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    30 November 2015, Volume 19 Issue 49 Previous Issue    Next Issue
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    Effect of warm needling on histomorphology of compressed lumbar nerve root in rats
    Xie Yan-yan, Wu Yao-chi
    2015, 19 (49):  7878-7878.  doi: 10.3969/j.issn.2095-4344.2015.49.001
    Abstract ( 388 )   PDF (628KB) ( 523 )   Save

    BACKGROUND: 80% of patients with early-stage lumbar disc herniation can be alleviated or cured by non-surgical treatment. Acupuncture is one of the very important non-surgical treatments, and has a definite therapeutic effect. Warm needling is an important part of acupuncture and moxibustion and exhibits the dual effects of acupuncture and moxibustion, including promoting coordination between Qi and blood, dredging the meridian passage and removing blood stasis to alleviate pain.
    OBJECTIVE: To observe the effect of warm needling on histomorphology of compressed lumbar nerve root compression in rat models.
    METHODS: Sixty Sprague-Dawley rats were divided into five groups: normal, model, meloxicam, acupuncture and warm needling groups (n=12 rats/group). A special silicone sheet was placed on the right side of the dural sac at the L5 nerve root to establish rat models of lumbar nerve root compression. Rats in normal group were not subjected to model establishment and treatment. Rats in model group were subjected to model establishment, but without treatment. Rats in meloxicam group were given the meloxicam treatment for 2 weeks after modeling. Rats in acupuncture group were given 2 weeks of acupuncture treatment after modeling. Rats in warm needling group were given 2 weeks of warm needling treatment after modeling. At 2 weeks after modeling, compressed lumbar nerve root tissue was harvested and stained with hematoxylin-eosin for histomorphological observation.
    RESULTS AND CONCLUSION: At 2 weeks after rat models of lumbar nerve root compression were successfully established, as confirmed by pathological observation of nerve root section. Inflammatory response in the tissue surrounding the nerve fibers and dorsal root ganglia in the warm needling group better improved than in the meloxicam and acupuncture groups. These results suggest that compared with meloxicam and acupuncture, warm needling can more effectively improve the damaged rat nerve root structure and reduce the inflammatory response in rats with lumbar nerve root compression.  

     

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    Type II collagen expression in a rabbit model of developmental dysplasia of the hip
    Zhang Xiang-xin, Chen Guang-xiang, Cheng Liang, Xu Ren-jie, Zou Tian-ming
    2015, 19 (49):  7879-7884.  doi: 10.3969/j.issn.2095-4344.2015.49.002
    Abstract ( 395 )   PDF (440KB) ( 565 )   Save

    BACKGROUND: Mild developmental dysplasia of hip causes high morbidity of osteoarthritis on adults. It is thought that change of collagen and proteoglycans in cartilage may be the direct reasons for osteoarthritis.
    OBJECTIVE: To observe the changes in the expressions of type II collagen of acetabular cartilage in a rabbit model of early developmental dysplasia of hip. 
    METHODS: Left lower extremity served as experimental side and right one as control. The rabbit model of developmental dysplasia of hip was successfully established by applying the method of knee extending and fixing with cylinder cast.
    RESULTS AND CONCLUSION: Local chondrocyte clusters were visible in acetabular cartilage on the experimental side. Toluidine blue staining loss was observed in extracellular matrix. Moreover, the number of type II collagen-positive cells and type II collagen expression were higher in the experimental side than in the control side. These results indicate that type II collagen expression may be associated with cartilage degeneration.
     中国组织工程研究杂志出版内容重点:肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程

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    Establishment of rabbit models of chronic femoral osteomyelitis
    Chen Jin-shui, Liao Zhao-shan, Lin Song-qing, Lu Qiao-ping, Wang Ben-hai, Huang Wen-qi, Lin Yong-yang, Zhang Hui-hao
    2015, 19 (49):  7885-7889.  doi: 10.3969/j.issn.2095-4344.2015.49.003
    Abstract ( 489 )   PDF (534KB) ( 430 )   Save

    BACKGROUND: Establishment of an effective and stable animal model of chronic osteomyelitis is the basis of the experimental studies on treatment of chronic osteomyelitis. Previous chronic femoral osteomyelitis models are always instable and therefore not successful.
    OBJECTIVE: To investigate the experimental method of establishing rabbit models of chronic femoral osteomyelitis with Staphylococcus aureus .
    METHODS: Forty healthy adult New Zealand rabbits were anesthetized. After stripping periosteum located at 2 cm below the greater trochanter, two bone holes till the medullary cavity, which were connected longitudinally and overlapped partially, were drilled with a drill of 3.5 mm bore bit. Holes were sealed with sterile bone wax. Twenty rabbits from the experimental group were intramedullarily injected first with 5% sodium morrhuate (0.1mL), and then with 0.5 mL Staphylococcus aureus (1×106 cfu). Twenty rabbits from the control group were intramedullarily injected first with 5% sodium morrhuate (0.1 mL) and then with 0.5 mL normal saline. After 4 weeks, infected regions were observed, rabbit body weight and body temperature were measured and X-ray radiographs of the damaged regions were taken, bacterial culture was performed and femoral osteomyelitis lesion degree was 
    evaluated using the modified osteomyelitis scoring system.
    RESULTS AND CONCLUSION: After 1 week of bacterial inoculation, 20 rabbits from the experimental group began to  present the symptoms of increased body temperature, loss of appetite, lameness, vaccination site reaction. After 2 weeks, exudate or pus discharge in the local area were present in 13 rabbits. After 4 weeks, 19 rabbits all presented the symptoms of local swelling, pus discharge and fever. At this time period, rabbit body temperature was noticeabley increased, body weight was remarkably decreased, and modified Norden osteomyelitis scores were obviously higher in the experimental group compared with the control group. The secretion cultures were positive for Staphylococcus aureus in 14 rabbits. The success rate of modeling was 95%. These results suggest that stable and reliable rabbit models of chronic femoral osteomyelitis can be prepared by injection with Staphylococcus aureus and sodium morrhuate into marrow cavity. 
     

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    Quick establishment of nude mouse models of orthotopic osteosarcoma using tissue mass transplantation  
    Zhou Bin
    2015, 19 (49):  7890-7894.  doi: 10.3969/j.issn.2095-4344.2015.49.004
    Abstract ( 500 )   PDF (503KB) ( 628 )   Save

    BACKGROUND: Construction of animal models of osteosarcoma is difficult, and the progress is slow because osteosarcoma cells are from mesenchymal tissue.
    OBJECTIVE: To establish the nude mouse models of orthotopic osteosarcoma using tissue mass transplantation method, observe and evaluate its biological characteristics.
    METHODS: Fifteen nude mice aged 6 weeks of SPF grade were selected. The right forelimb armpit of nude mice was injected with the prepared MG-63 osteosarcoma cell suspension. After tumorgenesis, osteosarcoma cells were sub-cultured for 3 generations. When the tumor grew stably, tumor tissue block was transplanted into lower tibial marrow cavity of nude mice and its biological characteristics were evaluated. Tumor-forming rate and tumor characteristics were observed by X-ray, and 5 normal nude mice were taken as controls.
    RESULTS AND CONCLUSION: The modeling process in these 15 nude mice went smoothly. After modeling, only one nude mouse failed in tumor induction and the tumor-forming rate was 93%. Tumor-like tissue formation in the lower extremity of nude mice was visible at 3-4 weeks after tissue mass transplantation. After four or five weeks, X-ray observation showed that unobvious osteolytic bone tumor and tumor-like lesions were present in the upper and middle segments of the tibia of nude mice. The expression level of alkaline phosphatase in the osteosarcoma model group was significantly higher than in the control group (P < 0.01). These results show that tissue mass transplantation for modeling orthotopic osteosarcoma in nude mice is simple to operate and has a high tumor-forming rate. The induced tumors grow quickly and exhibit strong destructive power to cortical bone and surrounding soft tissue, which are close to the actual situation of patients with osteosarcoma in the clinic.
     

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    Construction of elderly SIRT1 gene knockout mouse models of knee osteoarthritis
    Yu Fei, Lei Ming, Zeng Hui, Li Ming-hua, Xiao De-ming
    2015, 19 (49):  7895-7901.  doi: 10.3969/j.issn.2095-4344.2015.49.005
    Abstract ( 711 )   PDF (702KB) ( 559 )   Save

    BACKGROUND: Scholars in China and countries outside China conducted a lot of researches on the pathogenesis and treatment of osteoarthritis, and recently found that SIRT1 gene has an important significance in the pathogenesis of osteoarthritis. However, little is reported on SIRT1 gene, and there are few studies on animal models of knee osteoarthritis used for SIRT1 research.
    OBJECTIVE: To establish specific SIRT1 gene knockout mouse models of knee osteoarthritis, in order to provide favorable conditions for the study of osteoarthritis.
    METHODS: Mice were randomly divided into four groups according to the method of randomized controlled grouping: SIRT1+/+ control group (Group A, n=6); SIRT1+/+ osteoarthritis model group (Group B, n=6); SIRT1-/- control group (Group C, n=6); SIRT1-/- osteoarthritis model group (Group D, n=6). Unilateral knee anterior cruciate ligament transection and medial meniscectomy were conducted to establish mouse models of osteoarthritis. SIRT1 gene knockout was determined by quantitative fluorescence polymerase chain reaction. Knee cartilage morphological changes were observed by hematoxylin-eosin and safranin O-fast green double staining. The degree of knee osteoarthritis was evaluated by Mankin score. 

     

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    Effects of remote ischemic postconditioning on inflammation related factors of cerebral ischemia/reperfusion injury
    Wang Ning, Li Xiao-ou, Ba Xiao-hong
    2015, 19 (49):  7902-7907.  doi: 10.3969/j.issn.2095-4344.2015.49.006
    Abstract ( 530 )   PDF (478KB) ( 516 )   Save

    BACKGROUND: Although some studies have suggested that remote ischemic postconditioning plays a neuroprotective role, but the specific mechanism is still unknown.
    OBJECTIVE: To investigate the protective effects of remote ischemic postconditioning on focal cerebral ischemia/reperfusion injury in rats.
    METHODS: Focal cerebral ischemia/reperfusion injury rat models were established by occlusion of the middle cerebral artery using suture method and were subjected to remote ischemic postconditioning. Meanwhile, sham and ischemia/reperfusion groups were established as comparison. After 24 hours of ischemia/reperfusion, neurological function was graded to determine cerebral infarction volume and brain water content. The mRNA expression of interleukin 1β, interleukin-6, tumor necrosis factor α and monocyte chemoattractant protein 1 in brain tissue around ischemic area was detected by RT-PCR. The protein expression of Bcl-2 and Bax was detected by western blot analysis.
    RESULTS AND CONCLUSION: Compared with ischemia/reperfusion group, the neurological function scores in the remote ischemic postconditioning group were slightly, but not significantly, decreased. Infarct volume and brain water content in remote ischemic postconditioning group were significantly decreased compared with those in ischemia/reperfusion group (P < 0.05). The mRNA expression of interleukin 1β, interleukin-6, tumor necrosis factor α and monocyte chemoattractant protein 1 and the protein expression of Bax in brain tissue around 
    ischemic area in ischemic postconditioning group were significantly decreased compared with those in ischemia/reperfusion group (P < 0.05). Bcl-2 protein expression significantly increased (P < 0.05). These results demonstrate that remote ischemic postconditioning can relieve focal cerebral ischemia/reperfusion injury in rats, the mechanism of which may be associated with reduction of inflammatory response. 

     

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    Effects of cyclooxygenase inhibitors on nuclear factor κB and CD4+/CD8+ expression in rat models of acute incomplete spinal cord injury 
    Xin Zhi-qiang, Pan Meng, Wang Jing-liang, Zheng Quan-xin, Xu Yi-dong, Pi An-ping
    2015, 19 (49):  7908-7913.  doi: 10.3969/j.issn.2095-4344.2015.49.007
    Abstract ( 270 )   PDF (647KB) ( 289 )   Save

    BACKGROUND: Celecoxib has advantages including fewer adverse reactions and low cost over the commonly used methylprednisolone in the treatment of acute spinal cord injury, but the effective dose cannot be well controlled.
    OBJECTIVE: To investigate different doses of cyclooxygenase inhibitors on nuclear factor κB and CD4+/CD8+ expression in rat models of acute incomplete spinal cord injury.
    METHODS: Rat models of incomplete spinal cord injury in T8 and T9 segments were established using modified Allen method. Sixty successful modeling rats were randomly and evenly divided into 4 groups: simple injury group and low, medium-and high-dose of celecoxib groups. Rats in simple injury group were not applied any drugs. Rats in low-, medium- and high-dose celecoxib groups were respectively intragastrically administered 25, 50 and 100 mg/kg celecoxib after 30 minutes of spinal cord injury. 
    RESULTS AND CONCLUSION: After celecoxib treatment, rat neurological function was significantly improved. Nuclear factor κB expression level in spinal cord was significantly decreased. CD4+ and CD8+ T cell numbers in peripheral blood were significantly decreased. Low dose of celecoxib treatment had a better outcome. These results suggest that different doses of celecoxib can treat acute traumatic spinal cord injury, and low-dose
    (25 mg/kg) celecoxib treatment can achieve a better outcome. 
     

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    Establishment of an asthma model in Balb/c and DO11.10 mice
    Deng Xue-quan, Yao Yin, Guo Jie-bo, Gao Wen-xiang, Sun Yue-qi, Fu Qing-ling
    2015, 19 (49):  7914-7919.  doi: 10.3969/j.issn.2095-4344.2015.49.008
    Abstract ( 640 )   PDF (636KB) ( 624 )   Save

    BACKGROUND: The incidence of asthma increases year by year. Therefore, establishment and improvement of optimized animal models of asthma are important for the investigation of asthma. 
    OBJECTIVE: To induce asthma method in Balb/c and DO11.10 mice and to compare their difference. 
    METHODS: A total of 24 Balb/c mice were equally and randomly divided into groups A-D and 24 DO11.10 mice were equally divided into groups E-H. Mice in the groups A and E were induced to sensitization using PBS on day 1, and atomization using PBS on days 8-11. Mice in the groups B and F were induced to sensitization using ovalbumin on day 1, and atomization using ovalbumin on days 8-11. Mice in the groups C and G were induced to sensitization using PBS on days 1, 7 and 14, and atomization by PBS on days 22-25. Mice in the groups D and H were induced to sensitization using ovalbumin on days 1, 7 and 14, and atomization using ovalbumin on days 22-25. 
    RESULTS AND CONCLUSION: One time of sensitization and four times of atomization only caused mild inflammatory reaction in the lungs in Balb/c mice. Three times of sensitization and 4 times of atomization caused clear airway hyperreactivity, inflammatory cell infiltration in lung, increased mucus secretion in bronchial goblet epithelium in Balb/c mice. Simultaneously, the number of inflammatory cells and eosinophils significantly increased in bronchoalveolar lavage fluid. Th2 type cytokine levels also increased in the bronchoalveolar lavage fluid. In DO11.10 mice, one time of sensitization and four times of atomization induced significant inflammatory response. Specially, the number of neutrophils obviously increased in the bronchoalveolar lavage fluid. However, after three times of sensitization and four times of atomization, inflammatory reaction significantly decreased in DO11.10 mice. These findings indicate that asthma model could be successfully induced with ovalbum in both Balb/c and DO11.10 mice. There were some differences in the induction and pathological features for asthma model between the two species of mice.
     

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    Behavioral and morphological changes of different spinal cord injury rat models 
    Song Wei, Duan Hong-mei, Rao Jia-sheng, Zhao Can, Yang Zhao-yang
    2015, 19 (49):  7920-7925.  doi: 10.3969/j.issn.2095-4344.2015.49.009
    Abstract ( 401 )   PDF (748KB) ( 454 )   Save

    BACKGROUND: Complete spinal cord transection models include cut-type injury and excision-type injury models. It is necessary to compare and discuss these two models, in order to select more appropriate animal models for studying spinal cord injury and repair.
    OBJECTIVE: To analyze the behavioral and pathological changes of different spinal cord injury models and to select the animal models of spinal cord injury which are more suitable for studying regeneration and treatment of injured spinal cord.
    METHODS: One hundred and sixty Sprague-Dawley rats were randomly divided into two groups to establish cut-type and excision-type spinal cord injury models. At 1, 2, 4, 8, 10, 20, 30 and 50 weeks after operation, rat hindlimb locomotor skills were evaluated using BBB scores; histochemical staining and immunohistochemical staining were conducted to observe the residual fibers in injured area; at 4, 10, 26 and 52 weeks after operation, BDA-FITC tracer agent was respectively injected into the motor area of the cerebral cortex and cervical cord of 
    rats, to further observe the continuity of residual fibers in injured area.
    RESULTS AND CONCLUSION: The BBB scores of rat hind limbs in cut-type spinal cord injury model group from 2 to 50 weeks after operation were significantly higher than those in excision-type spinal cord injury model group (P < 0.05). In cut-type spinal cord injury model group, there were a large amount of residual tissues, neurofilaments and glial fibrillary acidic protein-positive fibers in injured area, but there were almost no residual fibers in excision-type spinal cord injury model group. The corticospinal tract could pass through the injured area in neither cut-type spinal cord injury models nor excision-type spinal cord injury models and terminated at the cranial end of the injured area. In cut-type spinal cord injury models, the spinal cord propriospinal tract re-entered into the host spinal cord tissue at the caudal end, whereas the spinal cord propriospinal tract in excision-type spinal cord injury models terminated at the cranial end of the injured area. These results demonstrate that cut-type spinal cord injury models are more suitable for screening and evaluating the regeneration effect of treatments or drugs after spinal cord injury.  

     

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    Effect of breviscapine on protein kinase C-gamma expression in brain tissue of rat models of traumatic brain injury
    Yin Yan-qing, Fang Qi, Liao Zhuang-bin, Lin Heng, Huang Zi-xiong, Liang Yuan-sheng, Xu Cheng-jie
    2015, 19 (49):  7926-7931.  doi: 10.3969/j.issn.2095-4344.2015.49.010
    Abstract ( 370 )   PDF (547KB) ( 297 )   Save

    BACKGROUND: Preventing secondary injury after traumatic injury is of great importance, which is also the target of brain protective treatment.
    OBJECTIVE: To preliminarily investigate the protective effect of breviscapine on brain tissue of rat models of traumatic brain injury through observing the effect of breviscapine on protein kinase C-γ expression in injured brain tissue.
    METHODS: Sixty-five Sprague-Dawley rats were randomly divided into control (n=5), sham (n=20), model (n=20) and breviscapine (n=20) groups. Rat models of traumatic brain injury were established in the model and breviscapine groups by a falling heavy object. In the sham group, rat bone window was open through an incision made on the scalp, and then the incision was sutured immediately. Rats in the breviscapine group were intraperitoneally injected with 0.4 mg/kg breviscapine daily immediately after injury. Rats in the model and sham groups were intraperitoneally injected with 0.4 mL/kg normal saline. The expression of protein kinase C-γ protein in brain tissue was detected by immunohistochemistry.
    RESULTS AND CONCLUSION: After traumatic brain injury, the protein expression of protein kinase C-γ was significantly increased (P < 0.05), and reached the peak at 24 hours. Breviscapine could significantly decrease the protein expression of protein kinase C-γ in brain tissue after traumatic brain injury (P < 0.05). These results demonstrate that breviscapine has a protective effect on brain tissue after traumatic brain injury. 

     

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    Electrical stimulation of the perianal skin regulates neurogenic intravesical pressure in rabbit models of spinal cord injury
    Yan Peng, Zheng Wei-dong, Tan Yun-bing, Deng Xiao-ming, Yang Xiao-yu
    2015, 19 (49):  7932-7937.  doi: 10.3969/j.issn.2095-4344.2015.49.011
    Abstract ( 294 )   PDF (840KB) ( 435 )   Save

    BACKGROUND: Previous studies have shown that different frequencies of electrical stimulation of pudendal nerve and its branches can inhibit detrusor overactivity, thereby treating neurogenic bladder. Based on this, is it possible to inhibit detrusor overactivity through electrical stimulation of pudendal nerve branches?
    OBJECTIVE: To investigate the feasibility of electrical stimulation of the perianal skin regulating neurogenic intravesical pressure in rabbit models of spinal cord injury.
    METHODS: Ten adult female rabbits were selected and subjected to spinal cord transection at T9-T10 level. After 4 weeks of feeding, the experiment began. The skin around the anus was stimulated using a pair of hook-shaped electrodes. A double-lumen balloon catheter was inserted into the bladder. Normal saline was infused into the double-lumen balloon catheter. Intravesical pressure was measured.
    RESULTS AND CONCLUSION: When the perfusion volume of the bladder was higher than the threshold of voiding volume, detrusor overactivity was significantly inhibited after perianal electrical stimulations at frequencies below 10 Hz. When the perfusion volume of bladder was lower than the threshold of voiding volume, bladder 
    contractions were induced by electrical stimulations at frequencies between 20 and 50 Hz. The optimal inhibitory stimulation (6 Hz) could significantly increase the bladder capacity of (30±10)%. The optimal excitatory stimulation (25 Hz) could induce large amplitude (greater than 25 cm H2O, i.e. 24.5 kPa), long duration (greater than 20 seconds) of bladder contractions. These results demonstrate that electrical stimulation of the perianal skin can effectively regulate neurogenic intravesical pressure in rabbit models of spinal cord injury. 
     

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    Effects of ischemic postconditioning on related pathways and interleukin 8 expression in rat models of focal cerebral ischemia/reperfusion
    Li Zhi-xing, Lv Jing-lei, Chen Ping, Zhao Ren-liang
    2015, 19 (49):  7938-7944.  doi: 10.3969/j.issn.2095-4344.2015.49.012
    Abstract ( 374 )   PDF (712KB) ( 419 )   Save

    BACKGROUND: Ischemic postconditioning may motivate endogenous protective effect and inhibit inflammatory response after ischemia/reperfusion, but the specific mechanism is still unclear.
    OBJECTIVE: To investigate the effects of ischemic postconditioning on Toll-like receptor 4/nuclear factor кB signaling transduction pathway and interleukin 8 expression in rat models of focal cerebral ischemia/reperfusion, and further illustrate the neuroprotective mechanism of ischemic postconditioning.
    METHODS: One hundred and ten rats were randomly divided into sham group (n=10), model group (n=50) and ischemic postconditioning group (n=50). Focal cerebral ischemia/reperfusion rat models were established according to Zea-Longa method, and then subjected to ischemic postconditioning, i.e., middle cerebral artery occlusion for 2 hours followed by 3 cycles of 15-second reperfusion/15-second ischemia. Model group and sham group were set for comparison. In each group, rat neurobehavioral deficit scores were evaluated, and rat infarct volume was measured with TTC staining. The expression of Toll-like receptor 4, nuclear factor кB and interleukin 8 protein in brain tissue was detected by immunohistochemistry. Their mRNA expression was detected 
    by in situ hybridization.
    RESULTS AND CONCLUSION: Rats in the model and ischemic postconditioning groups all presented neurobehavioral deficits and cerebral hemisphere infarction on the ischemic side. At 6, 12, 24, 48 and 72 hours after reperfusion, the neurobehavioral deficit scores were significantly decreased in the ischemic postconditioning group compared with that in the model group (P < 0.05). The infarct volume of rats in the ischemic postconditioning group was significantly decreased compared with that in the model group (P < 0.05). The expression of Toll-like receptor 4, nuclear factor кB and interleukin 8 protein and mRNA was increased at 6 hours after reperfusion and reached the peak at 24 hours after reperfusion. At 6, 12, 24, 48 and 72 hours after reperfusion, the expression of Toll-like receptor 4, nuclear factor кB and interleukin 8 protein and mRNA was all significantly decreased in the ischemic postconditioning group compared with that in the model group (P < 0.05). The results confirm that ischemic postconditioning may inhibit inflammatory response induced by ischemia/reperfusion, and play its neuroprotective effect by inhibiting Toll-like receptor 4/nuclear factor кB signaling transduction pathway and down-regulating the expression of interleukin-8. 

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    Effect of nebivolol on pulmonary artery remodeling in rats with high pulmonary blood flow
    Chen Peng, Jiang Ping, Zhu Can-yang,Yang Mei, Ma Hong-jun, Li Yan-feng
    2015, 19 (49):  7945-7950.  doi: 10.3969/j.issn.2095-4344.2015.49.013
    Abstract ( 409 )   PDF (643KB) ( 395 )   Save

    BACKGROUND: It is still unclear whether nebivolol, a vasodilatory-type adrenergic β1 receptor antagonist, can effectively reduce pulmonary artery pressure and what is the effect on pulmonary vascular remodeling.
    OBJECTIVE: To investigate the effect of nebivolol on pulmonary artery remodeling of rats with high pulmonary blood flow.
    METHODS: Forty Sprague-Dawley rats were randomly divided into four groups: model, nebivolol, captopril and sham groups. Rats in the model, nebivolol and captopril groups were prepared to establish rat models of pulmonary hypertension and pulmonary vascular remodeling. Rats in the sham group were only subjected to separation of the abdominal aorta and inferior vena artery. After 5 days of modeling, the rats in the nebivolol group and captopril group were respectively intragastrically administered 1 mg/kg nebivolol solution and 5 mg/kg captopril solution daily. Rats in the model and sham groups were intragastrically administered the same amount of normal saline. After 8 weeks of administration, mean pulmonary arterial pressure, right ventricular hypertrophy index, pulmonary artery morphological changes, pulmonary artery ultrastructure and pulmonary arterial ring diastolic rate were compared among these four groups.
    RESULTS AND CONCLUSION: Compared with the sham group, the muscularization of pulmonary arterioles of 
    rats in the model group was obvious; mean pulmonary artery pressure and right ventricular wet weight/(left ventricle wet weight+interventricular septum wet weight) were significantly increased (P < 0.05 or P < 0.01); pulmonary arterial ring diastolic rate was decreased (P < 0.05 or P < 0.01). Compared with the model group, mean pulmonary artery pressure and right ventricular wet weight/(left ventricle wet weight + interventricular septum wet weight) were significantly decreased (P < 0.05 or P < 0.01); the muscularization of pulmonary arterioles was alleviated and pulmonary arterial ring diastolic rate was increased in the nebivolol and captopril groups (P < 0.05 or P < 0.01). These results demonstrate that nebivolol can effectively alleviate pulmonary artery reconstruction in rats with high pulmonary blood flow through protecting vascular endothelium and reducing pulmonary artery pressure.  

     

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    Establishing an animal model of ischemic stroke with photothrombosis method
    Ma Jun-ning, Gao Jun-wei, Hou Bo-ru, Ren Hai-jun, Liu Ji-xing, Chen Si-hua, Yan Gui-zhong
    2015, 19 (49):  7951-7957.  doi: 10.3969/j.issn.2095-4344.2015.49.014
    Abstract ( 833 )   PDF (698KB) ( 2292 )   Save

    BACKGROUND: Animal model of experimental cerebral ischemia stroke is an indispensable tool in the research of stroke pathology and the development of novel treatment. The animal model of cerebral ischemia stroke established with the photothrombosis method has simple operation, good stability and high repeatability, it can control the position and size of infarction loci, and therefore it is the preferred choice for studying the pathological mechanism of cerebral ischemic stroke.
    OBJECTIVE: To establish animal models of cerebral ischemic stroke with the photothrombosis model, and compare the infarct volume and behaviors of difference genders of mice after ischemic stroke.
    METHODS: Totally 62 Kunming mice were included in this study, 6 of them were used for the pathology detection, 16 of them for the measurement of infarction volume, and 40 for behavioral test. In the pathology detection, 6 mice were equally divided into sham group and model group; in the measurement of infarction volume, 16 mice were equally divided into female model group and male model group; in the behavioral test, 40 mice were equally divided into female model group, female sham group, male model group and male sham group. In the model groups (female model group and male model group), focal cerebral ischemic stroke model was established
    with the photothrombosis method; in the sham groups (female sham group and male sham group), mice were not injected with rose Bengal dye.
    RESULTS AND CONCLUSION: Nissl staining found neuronal degeneration, necrosis, and vacuolar pathology at the center of infarction loci in the model groups. Fluoro-Jade C staining shown neuronal degeneration at the center of infarction loci in the model groups. Infarct volume in female group was significantly smaller than that in male group. The motor function impairment in the model group was significantly greater than that in sham group, and female group showed less impairment of motor function than male group. These results suggest that the animal model of ischemic stroke can be successfully established with the phtothrombosis method. Male animals suffer more neurological damage from ischemic stroke than female under the same condition.
     

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    Construction of diabetic rat models: protective effect of Klotho gene transfer on coronary artery
    Shu Li-liang, Chen Mao-lin, Su Gang, Xu Jing
    2015, 19 (49):  7958-7963.  doi: 10.3969/j.issn.2095-4344.2015.49.015
    Abstract ( 301 )   PDF (585KB) ( 254 )   Save

    BACKGROUND: Klotho gene has an important correlation with organism aging, metabolism and disease. It is confirmed by mouse experiments that Klotho gene can slow down and inhibit atherosclerosis, and its underlying mechanism is involved in lipid metabolism.
    OBJECTIVE: To establish diabetic rat models and observe whether Klotho gene transfer has a protective effect on coronary artery of diabetic rats. 
    METHODS: Klotho gene was extracted from the kidney tissue of normal Sprague-Dawley rats. The target gene was amplified by polymerase chain reaction. Adenovirus was taken as a carrier. Sprague-Dawley rats were randomly divided into model, control and treatment groups to establish diabetic rat models. Klotho gene was transferred into the treatment group, and common adenovirus was transferred into the control group. Rats in model group did not receive any treatment. At the 12th week after modeling, the model animals were sacrificed. Serum low density lipoprotein, high density lipoprotein concentrations, coronary artery intima-media thickness ratio were determined.
    RESULTS AND CONCLUSION: The high density lipoprotein concentration in treatment group was significantly higher than that in the model and control groups (P < 0.05). There was no significant difference in high density lipoprotein concentration between model and control groups. The low density lipoprotein in each group was significantly increased, but the differences were not significant (P > 0.05). The intima thickness in treatment group 
    was significantly smaller than that in model and control groups (P < 0.01), but the differences between model and control groups were not significant (P > 0.05). There was no significant difference in media thickness between each group (P > 0.05). The intima-media thickness ratio in the treatment group was significantly lower than that in the model and control groups (P < 0.01). There was no significant difference in the intima-media thickness ratio between treatment and normal groups (P > 0.05). These results showed that after Klotho gene transfer, the intima-media thickness ratio was substantially close to the normal level, and lower than that in the the model and control groups, and the degree of intimal thickening reduced. These results suggest that Klotho gene has a protective effect on coronary artery after being transferred into diabetic rats.
     

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    Effects of acupuncturing Changqiang acupoint on the learning and memory function and gamma-aminobutyric acid type A receptor alpha1 subunit expression in the hippocampal CA1 region of the fragile X mental retardation 1 located on X chromosome knockout mice 
    Wu Qiang, Chen Li-yun, Zhang Xue-jun
    2015, 19 (49):  7964-7968.  doi: 10.3969/j.issn.2095-4344.2015.49.016
    Abstract ( 310 )   PDF (549KB) ( 480 )   Save

    BACKGROUND: γ-Aminobutyric acid type A receptor α1 subunit expression was lower in the hippocampus of mice of fragile X mental retardation 1 (FMR1) gene located on the X chromosome knockout than in wild-type mice.
    OBJECTIVE: To study the effect of acupuncturing Changqiang (DU1) point on the learning and memory function and γ-aminobutyric acid type A receptor α1 subunit expression in the hippocampal CA1 region of the FMR1 located on X chromosome knockout mice.
     
    METHODS: There were five male and five female homozygous mice of FMR1 located on X chromosome knockout. One female and one male were housed in a cage. Their offspring that was identified as homozygous mice of FMR1 located on X chromosome knockout were randomly divided into model group, Changqiang group, and non-acupuncture group (n=10). An additional 10 wild-type mice were considered as blank group. Mice in the Changqiang group received acupuncture at Changqiang acupoint for 1 minute by mild reinforcing and attenuating method, and lifting and inserting method, at the frequency of 160-200 times/minute, once a day, for 10 consecutive days. Mice in the non-acupuncture group were acupunctured at 1 cm above the lowest point of mouse right costal arch. Mice in the model and blank groups only received analog crawl.
    RESULTS AND CONCLUSION: Compared with the blank group, the escape latency in the water maze was significantly prolonged in the model group (P < 0.05). Compared with the model group, the escape latency in the water maze was significantly shorter in the Changqiang group (P < 0.05), and the γ-aminobutyric acid type A receptor α1 subunit expression in the hippocampal CA1 region was increased (P < 0.05). The escape latency in the water maze and γ-aminobutyric acid type A receptor α1 subunit expression in the hippocampal CA1 region were similar between the non-acupuncture and model groups (P > 0.05). Moreover, γ-aminobutyric acid type A receptor α1 subunit expression in the hippocampal CA1 region was negatively associated with the escape latency after acupuncturing the Changqiang acupoint (r=-0.554, P=0.001). Data suggested that acupuncture at Changqiang acupoint improved the learning and memory capabilities of mice with the FMR1 located on X chromosome knockout, and up-regulated γ-aminobutyric acid type A receptor α1 subunit expression in the hippocampal CA1 region.  

     

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    Matrix-metalloproteinase-9 expression in focal cerebral infarction models experiencing hemorrhagic transformation 
    Xiao Jie, Li Xiao-gang
    2015, 19 (49):  7969-7975.  doi: 10.3969/j.issn.2095-4344.2015.49.017
    Abstract ( 307 )   PDF (549KB) ( 443 )   Save

    BACKGROUND: Theoretically, hemorrhagic transformation can appear in any patients with cerebral infarction, and occurs in any process of natural evolution of cerebral infarction. Therefore, to further study the pathogenesis of hemorrhagic transformation after infarction from the angle of ischemia time and reperfusion can provide theoretical evidence for preventing and treating hemorrhagic transformation after infarction in the clinic.
    OBJECTIVE: To observe matrix-metalloproteinase-9 expression in rats with persistent infarction and cerebral ischemia-reperfusion.
    METHODS: Healthy male adult Wistar rats were randomly assigned to three groups: sham surgery group, ischemia-reperfusion group and persistent infarction group. Suture method was used to establish models of middle cerebral artery occlusion. In the ischemia-reperfusion group, suture was pulled out at 4.5, 6, 12, 24 and 48 hours after ischemia and reperfusion were conducted for 24 hours. In the sham surgery group, the operation was the same as the ischemia-reperfusion group, but suture was not inserted. In the persistent infarction group,
     
    models of middle cerebral artery occlusion were generated, but the suture was not pulled out. Whether the models were successful or not was evaluated at 2 hours after the surgery. Neurological disorders were scored at corresponding time points after the surgery. 2,3,5-Triphenyltetrazolium chloride staining was utilized to observe hemorrhagic transformation in brain tissue. Immunohistochemistry was applied to determine matrix-metalloproteinase-9 expression.
    RESULTS AND CONCLUSION: Within 4.5 hours after cerebral infarction, neurological score of blood recanalization was obviously elevated. 6 hours later, especially, 12 hours later, blood recanalization aggravated neurologic impairment. In the ischemia-reperfusion group, neurological score gradually decreased with prolonged time of ischemia. At 48 hours of ischemia, neurological score was lowest. 2,3,5-Triphenyltetrazolium chloride staining revealed that at 12, 24 and 48 hours after ischemia, hemorrhagic transformation appeared to different degrees in rats. Matrix-metalloproteinase-9 as a negative factor participates in reperfusion injury and hemorrhagic transformation, aggravated ischemia-reperfusion injury and promoted the onset of hemorrhagic transformation.  

     

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    Establishing rat models of sciatic nerve injury: repair effect of spinal cord electrical stimulation versus local nerve electrical stimulation
    Pei Bao-an, Zi Jin-hua, Wu Li-sheng, Zhang Cun-hua, Chen Yun-zhen
    2015, 19 (49):  7982-7987.  doi: 10.3969/j.issn.2095-4344.2015.49.019
    Abstract ( 462 )   PDF (669KB) ( 341 )   Save

    BACKGROUND: After peripheral nerve injury, the nerve fibers distal to the injury site exhibited Wallerian degeneration, and neuronal apoptosis also appeared in the spinal cord segments proximal to the injury site, which inhibit axonal regeneration and affect the repair effect of nerve injury. At present, most researches of peripheral nerve injury are limited to the repair and stimulation of local injured nerve, and little is reported on neuron cell body in the nerve fibers proximal to the injury site.
    OBJECTIVE: To compare the efficacy on the treatment of peripheral nerve injury between spinal cord electrical 
     
    stimulation and local nerve electrical stimulation, and investigate the mechanisms of promoting peripheral nerve injury repair by spinal cord electrical stimulation.
    METHODS: Rats were divided into 3 groups according to the random number table: spinal cord electricalstimulation, local nerve electrical stimulation and control groups (n=15 rats/group). Rat models of sciatic nerve injury were established in three groups. Sciatic functional index, triceps wet weight, spinal cord neuron count and ultrastructure, myelin sheath thickness of regenerating nerve fibers and conduction velocity of rats in three groups were measured at different time points after modeling.
    RESULTS AND CONCLUSION: At 2 weeks after modeling, rat sciatic functional index was significantly higher in the spinal cord electrical stimulation group or local nerve electrical stimulation group than in the control group (P < 0.05). At 4, 6 and 8 weeks after modeling, rat sciatic functional index was highest in the spinal cord electrical stimulation group, followed by the local nerve electrical stimulation group, and lowest in the control group (P < 0.05). At 2 weeks after modeling, rat triceps wet weight was significantly greater in the spinal cord electrical stimulation group or local nerve electrical stimulation group than in the control group (P < 0.05). At 4 and 8 weeks after modeling, rat triceps wet weight was highest in the spinal cord electrical stimulation group, followed by the local nerve electrical stimulation group, and lowest in the control group (P < 0.05). At 2, 4 and 8 weeks after modeling, spinal cord anterior horn neuron counts were highest in the spinal cord stimulation group, followed by the local nerve electrical stimulation group, and lowest in the control group (P < 0.05). At 4 and 8 weeks after modeling, myelin sheath thickness of regenerating nerve fibers and conduction velocity of sciatic nerve were highest in the spinal cord electrical stimulation group, followed by the local nerve electrical stimulation group, and lowest in the control group (all P < 0.05). These results suggest that electrical stimulation to the corresponding spinal cord segments after peripheral nerve injury can effectively prevent neuronal apoptosis in the central nervous system, promote axonal regeneration and neurological function recovery, and the results are better than those produced by local nerve electrical stimulation.  

     

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    Construction of rat models of experimental tooth movement: time variation of chewing-like jaw movements after tooth movement
    Li Hong-shi, Chen Xin, Feng Yan, Liu Hong-chen
    2015, 19 (49):  7988-7993. 
    Abstract ( 381 )   PDF (521KB) ( 287 )   Save

    BACKGROUND: Tooth movement induced by interactive traction during orthodontic treatment causes regular spontaneous pain, however, the central nervous system conduction pathway and the basic mechanisms of pain are not clear.
    OBJECTIVE: To establish rat models of experimental tooth movement and observe the behavioral variation of rat chewing-like jaw movements in unit time after tooth movement.
    METHODS: Rats were randomly divided into blank, negative control and model groups. According to modified Colin. K. method, rat models of experimental tooth movement were established in the model group through interactive traction of the rat maxillary anterior and posterior teeth by orthodontic nickel-titanium wires. Rats in the blank group were not subjected to interactive traction treatment. Rats in the negative control group were not imposed corrective force. The behavioral variation of chewing-like jaw movements of rats was detected respectively at 4, 12 hours and at 1, 3, 5, and 7 days after tooth movement. At 1 day after tooth movement, relative variation of chewing-like jaw movements of rats was observed after imposing orthodontic force of 30, 60 and 90 g on movable tooth. 
    RESULTS AND CONCLUSION: Beginning from 4 hours after tooth movement, the total time of chewing-like jaw movements began to increase, and the time of chewing-like jaw movements was significantly increased at 12 
    hours after tooth movement, reached the peak at 1 day after tooth movement, and then was slowly declined until 7 days in the model group compared with that in the negative control and blank groups. At 1 day after tooth movement, there was significant difference in the time taken for chewing-like jaw movements in the model group among orthodontic force of 30, 60 and 90 g (P < 0.05). These results confirm that the time variation of chewing-like jaw movements after tooth movement is consistent with the rule of pain induced by tooth movement during clinical orthodontic treatment, which can be used as one of the related behavioral responses of oral and maxillofacial pain after tooth movement in rats.  
     

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    Molecular mechanism of hypoxia-reoxygenation injury to hepatocyte membrane F-actin microfilaments
    Zhang Ya-fei, Wang Jia-zhong, Ji Hong, Lu Hong-wei, Lu Le, Wang Jin-long, Shang Hao, Li Yi-ming
    2015, 19 (49):  7994-7999.  doi: 10.3969/j.issn.2095-4344.2015.49.021
    Abstract ( 337 )   PDF (610KB) ( 279 )   Save

    BACKGROUND: Human hepatocyte models of hypoxia-reoxygenation injury are established to simulate the ischemia/reperfusion injury of transplanted organ. There have been no research reports addressing the molecular mechanism underlying hypoxia-reoxygenation injury to hepatocyte membrane F-actin microfilaments.
    OBJECTIVE: To analyze the molecular mechanism of hypoxia-reoxygenation injury to hepatocyte membrane F-actin microfilaments.
    METHODS: Rat hepatocyte models of hypoxia-reoxygenation in vitro were established and then randomly divided into control and hypoxia-reoxygenation groups. The hypoxia-reoxygenation group was also subdivided into three subgroups: hypoxia-reoxygenation-2, 4 and 6 hours (after 3 hours of hypoxia, hepatocytes were given oxygen for 2, 4 and 6 hours respectively). Cell morphology was observed by light microscope, ultrastructural 
    changes by transmission electron microscope and the change in F-actin microfilament content by confocal laser microscopy. HSP27 and cofilin gene and protein levels were determined by real-time polymerase chain reaction and western blot assay respectively.
    RESULTS AND CONCLUSION: Light microscope observations showed that spindle cells and exfoliated cells significantly increased in hypoxia-reoxygenation group. Transmission electron microscope observations showed that in the hypoxia-reoxygenation group, the amount of endoplasmic reticulum significantly decreased, mitochondrial density increased and glycogen disappeared compared with the control group. Confocal laser microscopy observations showed that in the hypoxia-reoxygenation group, F-actin microfilament fluorescence disordered, F-actin microfilament morphology significantly changed, staining intensity significantly attenuated and mean fluorescence intensity was significantly lower than that in control group (P < 0.05). Real-time PCR and western blot detection results showed that HSP27, cofilin gene and protein expression levels in the hypoxia-reoxygenation group were significantly lower than in the control group (P < 0.05). These results demonstrate that hypoxia-reoxygenation affects the correct assembly of F-actin microfilaments and weakens the normal cycle of F-actin microfilaments through inhibiting the protein expression of HSP27 and cofilin, and gene transcription in hepatocytes, thereby changing the skeleton of F-actin microfilaments.
     

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    Effect of estradiol and progesterone on the number of macrophages and the expression of related factor in the endometrium of ovariectomized mice
     
    Zhao Ying-jian, Han Jun-yan
    2015, 19 (49):  8000-8004.  doi: 10.3969/j.issn.2095-4344.2015.49.022
    Abstract ( 342 )   PDF (605KB) ( 279 )   Save

    BACKGROUND: Studies have shown that macrophage function and macrophage colony-stimulating factor expression may be directly or indirectly affected by regulation of estrogen and progestational hormone, but the specific impact has not been studied in depth.
    OBJECTIVE: To observe the effect of estradiol and progesterone on the number of macrophages and the expression of macrophage colony-stimulating factor in the endometrium of bilaterally ovariectomized mice.
    METHODS: The bilaterally ovariectomized mice were randomly divided into estrogen, progestational hormone, and control groups. The number of macrophages and the expression of macrophage colony-stimulating factor in the endometrium of mice in each group were analyzed.
    RESULTS AND CONCLUSION: Immunohistochemical staining results showed that after 6 and 24 hours of intervention, the number of macrophages and the expression of macrophage colony-stimulating factor in the mouse endometrium in the estrogen and progestational hormone groups were significantly higher than those in the control group (P < 0.05). These results confirm that both estradiol and progesterone can increase the number of macrophages, and can increase the expression of macrophage colony-stimulating factors in the endometrium of bilaterally ovariectomized mice. 

     

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    Evaluation of Xiaozhiling injection effect on rectal internal mucosal prolapse based on the amount of displacement of mucosal nodes in a finite element model of rabbit anorectum
    Ke Min-hui, Ye Ling, Chen Li-wu, Niu Su-sheng, Zheng Ming-xiao,Zhang Zhi-xi
    2015, 19 (49):  8005-8009.  doi: 10.3969/j.issn.2095-4344.2015.49.023
    Abstract ( 330 )   PDF (740KB) ( 295 )   Save

    BACKGROUND: Previous studies have confirmed a scheme of establishing rabbit models of rectal internal mucosal prolapse within 2 months, which were similar to rectal internal mucosal prolapse in humans and had good stability. At present, Xiaozhiling injection is a preferred traditional Chinese medicine for treatment of rectal internal mucosal prolapse. Its short-term curative effect is positive, but rectal internal mucosal prolapse recurs after long-term administration of Xiaozhiling injection.
    OBJECTIVE: To evaluate the effect of Xiaozhiling injection in the treatment of rectal internal mucosal prolapse based on the amount of displacement of mucosal nodes in a finite element model of rabbit anorectum.
    METHODS: Thirty-two rabbit models of rectal internal mucosal prolapse were randomly divided into rectal internal mucosal prolapse group (n=8) and Xiaozhiling injection group (n=24). Rbbits in the rectal internal mucosal prolapse group were sacrificed and dissected to harvest the rectum for mechanical measurements. At the 3rd, 7th and 14th days after administration of Xiaozhiling injection, eight rabbits per time point from the Xiaozhiling 
    injection group were sacrificed to harvest the rectum for mechanical measurements. At last, the data from these two groups were input into a finite element model of the rabbit anorectum. The nodes in different coronal planes (ventral, middle and dorsal) were selected on three rectal mucosal cross-sections harvested from different areas (above, at and below the anorectal angle). The amounts of displacement of these nodes were measured under the condition of simulated defecation.
    RESULTS AND CONCLUSION: There was significant difference in the amount of displacement of each node between different time periods before and after administration of Xiaozhiling injection. There was significant difference in the amount of displacement between mucosal nodes from the same cross-section and between mucosal nodes from the same coronal plane in the same time period before and after administration of Xiaozhiling injection. These results suggest that Xiaozhiling injection is effective in the treatment of rectal internal mucosal prolapse, however, the difference in amount of displacement of different nodes in the rectal mucosa finite element after administration of Xiaozhiling injection may be one of the factors affecting the efficacy.  

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    Establishment of rat models of chronic rejection after heterotopic tracheal transplantation and the relationship with interleukin-17
    Wu Dong, Li Xiao-jun, Liu Mao-lin, Wang Jian-jun, Zhai Wei
    2015, 19 (49):  8010-8014.  doi: 10.3969/j.issn.2095-4344.2015.49.024
    Abstract ( 305 )   PDF (548KB) ( 265 )   Save

    BACKGROUND: Obliterative bronchiolitis is one of the major complications of lung transplantation. Establishing a stable and reliable animal model is a prerequisite to the study of obliterative bronchiolitis.
    OBJECTIVE: To establish rat models of chronic rejection after heterotopic tracheal transplantation and investigate the effect of interleakin-17 in this model. 
    METHODS: After making donor/recipient pairs according to body weight, Wistar and Sprague-Dawley rats were 
    randomly divided into 2 groups (n=30): Wistar (donor)→SD (recipient) group and SD (donor)→SD (recipient) group. The trachea was embedded by recipient’s cervical subcutaneous tissue. Tracheal tissue was harvested at different time points for histopathological analysis. The indicators including epithelial loss, lymphocyte count, the ratio of anuclear chondrocyte number to total chondrocyte number and the expression level of interleukin-17 in the transplanted trachea were determined.
    RESULTS AND CONCLUSION: Recipients from these two groups all survived. At the 7th,14th and 28th days after transplantation, there was significant difference in lymphocyte count between these two groups (P < 0.05). At the 14th and 28th days after transplantation, there was significant difference in the ratio of anuclear chondrocyte number to total chondrocyte number between these two groups (P < 0.05). At the 7th, 14th and 28th days after transplantation, the degree of epithelial loss of rats in Wistar (donor)→SD (recipient) group was respectively > 40%, 100%, 100%, however, there was no obvious loss in the SD (donor)→SD (recipient) group. At the 28th day after transplantation, the incidence of obliterative bronchiolitis was 100% in Wistar (donor)→SD (recipient) group, however, no occlusion was observed in rats from the SD (donor)→SD (recipient) group. At the 28th day after transplantation, the expression level of interleukin-17 in Wistar (donor)→SD (recipient) group was significantly higher than that in the SD (donor)→SD (recipient) group (P < 0.05) These results demonstrate that Wistar (donor)→SD (recipient) rat models of chronic rejection after heterotopic tracheal transplantation were successfully established in this study, which provide a platform for the study of bronchiolitis obliterans. Interleukin-17 may play an important role in this field.
     

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    Expression of interleukin-10 and cyclooxygenase 2 in keratitis rat models
    Lu Xia, Pan Xu-bin, Xie Yu-tao, Ji Qiu-di
    2015, 19 (49):  8015-8020.  doi: 10.3969/j.issn.2095-4344.2015.49.025
    Abstract ( 465 )   PDF (675KB) ( 291 )   Save

    BACKGROUND: Investigating the relationship between interleukin-10, cyclooxygenase-2 expression levels and the occurrence and development of keratitis is of great significance for the treatment of keratitis.
    OBJECTIVE: To detect the expression of interleukin-10 and cyclooxygenase 2 in keratitis rat models and analyze the relationship between interleukin-10, cyclooxygenase-2 expression and the occurrence and development of keratitis in rats.
    METHODS: Twenty-six healthy rats of either gender were randomly selected to establish keratitis rat models. The left eyes of rats were chosen as experimental group to establish keratitis models using epikeratophakia. The right eyes of rats were set as control group. At the 1th, 3rd, 7th and 14th days after modeling, the pathological changes of rat corneal tissue were observed in both eyes. The expression of interleukin-10 and cyclooxygenase 2 in the corneal tissue with different degrees of keratitis was determined using immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR).      
    RESULTS AND CONCLUSION: Histopathological observation showed that there were severe tissue necrosis and neutrophil infiltration in keratitis foci. PAS staining showed that the mycelium was mainly present in the corneal stroma. In the control group, the expression of interleukin-10 was hardly detected in rat cornea. After keratitis infection, the expression of interleukin 10 increased at first and then decreased, and there was significant difference between different time points (P < 0.01). In the control group, the expression of cyclooxygenase-2 was hardly detected in rat cornea. After keratitis infection, the expression of cyclooxygenase 2 was increased gradually and then decreased at the 14th day. There was significant difference between different time points (P < 
    0.01). PAS staining and pathologic tissue culture experiment suggest that rat models of keratitis were successfully established. These results show that the expression of interleukin-10 appeared in the late period of keratitis lesions, which indicate that interleukin-10 may be involved in the tissue damage repair in the late period of keratitis. The expression of cyclooxygenase-2 locates in the keratitis tissue and cyclooxygenase-2 is a sensitive inflammatory cytokine of keratitis.  

     

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    Expression of nestin in renal tissue of rat models of adriamycin nephropathy and the intervention of mycophenolate mofetil
    Pan Feng-qi, Jiang Hong
    2015, 19 (49):  8021-8025.  doi: 10.3969/j.issn.2095-4344.2015.49.026
    Abstract ( 340 )   PDF (707KB) ( 544 )   Save

    BACKGROUND: Studies have shown that podocyte injury is an important mechanism to generate glomerular proteinuria. However, it is not yet fully understood regarding the modulation method by which cytoskeletal proteins in the podocyte can maintain the unique morphology of podocytes. The construction and remodeling of podocyte cytoskeleton has become an increasing area of interest in the research field of proteinuria mechanism.
    OBJECTIVE: To establish rat models of minimal change adriamycin nephropathy, and detect the expression of nestin in rat renal tissue under the intervention of mycophenolate mofetil.
    METHODS: Thirty-six male Wistar rats were included and randomly divided into nephropathy model, mycophenolate mofetil and normal groups (n=12). Rats in nephropathy model and mycophenolate mofetil groups were intravenously injected with adriamycin via the tail vein at one time to establish models. Rats in normal group were intravenously injected with the same amount of normal saline via the tail vein. On the next morning, rats in mycophenolate mofetil group were intragastrically administered 20 mg/kg mycophenolate mofetil, once per day. Rats in nephropathy model group and normal group were given the same amount of normal saline daily. Four rats from each group were sacrificed at 14, 21 and 28 days after modeling. The renal cortex was harvested and subjected to hematoxylin-eosin staining and immunohistochemical staining. The pathological changes of renal tissue were observed and nestin expression in renal tissue was detected.
     
    RESULTS AND CONCLUSION: In the normal group, rat glomerular filtration membrane structure was complete and epithelial cell foot process was clear. In the nephropathy model group, glomerular epithelial cell foot processes were fused to a large extent, and basement membrane was normal. In the mycophenolate mofetil group, glomerular epithelial cell foot processes were partly fused, but the pathological changes were milder. Immunohistochemistry results showed that beginning from the 14th day of modeling, rat nestin expression in the nephropathy model and mycophenolate mofetil groups was significantly increased compared with the normal group (P < 0.05). Rat nestin expression in the mycophenolate mofetil group was significantly lower than that in the nephropathy model group (P < 0.05). These results suggest that after glomerular podocyte injury, nestin expression in podocyte increased, while the expression increased with the aggravation of the disease. Mycophenolate mofetil can alleviate podocyte injury, downregulate nestin expression, maintain the normal structure of podocytes, and thereby achieve the purpose of delaying kidney damage.
     

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    Effects of proanthocyanidins on free radical metabolism and apoptosis-related protein expression in rat splenocytes after heavy-load exercise
    Li Yuan, Liu Wen-jie, Yang Ning, Min Zhu
    2015, 19 (49):  8026-8031.  doi: 10.3969/j.issn.2095-4344.2015.49.027
    Abstract ( 257 )   PDF (773KB) ( 284 )   Save

    BACKGROUND: Studies have found that proanthocyanidins can inhibit cell apoptosis in cellular hypoxia and reoxygenation injury.
    OBJECTIVE: To investigate the effects of proanthocyanidins on free radical metabolism and the expression of apoptosis related proteins Bcl-2, Fas, Bax in rat splenocytes after heavy-load exercise.
    METHODS: Forty-eight Sprague-Dawley rats were randomly and evenly divided into placebo group and medication group. Rats in the medication group were intragastrically daily administered 15 mL/kg of proanthocyanidins solution. Rats in the placebo group were intragastrically administered equal amount of distilled water for 2 successive weeks. After 2 weeks of administration, rats in each group were evenly divided into three subgroups: static control group, immediately after exercise group, and 24 hours after exercise group. Rats in the 
    immediately after exercise and 24 hours after exercise groups were subjected to heavy-load treadmill exercise at a
    -10° gradient and at 20 m/min. The rats were sacrificed immediately after exercise and at 24 hours after exercise. Rats in the static control group were directly sacrificed. Bcl-2, Bax, Fas protein expression in splenocytes, and superoxide dismutase activity and malondialdehyde level in homogenate from spleen tissue were detected.
    RESULTS AND CONCLUSION: (1) Superoxide dismutase activity was highest in the static control group, followed by the immediately after exercise group and lowest in the 24 hour after exercise group. The superoxide dismutase activity in the immediately after exercise and 24 hours after exercise groups was greater than that in the placebo group at the corresponding time phrases. (2) In the placebo group, malondialdehyde level immediately after exercise and at 24 hours after exercise was higher than that in the static state. In the medication group, malondialdehyde level immediately after exercise and at 24 hours after exercise were lower than that in the static state (P < 0.01). Malondialdehyde level immediately after exercise and at 24 hours after exercise in the medication group was lower than in the placebo group
    (P < 0.01). (3) Bcl-2 expression was increased immediately after exercise and decreased at 24 hours after exercise in both placebo and medication groups. Bcl-2 expression immediately after exercise and at 24 hours after exercise in the medication group was higher than in the placebo group (P < 0.01). (4) Bax expression immediately after exercise and at 24 hours after exercise in the placebo group was higher than in the static state (P < 0.01). In the medication group, Bax expression immediately after exercise and at 24 hours after exercise was lower than in the static state (P < 0.01). Bax expression in the static state, immediately after exercise and at 24 hours after exercise in the medication group was higher than in the placebo group (P < 0.05, P < 0.01). (5) Fas expression was lowest in the static control group, followed by 24 hours after exercise group and highest in the immediately after exercise group. Fas expression immediately after exercise and at 24 hours after exercise in the medication group was lower than in the placebo group (P < 0.01). These results demonstrate that proanthocyanidins is helpful to alleviate the heavy-load exercise-induced spleen lipid peroxidation damage, effectively reduce sports fatigue induced by free radicals, and decrease the degree of splenocyte apoptosis.
     

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    Castor oil diet extract affects uterine contraction and the content of prostaglandin E2 in amniotic tissue 
    Liu Hai-bin
    2015, 19 (49):  8032-8036.  doi: 10.3969/j.issn.2095-4344.2015.49.028
    Abstract ( 351 )   PDF (472KB) ( 283 )   Save

    BACKGROUND: Studies have shown that castor bean extract has certain anti-fertility effect; however, there are few reports in China about anti-fertility aspect of castor bean.
    OBJECTIVE: To explore the effects of castor oil diet extract on uterine contraction of rats and the content of prostaglandin E2 in amniotic tissue.
    METHODS: Sixty Wistar unfertilized adult female rats were selected and randomly divided into normal saline, oxytocin and castor oil diet extract group (n=20 rats/group). Before and after drug administration, contraction intensity, frequency and motility of isolated uterus of rats in each group were observed and recorded. Ninety Wistar adult female rats of gestation for 18 days were selected and randomly divided into normal saline, oxytocin and castor oil diet extract groups (n=30 rats/group). Prostaglandin E2 level in amniotic tissues of rats was measured by radioimmunoassay method. The amniotic tissue of rats in normal saline group was obtained. The amniotic cells were cultured in vitro by adding different concentrations of ricinoleic acid in the culture medium for 18 hours. Prostaglandin E2 level in amniotic cells was measured by radioimmunoassay method.
    RESULTS AND CONCLUSION: Compared with before drug administration and normal saline group, contraction intensity, frequency and motility of isolated uterus of rats in castor oil diet extract group were significantly improved after administration. There were no significant differences in contraction intensity and frequency (t=2.321, P > 0.05) after drug administration between oxytoxin and castor oil diet extract groups, however, after drug administration, motility in the castor oil diet extract group was significantly lower (t=2.765, P < 0.05) than in the oxytoxin group. Compared with normal saline group, prostaglandin E2 level in amniotic tissues of pregnant rats in castor oil diet extract group was significantly higher (t=14.91, P < 0.01); however, prostaglandin E2 level in 
    amniotic tissue of pregnant rats in the castor oil diet extract group was significantly lower than in the oxytocin group (t=2.769, P < 0.05). Prostaglandin E2 levels in amniotic cells of rats were positively correlated with ricinoleic acid concentration and culture time. These results suggested that castor oil diet extract can enhance contractions and increase prostaglandin E2 levels in amniotic tissue. Ricinoleic acid may be the active ingredient of castor oil diet to induce aboration.
     

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