2.Experimental study of relationship on myocardial perfusion, regional contractile function and cell apoptosis in stunned myocardium by myocardial contrast echocardiography with computer-aided technique
Guoqing DU ; Jiawei TIAN ; Meng ZHANG ; Yang SU ; Yanhui GUO
Chinese Journal of Ultrasonography 2011;20(1):67-71
Objective To evaluate a new computer-aided technique applicable for myocardial contrast echocardiography(MCE) to quantitate automatically calibrated myocardial contrast intensity(CD and to test the feasibility of calibrated CI in assessing myocardial perfusion. To analyze the relationship on myocardial perfusion,regional contractile function and cell apoptosis in stunned myocardium. Methods According to coronary occlusion and reperfusion at different times, rabbits were divided into three groups: 15 min occlusion/30min reperfusion (group Ⅰ ),30 min occlusion / 60min reperfusion (group Ⅱ ) and 120 min occlusion / 60min reperfusion (group Ⅲ ). MCE was performed on all rabbits at baseline,occlusion and after reperfusion,and its images were analyzed by a new computer-aided technique. Myocardial calibrated CI of each segment was measured automatically by software. Percentage wall thickening (WT) of each risk segment at each stage were also measured by echocardiography. The apoptotic index(AI) in regional left myocardial dysfunction was calculated by terminal deoxynucleotidyl transferease-mediated biotinylated deoxyuridine triphosphate nick end labeling(TUNEL ). Results (1) During occlusion, WT in the areas at risk decreased to zero or negative and the calibrated CI values were significantly lower than those at baseline ( P <0.01 ). After reperfusion, WT in all risk segment remained depressed, but calibrated CI significantly improved in group Ⅰ and Ⅱ while those remained unchanged in group Ⅲ. (2)AI in risk myocytes were (13. 70 ± 5.48 ) %, (36.25 ± 5.55 ) % and ( 62.06 ± 6. 70 ) %, respectively, both statistically significant difference between the two groups ( P <0.05 or P < 0.01 ). AI were negatively correlated to WT and calibrated CI ( r = - 0. 87 and r = - 0. 77, P <0.05). Conclusions MCE with computer-aided technique can assess quantitatively myocardial perfusion and regional contractile function. Short-term ischemiareperfusion does not cause myocardial necrosis, but it will lead to myocardial cell apoptosis and the phenomenon of myocardial stunning. Prolonged ischemia, even if given sufficient reperfusion, can lead to apoptosis and necrosis simultaneously.
3.Ultrastructure changes after blocking hepatic perfusion using microbubble-enhanced non-focused ultrasound
Dawei ZHAO ; Meng TIAN ; Peng DU ; Jianzheng YANG ; Tao LI
Chinese Journal of Trauma 2015;31(11):1025-1029
Objective To investigate the effect of microbubble-enhanced non-focused ultrasound on liver uhrastructure during the treatment of hepatic trauma.Methods The model of hepatic trauma was established in 18 healthy New Zealand rabbits.The subjects were divided into trauma group,ultrasound group and ultrasound-microbubble group according to the random number table,with 6 rabbits each.Thicker region of the left hepatic lobe was treated by custom-made non-focused ultrasound for 5 min.Peak intensity (PI) was used to evaluate the blood reperfusion of the target region after treatment.Liver specimens were performed transmission electron microscope examination immediately and 24 h after treatment to analyze ultrastructure changes.Results PI ratio in ultrasound-microbubble group (15.1 ± 2.6) was significantly lower than that in trauma group (23.1 ± 1.1) and ultrasound group (23.4 ± 1.3) (P < 0.05).But the difference between trauma and ultrasound groups was insignificant (P > 0.05).Compared with trauma and ultrasound groups immediately after treatment,hepatic cells in ultrasoundmicrobubble group had obvious edema,sinusoids thinned,organelles arranged in disorder,mitochondrial edema was present,endothelia cells of interlobular hepatic artery,interlobular vein and bile canaliculi in the portal area damaged,and microvilli of bile canaliculi disappeared.Hepatic cells showed morphology of apoptosis 24 h after treatment.Conclusion Microbubble-enhanced non-focused ultrasound can be used to make rapid hemostasis by decreasing the blood perfusion,but it causes certain damage to the ultrastructure of hepatic cells and may induce apoptosis in the radiation zone and neighboring cells.
4.Study on HPLC Characteristic Chromatogram of Shenshitong Granules
Lili ZHOU ; Xinhua XIA ; Lin TANG ; Yang DU ; He DU ; Wei YUE ; Meng ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2015;(3):98-101
Objective To establish the HPLC characteristic chromatogram of Shenshitong Granules. Methods The chromatographie fingerprints were obtained through Thermo Hypersil GOLD-C18 column (4.6 mm×250 mm, 5μm) with the gradient elution solvent system composed of acetonitrile-0.2% phosphoric acid (0-20 min, 5%→10% acetonitrile;20-40 min, 10%→12%acetonitrile;40-60 min, 12%→14%acetonitrile;60-90 min, 14%→20%acetonitrile;90-120 min, 20%→28%acetonitrile). The detective wavelength was set at 280 nm;the flow rate was 1.0 mL/min;the column temperature was maintained at 30 ℃;the analysis time was 120 min. Results The HPLC characteristic chromatogram was built on basis of 10 batches of Shenshitong Granules, including 27 common peaks which contain the characteristic peaks of 6 Chinese herbal medicines, such as Radix Salvia Miltiorrhizae, Herba Lysimachiae, etc. Conclusion The established HPLC fingerprint has high sensitivity and good repeatability, and can be available for quality evaluation of Shenshitong Granules.
5.Research of different methods of stress-induced anxiety-like behavior in rats at different time points
Hongqing ZHAO ; Yuanshan HAN ; Qing DU ; Qin YANG ; Yuhong WANG ; Hui YANG ; Pan MENG
Chinese Journal of Comparative Medicine 2017;27(1):22-26
Objective To compare the biological characteristics of several different anxiety rat models established by different methods of stress at different time points and provide experimental basis for the most appropriate modeling methods .Methods 60 rats were randomly divided into normal , empty bottle stress , chronic emotional stress ( CES ) group, restraint stress for 3h, 6h, and modeling respectively .In the experimental 7 d, 14 d, 21 d, elevated plus maze and fear condition system was used to test anxiety-like behavior in rats , open field test to study anxiety or depression-like behavior , forced swimming test was used to detect depression-like behavior in rats , and using the Elisa test kit to detect the contents of 5-HT, DA in the hippocampus in rats .Results Anxiety-like behavioral test results showed that rats in empty bottle stress, CES, 6 h restraint stress group started to have anxiety-like behavior since 14 d, then anxiety-like behavior was becoming increasingly apparent .Forced swimming test results showed that immobility time in 6 h restraint rats was significantly increased in the first 7 d(P <0.05).Meanwhile, compared with control group, hippocampal 5-HT, DA contents in empty bottle stress and CES rats increased significantly since 14 d.Conclusions Among several stress methods established anxiety model , anxiety-like behavior in 3 h restraint stress was not obvious; 6 h restraint stress exhibited a depression-like behavior in the forced swimming test might be due to prolonged stress .Empty bottle stress and CES can successfully establish the anxiety rat model , and the anxiety behavior of the rats have some differences . Corresponding model methods can be selected according to different experimental purposes .
6.Changes of content of monoamine neurotransmitters and expression of neurotrophic factors in brain regions of rat models of anxious depression
Hongqing ZHAO ; Yuanshan HAN ; Zhuo LIU ; Qing DU ; Qin YANG ; Pan MENG ; Hui YANG ; Yuhong WANG
Acta Laboratorium Animalis Scientia Sinica 2017;25(4):373-379
Objective To study the content of monoamine neurotransmitters and neurotrophic factor in the hippocampus, amygdala and prefrontal cortex in anxious depression rats, and explore the possible pathogenesis.Methods 60 SD rats were randomly divided into normal group, vehicle group, anxiety group, depression group, and anxious depression group, 12 rats in each group.Chronic restraint stress combined with corticosterone injection was used to establish anxiety and depression model, the modeling time was 21 d.After modeling, elevated plus maze test, open field test, and forced swimming test were used to evaluate the anxiety and depression-like behavior, HPLC-ECD was used to detect the content of 5-HT, NE, and DA in the hippocampus, amygdala, and prefrontal cortex of rats.Western-blotting was used to detect the expression of BDNF and NT-3 in rats.Results Rats in anxious depression model group were comparable to the anxiety group in time and frequency entering open arm time, and number of locomotor activity in open field, and it had a significant difference when compared with the control and depression groups (P<0.01 or P<0.05).Immobile time in anxious depression model rats was increased significantly when compared with the control and anxiety groups (P<0.01).Meanwhile, compared with the control group, 5-HT in hippocampus and 5-HT, NE in amygdala or prefrontal cortex were significantly decreased in the depressive rats with anxiety (P<0.01 or P<0.05).Moreover, the content of BDNF and NT-3 was significantly decreased in each brain regions compared with the control group (P<0.01 or P<0.05), and BDNF levels were obviously decreased compared with the anxiety group (P<0.05).Conclusions Rats of anxious depression have significant anxiety and depression-like behaviors.Its mechanism may be associated with the down-regulation of monoamine neurotransmitters and neurotrophic factors BDNF and NT-3 in hippocampus, amygdala, and prefrontal cortex region.
7.Repairing rabbit's radial bone defect by using uncellular tissue engineered complex constructed by autologous red bone marrow wrapped by facial flap with vessels
Xinming YANG ; Wei SHI ; Yakun DU ; Xianyong MENG ; Yanlin YIN ; Huaguang LI
Chinese Journal of Tissue Engineering Research 2009;13(46):9050-9054
BACKGROUND:Foreign scholars have obtained a success to cure fracture by implanting the complex of red bone marrow and formation factor.Due to the in vitro culture process is not necessary,the complex of red bona marrow and scaffold formation factor is only required to be implant immediately,called uncellular tissue engineered bone.OBJECTIVE:This study innovatively constructs uncellular tissue engineered bone with autologous red bone marrow wrapped by fascial flap with pedicle,and validates the superiority of repairing bone defects.DESIGN,TIME AND SETTING:Homebody controlled animal experiment was performed in the Hebei North University and the Experiment Center of the Affiliated Hospital to Hebei North University from December 2007 to February 2009.MATERIALS:A total of 24 News Zealand albino rabbits,aged 4-5 months; uncellular tissue engineered bone was a mixture of autologous red bone marrow and osteoinductive absorbing materials containing bone morphogenetic proteins.METHODS:Bone defect models were induced on adult New Zealand rabbits' right radial bone,left side served as control group,only implanted with osteoinductive absorbing materials complex,while right side served as experiment group,which contained fascial flap with pedicle.A fascial flap prepared with capillary network containing nameless blood vessel pedicle was located to be adjacent to the bone defect using micro-surgical technique,to wrap the tissue engineered bone and to fill the bone defect.MAIN OUTCOME MEASURES:At 4,8,12,16 weeks postoperation,six rabbits were tested by radiograph,spectrodensitometry,gross morphology observation,histological inspection,quantitative analysis of bone morphometry in bone defect area and analysis of vessels image in the junctional zone.RESULTS:①X-ray determination:At 16 weeks,the implant surrounding bone defects formed bone shaft structure in the control group,cortical bone was not continuous and medullary cavity was obstructed; in the experiment group,normal bone shaft structure was formed and recanalization of medullary cavity was observed.②Histological observation:At 16 weeks,few vessels grew into implant in the control group,mature bone trabecular were observed,and medullary cavity was obstructed; in the experiment group,the implanting materials were completely degraded and substituted by new bone,mature bone structure formed and recanalization of medullary cavity was observed.③Quantitative analysis of bone morphometry in bone defect area:At 4,8,12,16 weeks postoperation,the volume of bone trabecula in the experiment group was more than that in control group (P < 0.05).④Analysis of vessels image in the junctional zone:The area of vessels in the unit area in the experiment group was greater than that in control group in every time stage (P<0.05).CONCLUSION:The uncellular tissue engineering complex constructed by autologous red bone marrow wrapped by fascial flap with pedicle shows double effects of hymeno-inducing regeneration of bone and the vascularization.It is feasible to repair large-segment bone defects.It has obvious therapeutic effect in the aspects such as reducing the bone defect reparation time and advancing the quantity and quality of the bone generation.
8.Animal models of laparoscopy assisted natural orifice translumenal endoscopic surgery
Gang SUN ; Wen LI ; Guohui SUN ; Xiangdong WANG ; Jiangyun MENG ; Hong DU ; Yunsheng YANG
Chinese Journal of Digestive Endoscopy 2008;25(5):225-228
Objective To explore the manipulation,feasibility and safety of laparoscopy assisted natural orifice translumenal endoscopic surgery(NOTES).Methods The technique was performed in 4 female porcine models.Diagnostic laparoscopy was performed first,and followed by transgastric access through the anterior gastric wall under the monitor of laparoscopy.The stoma was extended with the dilation ballon and the endoscope was sent to the abodominal cavity.Such procedures as endoscopic peritoneoseopy,fallopian tube ligation,oophorectomy,eholeeysteetomy and closure of gastrostomy were performed.The drainage tube and antibiotics of 3 days were used after operation according to the bleeding and potential risk of fistula and peritonitis.The animals were feeded on a semiliquid diet.Two weeks later,routine gastroseopy was pedbrmed to examine the healing of luminal incision.Subsequently,all models were sacrificed for the evaluation of the validity of NOTES.Results With the assistance of the laparoscope,endoscopic peritoneoseopy and,fallopian tube ligation,oophorectomy and cholecystectomy,twice for each,were successfully performed in all models.Abdominal drainage were used in two animals'after eholecystectomy.All porcines survived for 2 weeks postoperatively with no weight loss.Autopsic examination showed good healing of transgastric incisions,with little adhesion,no massive heinan'hage,abcesses,or injury to adjacent organs.Conclusion Combined laparoscopy and endoscopy for NOTES is feasible with relative safety.Laparoscopic assistance could facilitate the NOTES procedures at the present stage.
9.The experiment research on the repairment of bone defect by using fascial flap with vessels inducing the vas-cularization of uncellular tissue engingeering complex and the regenration of bone
Xinming YANG ; Wei SHI ; Yakun DU ; Chunyu NIU ; Xianyong MENG ; Yanlin YIN ; Huaguang LI
Clinical Medicine of China 2009;25(10):1013-1017
Objective To study the effect of fascial flap with vessels inducing the vascularization of uncel-lular tissue engingeering complex and the regenration of bone on the repair of bone defect, so as to provide the basis for the clinical application. Methods An animal model of bone defect on adult Newzland rabbits'right radial bone was established .and autologous red bone marrow were taken out and mixed into uncellulax tissue engineering comple-xes with OAM which contained BMP. The experiment animals were divided into two groups : experiment group and control group( n = 12 for each ). The control group was only implanted with complexes, meanwhile, the experiment group had fascial flap with vessels. By microsurgery technology,a non-named fascial flap with vessels was prepared, which belonged to capillary net,around the bone defect,and let it wrap tissue engineering complex,fill up bone de-fect. In a certian time, radiograph(X-ray) and light density measure was conducted, gross morphology and histological inspection was exmained. Bone shape measurement analysis and image of vessel analysis were conducted. All the sta-tistics were analyzed by the SPSS 11.5 software. Results Because of mechanically preventing fiber connective tis-sues and surrounding soft tissues from entering the areas of bone defect by fascial flap, it can keep bone defect having a relative stable environment ;The subfascial space itself, and also the shape and mass of filled-in subject had the de-cisive effect on the results of the regeneration of the bone; Owing to the establishment of blood supply during the con-structing tissue engineering complex. The experiment group was obviously superior to the control group. Compared with control group,the absor bance obviously increased in experiment group [(0. 732 ± 0. 021 ) vs (0. 651± 0.018)] (P < 0. 001 ) four weeks after the operation; also the bone trabecular body was significantly increased [(2.32±2.57)% vs(19.37±3.52)% ,(8.37±3.52)% vs(30.24±3.42)% ,(28.57±2.98)% vs(58.76± 4.62)% ,(47.24±3.42)% vs(88.72±5.84)%] ,and capillary area [(5.04±1.62)% vs(17.53±2.86)%, (10.37 ±2.96)% vs(35.24±1. 13)%,(18.20±2. 12)% vs(48.76±4. 62)%,(17.82 ±2. 74)% vs (57.72 ±5.84)%] (P <0.05) at each time period(4 weeks,8 weeks,12 weeks,and 16 weeks after operation). Despite of growth of implant's internal vessel, the number and speed of forming bone trabecula and cartilaginous tis-sue, even developing of mature bone structure, recreating of diaphysis structure, reconstructing of marrow cavity, ab-sorbing and decomposing of implant, the experiment group was obviously superior to the control group. Conclusions The induction of fascial flap with vessels shows double effects, one of which is the vascularization of uncellular tis-sue engineering complex and the other is membrane guided bone regeneration, So the method has a wonderful effect on the repair of bone defect.
10.Seeding fresh bone marrow aspirate directly on scaffolds to construct tissue-engineered ligament
Hong WANG ; Ming TANG ; Chunqing MENG ; Hanqi WANG ; Shuhua YANG ; Jingyuan DU ; Deyu DUAN ; Zengwu SHAO
Chinese Journal of Tissue Engineering Research 2014;(12):1870-1876
BACKGROUND:There are several reports about the application of fresh bone marrow aspirate being injected directly to repair partial ligament injury, but the application about fresh bone marrow aspirate directly being planted on scaffolds to build tissue-engineered ligament is rarely mentioned.
OBJECTIVE:To evaluate the feasibility of applying fresh bone marrow aspirate planted directly on scaffolds to construct tissue-engineered ligament
METHODS:We constructed fibroin fiber/smal intestinal submucosa composite scaffold, then planting fresh bone marrow directly to built bone marrow seeding group and planting seed cel s (bone marrow mesenchymal stem cel s) on the scaffold to built cel seeding group. The control group had no treatment. After that, we detected the density of cel adhesion, cel proliferation ability and extracel ular matrix secretion. Then, the composite in the bone marrow seeding group was implanted into the broken anterior cruciate ligament in rabbits, and material biocompatibility in vivo was evaluated after 12 weeks.
RESULTS AND CONCLUSION:After 4 hours of incubation, bone marrow seeding group was significantly higher than the cel seeding group in cel adhesion density and proliferation rate (P<0.05). Bone marrow seeding group and cel seeding group showed higher type I, III col agen secretion compared with the control group (P<0.05), but the col agen secretion of bone marrow seeding group and cel seeding group showed no significant difference. Composite cel scaffold implantation in vivo did not cause fatal immune rejection and severe inflammatory reaction, and no significant ligament regeneration and vascularization occurred. These findings indicate that fresh bone marrow aspirate can be seeded directly on scaffolds to construct tissue-engineered ligament, and the short-term biocompatibility in vivo is good.