1.Promoting effect of percutaneous electrical stimulation on nerve regeneration after end-to-side neurorrhaphy in rats
Qian SUN ; hua Qun CAO ; Wei ZHU ; dong Yuan MEI ; Chen GU
Chinese Journal of Comparative Medicine 2017;27(12):73-78
Objective To observe and explore the effect and clinical value of percutaneous electrical stimulation on nerve regeneration after end-to-side neurorrhaphy in rats. Methods Thirty-two SPF male S-D rats were randomly divided into four groups ( n = 8 ): group A, the normal control group; group B, with end to end neurorrhaphy of musculocutaneous nerve injury matched to the ulnar nerve; group C, with end to side neurorrhaphy of musculocutaneous nerve injury matched to the ulnar group; and group D, with end to side neurorrhaphy of musculocutaneous nerve injury matched to the ulnar nerve plus postoperative transcutaneous electrical stimulation ( 30 min per day for 6 weeks ) . Electromyography, postoperational nerve conduction velocity, the histological and ultrastructural changes of the nerve fibers were examined, and NF-200 expression in frozen sections was observed using imunohistological staining, to assess the recovery of muscle strength of the diseased side limb and the neuroregeneration in the rats after treatment. Results The amplitude and conduction velocity of the groups C and D were lower than that of the group A, the latency was higher than that of the group A, while the amplitude and conduction velocity of the group D were lower than that of the group C,and the latency was higher than that of the group C. The wet weight ratio of the biceps brachii muscle and the cross-sectional area of muscle fibers in the groups B, C and D were lower than those in the group A, and the recovery of muscle in the group C was the worst. The expression of NF-200 in the rats of groups B, C and D was significantly lower than that in the group A, and the expression of NF-200 in the group D was significantly higher than that in the group C, but still significantly less than that in the group B ( P < 0. 05 ) . Electron microscopy showed mature myelinated fibers in the group B, whereas unmyelinated fibers were the main component and the myelin sheath was poorly developed in the group C. The myelin regeneration in the group D was better than that in the group C, but still some unmyelinated nerve fibers were seen. Conclusions The percutaneous electrical stimulation can effectively promote nerve axonal regeneration and can delay the atrophy of the target muscle after end-to-side neurorrhaphy. Though there is difference compared with the end-to-end neurorrhaphy, the end-to-side neurorrhaphy is still an effective method in clinical repair of peripheral nerve injury.
2.Promoting effect of intermedin and adrenomedullin on cerebral microcirculation in rat models of cerebral ischemia
Xin GUO ; Jing WANG ; Meng WANG ; ying Yue LU ; na Na JIA ; gang Zhen LIU ; rui Jing TIAN ; Jie YUAN
Chinese Journal of Comparative Medicine 2017;27(12):66-72
Objective To explore the effect of intermedin ( IMD ) and adrenomedullin ( ADM ) on cerebral microcirculation in rats with cerebral ischemia. Methods Rat cerebral ischemia ( CI) model was established by middle cerebral artery occlusion. 40 SPF male adult Sprague-Dawley ( SD) rats were randomly divided into three groups:CI+NS ( normal saline) group, CI+ADM group and CI+IMD group, which were used to observe the changes of brain surface microcirculatory perfusion with a laser Doppler flowmeter. Results The differences of brain surface microcirculatory perfusion were statistically significant among the CI+NS group, CI+ADM group and CI+IMD group ( F=53. 426, P<0. 05 ) . Multiple comparison showed that the brain surface microcirculatory perfusion in the CI+IMD group was higher than that of the CI+NS group and CI+ADM group. Conclusions Intermedin can improve the cerebral microcirculation in rats with cerebral ischemia, and its therapeutic effect is better than adrenomedullin.
3.Effect of hypobaric hypoxia and cold exposure on formation of brown adipose tissue in mice
juan Yong TONG ; Duo YANG ; Na LI ; Hua SHU ; Gang MA ; na Na GAO ; ying Guo ZHANG ; di Xiao ZHANG
Chinese Journal of Comparative Medicine 2017;27(12):61-65
Objective To investigate the effect of hypobaric hypoxia and cold exposure on brown adipose tissue in mice. Methods Twenty-four 6-week old SPF C57BL/6 male mice were randomly divided into 4 groups with 6 mice in each group: normal atmospheric pressure and temperature group ( 18~22℃, 20~60 m ) ( NTNP ) , low atmospheric pressure and normal temperature group ( 18~22℃, altitude of 5000 m ) ( NTLP ) , normal atmospheric pressure and cold exposure group(0~6℃, altitude of 20 ~60 m)(LTNP), low atmospheric pressure and cold exposure group(0 ~6℃, altitude of 5,000 m)(LTLP). The experimental period was 4 weeks. The body weight was measured at the beginning and end of the experiment. By the end of the four-week trial, the back and inguinal fat were dissected and observed by histology using HE staining. The expression of UCP-1 as the marker of brown adipose tissue in the back fat was detected by qPCR and western blot. Results The body weight gain of NTNP group was higher ( P< 0. 05 ) than the other three groups. Meanwhile, the color of the back and groin fat tissue of mice of LTNP and LTLP groups were darker, the blood supply in mice of these two groups was richer than the NTLP group. The volume of adipose tissue of NTNP group was higher than others. The histology showed that the back adipose cells of the mice were smaller and darker and full of multilocular lipid droplets, exhibiting a typical morphology of brown fat cells. Compared with the NTNP and NTLP groups, the mRNA and protein levels of UCP-1 were higher under cold exposure, while low atmospheric pressure had a tendency to reduce the mRNA expression of UCP-1. Conclusions The formation of brown fat is affected by the imitated conditions of low atmospheric pressure and cold exposure, and is more closely related to the decresed temperature.
4.Protective effects of Toddalia asiatica extract on oxidative damages in mice with myocardial ischemia and hyperlipidemia
Ming LIU ; Ying DENG ; Yang LIU ; Yun CHEN
Chinese Journal of Comparative Medicine 2017;27(12):51-55
Objective To explore the effects of Toddalia asiatica extract( TAE) on oxidative damages in mice with myocardial ischemia and hyperlipidemia. Methods Fifty SPF male KM mice were randomly divided into 5 groups:control group,model group,simvastatin group (5 mg/kg),high-dose TAE group (400 mg/kg) and low-dose TAE group (100 mg/kg) . Hyperlipidemia was induced by feeding milk for 4 weeks. At the same time,the corresponding drug was given by oral administration with 20 mL/kg for 28 d. At the end of four-week treatment, isoprenaline hydrochloride ( Iso ) was subcutaneously injected every 24 hours for three times. The body weight, electrocardiogram, heart and liver indexes, myocardial water content, and the serum levels of TC, TG, HDL, LDL, SOD, MDA and GPx were measured. Results TAE significantly improved the electrocardiogram changes induced by Iso and milk in the mice, weight lose, inhibited the heart and liver indexes, reduced the myocardial water content, decreased the levels of TC, TG, LDL and MDA, and increased the contents of HDL, SOD and GPx. Conclusions TAE may play an important role in cardiovascular protection by regulating oxygen free radical metabolism and improving oxidative damages.
5.Effect of sitostero-3-O-glucoside on proliferation, apoptosis and collagen synthesis of keloid fibroblasts
wei Jing LV ; jing Jia WANG ; Gang HU
Chinese Journal of Comparative Medicine 2017;27(12):46-50,114
Objective To explore the effect of sitostero-3-O-glucoside on proliferation, apoptosis and collagen synthesis of keloid fibroblasts ( KF) . Methods Cell viability was measured by MTT assay after cells were cultured with 0 (control), 3. 125, 6. 25, 12. 5, 25, 50, 100 μg/mL sitostero-3-O-glucoside. Cell proliferation was measured by EdU staining. Cell apoptotic rate and cell cycle were measured by flow cytometry. The level of COL I and COL III was detected by enzyme linked immunosorbent assay (ELISA). Results 3. 125, 6. 25, 12. 5, 25, 50, 100 μg/mL sitostero-3-O-glucoside reduced cell viability ( P< 0. 01 ) , increased inhibitory rate of cell proliferation ( P< 0. 01 ) , IC50 =27. 54 ± 2. 18 μg/mL. Compared with the control group, 12. 5, 25, 50μg/mL sitostero-3-O-glucoside reduced the cell proliferation rate (P< 0. 01), increased the cell early and late apoptotic rates (P< 0. 01), the cell cycle was arrested at G1 phase (P< 0. 01), and the levels of COL I and COL III were down-regulated (P< 0. 01). Conclusions The results show that sitostero-3-O-glucoside can significantly inhibit the proliferation, induce cell apoptosis and inhibit the synthesis of collagen of keloid fibroblasts.
6.Effect of storage time on serum biochemical test of rats
jun Ai KANG ; wei Yu LI ; wen Xue WANG ; fang Jun LIU ; guo Jian HU ; hui Zhen ZHENG
Chinese Journal of Comparative Medicine 2017;27(12):16-20
Objective To observe the effect of different storage time on 14 blood biochemical indexes in rats. Methods Randomly selected 40 adult SD rats were included in this study. Fasting venous blood samples were collected, serum was separated, sealed, and stored in the refrigerator (4℃ and -20℃). The serum parameters were detected at 0 h,4 h,24 h,96 h and 7 d, respectively, using an automatic biochemical analyzer. A total of 14 blood biochemical indexes were detected, including alanine aminotransferase ( ALT ) , aspartate aminotransferase ( AST ) , alkaline phosphatase (ALP), total protein (TP), albumin (ALB), creatinine (CREA-J), uric acid (UA), urea nitrogen (UREA), blood glucose ( GLU) , total cholesterol ( TC) , triglyceride ( TG) , low density lipoprotein cholesterol ( LDL-C) , creatine kinase ( CK) and lactate dehydrogenase ( LDH) . The effects of serum storage time on blood biochemical results were compared. Results The trends of blood biochemical data in male and female rats were consistent. C ompared with the indexes of serum preserved at 4℃ for 0 h, the ALP was significantly reduced after storage for 4 h, 24 h, 96 h, and 7 d (P< 0. 05), ALB were significantly increased after 96 h and 7 d (P< 0. 01), CREA-J was significantly increased after 96 h, 7 d (P<0. 05), UA was significantly increased after 24 h, 9 h, and 7 d (P < 0. 01), and no significant changes in other indicators ( P> 0. 05 ) . Compared with the values of 0 h serum, the serum preserved at -20℃ showed that ALT was significantly increased after 7 d (P < 0. 01), AST significantly increased after 96 h and 7 d (P< 0. 05), TP significantly decreased after 4 h and 24 h ( P< 0. 05 ) , ALB significantly increased after 4 h, 24 h, 96 h, and 7 d ( P< 0. 01 ) , CREA-J significantly increased after 24 h, 96 h, and 7 d (P< 0. 01), UA significantly increased after 4 h, 24 h, 96 h, and 7 d (P< 0. 01), TC significantly increased after 4 h, 24 h, 96 h, and 7 d (P< 0. 01), TG significantly increased after 96 h and 7 d (P< 0. 05), CK significantly increased after 96 h and 7 d (P< 0. 05), LDH significantly increased after 96 h and 7 d ( P < 0. 05 ) , and no significant changes in other indicators ( P > 0. 05 ) . Conclusions The biochemical tests of rat serum should be immediately performed as they were collected, especially for ALP test. For the sera stored at 4℃, the test should be finished in different times:UA test in 4 hours, ALB and CREA-J test in 24 hours, and ALT, AST, TP, UREA, GLU, TC, TG, LDL-C, CK, and LDH test in 7 days.
7.Current status of research on glucocorticoid-induced mouse models of osteoporosis
qun Xiao LI ; Jin QIAN ; can Jia SU
Chinese Journal of Comparative Medicine 2017;27(12):120-124
Glucocorticoid is a kind of steroids hormone secreted by the adrenal cortex zona fasciculate or artificially synthesized. It can mediate the synthesis and metabolism of carbohydrate, lipid and protein and has the function of inhibiting immune response and anti-inflammation, anti-toxic and anti-shock effects. However, long-term intake of corticostereoid hormone will result in bone loss, even severe osteoporosis, and increase the risk of fracture. As a result the research on the mechanisms of osteoporosis become more and more necessary. Now, according to the differences in establishment methods, there are three approaches, i. e. pellet implantation, drinking water and injection. As a result of the only partial similarity between different osteoporosis models and humans, there is a special application of each type of animal model of osteoporosis. The osteoporosis in glucocorticoid-induced model is mainly caused by modulating incretion, promoting bone absorption and inhibiting the osteoblast differentiation. In the meanwhile, compared with the other animal models, genes in mice are closer to humans, and they have many advantages in cost, gene and cell techniques. Therefore, glucocorticoid-induced mouse models of osteoporosis has a great significance in osteoporosis research. This article will review the establishment methods of glucocorticoid-induced mouse models of osteoporosis.
8.Research progress of bronchoalveolar lavage in rats and mice
Long LONG ; li Xian ZHAO ; Hong TAN ; hua Ai ZHANG ; bo Hong YANG ; lin Jin HE
Chinese Journal of Comparative Medicine 2017;27(12):115-119
Bronchoalveolar lavage is an important animal experimental technique in the study of respiratory system and its pathological changes. It can acquire a variety of biochemical factors, inflammatory mediators and immune cells from the respiratory tract and lungs, and provides an important evaluation index and reference for animal experiment. Bronchoalveolar lavage is an effective and reliable method for the diagnosis of respiratory diseases. It has been gradually standardized and widely used in clinical practice at present, however, there is no set of standard for bronchoalveolar lavage in rats and mice. The results of bronchoalveolar lavage fluid are affected by many factors, such as the lavage fluid, suction pressure, the amount of lavage and recovery, and the retention time of lavage fluid in the lungs. Successful and efficient acquisition of lavage specimens is the key to the study and evaluation of respiratory diseases. This paper summarizes the current lavage methods commonly used by domestic and foreign researchers, and provides a reference for further research in the this field.
9.Advances in research on the role of Siglec-1 in diseases
lei Ke LI ; Xiang LI ; Zhe CONG ; Jing XUE
Chinese Journal of Comparative Medicine 2017;27(12):109-114
Sialoadhesin ( Siglec-1 or CD169 ) is a sialic acid-binding Ig-like lectin expressed selectively on macrophage subsets. In inflammatory response, Siglec-1 can modulate the secretion of MIP-1 alpha / beta, MCP-1, MIP-2 and other cytokines, and promote the occurrence of inflammatory reaction. During viral infection, Siglec-1 can promote the infection and phagocytosis of virus by mediating the binding of pathogens and macrophages. In the regulation of immunity, Siglec-1 can regulate innate immunity and adaptive immunity, by inhibiting the excessive expression of IFN-α and the activation of DC cells. This review mainly focuses on the new advances of research on Siglec-1 in pathogen infections, inflammation and immunoregulation.
10.Establishment of a mouse model of IgA nephropathy induced by superantigen staphylococcal enterotoxin B
qiang Li SHI ; Qiang WAN ; sheng Yan WU ; wei Wei LIU ; dong Jian GAO
Chinese Journal of Comparative Medicine 2017;27(12):102-108
Objective To establish a mouse model of IgA nephropathy and to observe its biochemical and pathological characteristics. Methods Twelve BALB/c mice were randomly divided into the normal group and model group, with 6 mice in each group. Mice in the model group received an intravenous injection of 0. 8 mg/kg superantigen staphylococcal enterotoxin B (SEB) into the tail vein once a week for three weeks. At the end of the 4th week, the mice were sacrificed, and the 24 h-urinary protein, urinary microalbumin, the renal function indicators BUN, Scr and UA were measured, levels of liver function indicators ALT, AST, ALP, and the blood lipid levels of TC, TG, and LDL were determined, the renal morphological changes were examined by pathology using HE, PAS, PASM and Masson staining, and by electron microscopy, the IgA deposition in the renal tissue was observed with immunofluorescence, and the liver and small intestine were observed by pathology using HE staining. Results Compared with the normal group, the mice of model group showed increased 24-hour urinary protein and urinary microalbumin (P<0. 01), increased CREA and UA (P<0. 05), but not significantly changed BUN, TP and ALB. The liver function indicator AST was significantly increased (P<0. 05), but ALT and ALP were not significantly changed. The blood lipid TG was significantly decreased (P<0. 05) and LDL increased (P<0. 01), while the TC was not significantly changed. The kidney tissues had moderate histological changes, and immunofluorescence observation showed granular or massive IgA deposition in the renal glomerular mesangium. The liver tissue had some inflammatory cell infiltration and hepatocyte necrosis. The small intestine showed slender and shortened villi with widened inter-villous space and sloughed off epithelial cells, dilated central lacteal, and lymphocyte infiltration. Conclusions A mouse model of IgA nephropathy can be successfully established by tail vein injection of superantigen staphylococcal entrotoxin B.