1.Relationship between plasma ghrelin levels and insulin resistance and blood pressure in octogenarians.
Yintao, ZHAO ; Li, SHAO ; Lili, TENG ; Daifu, ZHANG ; Hua, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(3):307-11
The association between fasting plasma ghrelin levels and insulin resistance and blood pressure (BP) in octogenarians was investigated in this study. A total of 487 unrelated octogenarians (including 203 men and 284 women) were enrolled in this cross-sectional study at the Healthy Care Center of Shanghai East Hospital, Tongji University, China, from October 2008 to April 2009. Plasma ghrelin was determined by using the enzyme linked immunosorbent assay (ELISA). Insulin sensitivity was assessed using the homeostasis model of assessment-insulin resistance (HOMA-IR). The age of the participants ranged from 80 to 89 years (mean=83.9+/-4.8 years) with a body mass index (BMI) of 25.3+/-4.9 kg/m(2). Plasma ghrelin levels were 20.94+/-5.34 mug/L, being 20.89+/-5.53 mug/L in men and 21.38+/-3.73 mug/L in women respectively. Plasma ghrelin was not associated with systolic (P=0.981) or diastolic (P=0.724) BP, waist circumference (P=0.278), fasting insulin (P=0.246), fasting blood glucose (FBG) (P=0.693) and HOMA-IR (P=0.232). In the control cohort, no significant differences in plasma ghrelin were found between genders (P=0.489), and among subjects with hypertension (BP>140/90 mmHg) (P=0.284) and type 2 diabetes (P=0.776). In conclusion, fasting plasma ghrelin levels are not directly correlated with insulin resistance and BP among octogenarians.
2.The radiosensitizing effects of cryptotanshinone on H22 hepatoma-bearing mice
Yintao YE ; Chen WANG ; Xiaokun SONG ; Jie ZHANG ; Ning GAO
China Oncology 2014;(1):29-34
Background and purpose:Quinonoids can change the cell cycle distribution of tumor cells, and effect the radiosensitizing. This study aimed to investigate the radiosensitization effects, cell cycle and apoptosis of cryptotanshinone on H22 hepatoma-bearing mice. Methods:The mouse hepatoma H22 model was established, then divided into blank control group, irradiation alone group, high dose of cryptotanshinone group, cryptotanshinone (low, medium and high)+IR groups. After irradiated, observed the growth of tumor’s conditions, record epigenetic tumor irradiation time, calculated the delay time of tumor growth and enhancement factor (EF). After 22 days, mice were killed, stripped tumor, and calculated the inhibition rate. The cell cycle distribution and apoptosis were measured by lfow cytometry. Results:Cryptotanshinone (low, medium and high) groups inhibited the tumor growth better than the blank group, and had the signiifcant radiosensitizing effect. The enhancement factor was 1.22, 1.43,2.19, respectively. Cells were treated with cryptotanshinone which had significant effects on cell cycle, and induced apoptosis, which indicated signiifcant G2/M phase arrest and a decrease in S phase. Conclusion:Cryptotanshinone inhibited the tumor growth and had the radiosensitizing effects on H22 hepatoma-bearing mice. One of the mechanism may be that it might make significant G2/M phase arrest and S phase decreased, and induced apoptosis. So cells were more sensitive to radiation.
3.Analysis of the risk factors and angiographical characteristics of patients with metabolic syndrome and coronary heart disease
Lei YUAN ; Daifu ZHANG ; Yan LAI ; Yian YAO ; Yintao ZHAO
Chinese Journal of Endocrinology and Metabolism 2010;26(9):764-766
A total of 167 coronary heart disease (CHD) patients were divided into metabolic syndrome (MS) group (68 cases) and non-MS group (99 cases). There were significant differences in the disease-related metabolic indicators and coronary angiography (multivessel lesions, diffuse stenosis, occlusive lesions, Ginsini score) between MS group and non-MS group ( all P<0.05 ). When the patients with MS were divided into 3 groups according to the number of componernts of MS, three lesions, diffuse stenosis, and occlusive lesions were more frequent in five components group compared with three components group. Ginsini points rised with the increased risk factors. There existed differences in Ginsini score between three components group and four, five components group (P<0. 05 or P<0.01 ). Multivariable logistic regression analysis showed that obesity, hypertension,diabetes, high low-density lipoprotein-cholesterol, and low high-density lipoprotein-cholesterol were the predictors of CHD in patients with MS (P<0.05 or P<0.01 ).
4.Relationship between Plasma Ghrelin Levels and Insulin Resistance and Blood Pressure in Octogenarians
ZHAO YINTAO ; SHAO LI ; TENG LILI ; ZHANG DAIFU ; ZHANG HUA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(3):307-311
The association between fasting plasma ghrelin levels and insulin resistance and blood pressure(BP)in octogenarians was investigated in this study.A total of 487 unrelated octogenarians (including 203 men and 284 women)were enrolled in this cross-sectional study at the Healthy Care Center of Shanghai East Hospital,Tongji University,China,from October 2008 to April 2009.Plasma ghrelin was determined by using the enzyme linked immunosorbent assay(ELISA).Insulin sensitivity was assessed using the homeostasis model of assessment-insulin resistance(HOMA-IR).The age of the participants ranged from 80 to 89 years(mean=83.9±4.8 years)with a body mass index(BMI)of 25.3±4.9 kg/m2.Plasma ghrelin levels were 20.94±5.34 μg/L,being 20.89±5.53 μg/L in men and 21.38±3.73 μg/L in women respectively.Plasma ghrelin was not associated with systolic(P=-0.981)or diastolic(P=0.724)BP,waist circumference(P=0.278),fasting insulin(P=0.246),fasting blood glucose(FBG)(P=0.693)and HOMA-IR(P=0.232).In the control cohort,no significant differences in plasma ghrelin were found between genders(P=0.489),and among subjects with hypertension (BP>140/90 mmHg)(P=0.284)and type 2 diabetes(P=0.776).In conclusion,fasting plasma ghrelin levels are not directly correlated with insulin resistance and BP among octogenarians.
5.Study on the Effects and Mechanism of Couplet Medicines of Scutellaria baicalensis-Paeonia lactiflora on Improving Ulcerative Colitis in Mice
Yan LIU ; Liantai LI ; Xiaoqing JI ; Yintao ZHAO ; Siyuan XIN ; Xiumin DENG ; Mengjiao ZHANG ; Yingjun ZHANG
China Pharmacy 2018;29(3):356-360
OBJECTIVE: To study the effects and mechanism of couplet medicines of Scutellaria baicalensis-Paeonia lactiflora on improving ulcerative colitis (UC) in mice. METHODS: A total of 70 mice were randomly divided into blank group, model group, S. baicalensis group (1. 5 g/kg), P. lactiflora group (1. 5 g/kg), S. baicalensis-P. lactiflora (2:1, 1:1, 1:2, m/m) groups (total amount of 1. 5 g/kg), with 10 mice in each group. Except for blank group, UC model of mice was induced in each group. The next day after modeling, treatment groups were given relevant medicine liquid 0. 2 mL/10 g (75 mg/mL, calculated by crude drug mass concentration), while blank group and model group were given constant volume of normal saline intragastrically, once a day, for consecutive 7 d. After administration, disease activity indexes (DAI) of rats were scored, and the serum levels of TNF-α, IL-1β, IL-6, D-LA and myeloperoxidase (DAO) were determined. The length of the colon was measured and the intestinal mass index was calculated in mice. The activities of medullary peroxide (MPO) and SOD, the levels of NO and MDA were determined in colon tissue. RESULTS: Compared with blank group, DAI score, serum levels of TNF-α, IL-1β, IL-6, D-LA and DAO, the levels of MPO, NO and MDA in colon were increased significantly in model group, while the length of colon, intestinal mass index and SOD level of colon tissue were decreased significantly, with statistical significance (P<0. 05 or P<0. 01). Compared with model group, DAI score, serum levels of TNF-α, IL-1β and DAO, the level of MDA in colon were decreased significantly in S. baicalensis group (P<0. 05 or P<0. 01). The serum levels of TNF-α and IL-1β, the level of MDA in colon were decreased significantly in P. lactiflora group (P<0. 05 or P<0. 01). Above indexes of S. baicalensis-P. lactiflora (2:1) group were improved significantly except for the length of colon (P<0. 05 or P<0. 01). Above indexes of S. baicalensis-P. lactiflora (1:1) group were improved significantly except for serum level of IL-6 and the level of SOD in colon (P<0. 05 or P<0. 01). Above indexes of S. baicalensis-P. lactiflora (1:2) group were improved significantly except for serum level of NO (P<0. 05 or P<0. 01). CONCLUSIONS: The couplet medicines of S. baicalensis-P. lactiflora can reduce the expression of proinflammatory factors, enhancing antioxidant activity of the body and decrease intestinal mucosal permeability so as to improve UC symptom of mice; and the effect of S. baicalensis-P. lactiflora (2:1) group is the best.
6.Effect of miR-125a-5p targeting Scarb1 gene on hypoxia/reoxygenation injury of cardiomyocytes and its mechanism.
Yintao ZHAO ; Jingjing SUN ; Yuan LIU ; Lu ZHENG ; Xiangqin ZHANG ; Haibo YANG
Chinese Journal of Medical Genetics 2020;37(9):980-986
OBJECTIVE:
To explore the effect and mechanism of miR-125a-5p targeted regulation of scavenger receptor B1 (Scarb1) gene on anoxia/reoxygenation injury of rat cardiomyocytes.
METHODS:
H9c2 rat cardiomyocytes were randomly divided into blank control group, hypoxia/reoxygenation group, transfection control group and mir-125a-5p transfection group. The expression of miR-125a-5p, cardiomyocyte viability, apoptosis rate, ATP content and the expression of Scarb1, Cyt C, Bax, Bcl-2 and NF-κB signaling pathway related proteins were determined. Target gene of miR-125a-5p was predicted with Targetscan software, and the targeting of miR-125a-5p on Scarb1 was verified by double luciferase reporter gene experiment.
RESULTS:
Compared with the blank control group, the expression of miR-125a-5p, Bax, Cyt C and the apoptotic rate of cardiomyocytes in the hypoxia/reoxygenation group were significantly increased (P<0.05), while the expression of Scarb1, Bcl-2 and the content of ATP were significantly decreased (P<0.05). Compared with the control group, the situation of mir-125a-5p transfection group was just the opposite. Double luciferase reporter gene experiment has confirmed Scarb1 to be the target of miR-125a-5p. Hypoxia/reoxygenation can promote the expression of NF-κB p65, C-myc and Cyclin D1 in cardiomyocytes, while down-regulating the expression of miR-125a-5p can inhibit the expression of such proteins.
CONCLUSION
Hypoxia/reoxygenation can induce the expression of miR-125a-5p in rat cardiomyocytes. Inhibition of miR-125a-5p can protect cardiomyocytes from hypoxia/reoxygenation by up-regulating the expression of Scarb1. The mechanism may be related to the inhibition of activation of NF-κB signaling pathway.
7.Sequential transcatheter arterial chemoembolization and selective portal vein embolization before major hepatectomy for large hepatocellular carcinoma: a pilot study
Wenchao ZHAO ; Yintao WU ; Yingxiang YANG ; Yang AN ; Nianxin XIA ; Peng LIU ; Jianyong ZHU ; Che LIU ; Hong ZHANG ; Jingbo LI ; Baoan QIU
Chinese Journal of Hepatobiliary Surgery 2021;27(3):164-168
Objective:To preliminarily study the feasibility, safety and efficacy of transcatheter arterial chemoembolization (TACE) combined with selective portal vein embolization (SPVE) before surgical resection in the treatment of large liver cancer.Methods:A retrospective study was conducted on the clinical data of 17 patients with large liver cancer treated with TACE combined with SPVE from January 2016 to December 2019 at the Department of Hepatobiliary Surgery, the Sixth Medical Center of PLA General Hospital. The study included 15 males and 2 females, aged (59.17±10.30) years. The levels of alanine aminotransferase, tumor changes and patient survival were analyzed before operation, after TACE, and after SPVE.Results:Among the 17 patients, the levels of alanine aminotransferase on the 1st and 3rd day after SPVE was significantly higher than those after TACE [191.4 (30.5-1966.4) IU/L vs 125.3 (35.7-846.2) IU/L on the first day, and 298.5 (24.6-1334.2) IU/L vs 208.6 (21.6-775.6) IU/L on the 3rd day], all P<0.05. One month after the two combined embolism, among the 6 patients with a tumor diameter of 5-10 cm, 2 patients (33.3%) had complete remission, 3 patients (50.0%) had partial remission, and 1 patients (16.6%) had stable disease. For the tumor’s longest diameter, among the 11 patients with tumors >10 cm, 1 patient had complete remission (9.1%), 4 patients had partial remission (36.4%), 5 patients had stable diseases (45.5%), and 1 patient had disease progression (9.1%). Eventually, 11 patients underwent surgical exploration. The median residual liver volume before treatment was 329.5 (284.9-365.7) ml, and after the combined procedure 415.6 (354.7-718.8) ml. The median hyperplasia ratio was 28.1% (14.1%-51.3%). Eight patients finally underwent surgical resection. There was no death in the perioperative periods. The median tumor-free survival time was 17 (7-42) months, and the median survival time was 27 (7-42) months. Conclusion:For patients with large liver cancer with insufficient remnant liver volume, preoperative TACE+ SPVE has certain value in controlling tumor progression, promoting remnant liver hyperplasia, increasing surgical resection rate and improving prognosis.
8.Effects of local vibration and extracorporeal shock wave therapy on triceps spasticity and the walking ability of hemiplegic stroke survivors
Wei ZHANG ; Reiqing LI ; Yujing GU ; Yintao GUO ; Xiangzhe LIU
Chinese Journal of Physical Medicine and Rehabilitation 2022;44(4):318-323
Objective:To compare the effect of local vibration and extracorporeal shock wave therapy (ESWT) on triceps spasticity and the walking ability of hemiplegic stroke survivors.Methods:Sixty-nine stroke survivors with hemiplegia were randomly divided into a control group, a vibration group and an ESWT group. The control group received 60 minutes of conventional Bobath rehabilitation treatment and motor relearning from Monday to Saturday for 4 weeks. For the vibration and ESWT groups, 10 minutes of that traditional therapy were replaced by either local vibration or extracorporeal shock wave treatment every Tuesday, Wednesday and Saturday. Before and after the treatment, the three groups were evaluated using the Comprehensive Spasticity Scale (CSS) and in terms of passive joint range of motion (PROM), ankle plantar flexion angle, 10m maximum walking speed, stride frequency, and stride length.Results:After the intervention the average CSS, PROM, and ankle plantar flexion angle were significantly better for all three groups than before the treatment. At that point the ESWT group′s averages were significantly better than those of the vibration group, and the vibration group′s averages were significantly superior to those of the control group. Walking speed, stride frequency and stride length had also improved significantly in all three groups, with those in the vibration and ESWT groups significantly outperforming the control group. There was no significant difference between the vibration and ESWT groups in terms of walking ability.Conclusions:Both local vibration and extracorporeal shock wave therapy improve triceps spasticity and the walking ability of hemiplegic stroke survivors. Shock waves are more effective for improving spasticity, but there is no significant difference between the therapies in terms of improving walking ability.
9.Expression and significance of HERG protein in Cajal interstitial cells of gallbladder tissue in patients with gallbladder stones
Peng LIU ; Che LIU ; Yintao WU ; Hangyu ZHANG ; Zhuzeng YIN ; Yong XU ; Yongwei CHEN
Chinese Journal of Hepatobiliary Surgery 2022;28(11):848-853
Objective:To investigate the expression and significance of human ether-a-go-go related gene (HERG) protein in interstitial cells of Cajal (ICC) in patients with gallbladder stones.Methods:The gallbladder tissues of 60 patients with gallbladder diseases who underwent cholecystectomy from January 2018 to December 2020 in the Faculty of Hepato-Pancreato-Biliary Surgery, Chinese PLA General Hospital were collected, including 36 males and 24 females, aged (46.0±14.0) years. They were divided into two groups according to whether there were gallstones: gallstone group and control group (patients with gallbladder polyps and gallbladder adenomyosis), with 30 cases in each group. Color ultrasound was used to detect and calculate the gallbladder contraction rate. The neck, body and bottom tissues of the gallbladder were excised and sectioned. The expression of HERG protein and CD117 ( marker of ICC) was detected by immunofluorescence staining, immunohistochemistry and Western blot.Results:The gallbladder contraction rate in the gallstone group was (65.8±4.1)%, lower than that in the control group (73.8±5.3)%, with a statistically significant difference ( t=4.14, P<0.001). Immunohistochemistry showed that HERG protein was mainly distributed in the mucosal layer of gallbladder tissue, which was pale brown. The relative expression of HERG protein at the bottom of gallbladder in the gallstone group was (0.293±0.102), lower than that in the control group (0.694±0.059), with a statistically significant difference ( t=3.38, P=0.027). Immunofluorescence staining showed that HERG protein was mainly distributed in ICC of gallbladder epithelium. HERG protein expression in ICC at the bottom of gallbladder in gallstone group was lower than that in control group, while HERG protein expression at the neck and body of gallbladder had no significant difference. Conclusion:There are ICC and HERG protein in gallbladder tissue of patients with gallstone. The decrease of gallbladder contraction rate may be related to the decrease of HERG protein expression in ICC in gallbladder bottom tissue.
10.Construction of THP-1 cell line with GPR108 gene deletion by CRISPR / Cas9 system and exploration of its function
Wenwen Wang ; Zhenzhen Tu ; Dandan Zang ; Jing Wang ; Yintao Zhang ; Haisheng Zhou
Acta Universitatis Medicinalis Anhui 2023;58(4):528-533
Objective:
To establish a Tohoku hospital pediatrics-1 (THP-1) cell line with G protein-coupled recep- tor108 ( GPR108) deletion and explore its functions.
Methods:
According to the requirements of the clustered regularly interspaced short palindromic repeats ( CRISPR) / CRISPR-associated protein 9 ( Cas9 ) system ,two single guide RNAs (sgRNA1 and sgRNA2) paring to the flanking fragments of human GPR108 gene were designed and synthesized.The two oligonucleotides were inserted in the pL-CRISPR. EFS.GFP vector to generate the new recombinant vectors ( pL-CRISPR. EFS.GFP-sgRNA1 and pL-CRISPR. EFS.GFP-sgRNA2 ) .The recombinant vectors and packaging plasmids (pMD2. G and psPAX2) ,were co-transfected into 293T cells to generate virus for infecting THP-1 cells.The GFP + cells were screened and isolated in 96-well culture plates by flow cytometry to obtain single-cell clones.PCR and Western blot were used to detect whether GPR108 was successfully knocked out in THP- 1 cells.Both GPR108 + / + and GPR108 -/ - THP-1 cells were treated with lipopolysaccharide (LPS) .Interleukin 8 (IL-8) derived from the THP-1 cells,which were treated by LPS,was detected with Western blot and cytometric bead array ( CBA) analysis.
Results:
The recombinant lentiviral vector pL-CRISPR. EFS.GFP-sgRNA was successfully constructed and single-cell clone F9 was obtained by flow cytometric sorting after transfection of THP-1 cells.PCR and Western blot both confirmed that F9 was a GPR108 -/ - THP-1 single-cell clone. LPS stimulated GPR108 -/ - and GPR108 + / + THP-1 cells,both Western blot and CBA results showed a significant decrease in IL- 8 synthesis and secretion in GPR108 -/ - THP-1 cells.
Conclusion
The GPR108 -/ - THP-1 cell clone is success- fully obtained based on the CRISPR / Cas9 system.GPR108 deletion in THP-1 cells treated by LPS leads to a decrease of IL-8 expression and secretion.It lays the foundation for further research on the molecular mechanisms of GPR108 in the immune inflammatory response.