety and efficacy of umbilical cord-derived mesenchymal stem cells (MSCs) infusion in patients with steroid-resistant severe acute graft-versus-host disease (aGVHD).
METHODSA total of 19 patients with steroid-resistant severe aGVHD received MSCs infusion treatment. The treatment response, transplantation-related mortality, events associated with infusion and relapse rate were analyzed.
RESULTSTwo patients with grade II, 5 patients with grade III and 12 patients with grade IV aGVHD received a total of 58 infusions of MSCs. The mean total dose of MSCs was 2.13 (range 0.60 - 7.20)×10(6) cells per kg bodyweight. Seven patients received one infusion, 2 patients received two infusions, and 10 patients received three or more infusions. Eleven patients had a complete response and 4 had a partial response and 4 had no response. No patients had side-effects during or immediately after infusions, and no MSCs related tumorigenesis was detected to date. Eleven patients survived and 8 died, 4 for aGVHD, 1 for infection and 2 for aGVHD with concomitant infection and 1 for underlying leukemia relapse. The cell viability of freshly prepared MSCs is 93% (92% - 95%) by trypan blue staining. The cell viability of programmatically frozen and thawed MSCs is 72% (70% - 74%).
CONCLUSIONInfusion of umbilical cord-derived MSCs expanded in vitro is an effective therapy for patients with steroid-resistant severe aGVHD without negative impact on relapse. Freshly prepared MSCs are superior to frozen and thawed cells in terms of cell viability.
Adolescent ; Adult ; Cord Blood Stem Cell Transplantation ; Female ; Graft vs Host Disease ; etiology ; surgery ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; Middle Aged ; Steroids ; pharmacology ; Survival Rate ; Umbilical Cord ; cytology ; Young Adult
9.The assemblage, purification and characterization of EV71 VLPs expressed in baculovirus.
Lei CAO ; Yao YI ; Jing-Dong SONG ; Miao-Miao TIAN ; Rui-Guang TIAN ; Qing-Ling MENG ; Feng QIU ; Zhi-Yuan JIA ; Sheng-Li BI
Chinese Journal of Virology 2012;28(3):201-206
To construct a recombinant expression plasmid Bacmid-P1-3CD containing the P1 and 3CD genes of enterovirus 71(EV71), the P1 and 3CD genes were cloned into the same baculovirus shuttle vector (Bacmid). Recombinant AcMNPV-P1-3CD was obtained by transfecting the Bacmid-P1-3CD into the insect cell line of S f9. With the IFA and Western-blot methods for identification of expression products confirmed that the target protein was expressed in interior of infected S f9 cells. Electron microscopy showed that the structural protein capsid P1 was cut by virus-encoded protease 3CD and assembled into EV71 virus like particles (VLPs) about 27nm diameter. Different values of MOI and time points of expression were compared to explore the optimal expression condition, and the results showed that the time point could be a more important factor. Then we used S f9 cells with serum-free medium in CellSTACK-10 Culture Chambers to produce EV71 VLPs in the confirmed condition. After purification of VLPs by density gradient centrifugation, we observed on SDS-PAGE profile the purified sample contained three major proteins whose molecular masses corresponded to those of VP1 (39kD), VP0 (34kD) and VP3 (26kD) as well as the intact structure, which can be greatly used for further study in protein structure and genetic engineering vaccine research.
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Baculoviridae
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genetics
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metabolism
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Cell Line
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Enterovirus A, Human
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genetics
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isolation & purification
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physiology
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ultrastructure
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Gene Expression
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Spodoptera
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Viral Proteins
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genetics
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metabolism
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Virion
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genetics
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isolation & purification
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physiology
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ultrastructure
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Virus Assembly
10.Expression level and clinical significance of MEF2C gene in adult acute myeloid leukemia.
Ling Zhi YAN ; Su Ning CHEN ; Xue Feng HE ; Yun ZHAO ; Xiu Yan ZHANG ; Li Li WU ; Na Na PING ; Xiao Yu XU ; Ai Ning SUN ; Hui Ying QIU ; Xiao Wen TANG ; Yue HAN ; Cheng Cheng FU ; Zheng Ming JIN ; Miao MIAO ; De Pei WU
Chinese Journal of Hematology 2018;39(8):682-685