1.Effect of Electroacupuncture on Learning and Memory Ability of Cerebral Ischemia/Reperfusion Injured Rats
Yukun LIN ; Jing TAO ; Bin CHEN ; Ruhui LIN
Chinese Journal of Rehabilitation Theory and Practice 2014;(9):831-834
Objective To investigate the effect of electroacupuncture at Shenting (DU24) and Baihui (DU20) on learning and memory ability, as well as the nerve cell and the expression of mRNA of Bcl-2 and Bax in the hippocampus of rats with cerebral ischemia/reperfusion (I/R) injury. Methods 45 male Sprague-Dawley rats were randomly diveded into sham group (n=15), control group (n=15) and electroacupuncture group (n=15). The latter 2 groups were modeled with middle cerebral artery occlusion for 2 h and reperfusion. The electroacupuncture group received electroacupuncture at Shenting and Baihui for 7 days. They were tested with Morris water maze, and then, their hippocampus was observed under Nissl staining and the level of Bcl-2 and Bax mRNA were determined with RT-PCR. Results The escape latency decreased (P<0.001) and the frequence the platform was crossed increased (P=0.001) in the electroacupuncture group compared with the model group, with less damage of the nerve cell and the expression of Bax mRNA, and more of Bcl-2 mRNA (P<0.05). Conclusion Electroacupuncture can prevent learning and memory ability from cerebral ischemia/reperfusion injury in rats, which may associate with adjustment of expression of apoptosis-related gene to inhibit apoptosis of nerve cell.
2.Effects of silencing of iASPP gene on human bladder cancer cells
Tao LIU ; Lin LI ; Hui JIA ; Hongwei JING ; Chuize KONG
Chinese Journal of Urology 2011;32(4):239-243
Objective To discuss the effects of silencing of iASPP gene on human bladder cancer cells. Methods RNAi silencing of iASPP gene in bladder cancer cell 5637 and T24 cells were used by lentiviral mediated interfering short hairpin RNAs. Cell proliferation was tested by MTT assay, and rate of colony was tested by colony formation assay. Cell cycles were tested by using fluorescence-activated cell sorting. Results Down-regulation of iASPP could inhibit the growth and proliferation of human bladder cancer cells (P<0.05). iASPP know-down could decrease the colony formation of 5637 and T24 cells (P<0, 05). Knocking down of iASPP in 5637 and T24 cells showed cell arrested at G1. Conclusions Silencing of iASPP gene could inhibit proliferation and colony formation of bladder cancer, iASPP might be an important target for gene therapy of bladder cancer.
3.Effects of Electroacupuncture on Learning and Memory Ability and RhoA expression in Rats after Cerebral Ischemic-reperfusion
Yunan WU ; Yun ZHANG ; Ruhui LIN ; Lidian CHEN ; Jing TAO
Chinese Journal of Rehabilitation Theory and Practice 2015;(1):17-21
Objective To explore the effects of electroacupuncture at Shenting (DU24) and Baihui (DU20) on learning and memory ability in the cerebral ischemia-reperfusion rats and its possible mechanism. Methods 45 male Sprague-Dawley rats were randomly divided into sham group (n=15), ischemia group (n=15) and electroacupuncture group (n=15). The latter 2 groups were modeled as focal cerebral ischemia-reperfusion injury, and the electroacupuncture group received electroacupuncture at Shenting (DU24) and Baihui (DU20) for 7 days. They were tested with Morris Water Maze, observed with Nissl's staining. The protein expression of RhoA was detected with Western blotting. Results The learning and memory ability improved in the electroacupuncture group (P<0.05), the injury of the neurons reduced (P<0.05) and the expression of RhoA in hippocampus decreased compared with the ischemia group (P<0.05). Conclusion Electroacupuncture could ameliorate the learning and memory ability in ischemia- reperfusion rats, which may relate with the inhibition of the expression of RhoA in hippocampus.
4.Central Mechanism of Acupuncture at Baihui for Memory Impairment after Stroke: A Resting- state Functional Magnetic Resonance Imaging Study
Zhicheng LIN ; Shanli YANG ; Xiehua XUE ; Jing TAO ; Lidian CHEN
Chinese Journal of Rehabilitation Theory and Practice 2015;21(2):184-188
Objective To explore the central mechanism of acupuncture at Baihui (GV20) for memory impairment after stroke. Methods 32 stroke patients were randomized to receive acupuncture at Baihui (GV20) (as observation group) and acupuncture at Yanglingquan (GB34) (as control group) for 8 weeks. At the meantime, all participants received routine treatment including physical and occupational therapies. They were scanned with resting-state functional magnetic resonance imaging (fMRI) to find functional connectivity and assessed with Wechsler Memory Scale (WMS) before and after treatment. The results of WMS and functional connectivity were analyzed with Pearson's correlation. Results The memory scores and memory quotient improved significantly after treatment in the observation group (P<0.05). The functional connectivity significantly increased occurred in the left hippocampus to right inferior frontal gyrus, right middle frontal gyrus and left inferior frontal gyrus; and right hippocampus to left middle frontal gyrus, left superior frontal gyrus and left parietal lobe. Significant correlations were found between memory quotient and functional connectivity of hippocampus to frontal lobe and left parietal lobe in the observation group. There was no statistical significance in memory scores and memory quotient in the control group. The functional connectivity significantly decreased in left hippocampus to right middle occipital gyrus, and right hippocampus to right superior temporal gyrus and right posterior lobe of cerebellum. There was no statistical correlation between functional connectivity and results of WMS. Conclusion The acupuncture at Baihui can improves memory ability of stroke patients, which may associate with the increase of functional connectivity of hippocampus with frontal lobes and parietal lobes.
5.Expression of angiotensin Ⅱ and its type 1 receptor in circulation, placenta and kidney of the preeclampsia rat model
Lin TAO ; Jianying NIU ; Li YOU ; Jing CHEN ; Yong GU
Chinese Journal of Nephrology 2008;24(9):663-668
Objective To investigate the expression of angiotensinⅡ (Ang Ⅱ ) and its type 1 receptor (AT1) in circulation, placenta and kidney of the rots preeclampsia. Methods Preedampsia rat model was developed by inhibitor of nitric oxide synthase (L-NAME). The systolic blood pressure (SBP), 24 h urine protein, hepatic and renal function were compared among the precelampsia group, the normal pregnant group and nonpregnant control group. The kidney tissue was observed by light microscopy. ELISA and radioimmunoassay were used to detect Ang Ⅱ in rat plasma and kidney homogenate respectively. Placental AT1 was measured by Westem blot. The level of kidney AT1 was evaluated by immunohistochemistry and Western blot. Results In preeclampsia rats, SBP and 24 h urine protein were significantly higher compared with control groups. Compared to normal pregnant group, plasma Ang Ⅱ of preeclampsia rats was much higher [(0.706±0.086) ng/L vs (0.540±0.085) ng/L, P<0.05]; placental AT1 was increased by 46%(P< 0.05); kidney Ang Ⅱ was decreased signigicantly [(65.543±40.634) ng/g vs (165.543±33.078) ng/g, P<0.05]. The expression of ATI in kidney of preeclampsia rats was reduced evidently,which was only 33% of normal pregnancy group and 59% of nonpregnant control greup,respeetively (P<0.05). Conclusions In preeclampsia rat model, the circulating Ang Ⅱ is increased, the placental RAS isactivated, while the kidney BAS is suppressed. The underlying mechanism of proteinuria and kidney damage associated with this phenomenon in preeclampsia needs further research.
6.Optimal animal model of osteoarthritis
Mingjiang HE ; Hongmei ZHANG ; Lin JING ; Zhikun TAO ; Rui HU
Chinese Journal of Tissue Engineering Research 2013;(46):8069-8074
BACKGROUND:There are a variety of methods for establishing osteoarthritis animal model, and different methods and different animals have their own characteristics.
OBJECTIVE:To review the status of the research on osteoarthritis animal models.
METHODS:The relevant articles to the osteoarthritis animal models were searched in Medline database from January 2002 to October 2011, with the key words of“animal model, osteoarthritis”in English by the computer. Similarly, Chinese Journal Ful-text Database was retrieved for related articles published in Chinese from January 2002 to October 2011, with the key words of“animal model, osteoarthritis, research development”in Chinese. Total y 423 relevant articles were col ected and 128 of them conformed the standard. At last 40 articles were included to review after ful-text reading.
RESULTS AND CONCLUSION:The artificial induced animal models are the principal means to establish animal models of osteoarthritis and they have been widely used in the study of osteoarthritis pathogenesis, drug efficacy, cartilage physiology and pathology. Spontaneous osteoarthritis animal models have great advantages in the study addressing the initial mechanism of osteoarthritis, the biochemical changes of articular cartilage and the comparison of prevention and cure effects. Transgenic animal models have great application prospects on studying cartilage repairing mechanisms and testing a gene to prevent, delay or reversal the morphological changes. Accordingly the choice of models should be based on research needs.
7.Expression of cellular FLICE-inhibitory protein in peripheral blood and lesions of patients with psoriasis
Juan TAO ; Yan LI ; Jing DONG ; Lin WANG ; Yeqiang LIU ; Jing YANG ; Yating TU
Chinese Journal of Dermatology 2008;41(6):377-379
Objective To investigate the expression and distribution of cellular FLICE-inhibitory protein (c-FLIP) in peripheral blood and lesions of psoriatic patients. Methods Peripheral blood and skin samples were obtained from 30 patients with psoriasis vulgaris and 20 normal controls. Flow cytometry was used to detect intracellular c-FLIP protein in peripheral T and B lymphocytes, immunohistochemistry to examine the expression of c-FLIP in lesional tissue. Results Based on the positivity rate of c-FLIP, there was a significant increase in T lymphocytes in active psoriasis compared with regressive psoriasis and normal controls (6.32%±1.17% vs 2.64%±0.74% and 2.28%±0.54%, P<0.01 and 0.05, respectively), while no significant difference was found in B lymphocytes among these three groups (0.78%±0.16%, 0.71%±0.32%, 0.69%±0.18%, respectively, P>0.05). The expression intensity of c-FLIP in keratinocytes was also higher in active psoriasis than in regressive psoriasis and normal controls (89.73±5.24 vs 117.40±7.50,121.58±7.93, P<0.01 and 0.05 respectively), and there was no difference between regressive psoriasis and normal controls (P>0.05). Conclusions c-FLIP is highly expressed in lesions and peripheral T lymphocytes of patients with active psoriasis, suggesting the possible involvement of c-FLIP in the proliferation of T lymphocytes in psoriasis.
8.Identification and genetic characterization of Coxsackievirus B5 isolated from an outbreak of aseptic meningitis
Peng CHEN ; Zexin TAO ; Haiyan WANG ; Guifang LIU ; Jing YANG ; Xiaojuan LIN ; Lizhi SONG ; Aiqiang XU
Chinese Journal of Infectious Diseases 2012;30(3):146-151
ObjectiveTo identify the pathogen of an aseptic meningitis outbreak which occurred in Linyi City of Shandong Province during the summer of 2009,and to analyze the genetic variations of Coxsackicvirus B5 (CVB5) isolates.MethodsForty-two cerebrospinal fluids (CSF) specimens were collected from aseptic meningitis cases and virus isolation was performed. The viral RNA was extracted and amplified from the positive specimens using reverse transcription polymerase chain reaction (RT-PCR).The partial VP1 coding region was purified and sequenced. The phylogenetic trees based on VP1 sequences were constructed among CVB5 isolates and those in GenBank.ResultsSeventeen enteroviruse strains were isolated from 42 CSF samples with 40.5% isolation positive rate. All these strains were identified as CVB5 using both microneutralization test and molecular typing methods. Homology comparisons indicated that the nucleotide acid identities and amino acid sequence identities were 92.3 %- 100.0% and 98.7 %- 100.0%,respectively among these CVB5 isolate.s,and compared with the Faulkner prototype strain,which were 81.0%-82.4% and 96.6%97.0%,respectively.Phylogenetic analysis on VP1 sequences showed that all CVB5 could be separated into four genogroups of A,B,C and D.Isolates of this outbreak belonged to genogroup D.Interestingly,two distinct genogroups in the phylogenetic tree were observed among the 17 isolates.Conclusions CVB5 is responsible for the outbreak of aseptic meningitis in Linyi City of Shandong Province,China. The genetic diversity is high among the isolates and all belong to genogroup D.
9.Diagnosis and treatment of thyroglossal duct cyst in infants
Jing MA ; Tao LU ; Zhengcai LI ; Ken LIN ; Lijuan ZHOU ; Tiesong ZHANG
International Journal of Surgery 2012;39(9):602-605
Objective To explore the imageology features and curative effect of operation for thyroglossal duct cyst in infants.Methods CT and B-mode ultrasound features of 23 infant cases diagnosed as thyroglossal duct cyst were retrospectively analysed.The patients underwent classic Sistrunk operation.And ampliate Sistrunk operation was performed for those with postoperative recurrence.Results CT showed a cystic mass in the anterior triangle of neck,relevant to the hyoid bone and thyroid.B-mode ultrasound showed hypoechoic and heterogeneous image companied with posterior echo enhancement.None cyst had a solid component.Twenty-three cases got satisfied curative effect after repeat operation.There were no complications such as trachyphonia,bucking and dyspnea.Following up more than 2 years,no recurrence case occurred.Conclusions CT and B-mode ultrasound scan can help to make accurate diagnosis of the thyroglossal duct cyst,based on the special location and the typical manifestations.Thyroglossal cyst had high recurrence rate after operation.Sistrunk operation is the basic procedure for the complete removal of the thyroglossal cyst in infants.
10.Study on Rac1/MAPK/ERK pathway mediated mechanism and role in rats with ventilator induced lung injury
Guanghua TAO ; Linghui PAN ; Ren JING ; Fei LIN ; Huijun DAI ; Wanyun GE
Chinese Critical Care Medicine 2017;29(3):249-254
Objective To investigate the role of Ras-related C3 botulinum toxin substrate 1/mitogen-activated protein kinase/extracellular signal-regulated kinase (Rac 1/MAPK/ERK) signal pathway in rats with ventilator induced lung injury (VILI) and its mechanism.Methods Thirty Sprague-Dawley (SD) rats were randomly divided into spontaneous respiration group,normal tidal volume (VT) group and high VT group with 10 rats in each group.The rats in spontaneous respiration group were kept their spontaneous breathing.The rats in normal VT group and high VT group were performed tracheal intubation after tracheostomy,and underwent mechanical ventilation on bilateral lungs with 6 mL/kg and 40 mL/kg VT respectively with maintenance anesthesia.After 4-hour ventilation,heart blood,bronchoalveolar lavage fluid (BALF) and lung tissues were harvested.The levels of interleukins (IL-1β,IL-6),tumor necrosis factor-α (TNF-α),myeloperoxidase (MPO) and macrophage inflammatory protein-2 (MIP-2) in serum and BALF were determined by enzyme linked immunosorbent assay (ELISA).Lung wet/dry radio (W/D) was determined.The lung tissues were stained with hematoxylin and eosin (HE),and pathological changes were observed,and pathological scores were evaluated.The ultra structure changes in type Ⅱ alveolar epithelial cells (AEC Ⅱ)were observed with transmission electron microscope.The positive expressions of phosphorylation of extracellular signal-regulated kinase (p-ERK) were determined by immunohistochemistry,and those of Racl and F-actin were determined by immunofluorescence.The mRNA expressions of ERK and Rac1 were determined by real-time fluorescent quantitation reverse transcription-polymerase chain reaction (RT-qPCR),and protein expressions of Rac-1,p-ERK and F-actin were determined by Western Blot.Results ① Compared with spontaneous breathing group,lung W/D in both mechanical ventilation groups was significantly increased,with more significant increase in the high VT group (6.64 ± 0.88 vs.1.79 ± 0.36,P < 0.01).② There was no obvious pathological changes in the lung tissue and AEC Ⅱ of the spontaneously breathing group.In the normal VT group,there was slight edema and infiltration of inflammatory cells;AEC Ⅱ had less lamellar bodies and uniform distribution of the villi of the alveolar epithelium.In the high VT group,the edema of the lung tissue,the widening of the pulmonary septum,the alveolus congestion,the infiltration of inflammatory cells,and alveolar structure disorder were found;and AEC Ⅱ was irregular,the number of lamellar bodies in the plastids was decreased and was unevenly distributed.The pulmonary histopathological score in the high VT group was significantly higher than that in the spontaneous breathing group and the normal VT group (12.00 ± 2.00 vs.6.00 ± 1.51,8.50 ± 0.53,both P < 0.01).③ Compared with spontaneous breathing group,IL-1β,IL-6,TNF-α,MPO,and MIP-2in serum and BALF in both mechanical ventilation groups were significantly increased,with more siguificant increase in the high VT group [serum IL-1 β (ng/L):104.2 ± 15.1 vs.20.3 ± 8.3,IL-6 (ng/L):46.6 ± 11.5 vs.22.7 ± 7.5,TNF-α (ng/L):39.4±6.5 vs.5.4± 1.9,MPO (ng/L):0.66±0.24 vs.0.06±0.03,MIP-2 (ng/L):109.2±25.8 vs.22.8±8.4;BALF IL-1 β (ng/L):121.5 ± 25.6 vs.24.0 ± 7.5,IL-6 (ng/L):136.7 ± 32.7 vs.31.4 ± 10.5,TNF-α (ng/L):98.0 ± 14.8vs.10.1 ±2.6,MPO (ng/L):0.80±0.31 vs.0.08±0.04,MIP-2 (ng/L):144.4±28.9 vs.41.2±20.7;all P < 0.01].④ There were only a few p-ERK,Rac1 and F-actin positive expressions in the spontaneous breathing group.The positive expressions in normal VT group were increased.In high VT group,the positive expression of p-ERK was significantly increased;Rac1 and F-actin were mainly distributed in the cell membrane and cytoplasm respectively,the positive expressions were further enhanced.⑤ The gene expressions of ERK and Rac1,and protein expressions of p-ERK,Rac1 and F-actin in the high VT group were significantly higher than those in the spontaneous breathing group and normal VT group [ERK mRNA (2-△△Ct):8.23±2.83 vs.1,3.02± 1.38,p-ERK protein (gray value):1.15±0.36 vs.0.61 ±0.23,0.88±0.22;Rac1 mRNA (2-△△Ct):4.45 ±2.26 vs.1,1.22±0.39,Rac1 protein (gray value):0.91 ±0.16 vs.0.48±0.11,0.55 ± 0.10;F-actin protein (gray value):0.70± 0.09 vs.0.49 ± 0.08,0.55 ± 0.04;all P < 0.01].Conclusion F-actin expression in lung tissue was up-regulated in rats with VILI,which resulted in reconstruction of AEC Ⅱ cyto-skeleton,and variation of cell membrane permeability through Rac 1 /MAPK/ERK sigualing pathway during VILI.