1.CHARACTERIZATION,ISOLATION AND CULTURE OF OSTEOCLASTS FROM NEONATAL RAT LONG BONES
Weizhen ZHANG ; Shifeng YU ; Linfan ZHENG
Acta Anatomica Sinica 1955;0(03):-
Osteoclasts were isolated mechanically from neonatal rat long bones and cultured on glass or devitalized human cortical bone slices. Both morphological and functional studies showed that the isolated cells were of typical osteoclasts, because; 1. they attached rapidly on the matrix and had pseudopodial activity when cultured on glass;2. they had multinuclei,most of them contained 3-10 nuclei;3. calci-tonin and clacitonin gene related peptide directly inhibited the cellular activity; 4. they showed high acid phosphatase activity; 5. resorption lacunae could be found when the cells cocultured with devitalized bone slices.
2.Biological functions and drug development of Pyk2
Zhao ZHANG ; Shifeng CHU ; Naihong CHEN
Chinese Pharmacological Bulletin 2014;(6):744-747,748
Pyk2(Proline-rich tyrosine kinase 2) is an important member of focal adhesion kinase family. Pyk2 is highly ex-pressed in the central nervous system and the hematopoietic sys-tem . Pyk 2 can trigger a variety of SH 2 domain-containing pro-
teins to phosphorylate their substrates, thus it can participate in the regulations including ion channel activation, stress response, cell adhesion, cytoskeleton reorganization, vesicle transport and so on. Through the regulations above, the ability of cell migra-tion, survival, proliferation changes accordingly, suggesting that Pyk2 in many important regulatory processes may become a tar-get for clinical effects. Current drug development for Pyk2 main-
ly focuses on cancer, osteoporosis and immune response. This review illustrates the domain structure, regulatory mechanisms, potential drug targets, and the drug development of Pyk2 based on the above three fields, which will provide a theoretical basis for clinical treatment.
3.The dynamic expression profile of forkhead transcription factor genes in normal Balb/c mice and after bile duct ligation
Shuangshuang YANG ; Shifeng HUANG ; Liping ZHANG
International Journal of Laboratory Medicine 2010;31(6):533-535
Objective The present study was initiated to determine the expression profiles of the Fox genes in normal Balb/c mouse liver and their dynamic expression changes during fibrogenesis induced by experimental bile duct ligation(BDL).Methods RT-PCR was employed to detect 18 Fox family members including Foxo1,Foxo3,Foxm1,and Foxl1 in normal mouse liver.After mice were bile-duct-ligated,real time fluorescence quantitative PCR(FQ-PCR)was performed to ananlyze the dynamic mRNA expression changes of 9 inflammation-or proliferation-related Fox family genes.Results All the 18 Fox genes were found to be exDressed in the normal mouse liver and bile duct ligation profoundly influenced the expression of Fox transcriptional factor family genes.The expression of Foxol was significantly reduced by BDL,while FoxoL1 and Foxom1 expression were enhanced in this roodel.Moreover,the expression of Foxo1 and Foxo3 were the highest among the Fox members.Conclusion The Fox family genes-related to inflammation and proliferation were dynamically changed during BDL-induced liver injury,it indicates these genes were involved in the pathogenesis of liver fibrosis.
4.Expressions of fibroblast growth factor-1, mitogen activated protein kinase-1 and CD34 in the lesions of patients with psoriasis vulgaris and their significance
Jian JIAO ; Chunhong ZHANG ; Shifeng Lü
Chinese Journal of Dermatology 2013;46(11):784-787
Objective To estimate the relationship of fibroblast growth factor-1 (FGF-1),mitogen activated protein kinase-1 (MAPK-1) and CD34 expressions with clinicopathological findings in lesions of psoriasis vulgaris.Methods Skin specimens were collected from 30 patients with psoriasis vulgaris,whose general information and clinical findings were recorded.The protein expressions of FGF-1,MAPK-1 and CD34 in these specimens were detected by tissue chip technology and immunohistochemistry,and the mRNA expression of FGF-1 by reverse transcription-PCR.The correlations of FGF-1 protein expression with patients' gender,age,clinical stage,psoriasis area and severity index (PASI) score,angiogenesis and abnormal epidermal proliferation were statistically analyzed.Results The protein and mRNA expressions of FGF-1 were significantly higher in the lesions than in the control skin (both P < 0.05).The protein expressions of CD34 and MAPK-1 were significantly increased,and positively associated with the protein expression of FGF-1 in psoriatic lesions.The up-regulation of FGF-1 protein was unrelated to the age or gender of patients,but associated with clinical stage and PASI score.The proportion of patients with increased FGF-1 expression was significantly larger in patients at active stage and with high PSAI score than in those at quiescent stage and with low PSAI score (P < 0.05).Conclusions In psoriasis vulgaris,FGF-1 may participate in the abnormal epidermal proliferation and vascular dysplasia in dermal papilla,and the upregulation of FGF-1 appears to be associated with the progression of disease and aggravation of lesions.
5.Regular monitoring plasma CMV-DNA level and risk factors analyzing after allogenic hematopoietic stem cell transplantation
Huan LI ; Sanling ZHANG ; Jianchuan DENG ; Ying ZHANG ; Shifeng LOU
Chongqing Medicine 2015;(29):4036-4038,4041
Objective To observe the positive rate of plasma cytomegalovirus DNA(CMV‐DNA) level after allogenic hema‐topoietic stem cell transplantation (allo‐HSCT) ,analysis and explore the risk factors related to CMV infection .Methods Choose 30 patients who had performed allo‐HSCT in our department from July 2012 to September 2014 .PCR were used regularly to detect the plasma CMV‐DNA levels in these patients .The regular monitoring times were as follow :the first month(once a week) ,the second to third month(twice a week) ,the fourth to sixth month(once a month) after allo‐HSCT respectively .The positive rates were coun‐ted in every period .Results Thirteen patients had CMV infection ,and the infection rate were 43 .3% .In the first month ,there were 4 cases (13 .3% )whose plasma CMV‐DNA levels were positive ,however ,the positive cases in the second month ,the third month , the fourth month ,the fifth month and the sixth month were 11(36 .7% ) ,2(6 .7% ) ,0 ,2(6 .7% ) and 0 respectively .Statistical data showed that it was in the second month after allo‐HSCT that the CMV‐DNA positive rate was higher than other periods .The anal‐ysis suggested that the positive rate of CMV‐DNA related to the administration of rabbit anti‐human thymocyte globulin(ATG) ,ba‐siliximab ,and the occurrence of acute graft versus host disease(GVHD) ,there were no relationship among gender ,age ,risk stratifi‐cation of primary disease ,HLA condition ,preparative project ,recovery time of neutrophile granulocyte .Conclusion It is necessary and beneficial to monitor blood CMV‐DNA level regularly and take treatment early to avoid CMV related comobidity after allo‐HSCT .
6.Preparation of LEM-GM-CSF and its inhibition effect on HL60/ADM cells in vitro
Shu CHEN ; Shifeng LOU ; Ying ZHANG ; Ping ZHANG
Journal of Third Military Medical University 2003;0(10):-
Objective To prepare liposome-entrapped mitoxantrone (LEM)-GM-CSF and observe the cytotoxicity of HL-60/ADM cells treated with LEM-GM-CSF, LEM and dihydroxyanthraquinone (DHAQ) in vitro. Methods LEM was prepared by reverse phase evaporation (REV). High speed centrifugation was applied to separate LEM and dissociate DHAQ. Colorimetry was employed to determine encapsulation efficiency. The liposome structure and particle size were determined by transmission electron microscopy. GM-CSF was coupled to LEM by glutaraldehyde method. UV-spectrophotometric analysis was applied to measure the coupled efficiency. Flow cytometry was applied to determine the immunoconjugate retained efficiency. The cytotoxicity of HL60/ADM cells and interdiction efficiency of GM-CSF were investigated by MTT test. Results The encapsulation efficiency of LEM was 80%. Most liposomes were monolayer, and the particle size was 170-220 nm. Its coupled efficiency with GM-CSF was 42.3%, and the immunoconjugate retained efficiency was 74.6%. All LEM-GM-CSF, LEM and DHAQ had cytotoxicity on HL60/ADM, their cytotoxic power in decrement sequence: LEM-GM-CSF, LEM, DHAQ. After treated with LEM-GM-CSF, LEM and DHAQ for 24 h, the IC50 of HL-60/ADM was 8.73, 12.42, 27.31 ?g/ml respectively and for 48 h the IC50 were 0.62, 8.25, 12.44 ?g/ml. The inhibition rate increased in a dose-dependent manner. Conclusion The encapsulation efficiency, the coupled efficiency and the immunoconjugate retained efficiency of LEM-GM-CSF prepared by our method were satisfying. LEM-GM-CSF representing anti-leukaemia efficiency in vitro had cytotoxicity on HL60/ADM cells.
7.The relationship between NO and gossypol in decreasing sperm quality in male rats
Shifeng CHU ; Yuzhu WANG ; Lixiang ZHENG ; Juntian ZHANG
Chinese Pharmacological Bulletin 2003;0(11):-
Aim To elucidate the biochemical mechanism of gossypol in inducing the decline of sperm quality.Methods Gossypol was administered orally at the dose of 50 mg?kg-1/2 d for two weeks.Then,the sperm was collected from the left caudal epididymis and was analysed by CASA.The morphological changeand the concentration of nitric oxide(NO)in testes as well as the level of hormone〔follicle-stimulating hormone(FSH)〕,luteotrophic hormone(LH),testosterone(T)in serum were assayed.Results Gossypol could induce the decrease of sperm number and sperm quality.The concentration of NO in testes increased significantly.Among the three kind of hormone,only the concentration of T showed decrease after the oral administration of gossypol.NA1108 could antagonize the decline of sperm quality damaged by gossypol and decrease the content of NO in testes.Conclusions The concentration of NO in testes beyond normal value was one of the toxic mechanism of gossypol that contributed to the inhibition of spermatogenesis.Some drugs with the ability to reduce NO content in testes could also increase sperm quality.
8.Acupuncture for aphasia: a retrospective analysis of clinical literature.
Jie TAN ; Hong ZHANG ; Guodong HAN ; Kun AI ; Shifeng DENG
Chinese Acupuncture & Moxibustion 2016;36(4):431-436
With the Meta-analysis method, the clinical efficacy of acupuncture and other regular methods for aphasia was evaluated, and the acupoints selection for aphasia was explored. The acupuncture literature of clinical randomized control trials for aphasia published in CNKI, WANFANG, VIP and CBM database was searched; the statistical analysis of clinical efficacy of acupuncture and other regular methods for aphasia was performed by using software Revman 5. 2 provided by Cochrane library. A file of Microsoft Excel was established to perform the analysis of acupoints selection based on frequency analysis method, so as to summarize the characteristics and rules. Totally 385 articles were searched, and 37 articles those met the inclusive criteria was included, involving 1,260 patients in the acupuncture group and 1 238 patients in the control group. The Meta-analysis results showed OR = 3.82, 95% Cl [3.01, 4.85]; rhombus was located on the right side and the funnel plot was nearly symmetry, indicating the treatment effect of the acupuncture group for aphasia was superior to the control group (Z = 11.04, P < 0.000 01). The frequency-analysis results showed that the frequency of acupoints from top to bottom was Lian-quan (CV 23), Tongli (HT 5), Yamen (GV 15), Jinjin (EX-HN 12), Yuye (EX-HN 13), Baihui (GV 20), Yuyan II, Yuyan I and Yuyan III. The frequency of meridians from top to bottom was the governor vessel, extra channels, conception vessel, heart meridian and large intestine meridian. It is concluded that the clinical efficacy of acupuncture combined with speech rehabilitation training and medication treatment for aphasia is superior to that of speech rehabilitation training and medication treatment alone. The clinical treatment for aphasia focuses on its local effect; the main acupoints are in the head and face, and the meridians are governor vessel, extra channels and conception vessel.
Acupuncture Therapy
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Aphasia
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therapy
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Databases, Bibliographic
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Humans
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Retrospective Studies
9.Relationship among blood lipid,blood glucose,peripheral blood white cell levels and slow coronary flow
Lihua SUN ; Gang LIU ; Ying ZHANG ; Shifeng XING ; Suxia HAN
Chinese Journal of cardiovascular Rehabilitation Medicine 2014;23(4):385-388
Objective:To explore the relationship among blood glucose,blood lipid levels,peripheral blood white cells and slow coronary flow (SCF).Methods:Clinical data of 183 patients receiving angiography in our hospital from Apr 2010 to Apr 2013 were retrospectively analyzed.Patients with TIMI grade 2 or lower coronary blood flow were defined as SCF;patients were divided into SCF group (n=93)and normal control group (n=90).Levels of blood lipid,glycosylated haemoglobin (HbA1c),hematocrit,and peripheral blood white cell levels were measured and compared between two groups.Results:Compared with normal control group,there was significant reduction in high density lipoprotein cholesterol (HDL-C) level [(1.21 ± 0.26)mmol/L vs.(1.12 ± 0.28)mmol/L,P =0.043],and significant rise in HbA1c level [(5.41±0.50)% vs.(5.83±0.45)%,P =0.01],hematocrit [(0.41 ±0.04)vs.(0.43±0.07),P =0.01]and white blood cell count [(6.1±1.6)109/L vs.(6.7±1.7)109/L,P <0.05]in SCF group.Logistic analysis indicated that white cell count was the risk factor of SCF (OR 1.920,95% CI 1.234~2.987,P =0.004).Conclusion:Levels of high density lipoprotein cholesterol,glycosylated haemoglobin, hematocrit and white blood cell count are related to slow coronary flow,and elevated white blood cell count may be a risk factor aggravating slow coronary slow.
10.Influence of Rapid Reduction of Physical Activity on Bone Mass and Biochemical Markers of Bone Metabolism in Growing Rats
Shifeng KAN ; Bo YU ; Wenhua CHEN ; Wen ZHANG ; Qi QI
Chinese Journal of Rehabilitation Theory and Practice 2012;18(3):222-225
Objective To observe the effect of rapid reduction of physical activity on bone mass (BMC), bone mineral density (BMD)and bone metabolism in growing rats. Methods 48 male SD rats were divided into the high-intensity exercise group (T1, n=12), high tolow-intensity exercise group (T2, n=12), low-intensity exercise group (T3, n=12) and control group (Con, n=12). During the first 6 weeks,the rats in T1 and T2 group ran on treadmill with high intensity, while rats in T3 group took low-intensity treadmill exercise. 6 rats were sacrificedfrom each group at the end of 6th week. BMC, BMD of femur were measured with DEXA. Alkaline phosphatase (ALP), osteocalcin(BGP), tartrate-resistant acid phosphatase (TRACP) were detected with ELISA. The exercise intensity of T2 group was reduced rapidly to alow level as in T3 group, but remained in the other 2 groups. At the end of the second 6 weeks, the other rats were investigated again. ResultsCompared with the first measurements, ALP and BGP of the T2 group decreased significantly (P<0.05), and TRACP increased (P<0.05). There was no significant difference in BMC and BMD of femur (P>0.05). Conclusion Rapid reduction of physical activity can suppressbone formation and stimulate bone resorption of long bone among growing rats.