2.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.
3.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.
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.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.
6.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 .
7.Effects of Zuogui Jiangtang Jieyu Formula on Hippocampal Neuronal Apoptosis in Diabetic ;Rats with Depression
Pan MENG ; Hongqing ZHAO ; Qing DU ; Yuhong WANG ; Hui YANG ; Xuli ZHANG ; Yalan XU
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(9):78-81
Objective To observe the effects of Zuogui Jiangtang Jieyu Formula (ZJJF) on the ability of learning and memory and the expressions of JNK, Bcl-2 and Bax in hippocampus in diabetic rats with depression; To explore the protective mechanism of hippocampal damage in diabetic rats with depression. Methods High-fat gavage combined with intravenous injection of STZ was used to establish the model of diabetic rats. 28 days of chronic stress was given continuously and diabetic rats complicated with depression were built successfully. Then rats were randomly divided into 6 groups, including normal, model, positive medicine, high-, medium-, and low-dose of ZJJF groups. After the last administration, Morris water maze was used to detect escape latency time;Western blot was used to disclose the protein expressions of JNK, Bcl-2 and Bax in rat hippocampaus;RT-PCR was used to test the gene expressions of JNK and Bcl-2 and Bax. Results Compared with the normal group, escape latency time in model rats was significant longer (P<0.01), the protein and gene expression of JNK and Bax in rat hippocampaus significantly increased, Bcl-2 was markedly decreased (P<0.05, P<0.01);Compared with the model group, escape latency time in positive medicine group and high-dose of ZJJF group was significant shorter (P<0.01), the protein and gene expressions of JNK and Bax significantly decreased, and Bcl-2 markedly increased (P<0.05, P<0.01). Conclusion ZJJF can significantly improve the ability of learning and memory in diabetic rats with depression, which might be associated with preventing neuronal apoptosis in hippocampus.
8.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.
9.Effects of MK801 on the depression-like behaviors and damaged hippocampus in diabetes with depression rats
Hui YANG ; Lin LIU ; Qing DU ; Wei LI ; Hongqing ZHAO ; Jian LIU ; Pan MENG ; Yuhong WANG
Chinese Journal of Behavioral Medicine and Brain Science 2017;26(8):684-688
Objective To investigate the effects of dizocilpine(MK801) on depressive-like behaviors and damaged hippocampus in rats with diabetes-related depression.MethodsThe animal models of diabetes-related depression were established and they were randomly divided into two groups based on random number table: model group and MK801 group,while 6 rats were included in each group.And another six health rats were regarded as control group.The Open-field test was used to detect the activities.The damage of hippocampus was valued by HE staining,Nissl staining,and Tunel staining.The protein expressions of Bax,Bcl-2 in hippocampus were detected by Western blot.ResultsThe number of activities was significant decreased in Open-field test in model group when compared with control.Hippocampal neurons vacuoles,Nissl bodies were decreased and apoptotic cells were increased in hippocampus in model group as well.Furthermore,the expression of Bax was significant up-regulated(94.57±7.97,P<0.01),while the Bcl-2 was declined(24.65±5.26,P<0.01).Compared with the model group,the animals in MK801 group exhibited increased activities(12.50±4.42,P<0.01),which accompany with an increased Nissl body(133.55±16.74,P<0.01) and a decreased apoptosis(22.50±6.35,P<0.01).Moreover,the expression of Bax was decreased and the Bcl-2 was increased in MK801 group when they were contrasted to model(33.00±4.57,P<0.01).Conclusion MK801 is a significant element to regulate the expression of apoptosis protein including Bax,Bcl-2,and to protecte the hippocampal neuron in rats with diabetes-related depression effectively.
10.Effects of Zuogui Jiangtang Jieyu Prescription on Neurotrophic Effects of Astrocytes in Diabetes Mellitus Rats with Depression
Hui YANG ; Yuhong WANG ; Qing DU ; Yuanshan HAN ; Pan MENG ; Jian LIU ; Lin LIU ; Hongqing ZHAO
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(9):43-47
Objective To investigate the effects of Zuogui Jiangtang Jieyu Prescription on neurotrophic effects of astrocytes in diabetes mellitus rats with depression. Methods The diabetes mellitus with depression rat models were established by composite method, and then were randomly divided into 5 groups: model group, positive group, Zuogui Jiangtang Jieyu Prescription high-, medium-, low-dose groups, with 12 rats in each group. 12 normal rats were set as control group. Each administration group was given relevenat medicine for gavage for continuous 4 weeks. Open-field test was used to evaluate the depression-like behavior. The expression of GFAP in astrocyte and MAP2 in neuron were tested by immunohistochemistry. The expression of protein and mRNA of BDNF, GDNF, and NGF were tested by Western blot and RT-PCR. Results Compared with the control group, the activities of rats in model group were significantly reduced. The expression of GFAP increased, while the expressions of MAP2, BDNF, GDNF and NGF decreased. Compared with the model group, the depression-like behavior of rats in model group were significantly improved. The expression of GFAP decreased, while the expressions of MAP2, BDNF, GDNF and NGF increased, and GFAP decrseaed significantly. Conclusion The secretion function of astrocyte can be improved by Zuogui Jiangtang Jieyu Prescription. Its anti-depression and neuron-protection function might be correlated with the enhancement of neurotrophic effects of astrocytes.