2.Preliminary screening of RPL5 interacting with HSPC238
Jinglin CHEN ; Xiang HUANG ; Jiayu TAN ; Yuheng ZHONG ; Zhidan WAN
Chinese Journal of Immunology 2015;(11):1494-1497
Objective:To construct a bait vector for HSPC238, and to screen the target proteins which interact with the HSPC238.Methods:Gene synthesis method was used to synthetic gene HSPC238, then connected with the pGBKT7 vector after digesting by the sfiIA and sfiIB,to obtain the bait plasmid pGBKT7-HSPC238,then transferred into the yeast strains AH109 with the empty plasmid pGBKT7after sequencing,to observe its self-activating effect in the nutrient deficiencies medium,and further to screen the target proteins which interact with HSPC238 from the human fetal liver cDNA library.Results:The bait vector pGBKT7-HSPC238 was successfully constructed,and it had no self-activating effect through the phenotypic screening,after the yeast two-hybrid technology with literature analysis,we preliminary screened and found that the ribosomal protein L5(RPL5) may be the one of the target proteins which interacted with HSPC238 from the human fetal liver cDNA library.Conclusion: We successfully constructed the bait plasmid vector PGBKT7-HSPC238,and after the yeast two-hybrid technology with literature analysis,we preliminary screened and found that the ribosomal protein L5( RPL5) may be the one of the target proteins which interacted with HSPC238.
3.A preliminary study on regulation of HMOX1, RPS27a, MT2A and UBB by HSPC238
Jiayu TAN ; Xiang HUANG ; Jinglin CHEN ; Yuheng ZHONG
Chinese Journal of Immunology 2016;32(4):509-512
Objective:To investigate the effect of HSPC 238 overexpression or low expression on the regulation of the target protein ( HMOX1, MT2A, UBB, RPS27a ) and the regulation pathways.Methods: To construct the interference vector and overexpression vector of HSPC238 respectively,transfected the HEPG2 cell lines by the liposome method,detect the expression of mRNA and protein of the HSPC 238 and the target proteins by the RT-PCR and Western blot , further to transfer the overexpression plasmids of the target proteins into the HEPG 2 cell lines which had been transfected with interference vector and overexpression vector of HSPC238,the experimental group cell lines were treated with proteasome inhibitor MG 132,to purify the target proteins with nickel column,do immunoblotting with HSPC238 to detect the accumulation situation of the target proteins.Results: The HMOX1,MT2A, RPS27a were downregulated obviously when the HSPC 238 was interfered exogenous;and the MT2A,UBB,RPS27a were up-regulated after the HSPC238 overexpressed.The overexpression plasmid of target proteins were transfected into the HEPG 2 cell lines which have been transfected with interference vector and overexpression vector of HSPC 238 ,compared with the control group ,the target protein band in the experimental group was significantly increased after treatment with the proteasome inhibitor MG 132.Conclusion:The HSPC238 overexpression can upregulate the expression of MT 2A,UBB and RPS27a,and interfering HSPC238 can downregulate the expression of HMOX1,MT2A and RPS27a;the HSPC238 target protein may play a regulatory role through the UPP pathway;the HSPC238 may play a regulatory role on the target proteins through the UPP pathway.
4.Anti-oxidative stress effects of miR-125b on lens epithelial cells and its mechanism
Xiang LI ; Ying ZHANG ; Yalin HUANG ; Zhong WU ; Haoyi GUO
Chinese Journal of Experimental Ophthalmology 2021;39(4):280-288
Objective:To investigate the anti-oxidative stress effects of microRNA 125b (miR-125b) on lens epithelial cells (LECs) and its possible mechanism.Methods:Twenty-four anterior capsule specimens were collected from 24 eyes of 24 age-related cataract patients during phacoemulsification and 20 normal anterior capsule specimens were obtained from 20 eyes of 20 donors in Henan Eye Hospital from July 2018 to March 2019 under the approval of a Medical Ethics Committee of Henan Eye Hospital (No.YKYY20193151).The reverse transcription PCR and Western blot assay were employed to detect and compare the relative expression levels of miR-125b and nuclear factor E2-related factor 2 (Nrf2) in different specimens.The human lens epithelial cell line HLEB-3 was divided into control group and oxidative stress model group.The oxidative stress models were established by coculture with different concentrations (100, 200, 400 μmol/L) of H 2O 2 for 24 hours, and the cells were cultured with normal medium without H 2O 2 in the control group.The reactive oxygen species (ROS) content was detected by DCFH-DA fluorescent probe, and the activities of total-antioxidative capability (T-AOC), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) as well as malondialdehyde (MDA) concentration were detected by ELISA, and compared among the groups.The expression levels of miR-125b and Nrf2 were detected by reverse transcription PCR and Western blot assay, respectively.The cells were transfected with miR-125b mimics, miR-125b control and miR-125b inhibitor for 24 hours, respectively, and ROS content was detected by DCFH-DA fluorescent probe and T-AOC, SOD and GSH-Px activities as well as MDA concentration were detected by ELISA and compared among different transfected groups.A dual luciferase reporter assay was used to assess an association between miR-125b and Nrf2.The expression level of Nrf2 protein was detected by Western blot assay and the expression levels of Nrf2 and Keap1 were assayed and located by immunofluorescence double staining. Results:The relative expression levels of miR-125b and Nrf2 in the normal lens anterior capsule specimens were 0.21±0.03 and 0.27±0.06, which were significantly lower than 0.89±0.05 and 0.84±0.12 in the cataract specimens, respectively ( t=15.355, P<0.05; t=18.647, P<0.05).The relative expression levels of miR-125b and Nrf2 were significantly increased in various H 2O 2 treated groups in comparison with the control group and were gradually elevated with the increase of H 2O 2 concentration (all at P<0.05).Compared with the control group, the T-AOC, SOD and GSH-Px activities were reduced, and ROS content and MDA concentration were significantly ascended (all at P<0.05).Compared with the miR-125b control group, the T-AOC, GSH-Px and SOD activities were increased, and ROS content and MDA concentration were decreased in the miR-125b mimics group (all at P<0.05).In addition, the T-AOC, GSH-Px and SOD activities were significantly weakened, and ROS content and MDA concentration were significantly increased in the miR-125b inhibitor group in comparison with the miR-125b control group (all at P<0.05).Dual luciferase reporter assay showed that miR-125b targeted to the expression of Nrf2 in the H 2O 2 model cells.The fluorescence of Nrf2 in the cytoplasm was the strongest with more nuclear transfer in the miR-125b mimics group, and the expression intensity of Keap1 in the cytoplasm was weaker.The expression of Nrf2 was the weakest with less nuclear transfer in the miR-125b inhibitor group, and the expression level of Keap1 in the cytoplasm was stronger. Conclusions:MiR-125b can enhance the anti-oxidative stress of LECs in age-related cataractous eyes probably by upregulating the expression of Nrf2 and activating the Keap1/Nrf2 signaling pathway.
5.Chemical constituents from Perovskia atriplicifolia.
Jun ZHONG ; Chao-guan HUANG ; Yi-Jiang YU ; Zhong-qiu LI ; Wei WANG ; Xiang-zhong HUANG ; Wen-xing LIU ; Yan YUAN ; Zhi-yong JIANG
China Journal of Chinese Materia Medica 2015;40(6):1108-1113
An investigation on the chemical constituents of the 90% EtOH extract of Perovskia atriplicifolia led to the isolation of fifteen compounds from the EtOAc fraction. Based on the detailed spectral analysis (MS, 1D and 2D NMR), as well as comparison with the literatures, the structures of compounds 1-15 were determined as cirsimaritin (1), salvigenin (2), syringaldehyde (3), vinyl caffeate (4), 2α, 3α-dihydroxyolean-12-en-28-oicacid (5), 2α, 3α-dihydroxyurs-12-en-28-oicacid (6), niga-ichigoside F1 (2α, 3β, 19α, 23- tetrahydroxyurs - 12-en-28-oicacid- O-β-D- glucopyranoside, 7), sericoside (8), 4-epi-niga-ichigoside F1 (2α, 3β, 19α, 24-tetrahydroxyurs-12-en-28-oicacid O-β-D-glucopyranoside, 9), 2α, 3β, 24-trihydroxyolean-12-en-28-oicacid O-β-D-glucopyranosyl-(1 --> 2) - β-D-glucopyranoside (10), pruvuloside A (11), asteryunnanoside A [2α, 3β, 23-trihydroxyolean-12-en-28-oicacid O-β-D-glucopyranosyl-(1 --> 2)-β- D- glucopyranoside,12], rosmarinic acid methyl ester (13), β-sitosterol (14), and daucosterol (15), respectively. Compounds 1-13 were isolated from the Perovskia genus for the first time. All the compounds were obtained from P. atriplicifolia for the first time.
Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Lamiaceae
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chemistry
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Magnetic Resonance Spectroscopy
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Molecular Structure
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Spectrometry, Mass, Electrospray Ionization
6.Studies on chemical constituentsfrom leaves of Acer truncatum.
Xiang-zhong HUANG ; Li-xiang TAN ; Kun GU ; Cong LI
China Journal of Chinese Materia Medica 2007;32(15):1544-1604
OBJECTIVETo study the chemical constituents from the leaves of Acer truncatum.
METHODVarious chromatographic techniques were used to isolate and purify the constituents. The structures of these compounds were elucidated on the basis of spectral analysis.
RESULTSeven compounds were isolated and identified as p-sitosterol (1), beta-amyrin (2), beta-amyrin acetate (3), 3, 5-dihydroxy-4-methoxybenzoic acid (4), astragalin (5), quercetin-3-O-beta-D-galactoside (6), and quercetin-3-O-alpha-L-rhamnoside (7).
CONCLUSIONAll of compounds were isolated from this plant for the first time.
Acer ; chemistry ; Gallic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Kaempferols ; chemistry ; isolation & purification ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry
7.Antiviral therapy for hepatitis B virus-related hepatocellular carcinoma after radical hepatectomy
Yang KE ; Jianhong ZHONG ; Xuemei YOU ; Shengxin HUANG ; Yongrong LIANG ; Bangde XIANG ; Lequn LI
Chinese Journal of Clinical Oncology 2013;(19):1184-1188
Objective:The effect of antiviral therapy for hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) after radical hepatectomy was assessed. Methods:A total of 478 HBV-HCC patients treated by radical hepatectomy were retrospectively col-lected. Patients in the treatment group (n=141) received postoperative lamivudine therapy (100 mg/d), whereas patients in the control group (n=337) did not. Recurrence-free survival rates, overall survival rates, treatments for recurrent HCC and cause of death were com-pared between the two groups. Propensity score matching was also conducted to reduce confounding bias between the groups. Results:The one-, three-, and five-year recurrence-free survival rates didn't significantly differ between the two groups (P=0.778);however, the one-, three-, and five-year overall survival rates in the treatment group were significantly higher than those in the control group (P=0.002). Similar results were observed in the matched data. Subgroup analysis showed that antiviral treatment conferred a significant sur-vival benefit for Barcelona Clinical Liver Cancer stage A/B patients. Following HCC recurrence, more people in the treatment group were able to choose curative treatments than those in the control group (P=0.031). For cause of death, fewer people in the treatment group died of liver failure than those in the control group (P=0.041). Conclusion:Postoperative antiviral therapy increases chances of receiving curative treatments for recurrent HCC and prevents death because of liver failure, thereby significantly prolonging overall sur-vival, especially in early-or intermedian-stage tumors.
8.Short-term results of cutting balloon and intravascular brachytherapy for the treatment of coronary in-stent restenosis
Fei YE ; Shao-Liang CHEN ; Bao-Xiang DUAN ; Jin HUANG ; Zhi-Zhong LIU ; Jie SONG ;
Chinese Journal of Clinical Pharmacology and Therapeutics 1999;0(04):-
0.05); the levels of LL, LI, RRS in CBA group and CBA+IBT group were significantly lower than those in control group(P
9.Comparison of holmium laser enucleation and plasmakinetic enucleation of the prostate for benign prostatic hyperplasia
Zifeng WANG ; Hong ZHANG ; Qun YANG ; Zhong HUANG ; Xinqi ZHANG ; Xinhui NIU ; Yanqing XIANG
Clinical Medicine of China 2015;31(9):831-834
Objective To evaluate and compare the efficacy of holmium laser enucleation of the prostate(HoLEP) and plasmakineticenucleation of the prostate(PKEP) for treating benign prostatic hyperplasia (BPH).Methods A total of 86 cases of BPH were selected from the Shandong Energy Zibo Mining Group Co.Ltd Central Hospital from January 2013 to June 2014.The patients received either HoLEP (40 cases) or PKEP(46 cases) treatment.Clinical data including postvoid residual volume (PVR),international prostate symptom score(IPSS),quality-of-life score(QOL score),maximum urinary flow rate (MFR) were evaluated before and 3 months after operation.Blood loss in operation,operation time,weight of resected prostate tissue,bladder irrigation time,catheterization time and length of hospital stay were also collected from the perioperative period.Results Three months after surgery,PVR,IPSS,QOL and MFR were all significantly improved in both HoLEP and PKEP groups compared with the data before operation (P =0.000),while there was no significant difference between the two groups(P>0.05).Patients from the HoLEP group showed markedly lower values of bone loss in operation than that of PKEP group((69.5±23.6) ml vs.(87.5±38.0) ml,P=0.011).There were no significant differences in terms of other indexes (P> 0.05).Conclusion HoLEP and PKEP have similar efficiency in treating BPH,while HoLEP is suggested superior in regard of bone loss.
10.Long-term effects of vascularized pisiform transfer for Kienb(o)ck's disease
Cong XIAO ; Guoming LIU ; Lin TENG ; Zhou XIANG ; Gang ZHONG ; Shiqiang CEN ; Fuguo HUANG
Chinese Journal of Orthopaedics 2011;31(3):238-242
Objective To evaluate the long-term efficacy of vascularized pisiform transfer for patients with Kienb(o)ck's disease in Lichtman stages Ⅲ-Ⅳ. Methods Eleven patients were reviewed to analyze results after lunate resection and vascularized pisiform transfer for Lichtman stages Ⅲ and Ⅳ. There were six men and five women. Age ranged from 20 to 67 years with a average of 41.0±14.3 years. According to Lichtman stage. There were 4 cases in stage Ⅲa, 5 cases in stage Ⅲb, and 2 cases in stage Ⅳ. Assessment criteria included subjective assessment of pain, visual analogue scale (VAS), range of motion (ROM), grip power,Cooney wrist score and radiographic changes on each follow-up visit. The radiographic changes including pis iform bone location, shape, sclerosis change, osteoarthritis, carpal height ratio, Nattrass index, Radioscaphoid angle and ulnar variance were recorded. Results The follow-up periods of all of cases were 61-202 months,with an average of 104.1 months. Pain had improved in 10 patients and disappeared in 7 cases. The VAS score was 2.2±1.9 at follow-up visit. Range of motion of injured wristw as only 65.3% of opposite side. Grip power was 84.3% of the contralateral hand. According to Cooney score, the results were excellent in 1 case, good in 7cases, fair in 2 cases and poor in 1 case, with the excellent and good rate of 72.7%. Radiologically, 8 cases had normal position of the pisiform bone, 2 had volar displacement and 1 had ulnar displacement which leaded to widen scaphopisiform space. Six pisiform bones had normal trabecular structure, three had degenerative changes. Bone sclerosis was seen in 2 cases and osteoarthritis was found in 3 patients. Compared with radiographic parameter before surgery, carpal height ratio and Nattrass index significantly lowered and radioscaphoid angle significantly increased. Conclusion Lunate resection and vascularized pisiform transfer is an effective method for Kienb(o)k′s disease in stages Ⅲ-Ⅳ. Although carpal collapse appeared postoperatively,the results show high patient satisfaction and good function after vascularized bone transplantation.