1.Establishment and research progress of retina ischemic animal model
Yongjie ZHANG ; Hong XU ; Xuesong TIAN
Chinese Journal of Ocular Fundus Diseases 2021;37(5):408-413
Retinal ischemia is the common pathologic process in many ophthalmic diseases, including ischemic optic neuropathy, retinal artery and vein occlusion, carotid artery obstructive disease, retinopathy of prematurity, chronic diabetic retinopathy and glaucoma. It is very important to establish animal models to investigate pathology mechanism and explore the treatment of retinal ischemia disease. At present, the commonly used methods for establishing retinal ischemia animal models include increasing intraocular pressure, ligating of blood vessels, suture method, photochemical method, and drug injection etc. This article summarizes the methods to establish the animal models and analyzes the indication for each animal model. It is expected that the method of establishing a retinal ischemic animal model will be helpful to the experimental design of follow-up retinal ischemia studies.
2.Effects of Ramulus Uncariae et Uncus on body mass, food intake,serum glucose and total antioxidative ability of high-fat-fed obese rats
Rong LUO ; Long JIN ; Xuesong TIAN ; Yuling WEI ; Wei LI ; Tianzhen ZHENG ; Songyi QU
Chinese Journal of Tissue Engineering Research 2005;9(31):246-248
BACKGROUND: Ramulus Uncariae et Uncus can be used to treat many diseases of cardiovesculer and neurosystem by calming and protecting endothelium and neuron.OBJECTIVE: To study the effects of drinking water extracts of Ramulus Uncariae et Uncus on body mass, food intake, energy intakes, serum glucose, insulin, total antioxidative ability of high-fat-fed rats.DESIGN: A completely randomized and controlled experiment.SETTING: Institute of Physiology, Foundational Medical College of Lanzhou University.MATERIALS: The experiment which is Gansu Province Key Labor of Pre-clinical Research for Chinese Herbs & New Drugs was carried out at the Physiological Laboratory of Institute of Foundational Medical College of Lanzhou University from March 2003 to May 2003. Twenty-seven healthy male SD rats were randomly divided into the following three groups (9 rats each group): namely, the normal control group, the high-fat-fed group and the Ramulus Uncariae et Uncus group.METHODS: [1] The rats of control group were fed with referenced diets(Protein, fat and carbohydrate was made up fore18.2%, 4.5%, 55.2% respectively), while the rats of high-fat-fed and the Ramulus Uncariae et Uncus group were fed with high-fat diets in which protein, fat and carbohydrate accounted for 23.7%, 21.6%, 39.0% respectively. [2]The rats in control and high-fat group could drink water freely every day while the rats in Ramulus Uncariae et Uncus group drank freely one percent water extract of Ramulus Uncariae et Uncus at the first day of the experiment, later the concentration of Ramulus Uncariae et Uncus increased gradually to six percent at the sixth day, which lasted until the end of the experiment. [3]Food intake was monitored daily, and body mass weekly at set time, the energy intake was calculated on basis of food intake. We calculated the Lee's Index [(body mass)1/3 (g) × 103 / body length (cm)]. [4] At the end of the seventh week, all rats were fasted for twelve hours to collect blood for the measure of some serum items. The serum levels of glucose were determined with hexokinase. The levels of insulin was measured by Radioimmunology methods. Malondiadehyde was measured by thiobarbituricbased colorimetric assay. Total antioxidative ability and free fatty acid were evaluated by colorimetric assay. [5] The difference of quantity data among the tree groups were statistically analyzed by F analysis, while the difference between two groups was analyzed by LSD methods. The main indexes of the experiment were body mass, food intake, energy intake, serum glucose after fasting, insulin, free fatty acid, malondialdehyde and total antioxidative ability respectively.RESULTS: Twenty-seven rats date was analyzed and nine rats in every group. [1] Body mass: Compared with the high-fat-fed group, body mass in rats of the control group and the Ramulus Uncariae et Uncus group were less from the end of the third week to the end of the experiment (P < 0.05-0.01).As compared between the Ramulus Uncariae et Uncus group and the control group, there was no significant difference from the first week to the sixth week (P > 0.05), while the body mass of the former group was bigger than that of the latter at the end of the seventh week (P < 0.01). [2] Lee's Index of the rats in high-fat-fed group was significantly higher than those of the rats in the control and Ramulus Uncariae et Uncus group (P < 0.01).[3] Food intake: From the first week, compared with the control group, the food intake of rats in Ramulus Uncariae et Uncus group was less (P < 0.01)and lower than that in high-fat-fed group at the end of the third, fifth, sixth and seventh week, too (P < 0.05-0.01). [4] Energy intake: As the food was high-fat, the Energy intake of rats in the Ramulus Uncariae et Uncus group was higher than that of the control group at the end of the first, second and forth week (P < 0.05-0.01), while less than that of the high-fat-fed group at the end of the third, fifth, sixth and seventh week (P < 0.01). [5] The serum glucose: There was no statistical significance among the three groups. [6]The level of serum insulin and malondiadehyde of the rats in high-fat-fed group were higher than that of the control group and the Ramulus Uncariae et Uncus (P < 0.01). [7] The level of free fatly acid of rats in high-fat-fed group was higher than that of the control group (P < 0.05), while there wasno statistical difference compared with the Ramulus Uncariae et Uncus group. [8] The level of total antioxidative of the rats in high-fat group was lower than that of the Ramulus Uncariae et Uncus group (P < 0.01).CONCLUSION: [1] Drinking water extracts of Ramulus Uncariae et Uncus could inhibit the weight gain and food intake, [2] reduce level of free radical and insulin of serum, which could be caused by the dec lining of free fatty acid. [3] In the same time, increase the total antioxidative ability of the obese rats caused by high-fat diets.
3.δ-opioid receptors protect neurons against neuronal injury induced by oxygen-glucose deprivation
Mingwei LI ; Min ZHU ; Xuesong TIAN ; Xiaomin OU ; Ying XIA ; Jingchun GUO
Fudan University Journal of Medical Sciences 2009;36(4):389-393
Objective To investigate the effect of cortical 8-opioid receptor (DOR) on oxygen-glucose deprivation-induced (OGD-induced) neuronal injury. Methods Primary cultured cortical neurons incubated with selective DOR agonist (TAN-67) and antagonist (naltrindole) or PKC inhibitor (chelerythrine, CHE) were exposed to OGD. Lactate dehydrogenase (LDH) release was detected after 24 h reperfusion. The expression levels of DOR were measured by Western blot. Results Compared with OGD group, TAN-67 significantly decreased OGD-indueed LDH release, and increased the expression levels of DOR, while nahrindole aggravated neuronal injury and decreased the DOR protein expression. CHE could abolish the LDH down-regulation induced by TAN-67 plus OGD (P< 0.05, compared with TAN-67 treated group). Conclusions DOR activation protects neurons against OGD injury. PKC might take part in the neuroprotection pathways of DOR.
4.Electroacupuncture protects the brain against acute ischemic injury via up-regulation of delta-opioid receptor in rats
Xuesong TIAN ; Fei ZHOU ; Ru YANG ; Ying XIA ; Genchen WU ; Jingchun GUO
Journal of Integrative Medicine 2008;6(6):632-8
OBJECTIVE: To explore the effect of delta-opioid receptor (DOR) in electroacupuncture (EA) protecting the brain against acute ischemic injury. METHODS: Fifty-one rats were randomly divided into sham ischemia group, ischemia group, sham EA group, EA group, and EA+DOR antagonist (naltrindole) group. Transient focal cerebral ischemia (1 hour) was induced in rat brain by middle cerebral artery occlusion (MCAO) method. EA was applied on Shuigou (GV 26) and Neiguan (PC 6) for 30 min, starting immediately after the onset of reperfusion. Neurological deficit scores and volume of cerebral infarction were detected after 24-hour reperfusion. Other 12 rats were randomly divided into sham ischemia group, ischemia group, EA group and EA + naltrindole group. DOR protein expressions were assessed by Western blotting after 24-hour reperfusion. RESULTS: In comparison with the ischemia group and sham EA group, EA significantly reduced ischemic infarction and neurological deficits (P<0.05); EA significantly increased the expression of 60 kD DOR protein (P<0.05) and tended to increase that of 36 kD DOR protein (P>0.05). When naltrindole was combined with EA, the naltrindole completely abolished the EA-induced protection in ischemic infarction and neurological deficits, and also arrested the expression of DOR. CONCLUSION: EA can up-regulate DOR expression and protect the brain from ischemia-reperfusion injury.
5.Protective effects of ferulic acid on mice bone marrow hematopoietic damage induced by irradiation
Shuai SHAO ; Mei TIAN ; Jianxiang LIU ; Qiao GOU ; Xuesong QI ; Chunyan WANG ; Gonglin QU ; Chen LI ; Xu SU
Chinese Journal of Radiological Medicine and Protection 2017;37(8):570-575
Objective To observe the effect and the mechanisms of ferulic acid on radiationinduced damage of mice peripheral blood and bone marrow hematopoietic function.Methods Ninety-six mice were randomly divided into sham irradiation group,irradiation group,positive drug group and 10,30,90 mg·kg-1 ·d-1 ferulic acid group,16 mice per group.Mice were exposed to 3.5 Gy γ-rays 24 h after first drug taken.Then,mice were given drugs for 7 d after irradiation.White blood cells in peripheral blood of 10 mice per group were counted 2 d before irradiation and 3,7,10,15 and 22 days after irradiation.The bone narrow of the other six mice was taken to detect the micronuclei frequency of polychromatic erythrocyte,the hematopoietic progenitor cell colony formation capacity,Thbd and HMGB1 protein expressions in mice bone marrow on the seventh day after irradiation.Results Compared with the irradiation alone group,the treatment of mice with ferulic acid 90 mg· kg-1 · d-1 increased the number of white blood cells in peripheral blood at 3,10,15 and 22 d after irradiation (t =2.267,2.399,1.945,2.828,P < 0.05).Treatment with mice with ferulic acid 90 mg· kg-1 · d 1 decreased the micronuclei rate of erythrocytes in irradiated bone marrow (t =4.013,P < 0.05),increased the clone numbers of CFU-E,BFU-E and CFU-GM of hematopoietic progenitor cells (t =2.366,2.953,3.115,P <0.05),improved the relative expression of the Thbd protein in bone marrow and the HMGB1 protein in nuclear (t =17.75,23.39,P < 0.01).Conclusions Ferulilc acid could protect the bone marrow hematopoietic of mice exposed to irradiation by regulating the expressions of Thbd and HMGB1 protein,and then accelerate the peripheral cells recovery.
6.Application of Virtual Reality in the Behavior Analysis of Laboratory Animal
Laboratory Animal and Comparative Medicine 2023;43(1):73-78
Virtual reality (VR) is an emerging technology for computer scene simulation that has become a reliable tool for animal behavior analysis in recent years. The behavior of an animal is crucial forits survival. Animal behavior analysis provides useful information for scientific progress in genetics, ecology, neuroscience, economics, and robotics. Animal behavior analysis can be divided into open-loop studies, in which the stimulus is independent of the animal's response to the stimulus, and closed-loop studies, in which the stimulus is adjusted according to the real-time movement of the animal. In this paper, we have discussed the origin, concept, and applications of animal behavior analysis, and reviewed the shortcomings of open-loop research on animal behavior represented by visual stimulation using traditional technology. Then taking visual, olfactory, tactile, and auditory stimulation as examples, we analyzed that the animal behavior analysis system using VR technology could combine precise experimental control with the complex behavior of laboratory animals, and complete the closed-loop research that could not be achieved using the experimental methods of the real physical world. The prospect of VR in developing superior simulations and virtual animals in the future was also discussed.
7.Role of connexin 43 in X-ray induced apoptosis in human umbilical vein endothelial cells and its mechanism
Chen LI ; Mei TIAN ; Qiao GOU ; Jianxiang LIU ; Xuesong QI ; Chunyan WANG ; Xu SU
Chinese Journal of Radiological Medicine and Protection 2018;38(10):728-733
Objective To study the role of Cx43 in X-ray induced apoptosis of HUVEC cells and its mechanism.Methods Flow cytometry was used to detect the apoptosis of HUVEC cells at 48-96 h after 10 Gy X-ray irradiation and at 72 h after irradiation of different doses.Western blot was used to detect the protein expressions of Cx43 and cleaved caspase-3 in HUVEC cells at 72 h after 0,5,10 and 20 Gy irradiation.Small interfering RNA was transfected into HUVEC cells to silence Cx43 expression,the Cx43 bearing plasmid was transfected into cells to overexpress Cx43.The effect of Cx43 knockdown or overexpression on apoptosis induction and cleaved caspase-3 protein expression were detected by flow cytometry and Western blot,respectively.Results The apoptosis of HUVEC increased significantly from 48 h to 96 h after X-ray irradiation and in a dose-dependent manner at 72 h after irradiation.The expression of Cx43 protein was negatively correlated with the dose but the expression of cleaved caspase-3 was positively correlated with the dose in the range of 0-20 Gy.After Cx43 silencing,the proportion of early apoptosis and apoptosis combined with dead cells were significantly higher than that of the siRNA control group(t =3.674,6.375,P < 0.05).After Cx43 overexpression,the proportion of early apoptosis and apoptosis combined with dead cells were significantly lower than that of vector control group(t =9.399,11.190,P < 0.05).The expression of cleaved caspase-3 in the Cx43 silencing group was higher than that in the siRNA control group,but this protein in the Cx43 overexpressed group was lower than that in the vector control group.Conclusions Cx43 may protect X-ray irradiated HUVEC cells from apoptosis by down-regulating the activation of caspase-3.
8. Effect of treatment of infant parotid hemangioma with no response to oral propranolol
Yu TIAN ; Xuesong GUO ; Jiang NAN ; Xiaoli WANG ; Gang SHEN
Chinese Journal of Stomatology 2018;53(1):49-51
Hemangioma is the most common vascular tumor in infantile period, and propranolol is the first choice, but there are still a few patients with poor curative effect. Seven cases of infant parotid hemangioma with no response to oral propranolol were treated with transcatheter arterial sclerosing embolization combined with cortisol and satisfactory results achievod. The treatment and efficacy are disccused in this paper.
9.Changes of connexin 43 in irradiated human vascular endothelial cells and its influence on cell stiffness
Chen LI ; Mei TIAN ; Qiao GOU ; Xuesong QI ; Xu SU
Chinese Journal of Radiological Medicine and Protection 2021;41(6):418-425
Objective:To investigate the changes of connexin 43 (Cx43) in human umbilical vein endothelial cells (HUVEC) after X-ray irradiation and its influence on the stiffness of irradiated cells.Methods:Western blot was used to detect the expression of Cx43 in HUVEC cells at different time points (0, 6, 12, 24 and 48 h) after different doses of X-ray irradiation (0, 2.5, 5, 10 and 20 Gy), and the phosphorylation levels of three phosphorylation sites (Ser279/282, Ser368 and Tyr265) of Cx43 at different time points (3, 6, 24 and 48 h) after 0, 5 and 10 Gy irradiation. The distribution of Cx43 protein in the irradiated HUVEC cells was detected by immunofluorescence. The stiffness changes of cells were detected by atomic force microscopy (AFM) at the depths of 50, 100 and 200 nm.Results:The expression of Cx43 in HUVEC cells was reduced at 6, 12, 24 and 48 h after 10 Gy X-ray irradiation( t=3.262, 3.708, 3.686, 6.825, P<0.05)and this decrease had a dose dependent manner at 24 h after 2.5, 5, 10 and 20 Gy irradiation ( t=3.034, 10.720, 13.130, 13.650, P<0.05). At 24 h after 5, 10 and 20 Gy X-ray irradiation, the distribution of Cx43 in HUVEC cells was transported from intercellular gap junctions to nucleus and perinuclear region. At 24-48 h after irradiation, the phosphorylation level of Ser368 at Cx43 increased and in a dose dependent manner. At 24 h after irradiation, the stiffness of the irradiated cells decreased significantly under the conditions of 100 and 200 nm ( t=3.362, 5.122, P<0.05), and recovered with overexpression of Cx43 ( t=2.674, 4.398, P<0.05). Conclusions:X-ray irradiation leads to the phosphorylation of Ser368 at Cx43, which promotes the degradation and nucleus/perinuclear translocation of Cx43 and reduces the stiffness of HUVEC. Increasing the expression level of Cx43 is helpful to the stiffness recovery of irradiated vascular endothelial cells, suggesting that Cx43 may be a potential target for regulating radiation injury of vascular endothelial cells.
10.Laparoscopic limited anatomical hepatectomy for hepatocellular carcinoma within the right anterior section: a propensity score matched study
Yuewen KUANG ; Xuesong LI ; Jianwei LI ; Xiaojun WANG ; Feng TIAN ; Li CAO ; Renjie LI ; Kexi LIAO ; Bowen ZHENG ; Yue WANG ; Shuguo ZHENG
Chinese Journal of Hepatobiliary Surgery 2023;29(11):826-831
Objective:To study the efficacy of laparoscopic limited anatomical hepatectomy (LLAH) for hepatocellular carcinoma (HCC) within the right anterior section.Methods:The clinical data of 144 patients with HCC confined in the right anterior section undergoing hepatectomy at the First Affiliated Hospital of Army Medical University from January 2015 to December 2022 were retrospectively analyzed, including 122 males and 22 females, aged (54.5±9.7) years. Patients were divided into LLAH ( n=27), laparoscopic anatomical hepatectomy (LAH, n=69), and laparoscopic non-anatomical hepatectomy (LNAH, n=48). Propensity score matching was used to compare the operative time, postoperative hospital stay, postoperative complications, serum total bilirubin and albumin, and the prognostic indicators such as tumor-free survival (DFS) rate and cumulative survival rate between the groups. Results:After propensity score matching, there were 26 cases each in LLAH and LNAH group. There was no significant difference in operative time, intraoperative blood loss and postoperative hospital stay between LLAH group and LNAH group (all P<0.05). The total bilirubin and albumin in LLAH on the third day after operation were [ M( Q1, Q3)] 24.1(20.9, 29.1) μmol/L and (35.8±2.9) g/L, better than those in LNAH group 39.3(33.2, 57.0) μmol/L and (33.9±2.5) g/L, respectively. The 1- and 3-year DFS rates in LLAH group were 92.3% and 57.7%, higher than those in LNAH group (80.8% and 19.2%) (all P<0.05). After propensity score matching, there were 25 patients each in LLAH and LAH group. The operative time, postoperative hospital stay and postoperative complications of LLAH group were lower than those of LAH group, and the liver function parameters of LLAH group was also better than those of LAH group (all P<0.05). There was no significant difference in DSF rate between the two groups LLAH group and LAH group ( χ2=0.10, P=0.800). Conclusions:The perioperative outcome of LLAH for HCC within the right anterior section are similar to that of LNAH and better than that of LAH. The DFS of LLAH were better than that of LNAH and similar to that of LAH.