1.Particle gun-mediated bone morphogenetic protein-2 gene transfection for treatment of chronic bone defects
Xiaomeng ZHU ; Chong WANG ; Xinghua SONG ; Yulin ZHAN ; Wenju LI
Chinese Journal of Tissue Engineering Research 2014;(11):1680-1686
BACKGROUND:Both in vitro and in vivo studies have confirmed that, bone morphogenetic protein (BMP) regulates the differentiation of osteoblasts and chondroblasts, induces heterotopic bone formation, promotes fracture healing, and controls the morphology of skeleton in mammals.
OBJECTIVE:To treat chronic bone defects using particle gun containing BMP2 gene eukaryotic expression plasmid via local injection.
METHODS:A total of 72 healthy New Zealand white rabbits were applied to establish chronic bone defect model in the rabbit radius. According to the length of bone defect, the rabbits were divided into three groups:1.5 cm group, 2.0 cm group, 2.5 cm group. Each group was further randomly assigned into two subgroups:treatment group (BMP-2 gene transfection) and control group (natural y healing). X-ray examinations were performed at 1, 3, 8 and 9 weeks after transfection, and soft tissue between the bone defects was harvested to detect BMP-2 using western blot analysis;and radius specimens were taken for gross observation at the same time points, to evaluate the healing.
RESULTS AND CONCLUSION:(1) Gross specimen observation:bone cal us formation in treatment group was general y more than that in control group. (2) Lane-Sandhu X-ray score in treatment group was significantly higher than that in control group at 1, 3, 8, 9 weeks after transfection (P<0.05). (3) BMP-2 concentration in treatment group was significantly higher than that in control group at each time point (P<0.05). The local transfer of particle gun-mediated BMP-2 gene is an effective therapy of chronic bone defect.
2.Basic study on Tuhuai decoction and its disassembled prescriptions on the amount of water lose in mice with epidermal hyperplasia
Yuejun SHI ; Bo LI ; Shunpeng SONG ; Cheng ZHAN ; Yinghua ZHU ; Qiao LI
International Journal of Traditional Chinese Medicine 2011;33(1):18-22
Objective To study the topical application of Tuhuai decoction and its disassembled prescriptions on skin function of mice with epidermal hyperplasia, and to provide a theoretical basis for the treatment of psoriasis. Methods According to the Denda method, skin barrier function in mice was destructed with tape, to produce epidermal hyperplasia model. Tuhuai decoction and its disassembled prescriptions with different concentrations of water and alcohol extracts were external applied on the mice. Since the beginning of the treatment, loss of skin moisture of both sides the trunk was detected with Multi-functional instrument at every day before using the medicine. Results Tuhuai decoction had function of reducing the amount of water loss of skin. The disassembled prescriptions, such as Angelica group, safflower group, Danshen root group, and water and ethanol extracts of Prtmella group can reduce the amount of water loss of skin effect. Conclusion Water and ethanol extracts of Tuhuai decoction has the functions of reducing water loss of skin and resisting hyperplasia,suitable for treating psoriasis by external application, setting a foundation for clinical treatment of psoriasis.
3.Prognostic analysis of 525 Chinese patients with diffuse large B cell lymphoma
Zhiying FU ; Jun ZHU ; Yuqin SONG ; Weiping LIU ; Xinqiang JI ; Siyan ZHAN
Journal of Peking University(Health Sciences) 2014;(3):405-411
Objective:To describe the clinical characteristics , overall survival as well as to evaluate the prognostic factors in Chinese diffuse large B cell lymphoma ( DLBCL) patients.Methods: DLBCL pa-tients who were initially diagnosed and treated in Peking University Cancer Hospital from January 1995 to December 2008 were identified and analyzed ,retrospectively .The 5-year OS rates were estimated with Ka-plan-Meier.Log-rank test was used to compare the survival curves of the different groups .The multivari-ate analysis of prognostic factors was conducted with Cox regression model , which included all statistically significant prognostic factors in the univariate analyses .Results:A total of 525 DLBCL patients were in-cluded in this retrospective analysis , of whom , 294 were male and 231 female ( male∶female=1 .27∶1 ) . The median age at the initial diagnosis was 55 (range 16-90) years, and 37.0% (n=194) were 60 years and above .Regarding the clinical staging at the initial diagnosis , 54 patients (10.3%) were diag-nosed as Stage Ⅰ of the disease, 152 (28.9%) as Stage Ⅱ, 117 (22.3%) as Stage Ⅲ and 202 (38.5%) as Stage Ⅳ.The ‘B symptoms’ and increased serum LDH were presented in 206 (39.2%) and 192 (36.6%) patients, respectively.A total of 197 (37.5%) patients were treated with rituximab (R).The survival follow-up continued till 31 January 2014 with a median follow-up time of 77.5 ( range:0-205) months.A total of 267 patients (50.9%) died during the follow-up period.The medi-al overall survival ( OS) time was 84 months, and 5-year OS rate was 52.3%.There were six statistically significant prognostic factors that were identified in both univariate and multivariate analyses : gender, Ann Arbor stage, B symptom, serum LDH, age at initial diagnosis and rituximab treatment .The relative risk ( RR) of these prognostic factors in the multivariate analyses were: age >60 years /≤60 years=1.380 (95%CI 1.078 -1.765), male /female =1.315 (95%CI 1.025 -1.687), stage Ⅲ/stageⅠ=3.034(95%CI 1.667-5.522), stage Ⅳ/Ⅰ=3.748(95%CI 2.102 -6.681), with B symp-toms/without B symptoms=1.278(95%CI 0.999-1.636),serum LDH increased/LDH not increased=1.351(95%CI 1.057 -1.726), without R treatment /with R treatment =1.543(95%CI 1.182 -2 .015 ) .Compared with the IPI , age >50 years/≤50 years was a statistically significant factor in both univariate and multivariate analyses RR =1.478 (95%CI 1.148-1.902), P=0.002.Conclusion:Six factors were related to DLBCL survival:gender, Ann Arbor stage, B symptom, serum LDH, age at initial diagnosis and rituximab treatment .Compared with the IPI , several specific factors may predict a poor prognosis in Chinese DLBCL patients:male , age>50 years and the presence of ‘B symptoms ’ .But this result is not conclusive until these factors are further tested .
4.Preincubation with low dose of hydrogen peroxide enhances anti-oxidative stress potential of mouse BMSCs
Yuqing SONG ; Minjia CHEN ; Zhan LI ; Ming ZHU ; Wei QIU ; Hong HUANG ; Xiang XU
Basic & Clinical Medicine 2017;37(3):313-319
Objective To investigate the effects of preconditioning with low-concentration hydrogen peroxide ( H2O2 ) on oxidative stress-induced bone marrow mesenchymal stem cells ( BMSCs ) apoptosis and its mecha-nism.Methods Mouse bone marrow mesenchymal stem cells ( BMSCs) were isolated and purified by differential centrifugation, and were treated with 0,200,250,300, 500 μmol/L H2O2 after preincubation with 50 μmol/L H2O2 or control medium.Apoptosis of these cells was measured by flow cytometry, and the expression of phos-phorylated PI3K, Akt and mTOR was analyzed by Western blot; BMSCs were also primed with PI3K inhibitor LY294002 for 30 min, then preincubated with 50 μmol/L H2O2 or control medium for 12 h before treatment with 300 μmol/L H2O2.Expression of apoptosis proteins Bcl-2, Bax, caspaase-3, cleaved-caspase-3 and the key pro-teins of the PI3K/Akt/mTOR pathway were detected by Western blot .Results H2O2 induced BMSCs apoptosis in a dose-dependent manner ,and pretreatment of BMSCs with low concentration of H2O2 significantly decreased H2O2-induced apoptosis of the BMSCs .Western blot results revealed that preconditioning with low-concentration H2O2 re-markably reversed the decrease in Bcl-2, total and phosphorylated PI3K, Akt and mTOR levels, and increased in Bax, cleaved-caspase-3 expression after high-dose H2O2 treatment.Such effects were antagonized by PI3K inhibitor LY294002 .Conclusions Preincubation with low-concentration H2O2 may indnce resistance of BMSC to oxidative stress, and such effect may be mediated by inhibition of pro-apoptotic proteins and activation of the PI 3K/Akt/mTOR pathway .
5.Combination of internal and external use of Chinese material medica with syndrome differentiation in treating 200 cases of acne vulgaris
Yuejun SHI ; Bo LI ; Yihong ZHENG ; Shunpeng SONG ; Cheng ZHAN ; Yinghua ZHU ; Qiao LI
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(06):-
Objective:To observe the effect of internal and external use of Chinese material medica with syndrome differentiation on acne vulgaris.Methods:200 patients with acne vulgaris were randomly divided into treatment group(110 cases)and control group(90 cases).Treatment group were given acne facial mask,oral medicine decoction,and the control group were treated with13-cis-Retinoic Acid Capsules,topical Clindamycin Gel,7 weeks for a course of treatment,the clinical effect were compared.Results:The effect on facial lesion in treatment group and control group had significant difference(P
6.Investigation of mental health education and service in community
Chuan-feng CHEN ; Xin-zhan HE ; Xiu-zhu SONG ; Bin TIAN
Chinese Journal of Rehabilitation Theory and Practice 2004;10(6):364-366
ObjectiveTo explore the effect of community education and service of mental health on improvement of people's diathesis.Methods483 subjects from 6 social strata were investigated with the questionnaire to find out the community resident's idea on mental health and need for psychological service.ResultsThere were significant differences in the concepts of criteria of mental health among people in different social strata and ages (P<0.001),and requirements for community education and service of mental health varied from one to another. Forms and contents of community education and service were multiplicate.ConclusionThe community education and service of mental health should be developed widely for improving people's diathesis.
7.Bone morphogenetic protein 2 expression in rabbit radial defect site with different lengths
Yulin ZHAN ; Zhiquan AN ; Luyuan SUN ; Changqing ZHANG ; Bingfang ZENG ; Feng XU ; Guozhu HOU ; Wenju LI ; Xiaomeng ZHU ; Xinghua SONG
Chinese Journal of Tissue Engineering Research 2013;(30):5511-5517
BACKGROUND:It has been studied that the distribution of bone morphogenetic protein 2 is regular under bone defect situation. OBJECTIVE:To observe the expression of bone morphogenetic protein 2 in rabbit radial defect site with different lengths. METHODS:Forty-eight New Zealand rabbits were divided into two groups randomly, 0.5 cm bone defect and 3.0 cm bone defect were made by wire saw at the middle part of radius bone after anaesthesia. RESULTS AND CONCLUSION:Western blot results showed that in the 0.5 cm bone defect group, the expression of bone morphogenetic protein 2 of the tissues in the bone defect site was increased gradual y at 1, 3, 4 weeks after operation, and the expression in each defect group was increased when compared with that immediately after injury (P<0.05). In the 3.0 cm bone defect group, the expression of bone morphogenetic protein 2 of tissues in bone defect site was increased gradual y and reached to its peak at 3 weeks after the operation (P<0.05), and the peak value in the 3.0 cm bone defect group was significantly higher than that in 0.5 cm bone defect group (P<0.05). The peak value was maintained in high level. The comparison of bone cal us formation showed that the bone cal us formation of 3.0 cm bone defect group was less than that of the 0.5 cm bone defect group at 3 and 4 weeks after operation (P<0.05). The results indicate that expression of the bone morphogenetic protein 2 in 3.0 cm bone defect site is increased significantly, but the expression level cannot make the bone defect heal itself.
8.The Relationships between cyclin D1 Expression and Prognosis of Non-small Cell Lung Cancer
ZHU JIPING ; YU LIKE ; ZHAN PING ; SONG YONG ; WANG QIN
Chinese Journal of Lung Cancer 2010;13(8):803-808
Background and objective cyclin D1 is a member of the cyclin family, and it has been proven that it plaied an important role in tumorigenesis, invasion and metastasis. We performed a retrospective study on the cyclin D1 expression in non-small cell lung cancer (NSCLC) according to the clinical characteristics. Methods One hundred fifteen postsurgical NSCLC patients were investigated. Immunohistochemistry was used to evaluate the cyclin D1 expression. Results Overall survival was significantly lower in patients with cyclin D1-high expression of tumors than those with cyclin D1 low expression of tumors (χ2=5.132, P=0.023). In early stage patients (stage I, II), the overall survival was significantly lower in patients with cyclin D1-high expression of tumors than those with cyclin D1-low expression of tumors (χ2=6.863, P=0.009). cyclin D1 status (hazard ratio=0.630; P=0.035), differentiation (hazard ratio=0.399; P<0.001), and pTNM (hazard ratio=1.576; P<0.001) to be independent prognostic factors for NSCLC patients. Specifically, the cyclin D1 status (hazard ratio=0.188; P=0.008) was a significant prognostic factor for patients with stage I NSCLCs. Conclusion cyclin D1 expression is an independent prognosis factor for postoperative patient in stage I, II NSCLCs.
9.Effect of PM2.5 exposure on in vitro fertilization (IVF) and embryo development in mice
Qiuhua LI ; Xiaoli ZHU ; Sida WEI ; Zheng WU ; Zehu ZHAN ; Ge SONG
The Journal of Practical Medicine 2017;33(20):3372-3375
Objective To investigate the effects of PM2.5 exposure on the quality of IVF eggs and the quality of embryos. Methods 40 healthy female Kunming mice and 10 male mice were used in this study. 40 female mice were randomly divided into 4 groups:low dose of PM2.5 group (1.5 mg/kg),middle dose of PM2.5 group (7.5 mg/kg),high dose of PM2.5 group (15 mg/kg) and the control group (to give the same volume of saline),with 10 mice in each group. PM2.5 was administered via tracheal instillation. The number of oocytes ,fertilization rate ,abnormal fertilization rate ,2PN cleavage rate ,2PN blastocyst rate and blastocyst formation rate were determined ,as well as levels of TNF-α and IFN-γ. Results The number of oocytes , fertilization rate and cleavage rate of 2PN were significantly higher in PM2.5 high dose of PM2.5 group than those in PM2.5 group. The rate of high quality embryos and blastocyviast formation were decreased ,but abnormal fertilization rate were decreased (P < 0.05). The expressions of TNF-α and IFN-γ in the high-dose of PM2.5 group were significantly higher than those in the middle-dose of PM2.5 group and in the low-dose of PM2.5 group(P < 0.05),while the expressions of TNF-α and IFN-γ in the middle dose group was significantly higher than those in the low dose group(P<0.05). Conclusions PM2.5 exposure can decrease the number of oocytes , fertilization rate ,2PN cleavage rate ,2PN blastocyst rate and blastocyst formation rate of in vitro fertilization in mice.
10.Analysis of Human Lung Squamous Carcinoma Cell Line NCI H520 Proteome by Two Dimensional Electrophoresis and MALDI TOF Mass Spectrometry
Quan Xian ZHAN ; Chu Zhu CHEN ; Jun Yong GUAN ; Cui LI ; Mei Chun HE ; Ping Song LIANG ; Yun Jin XIE ; Ping CHEN
Chinese Journal of Cancer 2001;20(6):575-582
Objective:This study was designed to establish and optimize the research methods for proteome,and to analyze the proteome components of human lung squamous carcinoma cell line NCI H520. Methods: A series of methods, including immobilized pH gradient two dimensional polyacrylamide gel electrophoresis(2DE), silver staining, PDQuest 2DE analysis software, peptide mass fingerprint based on matrix assisted laser desorption/ionization time of flying mass spectrometry (MALDI TOF MS) and SWISS PROT database searching, were used to separate and indentify the proteome of human lung squarmous carcinoma cell line NCI H520. Results: The good 2DE pattern including resolution and reproducibility was obtained. After silver staining, the 2DE image analysis by PDQuest 2DE software had detected average of 1146± 116 spots, and 851± 95 spots were matched. The average matching rate was 73.3% . There had a good reproducibility of spot position in 2DE map, with average deviation in IEF direction of 1.52± 0.22 mm, while in SDS PAGE direction it was 1.97± 0.13 mm. Sixty spots were incised from silver staining gel randomly and digested in gel by TPCKtrypsin. Fifty four peptide mass fingerprints (PMF) maps were obtained by MALDI TOF MS. The typic peptide masses were searched in the SWISS PROT database by PeptIdent software. Forty four proteins were preliminarily identified. Some of them were cell cycle related proteins such as Cyclin H, some were signal transduction related proteins such as mitogen activated protein kinase, protein kinase C and receptor protein tyrosine kinase ERBB 2, some were oncogene related proteins such as Ras related protein RAB 36, etc. Conclusions: The main proteome research system including IPG 2DE, image analysis, MALDI TOF MS derived PMFs and database searching has been established. The data of NCI H520 obtained by above methods will be useful for the establishment of human lung squamous cell proteome database.