4.THE EFFECT AND MECHANISM OF ASTROCYTE ON SYNAPTIC FORMATION IN RATS
Rong HU ; Xigui WU ; Zhong YANG ; Lu WEI ; Wenqin CAI
Acta Anatomica Sinica 1955;0(03):-
Objective To study the effect of astrocyte on synapse formation and the molecular mechanism. Methods Cortical astrocytes were isolated and purified from neonatal rats.On the 2h,7th day,14th day and 21th day after passage,we counted the number of astrocytes and the culture medium(astrocyte-conditioned medium,ACM)was harvested to measure the concentration of estrogen(E 2)by using ELISA techniques.Based on the model of pure culures of neonatal cortical neurons,the experimental groups were designed as follows:1.pure neuron cultures(group N);2.ACM cultures(group A);3.mixed cultures(group M);4.E 2 cultures(group E 2);5.ACM+Tamoxifen(estrogen receptor antagonist)cultures(group A+T);6.Tamoxifen cultures(group T).Then synaptic puncta in every group was stained and counted through immunofluorescence,and we also compared the differences in puncta number among those six groups(at 9th day in culture,number/per neuron). Results The numbers of astrocytes were:1?10 4/ml, 1.1?10 6/ml, 1.4?10 6/ml, 1.5?10 6/ml; The concentrations of E 2 were:(ng/L):0, 117?22, 266?22,252?27 respectively.No estrogen was detected in the primary culture medium.The concenteration of estrogen increased in correspondence with the culturing days and reached the peak around at the 14th day, then decreased gradually but kept at a certain high level,and the numbers of synaptic puncta of per neuron in group N,A,M,E 2,A+T,T were:14?3;79?5;83?8;80?6;32?3;29?3 respectively.The treatment of pure neuron culture with ACM increased the number of synapses on per neuron by up to 6 fold by comparison with pure neuronal culture.Exogenic estradiol added into pure neurons can mimick the effect of the ACM.Tamoxifen which is antagonist of estrogen receptor could decrease the effect of ACM by 75%.Conclusion The astrocytes of neonatal rat cortex do secrete E 2.Astrocyte-derived estrogen may be the molecule regulating the synaptic formation through estrogen receptors.
5.Construction of RNAi vectors for SmNAC1 transcription factors of Salvia miltiorrhiza using Gateway cloning technology.
Rong ZHAO ; Qi-Xian RONG ; Yu-Zhong LIU ; Ye SHEN ; Lu-Qi HUANG
China Journal of Chinese Materia Medica 2014;39(9):1569-1573
NAC transcription factors involved in plant growth and development, as well as responses to biotic and abiotic stress. RNAi Vectors for SmNAC transcription factors of Salvia miltiorrhiza was constructed by using Gateway cloning technology, in order to further study the function of SmNAC1 transcription factor. According to Gateway cloning technology, the specific fragments of SmNAC1 containing attB adapter was amplified by PCR using ultra-fideling phusion polymerase of NEB. By the BP recombination reaction, the PCR product containing attB was transferred to an donor vector (pENTR/SD/D-TOPO). Finally, SmNACi specific gene was cloned into pK7GWIWG2D plant expression vectors by LR recombination reaction. Experimental results showed that Gateway cloning technology provide a rapid and highly efficient way to clone the interested gene.
Cloning, Molecular
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methods
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Genetic Vectors
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genetics
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Plant Proteins
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genetics
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Polymerase Chain Reaction
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RNA Interference
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Reproducibility of Results
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Salvia miltiorrhiza
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genetics
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Transcription Factors
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genetics
6.Clinical study of ventricular tachycardia in children.
Zhong-he JIN ; Ning CHU ; Ze-rong WANG ; Jin ZHANG ; Yong-ri LU
Chinese Journal of Pediatrics 2003;41(10):778-779
7.Treatment with diazepanum and dimercaptopropansulfonate sodium for acute tetramine intoxication.
Chu-huan ZHAO ; Zhong-qiu LU ; Hui-ping LI ; Jing-rong LI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(1):68-69
Acute Disease
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Adolescent
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Adult
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Anticonvulsants
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therapeutic use
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Antidotes
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therapeutic use
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Bridged-Ring Compounds
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poisoning
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Diazepam
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therapeutic use
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Drug Therapy, Combination
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Electroencephalography
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Female
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Humans
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Male
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Poisoning
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drug therapy
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Treatment Outcome
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Unithiol
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therapeutic use
8.Treatment of radial neck fracture in children with elastic stable intramedullary nails.
Lu-feng YAO ; Zhao-ming YE ; Zhao-xu LI ; Zhao-ping ZHONG ; Rong-ming XU
China Journal of Orthopaedics and Traumatology 2013;26(12):1028-1031
OBJECTIVETo investigate effects of elastic stable intramedullary nails for the treatment of radial neck fracture in children.
METHODSFrom July 2006 to December 2011, 25 children with radical neck fractures, which included 16 males and 9 females aged from 7 to 15 years old (means 10.7), were treated with elastic stable intramedullary nails. According to Judet classification, 6 cases were type II, 17 cases were type III and 2 cases were type IV (including 1 case with type IVa and 1 case with type IVb). The fracture healing, pain, deformity and range of motion of elbow were recorded.
RESULTSAll patients were followed up for 6 to 24 months with an average of 14 months. Twenty-five patients were obtained bone healing. According to Tibone and Stoltz evaluation standard, 18 cases got excellet results, 4 cases in good and 3 cases in moderate.
CONCLUSIONElastic stable intramedullary nails for the treatment of radial neck fracture in children has advantages of simple operation,less trauma and good results.
Adolescent ; Bone Nails ; Child ; Female ; Fracture Fixation, Intramedullary ; Humans ; Male ; Radius Fractures ; physiopathology ; surgery ; Range of Motion, Articular ; Treatment Outcome
9.Evaluation of sphingosine kinase 1 expression and its clinical significance in breast invasive ductal carcinoma
Min YI ; Rong WANG ; Lu CHEN ; Tingting SHEN ; Lianchuan SONG ; Shengzhen ZHONG
Chinese Journal of Immunology 2014;(12):1683-1685,1691
Objective:To investigate the expression of sphingosine kinase 1 (SphK1) and its clinical significance in breast invasive ductal carcinoma ( BIDC).Methods: We detected SphK1 expression in 58 samples of surgically resected paired BIDC and normal tumor-adjacent tissues samples by using immunohistochemistry.The correlation between SphK 1 expression and clinicopathologic features was analyzed.Results: The positive expression rate of SphK 1 in BIDC tissues was 69.0% ( 40/58 ) , while its positive expression rate in normal tumor-adjacent was 17.2% (10/58),the difference was statistical significance (χ2=31.636,P=0.000). Clinicopathological evaluation suggested that SphK 1 positive expression was associated with ER negative (χ2=4.392,P=0.036),PR negative (χ2=7.920 , P=0.005 ) , lymph node metastasis (χ2 =5.033 , P=0.025 ) and tumor stage (χ2 =7.117 , P=0.008 ) . Conclusion:The high-expression of SphK1 is correlated with the poor clinicopathological features in BIDC ,suggesting SphK1 may play a key role in development and progression of BIDC.
10.Can PDA patient in early stage of Eisenmenger syndrome accept interventional surgery ?
Xiaoke SHANG ; Liang ZHONG ; Gangcheng ZHANG ; Ting PENG ; Dingyang LI ; Rong LU ; Xiaoxian DENG ; Hongmei ZHOU
Chinese Journal of Interventional Cardiology 2014;(9):553-557
Objective To explore whether the PDA patients in the early stage of Eisenmenger syndrome can accept interventional surgery. Methods Three patients were choose from the “National Multicenter Prospective Registration Study on Pulmonary Artery Hypertension”, Clinical assessment, as well as examinations and registrations associated with PAH, was conducted in accordance with relevant provisions of the registration study within 1 wk prior to surgery. The way of the interventional treatment were right heart catheterization and pulmonary vasodilator testing (a capsule of iloprost solution for inhalation), and measurement of the pulmonary arterial pressure, descending aortic pressure, ratio of pulmonary to systemic blood flow, ratio of pulmonary to systemic blood pressure, pulmonary capillary wedge pressure [mean≤15 mmHg(1 mmHg=0.133 kPa)], before and after testing. After clear the results of pulmonary vasodilator test all patients underwent transcatheter closure testing, if it was positive,the amplatzer was released if her family members’ approved;Conversely, the amplatzer was quickly withdrawn with negative results, and symptomatic treatment was conducted based on the patient’s clinical symptoms. Results The first cases pulmonary vasodilator test was negative, but positive in transcatheter closure testing. Satisfaction immediate effect of surgery, and the occluder was released with her family members’ approval. patients of Example 2 and Example 3 were negative in pulmonary vasodilator testing and transcatheter closure testing, can not released the occluder. Conclusions Some Patents Ductus Arteriosus (PDA) patients in early stage of Eisenmenger syndrome is feasible to accept interventional therapy , try plugging test is may be an important indicator of prognostic assessment.