2.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.
3.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.
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.Effect of Daidzein intravitreal injection on optic nerve injury in rats
Zhong-Jun, TANG ; Zhen-Ping, HUANG ; Wen-Jing, YANG ; Yong-Xiang, ZOU ; Ji-Ping, CAI
International Eye Science 2014;(8):1378-1381
AIM:To discuss Daidzein intravitreal injection whether has protective and recovery effects on acute nerve damages.
METHODS:After the crush models of acute optic nerve were set up, 72 males SD rats were divided into 4 groups randomly as common group without surgery, FBS negative control group, Daidzein treatment group ( 10μmol/L, 100μmol/L, 1000μmol/L ) and positive control group using rats nerve growth factor ( mNGF, 100ng/mL ). Three days after interference, all experimental animals were executed. HE staining was used to evaluate morphologic change of the retina, immunohisochemical staining and western-blot tests for identifying and quantifying the distinct expression of Caspase-3 and GAP-43 among the groups.
RESULTS: Compared with the normal group and negative control group, retinal morphology of different concentrations of each Daidzein treatment group and positive control group was more complete, the expression of Caspase-3 protein was relatively lower, the expression of GAP-43 protein was relatively higher, the differences have statistically significance (P<0. 05).CONCLUSION: Daizein injection in the vitreous cavity has the capacity of protection and restoration in rat's acute nerve damages.
8.X-ray diagnostic site selectivity studies of skeletal fluorosis
Heng-xiang, LI ; Xue-song, WANG ; Pei-zhong, CHEN ; Jie, GAO ; Ju-mei, HUANG
Chinese Journal of Endemiology 2013;32(5):565-568
Objective To study the X-ray signs of forearm and leg in skeletal fluorosis and its diagnostic value,aim at finding the easy examination parts.Methods One thousand four hundred and forty subjects were examined using developed shield,darkroom and other portable dedicated device combined with a small X-ray machine.A total of 384 cases were diagnosed skeletal fluorosis.All patients were divided into different groups and the time,degree and range of X-ray to the forearm and calf elbow,knee,and long bone were compared.Results The X-ray change in the forearm elbow was earlier than that of the leg knee,and trabecular bone change was the earliest indicator,197 cases and 157 cases,respectively,and the difference was statistically significant (x2 =28.006,P < 0.01).Membrane ossification of forearm backbone was earlier than that of the leg,and most of them were degree Ⅰ photos,213 cases and 126 cases respectively.The difference was statistically significant (x2 =17.626,P < 0.01).The direction of the interosseous membrane ossification was from the forearm radius to the ulna,then to the fibula and tibia,and was accompanied by changes in the aggravation of forearm.A variety of indicators were observed,especially the membrane ossification in bone and joint trabecular bone and the long bone was the most active,and the forearm was more sensitive,obviously than that of the calf.Conclusion In the X-ray screening or detection of endemic fluorosis,the forearm radiography is a simple,economical,and effective diagnostic method.
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.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.