1.Expression and significance of multidrug resistance-associated protein 1 in the liver tissues of rat obstructive jaundice models
Jianyong ZHU ; Yingguo CHEN ; Ping BIE
Journal of Third Military Medical University 2003;0(08):-
Objective To explore the role of multidrug resistance-associated protein 1 (Mrp1) on bilirubin metabolism in rat obstructive jaundiced (OJ) models. Methods Eighty Wistar male rats were randomly divided into three groups: sham operation group in which the common bile duct was educed but not ligated; common bile duct ligation (CBDL) group in which the common bile duct was ligated and cut off; bile reflow group in which on day 14 after common bile duct ligation operation, the internal drainage from common bile duct to duodenum by silica gel duct was performed. Serum prealbumin, serum total bilirubin and urine direct bilirubin were assayed routinely. The expressions of mrp1 mRNA and protein were detected in the liver tissues by semi-quantitative RT-PCR and indirect immunofluorescence respectively. Results Serum prealbumin was descending significantly (P
2.Effects of growth hormone on perioperative obstructive jaundice rats
Jianyong ZHU ; Ping BIE ; Yingguo CHEN
Journal of Third Military Medical University 2003;0(19):-
Objective To evaluate the effects of recombinant human growth hormone (rhGH) applied perioperatively on the surgical tolerance, safety, and recovery in obstructive jaundice (OJ) rats. Methods A total of 146 Wistar male rats were randomly divided into five groups: SHAM, CBDL, REF, CBDL-GH, and REF-GH. rhGH was injected subcutaneously at the dose of 0.5 u/kg per day. One-week mortality was observed. Liver function indices, prealbumin (PA), TNF-?, and urine DBIL were measured. Results One-week mortality in REF-GH group was much lower than that in REF group (P
3.Expression of bilirubin transporter Mrp2 in liver of obstructive jaundice rats and the significance
Yingguo CHEN ; Ping BIE ; Jianyong ZHU ;
Journal of Third Military Medical University 2003;0(16):-
Objective To study the changes of transport protein Mrp2 excreting bilirubin across the canalicular membrane in obstructive jaundice rats. Methods The Mrp2 transcription and protein levels of hepatocytes was measured by RT PCR and immunofluorescence microscopy in obstructive jaundice rats. A correlation analysis of the transcription and protein levels with TNF ? and serum bilirubin was conducted. Results The Mrp2 transcription and protein levels were significantly lower in hepatocytes following bile duct obstruction and negatively correlated with TNF ? and bilirubin in rat serum. Conclusion Overexpression of inflammatory factor TNF ? may cause downregulation of transcription and protein levels of Mrp2 gene in obstructive jaundice rats. Bilirubin secretion across canalicular membrane can be inhibited. These may lead to or exaggerate hyperbilirubinemia.
4.Mutation of DKC1 Gene in a Family of Dyskeratosis Congenita
Yingguo DING ; Wei JIANG ; Yong YANG ; Dingfang BU ; Xixue CHEN ; Ping TU ; Xuejun ZHU
Chinese Journal of Dermatology 1994;0(06):-
Objective To identify the mutation of DKC1 gene and its inheritance in a pedigree with dyskeratosis congenita (DKC). Methods The mutation was detected by polymerase chain reaction(PCR)and DNA sequencing, and restriction endonuclease digestion was performed to confirm the mutation. Results A transition mutation of C to T (1058C-T) in DKC1 gene was found in the proband and his brother. This mutation results in an amino acid change from alanine to valine (A353V) in dyskerin protein. The proband′s mother and sister were carriers of this mutation gene with no phenotype of DKC. Conclusion This pedigree is an X-linked form of DKC with 1058C-T mutation in DKC1 gene.
5.Clinical effect of internal fixation of Hoffa fracture
Yingguo YANG ; Bing GE ; Dongcheng ZHU ; Liang WANG ; Xiaomin WANG ; Zhisheng LI
Chinese Journal of Trauma 2012;28(4):339-342
Objective To approach the methods and effects of internal fixation for Hoffa fracture. Methods A total of 26 patients with 26 condylar Hoffa fractures ( medial condylar fractures in 13patients and lateral condylar fractures in 13) treated from August 1993 to February 2010 were retrospectively analyzed.According to the Letenneur classification,there were 16 patients with type Ⅰ fractures,four with type Ⅱ fractures and six with type Ⅲ fractures.Among them,two patients were with open fractures and 24 with closed fractures.Surgical approaches including screw fixation in 21 patients and lateral support plate fixation in five were selected based on the fracture types and affected sides. Results All patients were followed up for 12.5-48 months (average 18 months),which showed fracture healing in all the patients within 3-4 months (average 3.5 months).Two patients had slight shift together with knee joint pain,ie,one patient had ROM of knee for 95 °,and one failed the functional exercise because of pain and had ROM of knee for 60° during follow up.No complications including infection,delayed union or bone necrosis occurred.According to Letenneur' s functional assessment system,the postoperative outcomes were excellent and good in 24 condyles,fair in one and poor in one. Conclusions Surgical treatment for Hoffa fractures is safe and effective,but the key point is to choose correct screw fixation position and orientation according to the fracture types and fracture fragment size.
6.Feasibility of pre-scan size-specific dose estimate in coronary computed tomography angiography
Xiaolong HE ; Jian XU ; Feng ZHU ; Yingguo LYU ; Kuangnan LUO
Chinese Journal of Radiological Medicine and Protection 2020;40(9):717-721
Objective:To investigate the feasibility of optimization of coronary computed tomography angiography (CCTA) with pre-scan size-specific dose estimate(SSDE).Methods:Ninety patients from Quzhou People′s Hospital during March and May 2018 who underwent CCTA were defined as modelled patients, the regression equation of body mass index (BMI) and water-equivalent diameter ( dw) were conducted and upper quartile of CTDI vol values was calculated. Other 67 patients were divided into two groups: control group (CCTA scan protocol, based on CTDI vol) and study group (CCTA scan protocol using SSDE). The image quality and radiation dose including breast dose (Dose bre) in two groups were assessed. Results:The positive correlation was found between BMI and dw with a linear regression equation of dw=9.241 + 0.644×BMI( r=0.823, P<0.05), and the upper quartile of CTDI vol was 7.92 mGy. The proportions of high-level image quality were 94.10% (367/390) and 93.93% (402/428) in the control and study groups, respectively ( P>0.05). The median signal to noise ratio (SNR) and contrast to noise ratio (CNR) were 21.08/24.39, 17.24/19.94 in the control and study groups, respectively ( P>0.05). The CTDI vol, SSDE and Dbreof the study group were reduced by 37.04%, 35.77% and 37.37% compared with the control group, and statistically significant differences were found( z=-7.041, t=18.479, 15.079, P<0.05) among them. Conclusions:The CCTA scan protocol based on the pre-scan SSDE can significantly reduce the radiation dose and provide the diagnostic image quality.
7.Effects of high-energy shock and vibration on cortex and peripheral blood immune cells in goats
Yingguo ZHU ; Dongdong ZHANG ; Liangchao ZHANG ; Guanhua LI ; Ling GUO ; Zhaoxia DUAN ; Jing CHEN ; Jianmin WANG ; Guangming YANG
Chinese Journal of Trauma 2023;39(11):1029-1036
Objective:To investigate the effects of high-energy shock and vibration on cortex injury and peripheral blood immune cells in goats.Methods:Seventeen Boer goats without gender preference were selected. By using random number tables, the goats were divided into normal control group ( n=5) and shock and vibration injury group ( n=12). The goats in the normal control group were anatomized routinely and their brain was collected after being sacrificed without any other treatment. The goats in the high-energy shock and vibration model group were placed on a loading table (part of the BY10-100 instant shock and vibration simulation platform) in a restrained state, and made into a high-energy shock and vibration injury model induced by a vertical impact waveform generator. The intravenous blood samples were taken from the goats in the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury.Then, the goats were sacrificed and the following procedures were the same as the normal control group. At 24 hours after injury, the brain injury and the histopathological changes of the cerebral cortex in the normal control group and shock and vibration injury group were observed by gross pathological and anatomical examination and HE staining. The mRNA expression of zonula occludens 1 (ZO-1), tight junction protein 5 (Claudin-5), glial fibrillary acidic protein (GFAP), ionized calcium binding adaptor molecule 1 (IBA-1), interleukin (IL)-1β, IL-6 and cluster of differentiation antigen 177 (CD177) of the cerebral cortex in the normal control group and shock and vibration injury group were measured through fluorogenic quantitative polymerase chain reaction. The expression of ZO-1 and Claudin-5 proteins of the cerebral cortex in the normal control group and shock and vibration injury group were detected by Western blotting. Hematology analyzer and coagulation analyzer were used to detect white blood cell count, neutrocyte, lymphocyte, monocyte, prothrombin time 1 (PT-1), prothrombin time international normalized ratio (PT-INR), activated partial thromboplastin time (APTT), thrombin time (TT), prothrombin activity (PTA) and fibrinogen (FIB) levels in goats of the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury, respectively. Results:At 24 hours after injury, no visible contusion or necrosis was found in goat brain tissue in the shock and vibration injury group; the cerebral micro-vessels presented with a local dilation, hyperemia, edema, aggregation of inflammatory cells, disruption of vessel walls and leakage of red blood cells. These changes were not observed in the normal control group. In the shock and vibration injury group, ZO-1 and Claudin-5 mRNA expressions in the cerebral cortex were 0.25±0.10 and 0.09(0.04, 0.44) respectively, which were significantly lower than those of the normal control group [1.00±0.15 and 0.99(0.80, 1.20)]; GFAP, IBA-1, IL-1β, IL-6 and CD177 mRNA expression levels were 4.40(3.88, 6.75), 2.60±1.07, 3.04±0.51, 2.71±0.45 and 2.93±0.62 respectively, which were significantly higher than those of the normal control group [1.00(0.78, 1.22), 1.00±0.37, 1.00±0.27, 1.00±0.57 and 1.00±0.35]; ZO-1 and Claudin-5 protein expression levels were 0.41±0.06 and 0.42±0.11 respectively, which were significantly lower than those of the normal control group (1.08±0.12 and 0.91±0.23) (all P<0.01). In the shock and vibration injury group, the levels of white blood count, neutrocyte, and lymphocyte in peripheral blood were (13.7±3.3)×10 9/L, (35.3±14.8)% and (57.2±15.1)% respectively before injury, (19.4±3.1)×10 9/L, (60.5±12.5)% and (33.6±14.2)% respectively at 3 hours after injury, and (20.6±3.6)×10 9/L, (63.6±13.0)% and (30.9±15.0)% respectively at 6 hours after injury. By contrast, the levels of white blood count and neutrocyte were significantly increased but the level of lymphocyte was significantly decreased at 3 and 6 hours after injury ( P<0.05 or 0.01); the levels of the above indicators showed no significant changes at 0 and 24 hours after injury (all P>0.05); the level of monocyte did not change significantly at all time points before and after injury (all P>0.05). The levels of PT-1, PT-INR, APTT, TT, PTA and FIB in the shock and vibration injury group did not change significantly at each time point before and after injury (all P>0.05). Conclusion:Cerebral cortex microvascular injury and disruption of blood-brain barrier can be initiated in the early stage of high-energy shock and vibration injury in goats, accompanied by the presence of central and peripheral inflammatory response.
8. Involvement of intracellular organelle stress, autophagy and ferroptosis in cobalt chloride-induced vascular smooth muscle cell injury
Yan LEI ; Xiaoyong PENG ; Tao LI ; Liangming LIU ; Mengsheng DENG ; Dongdong ZHANG ; Yingguo ZHU ; Jianmin WANG ; Zhaoxia DUAN ; Guangming YANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(1):1-10
AIM: To investigate the relationship between vascular smooth muscle cell (VSMC) injury, organelle stress response and autophagic cell death (autophagy) and ferroptosis induced by the chemical hypoxia inducer cobalt chloride (CoCl2) through the bioinformatics analysis and in vitro cell experimentation. METHODS: The dataset GSE119226 of VSMC treated with cobalt chloride was acquired from the gene expression database (GEO). The R language was used to investigate the relationship between CoCl2 treatment and organelle stress response (Golgi stress, endoplasmic reticulum stress) and two forms of cell death (ferroptosis and autophagic cell death). With primary cultured rat VSMC (rVSMC) and CoCl2-induced anoxia model, the changes in cell viability were detected by CCK-8 method, and reactive oxygen species (ROS) levels were measured using DCFH-DA method. The expression levels of HIF-1α (a key molecule in hypoxia), Golgi stress markers GM130 and p115, endoplasmic reticulum stress markers GRP78 and CHOP, autophagy markers LC3-II / LC3-I and Beclin1, and ferroptosis markers GPx4 and xCT were detected by Western blot. The effect of inducing or inhibiting organelle stress and cell death on the CoCl2-induced cell damage was also observed. RESULTS: Differentially expressed genes analysis of GSE119226 dataset showed that CoCl2 treatment of VSMCs had significant effects on organelle function and stress response, autophagy and ferroptosis-related genes, in which endoplasmic reticulum stress, protein processing in endoplasmic reticulum, regulation of Golgi to plasma membrane protein transport, autophagy / autophagic cell death, and ferroptosis pathways were remarkably enriched. The results of in vitro experiment showed that compared with normal rVSMC, cell viability was significantly decreased after CoCl2 treatment, as well as HIF-1α protein expression and ROS levels in rVSMCs were increased. In rVSMC treated with Co-Cl2, the expression levels of Golgi structural proteins GM130 and p115 (reflecting the occurrence of Golgi stress) were decreased, while the markers GRP78 and CHOP (reflecting the occurrence of endoplasmic reticulum stress) were increased. At the same time, CoCl2 treatment also reduced the expression of autophagy markers LC3-II/LC3-I and Beclin1 (indicating the decrease levels of autophagy), while the expression of ferroptosis markers GPx4 and xCT were decreased (indicating the occurrence of ferroptosis). Compared with CoCl2 treatment group, induced Golgi stress, endoplasmic reticulum stress, or ferroptosis could further reduce cell viability, while inhibition of these processes could improve cell viability. On the other hand, increasing the level of autophagy can improve the cell viability. CONCLUSION: Hypoxia induced by cobalt chloride can lead to VSMC injury. Golgi stress, endoplasmic reticulum stress, ferroptosis, and the reduction of autophagy level play an important role in it. Inhibition of organelle stress response and ferroptosis, or increase of autophagy level can improve VSMC injury caused by cobalt chloride.