1.In vitro morphological observation on the effect of exogenous collagen on the construction of tissue-engineered tendon
Yue FANG ; Zhou XIANG ; Zhiming YANG
Chinese Journal of Tissue Engineering Research 2006;10(29):169-172
BACKGROUND: Collagen type Ⅰ possesses low antigenicity and provides special functions of scaffold structure for cell migration, proliferation and secretion.OBJECTIVE: To observe the morphological change after combination of transformed human embryonic tendon cells and artificial materials with collagen type Ⅰ, so as to provide the experimental basis for constructing tissue-engineered muscle tendon in vitro.DESIGN: Randomized and controlled observation.SETTING: Department of Orthopaedics, West China Hospital, Sichuan University.MATERIALS: This experiment was carried out at the Department of Transplantation Immunology, West China University of Medical Sciences from July to September 1998. Human hair, carbon fiber and polyglycolic acid were combined with collagen and then set corresponding groups respectively. Simply hair, carbon fiber and polyglycolic acid control groups were set respectively. Artificial materials: Healthy adult hair (30 cm in length, 80 g in weight), carbon fiber knitted strip (Lanzhou carbon factory )and polyglyolic acid fasciculi (Johnson , American) were used in this experiment.METHODS: ①In the group of human hair combined with collagen,nine healthy adult hairs were chosen . Three hairs as one bundle were knitted into tendon with 0.8 cm in length and 0.4 cm in diameter and a knot was tied respectively at the two ends. ②In the carbon fiber combined with collagen group, the purchased carbon fiber knitted strip was separated and made into artificial materials with the same diameter and length as those of human hair tendon. Two ends of the artificial materials were packed with 5/0 silk thread. ③In the polyglycolic acid combined with collagen group, three polyglycolic acid fasciculi were weaved into bundle with the same diameter and length as those of human tendon,and two ends were packed with 5/0 silk thread. ④The process of knitting scaffold materials was the same in control groups as in the corresponding experimental groups. ⑤ Five pieces were prepared in each scaffold material. After sterilized by 60Co, the scaffold materials were put in collagen type Ⅰ solution for 0.5 hour, then taken out and dried at roomtemperature for use in the experimental groups. ⑥transformed human embryonic tendon cell, density for 3×106/mm3, was used for combined culture in each group, and cell number and morphology were observed at different time points after culture under an inverted microscope and scanning electron microscope.MAIN OUTCOME MEASURES: ① Observation of cell growth and adherence under an inverted microscope ② Observation of cell morphology and material adherence under scanning electron microscope RESULTS: ① Observation of cell growth and adherenceunder an inverted microscope :In each experimental group, at two hours after culture, cells gathered around the materials, and some cells adhered to materials presented round; At three days after culture, the number of cells adhered to materials began to increase, morphology changed from round to oval, and the co-existence of round and oval cells was seen;While in each control group, at two and three hours after culture, few cells adhered to the three materials. On day five after culture, in each experimental group, cell morphology turned into fusiform, the number of cells increased further and a great many fusiform cells grew among carbon fiber materials; while in the control group, most cells presented round and ellipse, and the number of cells was less as compared with the corresponding experimental group. ② Cell morphology and material adherence under scanning electron microscope: On day 5 after in vitro culture, most tendon cells on the surface of materials presented spindle-shape in each experimental group. Apophysis stretched out from the tendon cells, grew along longitudinal axon of the materials. Silk and piece-like substance appeared among the apophysis and weaved each other. While there were fewer cells adhered in control group than in corresponding experimental group, cells also presented fusiform or round, but no silk or piece-like substance was found, also no apophysis appeared.CONCLUSION: Human hair, carbon fiber and polyglycolic acid combined with collagen type Ⅰ will facilitate cell adherence. Cells increase around the materials and co-existence of round and oval cells could be seen, and cells turned into fusiform, suggesting that exogenous collagen facilitate the cells to adhere onto the materials and proliferate.
2.Histone modification and neural stem cell differentiation
Xiang ZHOU ; Yang WANG ; Zhifeng DENG
International Journal of Cerebrovascular Diseases 2012;20(9):717-720
Neural stem cell (NSC) has self-renewal and differentiation potential.It can orderly differentiate into neurons,astrocytes and oligodendrocytes in the development of central nervous system.NSC differentiation was commonly regulated by a variety of factors,including cytokines,transcription factors and epigenetic factors.Epigenetic regulation is a heritable gene expression without changing the DNA sequence.Histone modification is an important regulation mode in epigenetic modifications,which is essential in the transcription process.It also plays a critical role in the development of central nervous system.This article briefly reviews the histone methylation and demethylafion in the regulation of NSC differentiation.
3.Effect of Acupuncture and Massage on Tiptoe in Children with Spastic Cerebral Palsy
Junlu XIANG ; Wenzhi ZHOU ; Ping TIAN ; Li YANG ; Xia YANG
Chinese Journal of Rehabilitation Theory and Practice 2015;21(3):334-337
Objective To observe the effect of acupuncture and massage on tiptoe in children with spastic cerebral palsy. Methods 49 children with spastic cerebral palsy were divided into treatment group (n=24) and control group (n=25) in accordance with the order of visiting. The control group accepted routine rehabilitation, and the treatment group accepted acupuncture and massage in addition. They were assessed with Composite Spasticity Scale (CSS), the angle of ankle passive dorsiflexion before and after treatment. Results The scores of CSS and the angle of ankle passive dorsiflexion improved in both groups after treatment (P<0.001), and improved more in the treatment group than in the control group (P<0.001). The rate of improvement was 95.6% in the treatment group, and 78.26% in the control group (P<0.05). Conclusion Acupuncture and massage may further improve the correcting of the tiptoe in children with spastic cerebral palsy.
6.Rosiglitazone upregulates Foxp3 mRNA expression of CD4+T cells in adults with latent autoimmune diabetes
Zhifang YANG ; Zhiguang ZHOU ; Can HUANG ; Jian PENG ; Xiang YAN
Chinese Journal of Microbiology and Immunology 2008;28(3):264-268
Objective To investigate the effect of rosiglitazone on the CD4+regulatory T cells in the patients with latent autoimmune diabetes in adults(LADA).Methods The CIM+T cells from IADA patients were isolated with anti-CD4-dynal magnetic beads.The expression of Foxp3 mRNA,along with peroxisome proliferators activator receptors gamma(PPARγ)mRNA and TGF-131 mRNA was determined.The effect of rosiglitazone on CD4+T cells was measured,after treated with rosiglitazone for 48 h.Cell viability was assessed by Mtit assay.The proliferation was assayed with 3 H-TdR.Two-color staining(anti-CD4,anti-CD25)flow cytometric analysis was employed to measure the percentage of CD4+CD25+T cells of Deriph eral blood.Resuits PPARγmRNA was expressed in peripheral CD4+T lymphocytes.RosiglitazoBe inhibited phytohemagglutinin(PHA)-induced human CD4+T cell proliferation in dose dependence.The percentage of CD4+CD25+T cells showed no significant change after the peripheral blood culture with 1 μmol/Land 10μmot/L rosiglitazone.10 μmol/L of rosiglitazone induced Foxp3 mRNA expression in vitro (3.27fold,P<0.05),whereas TGF-β1 mRNA expression did not change.Furthermore,only 1 μmol/L of rosiglitazone could promote Foxp3 mRNA expression if adding IL-2(10 U/m1)in cultures(3.48 fold.P
7.Analysis of prophylactic chemotherapy outcome and clinical characteristics in patients of high-risk hydatidiform mole
Shuo GENG ; Fengzhi FENG ; Yang XIANG ; Xirun WAN ; Ying ZHOU
Chinese Journal of Obstetrics and Gynecology 2011;46(1):24-27
Objective To analyze prophylactic chemotherapy outcome and clinical characteristics in patients of high-risk hydatidiform mole. Methods Twenty-three patients who were diagnosed as high-risk hydatidiform mole and undergone prophylactic chemotherapy in our hospital were retrospectively analyzed.After prophylactic chemotherapy, 11 patients didn't develop to gestational trophoblastic neoplasia (GTN),while the other 12 patients developed to GTN and needed a regimen change to combination chemotherapy.The clinical characteristics of these patients and outcome of prophylactic chemotherapy were compared between two groups. Results There was no significant difference between the two groups on patients' age,weeks of delayed menses, enlarged uterine size excessive for gestational age, and incidence of theca-lutein cysts of ovaries. However,the median levels of pre-evacuation serum β-hCG in two groups were 469 144 U/L and 768 044 U/L respectively, and median days needed for β-hCG declining to normal(≤2U/L) at the first time were 71 and 120 days respectively, which were both significantly different between two groups.Analyzed with receiver operating charactristic(ROC), the level of serum β-hCG could be a predictor for prognosis. Choosing 750 000 U/L as the cut-off value, we could expect the serum β-hCG to have a specificity of 91% and a sensitivity of 58% to predict whether prophylactic chemotherapy will be successful.Conclusions For those patients who have to receive prophylactic chemotherapy because of risk factors and unavailable hCG assessments for follow-up, it's better to use double-agent or combination chemotherapy if the level of serum β-hCG reached 750 000 U/L so as to reduce therapy duration and prevent relevant chemoresistance.
8.Anti-hepatocellular carcinoma effect of low dose cyclophosphamide combined with CIK cells in mice
Haijing ZHOU ; Liping YANG ; Liming WANG ; Xiang YAN
Journal of International Oncology 2016;43(11):812-816
Objective To explore the influence of low dose cyclophosphamide (CTX)acting on regu-latory T cells (Tregs)of hepatocellular carcinoma-bearing mice,and the anti-tumor effect of low dose CTX combined with cytokine induced killer cells (CIKs).Methods Models of tumor-bearing mice were established by subcutaneous inoculation with hepatocellular carcinoma cells.The mice were randomly divided into 4 groups (n =35),there were normal control group,single tumor group,single CTX group (1 time,1 00 mg/kg,intra-peritoneal)and cyclical CTX group (once every 6 days,3 times,1 00 mg/kg,intraperitoneal).The changes of the Treg/CD4 + in spleens were detected by flow cytometry.We isolated mononulear cells from spleen of mice to culture CIKs.To study the effect of anti-tumor in vitro,tumor-bearing mice were randomly divided into 6 treat-ment groups,there were single tumor group,single CTX group,single CIK group,cyclical CTX group,single CTX +CIK group and cyclical CTX +CIK group.The growth curves of tumor were drawn.At the end of the experiment,tumor was separated and weighted.The growth inhibition ratio of tumor was calculated.Results With time prolonged after tumor inoculation,the proportion of Treg/CD4 + increased gradually in spleen of mice in single tumor group.On the 1 0th day,this proportion of single CTX group was (8.95 ±1 .90)% and the sin-gle tumor group was (9.25 ±1 .74)%,and there was no difference between two groups (t =0.374,P =0.71 4).The proportion in the cyclical CTX group (8.99 ±2.1 1 )% was still lower than that in the single tumor group (1 6.76 ±2.02)% on the 1 9th day,and the difference was significant (t =8.544,P =0.000). Treatment for 21 days,the volume and weight of the single tumor group,single CTX group,single CIK group, cyclical CTX group,single CTX +CIK group and cyclical CTX +CIK group were (3 800.4 ±607.5 ), (3 764.8 ±537.7),(3 352.2 ±485.4),(2 076.6 ±620.2),(1 867.1 ±533.6),(970.7 ±1 35.3)mm3 and (4.01 ±0.66),(3.86 ±0.74),(3.80 ±0.42),(2.08 ±0.27),(1 .83 ±0.93),(0.86 ±0.25)g. The tumor volume and weight were minimum in the cyclical CTX +CIK group.The tumor inhibition effects were weaker in the cyclical CTX and single CTX +CIK groups.But tumor in single CIK and CTX groups were unsup-pressed.The differences among the 6 groups had statistically significance (F =21 3.750,P =0.000;F =27.1 42,P =0.000).Conclusion Cyclical administration of low-dose CTX combined with CIKs has an obvi-ous therapeutic effect on hepatocellular carcinoma bearing mice and can provide a new idea for anti-tumor therapy.
9.Application of parecoxib sodium for preemptive analgesia in patients undergoing cerebral angiography
Xiang ZHOU ; Mingchun WU ; Junzhe YANG ; Xiaoyang SONG
Chinese Journal of Postgraduates of Medicine 2011;34(12):3-5
Objective To evaluate the effects of parecoxib sodium for preemptive analgesia in patients undergoing cerebral angiography. Methods Sixty patients undergoing cerebral angiography were divided into two groups by random digits table with 30 cases in each : group P and group F. Patients in group P received parecoxib sodium 40 mg 30 min before operation, while group F received fentanyl 1 μ g/kg 2 min before operation. The changes of hemodynamics were observed before operation (To), immediately right internal carotid artery angiography during operation (T1), immediately right vertebral artery angiography (T2),immediately left internal carotid artery angiography(T3), immediately left vertebral artery angiography(T4), 10min after operation (T5). The mean arterial pressure (MAP), heart rate (HR), pulse oxygen saturation (SpO2)and visual analogue score (VAS) were recorded at different times during the whole operation. Results There were no significantly differences on MAP, HR and VAS between group P and group F (P > 0.05 ). But the SpO2 at T1 ,T2,T3 in group F (0.94±0.03,0.95±0.02,0.95±0.02) were significantly lower than those in group P (0.98 ± 0.01,0.98 ± 0.02,0.98 ± 0.02 )(P<0.05 ), and 2 cases SpO2 < 0.90 at Ti. Conclusion Parecoxib sodium administered preemptively provides a nice analgesic effect in patients undergoing cerebral angiography, and is more safe than fentanyl.
10.Quality of life changes after ischemia postconditioning in patients with acute ST segment elevation myocardial infarction
Shijie ZHENG ; Jingqun ZHOU ; Changqing XIANG ; Weihua YANG ; Bo ZHANG
Chinese Journal of Postgraduates of Medicine 2013;36(25):20-23
Objective To compare the quality of life before ischemia postconditioning and 6 months after ischemia postconditioning in patients with acute ST segment elevation myocardial infarction,and investigate the related risk factors.Methods One hundred and twenty-nine patients with acute ST segment elevation myocardial infarction were given the ischemie postconditioning.The quality of life was evaluated before ischemia postconditioning and 6 months after ischemia postconditioning by 36-item short-form health survey (SF-36) and seattle angina questionnaire (SAQ).Results In acute ST segment elevation myocardial infarction patients treated with ischemia postconditioning,the scores of SF-36 and SAQ were significantly lower at 6 months after ischemia postconditioning than those before ischemia postconditioning [SF-36:total score of body health (76.4 ± 17.3) scores vs.(56.3 ± 16.8) scores,physiologic function (75.3 ± 18.1) scores vs.(52.4 ± 19.2) scores,physiologic functional authority (75.6 ± 16.5) scores vs.(48.5 ± 20.3) scores,body pain (77.2 ± 15.4) scores vs.(58.7 ± 16.6) scores,total health state (73.6 ± 17.8) scores vs.(50.6 ± 14.7)scores,total score of mental status (77.5 ± 15.3) scores vs.(55.3 ± 17.3) scores,vitality (69.3 ± 18.1)scores vs.(43.2 ± 17.4) scores,society function (70.3 ± 17.5) scores vs.(41.3 ± 14.2)scores,affection functional authority (80.2 ± 17.4)scores vs.(63.5 ± 14.3)scores,mental health (77.6 ± 18.4) scores vs.(55.6 ± 20.1) scores,health change (76.3 ± 17.4) scores vs.(53.4 ± 16.6) scores;SAQ:body limitation of activity (78.46 ± 12.21) scores vs.(35.34 ± 15.33)scores,angina stable state (74.23 ± 8.53) scores vs.(30.12 ± 5.38) scores,angina episode state (72.34 ± 10.33) scores vs.(27.33 ± 9.12) scores,satisfaction degree of treatment (76.42 ± 12.13)scores vs.(30.56 ± 15.57) scores,knowledge of diseases (74.22 ± 9.35)scores vs.(37.25 ± 20.32) scores] (P < 0.05).The results of multiple linear regression equation showed that the older,more severe vascular lesions,and lower total score of body health score before ischemia postconditioning indicated lower total score of body health,and worse quality of life after ischemia postconditioning.Conclusions The quality of life of patients with acute ST segment elevation myocardial infarction after ischemia postconditioning is significantly improved,compared with that before ischemia postconditioning.Advanced age,vascular disease and lower total scores of body health before ischemia postconditioning are independent risk factors of total scores of body health after ischemia postconditioning.