1. Comparison of the marker effects of two different fluorescent dyes in labeling endogenous neural stem cells in the central nervous system
Medical Journal of Chinese People's Liberation Army 2012;37(9):673-678
Objective To observe the marker effects of two different fluorescent dyes, DIL and DAPI, in labeling endogenous neural stem cells (ENSCs) in rat central nervous system. Methods Thirty-six Sprague-Dawley rats were randomized into staining groups, comprising DIL group and DAPI group, and the corresponding control groups, including DMSO group for DIL group and PBS group for DAPI group. 0.2% DIL 10μl or 10μg/ml DAPI 10μl was stereotactically injected into the lateral ventricle of rats of DIL group or DAPI group, while DMSO or PBS 10μl was introduced into that of DMSO group or PBS group. Neurological severity score (NSS) was determined 2 hours and 24 hours respectively after the operation. Rats were sacrificed at day 1, 3, 7 after the injection. Serial coronal sections of the brain and spinal cord were carried out on a cryostat, and then they were observed under a confocal microscope. The fluorescence intensity of the targeted area, which highlighted by labeled ependymal cells, in the brain and spinal cord of cervical vertebrae, thoracic vertebrae and lumbar vertebrae were semi-quantified. Fluorescence intensity of each section was measured in triplicate, and a mean value was obtained. Statistical analysis was performed on 3 data sets, randomly selected from sections of brain and spinal cord obtained at day 1, 3, 7. Results Two hours after DIL injection, the rats showed no evident neurological defect. NSS value was very low, and there was no significant difference compared with the DMSO group (P>0.05). Twenty-four hours later, normal neurological function recovered in all the rats. Red fluorescence could be seen in the cytoplasm of ependymal cells in the lateral ventricle and each spinal cord segment at day 1 after the DIL injection, and it did not disappear until the 7th day. Nuclei of DAPI-labeled lateral ventricle cells were blue, with clear nuclear morphology. Choroid plexus cells of the ventricle were also labeled. However, there was no blue fluorescence in the medulla oblongata or any segment ofthe spinal cord. The picture at day 3 and day 7 was similar to that of day 1. No significant difference was found between fluorescence intensity in DIL or DAPI stained cells(P>0.05) at any time point. Conclusions DIL may serve as a marker of the cytoplasm of ependymal cells in the brain ventricle and spinal central canal. DAPI, which is often used in the nuclear staining, can label the ENSCs in the brain ventricle. Intraventricular injection of fluorescent dye is a relatively safety procedure.
2.Expression of caspase-3 and FasL in the hippocampns of the infantile rats with recurrent seizures
Chuanmei TAN ; Dingan MAO ; Tao BO ; Liqun LIU
Journal of Chinese Physician 2009;11(2):163-166
Objective To investigate the expression of caspase-3 and FasL in the hippocampus of the infantile rats with recurrent sei-zures. Methods 72 of 20-day-old Sprague-Dawley rats were randomly divided into two groups, control group and seizure group. Seizures in rats were induced by inhalant flurothyl daily in six consecutive days. Brain tissue was sampled at different time points (the 1st day, 3rd day and 7th day) after last seizure. The expressions of caspase-3 and FasL proteins in the hippocampus were detected by immunohistochemistry, and the expression of caspase-3 mRNA was measured by reverse transcription- polymerase chain reaction (RT-PCR). Results The caspase-3 protein, FasL protein and caspnse-3 mRNA levels were obviously increased at the 1st, 3rd and 7th day after recurrent seizure in the hippocampus of the rat(P<0.01). Conclusions Caspase-3 and FasL are participated in the infantile brain injury after recurrent seizures.
3.Effect and significance of Ginkgo biloba(银杏叶) extract on the expression of caspase-3 in the infant rats with brain injury following recurrent seizures
Dingan MAO ; Chuanmei TAN ; Liqun LIU ; Tao BO
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2006;0(02):-
Objective To investigate the effects of Ginkgo biloba(银杏叶) extract on the expression of caspase-3 in the hippocampus of the rats following recurrent seizures,the role of caspase-3 in the brain injury induced by seizures and the possible protective mechanism of Ginkgo biloba extract against brain injury.Methods One hundred and eight Sprague-Dawley(SD) rats,twenty-day-old,were divided randomly into three groups: the control group,the seizure group and the Ginkgo biloba extract treatment group.Seizures models in rats were produced by consecutive inhalation of flurothyl daily for six days.The rats in the Ginkgo biloba extract treatment group received 50 mg/kg of the Ginkgo biloba extract through intra-abdominal injection immediately after every seizure,and after 12 hours another injection was repeated,while the rats in seizure group and control group received volume-matched saline through intra-abdominal injections.Brain tissue was sampled at different time points(1 day,3 days,7 days) after last seizure.The expression of caspase-3 protein in the hippocampus was detected by immunohistochemistry,and the expression of caspase-3 mRNA was measured by reverse transcription-polymerase chain reaction(RT-PCR).Results The caspase-3 protein and caspase-3 mRNA levels at the time points of postseizure 1,3,7 days in the hippocampus of the rat pups in seizure group were much higher than those in control group,while they were highest on the post-seizure 3rd day(all P
4.Percutaneous balloon angioplasty combined with injectable coral artificial bone repairs femoral head necrosis
Kanghua ZHU ; Zhibin MENG ; Tao HUANG ; Haitao TAN ; Bo SUN ; Xueyi PANG
Chinese Journal of Tissue Engineering Research 2016;20(25):3687-3692
BACKGROUND: There are many methods for the treatment of femoral head necrosis, such as core decompression, bone graft, arthroplasty and joint replacement, and each of which has its own shortcomings. So, percutaneous bal oon angioplasty combined with coral artificial bone provides a new attempt for the treatment of femoral head necrosis. OBJECTIVE: To observe the effect of percutaneous bal oon angioplasty combined with coral artificial bone on femoral head necrosis repair. METHODS: Twenty-four Duroc piglets were enrol ed to establish bilateral femoral head necrosis models by liquid nitrogen freezing method. Then, model piglets were randomly treated with percutaneous bal oon angioplasty combined with injectable coral artificial bone (experimental group) or bone cement (control group) on one affected side, and meanwhile, given no treatment on the contralateral side (blank control group). At 2, 4, 8 and 16 weeks after surgery, X-ray examination, biomechanical test and histological detection were conducted. RESULTS AND CONCLUSION: X-ray showed that at 16 weeks after surgery, numerous new bones could be found in the experimental group and there was a fuzzy boundary between the artificial bone and surrounding tissues; no new bone formed in the control group, and the boundary was clear; in the blank control group, the surface of the femoral head col apsed, and bone trabeculae arranged disorderly, which were seriously destroyed. And in the histological detection at 16 weeks after surgery, there were numerous bone trabecula and osteoblasts around the coral bone in the experimental group, and the coral artificial bone almost dissolved; in the control group, bone cement was in an irregular shape and no bone trabecula formed; in the blank control group, bone trabecula were damaged in the col apsed area, whose structure was in disorder. Additional y, biomechanical changes in the experimental group were significantly better than those in the other two groups at different time points after surgery (P < 0.05). In conclusion, percutaneous bal oon angioplasty combined with coral artificial bone can repair femoral head necrosis by promoting new bone formation.
5.Researches on left atrial appendage occlusion by silk thread ligation during open heart operation for prevention of cerebral embolism in patients with rheumatic atrial fibrillation
Shengzhong LIU ; Jin TAN ; Bo XIANG ; Lu JIANG ; Tao YU ; Keli HUANG
Chongqing Medicine 2016;45(7):905-908
Objective To investigate the safety of left atrial appendage occlusion by silk thread ligation during open heart op‐eration in patients with rheumatic atrial fibrillation ,and to evaluate its effectiveness for prevention of cerebral embolism .Methods From April 2012 to March 2014 ,129 patients with rheumatic atrial fibrillation were undergone mitral valve replacement and left at‐rial appendage occlusion by ligation using two silk threads from the outside of the heart (ligation group) .The indexes related to the operation ,postoperative complications incidence ,and cerebral embolism incidence during the follow‐up period of ligation group were compared with the indexes of another 129 patients without ligation of left atrial appendage over the same period (control group) . Results The operation time ,the cardiopulmonary bypass time ,the clamp time ,the intensive care unit stay time ,the postoperative hospitalization time in ligation group were (235 ± 50)min ,(88 ± 24)min ,(57 ± 16)min ,(26 .5 ± 9 .3)h and (12 .4 ± 7 .5)d respective‐ly ,and significant difference was not found compared with control group (P>0 .05) .The thoracotomy for hemostasis(1 cases) ,low cardiac output syndrome(2 cases) ,acute renal failure(2 cases) ,pulmonary infection(3 cases) ,sternal wound dehiscence(2 cases) and other complications in ligation group had no significant difference ,compared with control group(P>0 .05);2 cases died in liga‐tion group ,3 patients died in control group ,the differences had no statistical significance(P>0 .05) .No cerebral embolism occurred in ligation group with 127 patients following‐up (23 .6 ± 11 .3) months ,but 5 patients suffered from cerebral embolism in control group with 126 patients following‐up (22 .9 ± 12 .1) months ,the difference had statistical significance(P<0 .05) .Conclusion Left atrial appendage occlusion by silk thread ligation during open heart operation in patients with rheumatic atrial fibrillation is simple and safe ,can reduce cerebral embolism incidence .
6.The early experience of robot-assisted thymectomy
Shaolin TAO ; Poming KANG ; Bin JIANG ; Cheng SHEN ; Bo DENG ; Ruwen WANG ; Jinghai ZHOU ; Qunyou TAN
Journal of Chinese Physician 2017;19(7):966-969
Objective To summarize the early experience of thymectomy by robot-assisted thoracoscopic surgery (RATS) with the da Vinci S Surgical System.Methods A total of 16 patients was underwent thymectomy from October 2016 to May 2017 by RATS.Among them,there were 10 cases of thymic hyperplasia,5 cases of thymoma and 1 case of thymic small cell carcinoma,including 11 cases of myasthenia gravis (MG).The Osserman types were as follows:type Ⅰ (5),type ⅡA (3),type ⅡB (2),and type Ⅲ (1).Clinical data were collected and analyzed.Results The operations were successfully accomplished by RATS in all the 16 patients without any conversion to open surgery and major complication.The docking time was 2-45 (11.69 ± 11.03) min.The operation time was 47-152 (84.06 ± 25.67) min.The blood loss was 0-150 (57.50 ± 37.32) ml.The diameter of thymus or tumor was 4-19.5 cm and (8.22 ± 3.64) cm.Fourteen cases were removed tracheal intubation in the operation day,and the other 2 cases taked off in 6th and 8th days.One case occurred myasthenia gravis crisis after surgery.All patients were followed up for 2-7 months after discharge without MG recurrence.Conclusions The da Vinci S Surgical System can be safely,accurately and stably used for thymectomy.
7.Correlation between overexpression of PC4 in lung adenocarcinoma with lymph node metastasis
Tianyu SUN ; Qunyou TAN ; Chunmeng SHI ; Ruwen WANG ; Bo DENG ; Jinghai ZHOU ; Shaolin TAO ; Poming KANG
Chongqing Medicine 2015;(11):1449-1451,1456
Objective To investigate the promotion effect of human transcriptional positive cofactor 4 (PC4) overexpression on lymphatic metastasis in lung adenocarcinoma .Methods 96 samples of lung adenocarcinoma tissue were collected .The immuno‐histochemistry(IHC) and real‐time quantitative polymerase chain reaction (qRT‐PCR) were adopted for detecting the expression levels of PC4 protein and mRNA .The correlation of PC4 expression with lymphatic metastasis and TNM stage was analyzed .Re‐sults The expression of PC4 protein was positively correlated mRNA in lung adenocarcinoma (r=0 .63 ,P<0 .01);the expression of PC4 protein was positively correlated with lymph node metastasis (χ2 =8 .29 ,P<0 .01) and TNM stage (χ2 =4 .71 ,P<0 .05);the expression of PC4 mRNA was also positively correlated with lymph node metastasis (χ2 = 8 .40 ,P< 0 .01) and TNM stage (χ2 =5 .10 ,P<0 .05) .Conclusion PC4 overexpression is found to be closely associated with the lymph node metastasis and TNM stage .PC4 may facilitate the lymph node metastasis of lung adenocarcinoma .
8.Comparafive study on arterial blood gas analysis and pulmonary functions of emphysematous rabbits after unilateral and bilateral lung volume reduction surgery
Qunyou TAN ; Bo DENG ; Dongliang LI ; Taiqian GONG ; Shaolin TAO ; Ruwen WANG ; Yaoguang JIANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2011;27(9):561-563,565
Objective Describes the changes of arterial blood gas analysis and pulmonary functions of emphysematous rabbits after unilateral and bilateral lung volume reduction surgery.Methods Sixty rabbits were randomly and equally divided into 6 groups:control group(A),emphysema group( B),unilateral sham operation group(C),unilateral lung volume reduction surgery group( D),bilateral sham operation group( E ) and bilateral lung volume reduction surgery group(F).Obstructive emphysema was induced with somg and papain in tie rabbits of the remaining groups except the control one.Eight weeks later,arterial blood gas analysis and pulmonary functions were detected.Results Compared with group B,pH,PaO2,PaCO2,SaO2,forced expirratory volume in 0.5 s,froced expiratory capacity,functional residual capacity,total lung capacity,dynamic compliance,and resistance in expiartory phase could be improved in group D ( P < 0.05 ).The amelioration of those results were more obvious in group F ( P <0.05).Two rabbits in group F died after opertation.Conclusion Bilateral lung volume reduction surgery could more markedly improve the arterial blood gas analysis and pulmonary functions of emphysematous rabbits than unilateral operation.
9. Comparison of the marker effects of two different fluorescent dyes in labeling endogenous neural stem cells in the central nervous system
Medical Journal of Chinese People's Liberation Army 2012;37(9):673-678
Objective To observe the marker effects of two different fluorescent dyes, DIL and DAPI, in labeling endogenous neural stem cells (ENSCs) in rat central nervous system. Methods Thirty-six Sprague-Dawley rats were randomized into staining groups, comprising DIL group and DAPI group, and the corresponding control groups, including DMSO group for DIL group and PBS group for DAPI group. 0.2% DIL 10μl or 10μg/ml DAPI 10μl was stereotactically injected into the lateral ventricle of rats of DIL group or DAPI group, while DMSO or PBS 10μl was introduced into that of DMSO group or PBS group. Neurological severity score (NSS) was determined 2 hours and 24 hours respectively after the operation. Rats were sacrificed at day 1, 3, 7 after the injection. Serial coronal sections of the brain and spinal cord were carried out on a cryostat, and then they were observed under a confocal microscope. The fluorescence intensity of the targeted area, which highlighted by labeled ependymal cells, in the brain and spinal cord of cervical vertebrae, thoracic vertebrae and lumbar vertebrae were semi-quantified. Fluorescence intensity of each section was measured in triplicate, and a mean value was obtained. Statistical analysis was performed on 3 data sets, randomly selected from sections of brain and spinal cord obtained at day 1, 3, 7. Results Two hours after DIL injection, the rats showed no evident neurological defect. NSS value was very low, and there was no significant difference compared with the DMSO group (P>0.05). Twenty-four hours later, normal neurological function recovered in all the rats. Red fluorescence could be seen in the cytoplasm of ependymal cells in the lateral ventricle and each spinal cord segment at day 1 after the DIL injection, and it did not disappear until the 7th day. Nuclei of DAPI-labeled lateral ventricle cells were blue, with clear nuclear morphology. Choroid plexus cells of the ventricle were also labeled. However, there was no blue fluorescence in the medulla oblongata or any segment ofthe spinal cord. The picture at day 3 and day 7 was similar to that of day 1. No significant difference was found between fluorescence intensity in DIL or DAPI stained cells(P>0.05) at any time point. Conclusions DIL may serve as a marker of the cytoplasm of ependymal cells in the brain ventricle and spinal central canal. DAPI, which is often used in the nuclear staining, can label the ENSCs in the brain ventricle. Intraventricular injection of fluorescent dye is a relatively safety procedure.
10.Recombined adenovirus expressing P53 in the treatment of malignant pleural effusion with lung cancer
Bo TANG ; Qunyou TAN ; Ruwen WANG ; Jinghai ZHOU ; Bo DENG ; Poming KANG ; Fuqiang DAI ; Bin JIANG ; Kai QIAN ; Shaolin TAO
Journal of Regional Anatomy and Operative Surgery 2015;(2):192-194
Objective To assess the therapeutic efficacy of a recombined adenovirus expressing p53 (rAd-p53) via intrapleural injec-tion in the treatment of lung cancer with malignant pleural effusion. Methods Thirty-six cases with lung cancer and malignant pleural effu-sion were randomly divided into two groups,which were given intravenous injection of Nedaplatin with (observation group,n=20) or without (control group,n=16) intrapleural injection of rAd-p53,respectively. Between the two groups,the efficacy in treatment of pleural effusion, the amelioration of maximal ventilatory volume ( MVV) ,Kamofsky scoring ( KPS) and quality of life were compared. Results The efficacy in treatment of pleural effusion in observation group are significantly higher than that in control group(17/20 vs. 50%,P<0. 05). The cases with KPS≥80 in observation group were significantly increased following treatment (5/20 vs. 11/20,P <0. 05). However,there was no difference with the cases in control group. Conclusion Intrapleural injection of recombinant adenovirus expressing p53 (rAd-p53) is effec-tive to reduce the occurrence of malignant pleural effusion and increase the quality of life remarkably.