1.Expression of nitric oxide synthase in the process of cerebral ischemia/reperfusion
Chao WANG ; Feng SUN ; Xiaojie DING
Chinese Journal of Tissue Engineering Research 2007;11(8):1589-1592
BACKGROUND: Nitric oxide synthase(NOS) is the key factor for the synthesis of nitric oxide (NO) . Because NO combines with oxygen, hemoglobin and other substances in vivo easily and deactivates quickly, and it is not exactly determined, so determining the activity of NOS is the important link for further studying the pathogenesis of NO in cerebral ischemia/reperfusion (I/R)injury.OBJECTIVE: To study the effect of different types of NOS in the process of cerebral I/R injury.DESIGN: Randomized and controlled animal experiment.SETTING: Instituteof Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University.MATERIALS: This experiment was carried out in the Shandong Key Laboratory of Prevention and Treatment for Encephalopathy from May to December 2005. Twenty-eight adult healthy male Wistar rats, of clean grade, weighing from 220 to 260 g, were provided by the Experimental Animal Center of Shandong University. The involved rats were randomly divided into sham-operation group (n =4) and cerebral ischemia group (n =24). Six time points were set in cerebral ischemia group: ischemia 1 hour reperfusion 6 hours, 12 hours, 1 day, 3 days, 7 days and 14 days, 4 rats at each time point.METHODS: Rat models of middle cerebral artery occlusion/reperfusion were established by suture-occluded method through inserting a suture into the left internal-external carotid artery. The expressions of different types of NOS at different time points after cerebral I/R were detected by immunohistochemical technique.MAIN OUTCOME MEASURES: ①Toluidine blue-stained two groups of nerve cells; ② The expression and distribution of neuronal NOS (nNOS), endothelial NOS (eNOS) and inducible NOS(iNOS) at different time points.RESULTS: ①Karyopyknosis and cell debris appeared in the nerve cells of the injured region of cerebral ischemia group,and there were no significant differences of cells among different time points. ② Six hours after reperfusion, the expressions of nNOS, eNOS and iNOS were found in the neurons of brain tissue and increased with the elongation of time of reperfusion. The regions in which different types of NOS in neurons of brain tissue were expressed were cortical area and corpora striata. nNOS and iNOS were highly expressed within 12 hours to 7 days after reperfusion in the brain, and eNOS was highly expressed within a short time period, i.e. 6 hours to 3 days after reperfusion. eNOS expression increasing and decreasing occurred earlier than nNOS and iNOS. But the expressions of three kinds of NOS all reached peak on the first day after reperfusion. The changing tendencies of the expression of three kinds of NOS in the cortical area and corpora striata were the same basically.CONCLUSION: After cerebral I/R injury, the high expression of eNOS occurs early and lasts for a short time, while that of nNOS and iNOS occurs late and lasts for a long time.
2.Role of Numb in tumors
Chao WANG ; Chuyao ZHANG ; Weiwei FENG
Journal of International Oncology 2012;39(7):541-544
Numb,one of the important cell fate determinants,plays a key role in regulation of nerve system development,and it is closely related to the occurrence of tumors.It is proven that Numb is implicated in the processes of occurrence,invasion and metastasis of tumors through regulating the way of cell division,cell polarity,epithelial-mesenchymal transitions and several signal pathways,e.g.,Notch,Hh,p53.The expression levels of Numb in cells are related to the malignant degree and prognosis of tumors.It is vital to explore the role of Numb in tumors for treating tumors at the standpoint of Numb.
3.Problems and Recommendations of Community Exercise Rehabilitation for Cardiovascular Disease
Min CHAO ; Feng LIANG ; Zun WANG ; Lei WANG
Chinese Journal of Rehabilitation Theory and Practice 2012;18(10):999-1000
Community rehabilitation of cardiovascular disease is of great significance. Exercise therapy of cardiovascular disease in communities is necessary and feasible. At present, the main problems China faced to carry out community exercise therapy of cardiovascular disease include: cardiovascular patients with a weak sense of community exercise therapy, lack of professional rehabilitation personnel, inadequate community-based exercise therapy practice and lack of community rehabilitation funding. To solve above problems, we should strengthen the community exercise therapy practice of cardiovascular disease, and promote the cooperation between community rehabilitation and hospital rehabilitation. In all, comprehensive efforts should be made in order to solve the problems.
6.Effect of insulin-like growth factor and its receptor on the neurologic deficit in rats with cerebral ischemia/reperfusion injury
Feng SUN ; Xiaojie DING ; Chao WANG ; Yunliang GUO
Chinese Journal of Tissue Engineering Research 2007;11(14):2780-2783
BACKGROUND: It has been demonstrated that insulin-like growth factor-1 (IGF-1) is a kind of neurotrophic factor and protects from cerebral ischemia/reperfusion injury, the expression of IGF-1 is associated with the attack of ischemic stroke. The effects of IGF-1 and its receptor (IGF-1R) on neurobehavioral function are to be further studied.OBJECTIVE: To observe the effects of IGF-1 and IGF-1R on neurobehavioral function in rat models of cerebral ischemia/reperfusion injury.DESIGN: A randomized controlled observation.SETTING: Institute of Cerebrovascular Diseases, Affiliated Hospital of Qingdao University Medical College.MATERIALS: The experiments were carried out in Shandong Key Laboratory for Prevention and Treatment of Brain diseases. Twenty-eight healthy adult Wistar rats of clean degree, weighing 220-260 g, were provided by the experimental animal center of Shandong University.METHODS: The rats were randomly divided into experimental group (n =24) and sham-operated group (n =4). The middle cerebral artery occlusion/reperfusion (MCAO/R) models were established by inserting a thread through left external-internal carotid arteries. The sham-operated rats were given the same treatments except inserting thread. ①Neurologic deficit test: The rats in the experimental group were assessed according to Bederson standard after 1-hour ischemia and 6, 12-hour, 1, 3, 7 and 14-day reperfusion respectively. The sham-operated rats were assessed at corresponding time points; Without neurologic deficit was marked as 0 point; flexion of anterior claws as 1 point; unable to act against the pushing from the contralateral side as 2 points; circling while walking as 3 points; shaking as 4 points;unconscious mind as 5 points. ② Sample collection and treatment: The samples in the experimental group were collected after 1-hour ischemia and 6, 12-hour, 1, 3, 7 and 14-day reperfusion, and those in the sham-operated group ere collected at 24 hours postoperatively. The rats were anesthetized, brain samples were got at about 5 mm posterior to optic chiasma after brains were removed completely, then serial coronal sections (5 μm) were prepared, and 1 from 10 sections was stuck to the cover glasses treated with poly-L-lysine. ③ Morphological observation of neurons: The neurons in brain were observed by toluidine blue staining. ④ Detection of IGF-1 and IGF-1R: The expressions of IGF-1 and IGF-1R in cortex and striatum were detected with immunohistochemical technique, 4 fields were randomly selected to count the positive cells under high-power microscope (×400).MAIN OUTCOME MEASURES: ① The neurologic deficit; ② Morphological changes of neurons in brain; ③ Expressions of IGF-1 and IGF-1R in cortex and striatum.RESULTS: All the 28 rats were involved in the analysis of results. ① The neurologic deficit: The scores of neurologic deficit were (1.50±058) and (1.50±0.78) in rats after 7 and 14-day reperfusion, which were lower than that in rats after 6-hour reperfusion [(3.00±0.00), P < 0.05]. ② Morphological changes of neurons in brain: The neurons in ischemic area appeared as paryopyknosis and became irregular in shape, there were obvious gaps around the cells, also deeply stained as purplish blue, nucleolus disappeared, and there were many scattered cellular fragments. ③ Expressions of IGF-1 and IGF-1R in cortex and striatum: The numbers of IGF-1 positive cells in cortex were (8.75±2.06), (11.13±1.14),(19.75±3.18), (17.38±3.11 ) and (11.23±2.28) respectively in rats after 6, 12-hours and 1, 3, 7-day reperfusion, which all were higher than that in sham-operated rats [(3.88±1.46), P < 0.05], the numbers of IGF-1 positive cells in striatum were(8.25±2.21), (11.34±2.21), (18.23±2.64), (18.56±2.34) and (11.31±2.14) respectively in rats after 6, 12 hours and 1, 3, 7days reperfusion , which were also higher than that in sham-operated rats [(4.12±2.24), P < 0.05]. The numbers of IGF-1R positive cells in cortex were (7.63±1.50), (10.50±2.34), (15.55±3.12), (15.37±3.01), (8.86±2.75) respectively in rats after 6, 12-hours and 1,3,7-day reperfusion, which all were higher than that in sham-operated rats [(4.13±1.81), P <0.05]. Those in striatum were (8.33±2.31), (10.24±2.09), (14.72±2.17), (14.24±2.77), (8.38±2.05), which were also higher than that in sham-operated rats [(3.76±2.35), P < 0.05].CONCLUSION: The neurological function is damaged after cerebral ischemia/reperfusion, but it has a trend of self-recovery. The expressions of IGF-1 and IGF-1R are mainly distributed in cortex and striatum. Higher expressions of IGF-1 and IGF-1R maintain during 12 hours to 7 days after reperfusion and have a peak value at 1-3 days, which suggests that early expression of IGF-1 and IGF-1R are certain related to the recovery of neurological function.
7.Neuronal apoptosis associated with basic fibroblast growth factor and its receptor following cerebral ischemia reperfusion
Xiaojie DING ; Feng SUN ; Chao WANG ; Yunliang GUO
Chinese Journal of Tissue Engineering Research 2007;11(14):2776-2779
BACKGROUND: Brain injury can induce the increased expression of basic fibroblast growth factor (bFGF) in brain,whereas FGFR is a very important player in the cell proliferation and differentiation, angiogenesis, skeletogeny, etc.OBJECTIVE: To observe the effect of bFGF and its receptor on neuronal apoptosis following cerebral ischemia/reperfusion injury in rats.DESIGN: A randomized grouping design and animal experiment.SETTING: Institute of Cerebrovascular Disease, Affiliated Hospital of Qingdao University Medical College.MATERIALS: Twenty-eight healthy adult Wistar rats of clean degree, weighing 220-260 g, were provided by the experimental animal center of Shandong University. Rabbit-anti-rat bFGF and fibroblast growth factor receptor-1(FGFR-1) monoclonal antibodies were provided by Wuhan Boster Biological Technology, Co.,Ltd.METHODS: The experiment was carried out in Shandong Key Laboratory for Prevention and Treatment of Brain diseases.① The rats were randomly divided into experimental group (n =24) and sham-operated group (n =4). Models of middle cerebral artery occlusion/reperfusion (MCAO/R) were established by thread occlusion via left external-internal carotid arteries, and 4 rats in the experimental group were sampled at 1-hour ischemia/6, 12-hour, 1, 3, 7 and 14-day reperfusion respectively. The rats in the sham-operated group were given the same treatment without inserting thread.After anesthesia, the brain was removed completely by cutting head, then the brain tissue at about 5 mm posterior to optic chiasma was cut down, then serial coronal sections (5 μm) were prepared. ② The brain tissues were stained with ematoxylin-eosin (HE), and the forms of neurons were observed under microscope. ③ TdT-mediated dUTP-biotin nick end labeling (TUNEL) method: there were buffy granules in nucleus which was positively stained (apoptosis). Four fields were randomly selected from cortex and striatum to count positive cells under high-power microscope (×400). ④ The sections were stained with rabbit-anti-rat bFGF and FGFR-1 monoclonal antibodies, 4 fields were randomly selected from cortex and striatum to count positive cells under high-power microscope (×400).MAIN OUTCOME MEASURES: Apoptosis and the expressions of bFGF and FGFR-1.RESULTS: All the 28 rats were involved in the analysis of results. ① In the experimental group, the neurons in the ischemic sites were obviously decreased, some neurons appeared as paryopyknosis and became irregular, also deeply stained as purplish blue, nucleolus disappeared, and there were many scattered cellular fragments. ② In the sham-operated group, there were a few apoptotic neurons in the brain tissue, and the apoptotic neurons were obviously increased after ischemia, which mainly observed in cortexes and striatums of frontal and paritetal lobes. In the experimental group, apoptotic cells in cortexes began to increase gradually at 6 hours, and there were more cells at 12hours and 3 days, which reached the peak value at 1 day, and began to decrease at 3 day, but there were still more apoptotic cells at 14 days than in the sham-operated group. The number of apoptotic neurons and the changing trend in striatums were generally the same as those in cortexes (P > 0.05). ③ In the sham-operated group, there were weak bFGF expression in the neurons of brain tissue, but there were fewer lightly stained positive cells. After cerebral ischemia, the bFGF expressions were increased, mainly observed in cortexes and striatums. The bFGF expression appeared at 6 hours after cerebral ischemia/reperfusion, and the number was increased gradually and deeply stained as the time of reperfusion prolonged (Figure 3), it reached the peak value at 1-3 days, and then weakened gradually, but it was still higher than in the sham-operated group at 14 days [(5.01 ±1.71), (5.21 ± 1.62) cells/visual field; (2.03± 1.73),(2.46± 1.38) cells/visual field, P < 0.05]. ④ In the sham-operated group, lightly stained FGFR-1 positive cells could be observed in brain tissue. At 6 hours after cerebral ischemia/reperfusion, the FGFR positive cells began to increased in cortexes and striatums, which were the most at 1-3 days, and gradually decreased after 3 days, and the number was still a little more than that in the sham-operated group at 14 days [(5.01± 1.41), (5.20± 1.33) cells/visual field; (2.25±1.67),(2.32± 1.61 ) cells/visual field].CONCLUSION: After cerebral ischemia/reperfusion, the expressions of endogenous bFGF and FGFR-1 may be activated in cortex and striatum, then inhibit the neuronal apoptosis, and play its neuroprotective role.
8.Effect and influencing factors on 131I treatment for cervical lymph node metastasis from papillary thyroid cancer
Yun DONG ; Fang FENG ; Chao MA ; Hongliang FU ; Hui WANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2016;36(4):330-333
Objective To assess the effect and influencing factors on 131I treatment for cervical lymph node metastasis (LNM) after operation in patients with PTC.Methods PTC patients (n =117;45 males,72 females;average age (45.17± 15.50) years) with postoperative cervical LNM from January 2010 to December 2014 were analyzed retrospectively.LNM was diagnosed by surgical pathology,imaging results and clinical follow-up.Single factor analysis was performed in age,gender,operation mode,TNM stage,131I treatment time and other factors.The results for 131I treatment effect included CR,PR,NC.Two-sample t test and x2 test were used.Results Among the 117 PTC patients with postoperative LNM,53 (45.3%) cases had non-131 I-avid metastasis.Fifty of the 64 (54.7%) patients with 131I-avid metastasis were treated with 131I.Nineteen patients (38.0%) achieved CR,22 (44.0%) achieved PR,and 9 (18.0%) showed invalid results.Fourteen of the 64 patients underwent another cervical lymph node dissection.Nine patients achieved CR,5 patients achieved PR,and 4 PR patients were then treated with 131I and finally achieved CR.Single factor analysis showed that the influencing factors of non-131I-avid lymph node included patients' age (t =3.459),serum Tg level (x2 =6.698) and metastasis with 1s F-FDG uptake (x2 =26.928;all P<0.05).The influencing factors of 131I treatment effect included lymph node dissection procedure (x2 =6.487),unilateral or bilateral lesion (x2=5.187) and LNM size (x2=8.099;all P<0.05).Conclusions 131I treatment is ineffective for nearly 50% of patients with non-131I-avid LNM.The influencing factors of 131I treatment effect include the lymph node dissection procedure,unilateral or bilateral lesions and LNM size.
9.Reconstruction Algorithms and Realization of Freehand 3D Ultrasonic
Chao JIANG ; Feng WANG ; Yun GE ; Sheng GAO
Chinese Journal of Medical Physics 2010;27(1):1621-1624
Objective:Three-dimensional (3D) ultrasound (US) is increasingly being introduced in the clinic,both for diagnostics and image guidance.Although dedicated 3D US probes exist,because of its expensive cost,3D US can also be acquired with the still frequently used two-dimensional (2D) US probes.Methods:Obtaining 3D volumes with 2D US probes is a two-step process.First,a positioning sensor must be attached to the probe for 2D image matching;second,a reconstruction of a 3D volume can be performed into a regular voxel grid.Results:This paper presents a way to realize the 3D US in irled-based Image Guided Radiotherapy using a homemade 2D US.Conclusions:The experiments demonstrate a method of saving costs and having advantages in clinic application.
10.The effect of daidzein on extracellular matrix of uremic rats
Hao WANG ; Bing FENG ; Chao ZHU ; Jianping YU ; Baozhuan HUANG
International Journal of Traditional Chinese Medicine 2017;39(1):43-47
Objective To observe the effect of daidzein on extracellular matrix of uremic rats and to discuss its mechanism.Methods Uremic rat model were established by 5/6 nephrectomized. Model rats were devided into daidzein group, control group. Rats with sham-operation were regarded as the normal control. At time of baseline, 4th and 8th week after operation, urinary protein and biochemical detection were measured. The pathologic changes, fibronectin (FN) and typeⅣcollagen (ColⅣ) were investigated at 8th week. The Western-Blot and RT-PCR were used to measure protein expression and mRNA transcription of TGF-β1. Results At 8th week after operation, the urinary protein (12.35 ± 2.13 mg/24 hvs. 19.93 ± 3.19 mg/24 h), serum urea (10.11 ± 0.65 mmol/Lvs.12.09 ± 0.78 mmol/L) and creatinine (68.10 ± 2.51μmol/Lvs.77.63 ± 3.20μmol/L) in the daidzein group decreased significantly than those in the control group (P<0.01). The deposition of ColⅣ (16.33% ± 2.14%vs. 24.68% ± 3.97%) and FN (19.17 ± 2.68 vs. 29.35 ± 4.15) in the daidzein group decreased significantly than those in the control group (P<0.01). Compared with control group, the pathological lesion in the daidzein group was less serious. The mRNA transcription (0.37 ± 0.06vs. 0.64 ± 0.08) and protein expression of TGF-β1 (0.28 ± 0.09vs. 1.40 ± 0.13) in the daidzein group were attenuated significantly than those in the control group (P<0.05). ConclusionsDaidzein had a beneficial effect on uremic rats. It may be associated with a decrease of extracellular matrix accumulation.