1.The changes of spinal cord pathological and hindlimbs motor function of rats after decompression on chronic spinal cord injury
Hongmei LIU ; Chunyang MENG ; Xiao LIANG
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(8):696-698
ObjectiveTo observe the spinal cord pathological change and hindlimbs motor function of rats after decompression on chronic spinal cord injury.Methods 30 Wistar rats were divided randomly into conrol group ( n =5 ),compression group ( n =5 ) and decompression group ( n =20 ),then the decompression group was subdivided into 4 parts as 1 week,2 weeks,4 weeks,6 weeks after decompression.Decompression was made after 30 days of chronic spinal cord copression injury,rats'motor function was detected by inclined plate experiment,the histopathological change,Nissl body and neural cells apoptosis in different spinal cord sections were assessed by the stainings of HE,Nissl and TUNEL.ResultsCompared with control group,the behavior testing showed all rats in compression group presented with weakness of muscle power in their hindlimbs (P < 0.01 ),but the hindlimbs motor functions recovered from the first week after decompression and the difference had statistical significance compared with the compression group(P< 0.01 ).Then the rats hindlimbs functions recovered gradually later on.The staining of HE,Nissl and TUNEL showed that the injured spinal cord section performed no improvement at the first 2 weeks after decompression,the neural apoptosis index(24.31 ± 4.73 )% decreased until the forth week after decompression(P<0.05).The spinal cord segments closed to the injured part recovered in early stage.At 1 week after decompression,lots of Nissl bodys were observed in spinal anterior horn,the neural apoptosis index in the 2 sections closed to the injured part were ( 15.21 ± 4.81 ) % and ( 16.21 ± 3.98 ) %,which declined observably compared with compression group(P < 0.05 ).ConclusionAfter decompression on chronic spinal cord injury,the recovery of rats'hindlimbs motor function in early stage is probably benefited by the functional compensation of the spinal cord segments closed to the injured section.
2.Separation, culture and identification of rabbit bone marrow mesenchymal stem cells by iliac puncture:operation details and techniques
Cong ZHANG ; Hongmei LIU ; Qingwei LI ; Guowu CHEN ; Xiao LIANG ; Chunyang MENG
Chinese Journal of Tissue Engineering Research 2014;(23):3639-3644
BACKGROUND:Bone marrow mesenchymal stem cells are considered as commonly used seed cells to construct tissue-engineered for repair of bone and cartilage defects. It is of great significance for cytology and tissue engineering experiments to study the common problems existing in the basic operation and how to avoid these problems in a timely manner. OBJECTIVE:To summarize the common problems existing in the process of operation in order to provide reliable methods about separation, culture and identification of bone marrow mesenchymal stem cells for beginners and researchers. These can reduce or avoid some errors and problems during operation. METHODS:Sixteen New Zealand white rabbits were selected as experiment objects, and bone marrow mesenchymal stem cells were separated from rabbits by iliac puncture, purified and augmented by using density gradient centrifugation combined with adherent culture method. Then cellmorphology was observed by inverted phase contrast microscope, growth curve detected by MTT method and cellphenotype identified by flow cytometry. RESULTS AND CONCLUSION:We encountered some problems in the process of separation and culture, when we operated the first five rabbits. After careful y summarizing and analysis of the reasons, the operation was successful y completed on the rest 11 rabbits. Bacteria pol ution and cellaging were not found in the process of cellculture. What is more, the cells at passage 3 appeared with high-expression of CD29, and CD44, but low expression of CD14 and CD34. The cellgrowth curve showed that the proliferation activity of cells at passages 3 and 5 was higher than that at passage 10. Although the technology of separation, culture and identification of bone marrow mesenchymal stem cells is mature, the failure wil be happen if we do not pay attention to the details of operation. By strictly carrying out normal operations, we can get high purity of bone marrow mesenchymal stem cells, which lays a good foundation for celland animal experiments in the future.
3.Recombinant bovine basic fibroblast growth factor gel prevents dry socket syndrome after toothextraction
Lingfa XUE ; Yaoxiang XU ; Jin YUE ; Shuangyi WANG ; Wenlin XIAO ; Chunyang ZHANG
Chinese Journal of Tissue Engineering Research 2013;(34):6097-6102
BACKGROUND:Recombinant bovine basic fibroblast growth factor is a manifold effect cytokine which can promote angiogenesis, wound healing, tissue repair and bone regeneration. Recombinant bovine basic fibroblast growth factor with good histocompatibility is easy to operate and has been widely used in oral and maxil ary surgery.
OBJECTIVE:To evaluate the effect of recombinant bovine basic fibroblast growth factor against dry socket syndrome after tooth extraction.
METHODS:A total of 160 patients who had been extracted mandibular third molar were selected and randomly divided into two groups. In the experimental group, recombinant bovine basic fibroblast growth factor was put into the sockets after mandibular third molars were extracted, while in the control group, we let the wounds to be healed natural y without any materials. The incidence of dry socket syndrome was observed and compared between two groups at 3 days, 5 days and 1 week after tooth extraction.
RESULTS AND CONCLUSION:One patient had dry socket after operation in the experimental group, and the incidence was 1.25%. In the control group, 10 patients suffered from dry socket, and the incidence was 12.5%. There was a significant difference in the incidence of dry socket between the two groups (P<0.01). There was visible granulation tissue within the tooth socket after tooth extraction in the experimental group, and extraction sockets narrowed and were fil ed with granulation tissues, which was 1-2 days earlier than the control group. No al ergies, tissue hyperplasia and other local and systemic reactions occurred in patients receiving implantation of recombinant bovine basic fibroblast growth factor gel. These findings indicate that local implantation of recombinant bovine basic fibroblast growth factor gel after mandibular tooth extractions can speed up the healing of dental extraction wounds.
4.Role of SphK1 in renal tubulointerstitial fibrosis and its mechanism
Chunyang DU ; Xia XIAO ; Xingui WANG ; Jiao FU ; Yiping FENG ; Fengli HU ; Enli CHEN ; Yunzhuo REN
Chinese Pharmacological Bulletin 2017;33(2):212-217,218
Aim To investigate the effect of sphingo-sine kinase 1 (SphK1 )on unilateral ureteral obstruc-tion(UUO)-induced tubulointerstitial fibrosis and ex-plore the possible mechanism.Methods The CD-1 mice were randomly divided into four groups:sham-op-eration group(Sham),PF-543 treatment control group (Sham +PF-543),model group(UUO)and PF-543 treatment group(UUO +PF-543).On 1 ,3,7 and 1 4 d after operation,eight mice were selected randomly from each group and sacrificed.The protein expressions of SphK1 ,mature TGF-β1 ,FN,ColⅠ,LC3,Beclin1 ,Atg5 and Atg1 2 were observed by Western blot.The histo-logical changes were examined by Masson′s trichrome stain.Immunhistochemistry was performed to measure the levels of expression of SphK1 ,FN and Col Ⅰ. Transmission electron microscope was used to observe the autophagic body.Results SphK1 expression and autophagy were both upregulated in a mouse model of kidney fibrosis induced by UUO. Meanwhile, in-creased mature TGF-β1 and deposition of extracellular matrix(ECM)were observed in tubulointerstitial areas compared with sham-operated mice.After intraperito-neal injection with the SphK1 specific inhibitor PF-543 in UUO mice,enhanced expression of SphK1 and acti-vated autophagy were significantly abrogated.Howev-er,aggravation of renal fibrosis was detected when SphK1 inhibitor PF-543 was applied to suppress SphK1 expression in UUO mice.Conclusion SphK1 activa-tion is renoprotective through the induction of autoph-agy in the pathogenesis of kidney fibrosis.
5.Comparative study of curative effect and blood glucose and lipid in schizophrenic patients who treated with ziprasidone and risperidone
Qiping LI ; Ao ZHAO ; Huiteng LI ; Chunyang LI ; Zaiping HUANG ; Huanyu XU ; Jieping GUAN ; Tanling XIAO ; Jinhua ZHANG ; Zhanhong FU
International Journal of Laboratory Medicine 2015;(17):2471-2472,2475
Objective To observe the effects of ziprasidone and risperidone on patients with schizophrenia and their influence on bloodglucoseandlipids.Methods 96patientswithschizophrenicenrolledinthestudywererandomlydividedintotwogroups,zi‐prasidone and risperidone group ,and both were treated for 8 weeks .Their blood glucose ,blood lipid of base line and at the end of the 4th ,8th week were determined respectively .Results The positive and negative symptoms scores of the two groups by using Positive and Negative Syndrome Scale(PANSS) before and after treatment were not statistically different(P> 0 .05) .Compared with the baseline scores ,scores at the end of 4th and 8th week in both ziprasidone and risperidone groups significantly decreased(P<0 .05) ,but there was no statistically significant difference between the two groups(P>0 .05) .After 8 weeks′ treatment ,the ef‐fective rate was 91 .7% in ziprasidone groups and 89 .6% in risperidone group .There were no significant differences between the two groups(P>0 .05) .The blood lipids and glucose levels were less increased after ziprasidone treatment ,but was not statistically significant(P>0 .05) .The blood lipids and glucose levels significantly increased after risperidone treatment(P<0 .05) .Conclusion Ziprasidone and risperidone had the same effect on schizophrenia .Ziprasidone had no effect on blood glucose and lipids in schizo‐phrenic patients ,while risperidone could increase blood glucose and lipids level ,we should pay attention to the side effects of long‐term use .
6.Effect of high glucose on cholesterol efflux in renal tubular cell and intervention of anthocyanins
Chunyang DU ; Yonghong SHI ; Yan ZHU ; Yunzhuo REN ; Haijiang WU ; Jinying WEI ; Ming WU ; Xia XIAO ; Huijun DUAN
Chinese Pharmacological Bulletin 2016;(1):114-118
Aim To investigate the effects of high glu-cose on cholesterol metabolism in renal tubular cells and the intervention of the anthocyanins. Methods HK-2 cells were grown in the DMEM medium supple-mented with 10% FBS and were divided into 5 groups:normal glucose group, high glucose group, mannitol group, C3G group and Cy group. Effect of anthocya-nins on cell viability was detected with MTT, and cho-lesterol accumulation was detected with Amplex Red Cholesterol Assay kit and Filipin staining. Expression of ABCA1 was detected with RT-qPCR and Western Blot. Results In compared with control groups, HG significantly promoted cholesterol mass inside the cell and decreased the cholesterol concentration in the me-dium after treatment for 24 h or 48 h. The levels of mRNA and protein of ABCA1 were detected with RT-qPCR and Western blot, and both were decreased in the presence of HG. Whereas treatment with C3G and Cy markedly attenuated HG-induced cholesterol mass inside the cell by up-regulating the expression of AB-CA1. Conclusions High glucose can reduce the ex-pressions of the ABCA1, and then decrease cholesterol efflux and increase the cholesterol accumulation in HK-2 cells. Anthocyanins can decrease cholesterol accu-mulation by up-regulating the expression of ABCA1.
7.Research advances in immune cellular pathogenesis in liver fibrosis
Journal of Clinical Hepatology 2015;31(9):1532-1536
Liver fibrosis is the common pathological consequence of all chronic liver diseases with various etiologies. The mechanism of liver fibrosis is associated with the activation and proliferation of hepatic stellate cells (HSCs). The interaction between immune cells and HSCs can regulate the production of extracellular matrix (ECM) and lead to the excessive deposition of ECM and subsequent liver fibrosis and cirrhosis. This article reviews the current understanding of the effects and action mechanisms of immune cells in the development of liver fibrosis and summarizes the regulatory functions of the innate and adaptive immune systems in liver fibrosis. Further study of the interactions between immune cells, cytokines, and HSCs and the regulatory mechanisms of the immune system will provide novel opportunity for the treatment of liver fibrosis.
8.Low-temperature condensation deposition method for 3D printing of bone tissue engineering poly-L-lactic acid/pearl powder composite scaffold
Fangli GANG ; Rui SHI ; Chunyang MA ; Yi XIAO
Chinese Journal of Tissue Engineering Research 2024;28(17):2702-2707
BACKGROUND:The repair of large-scale bone defects is still facing serious challenges.It is of great significance to develop personalized,low-cost,and osteogenic-inducing tissue engineering scaffolds for bone repair. OBJECTIVE:To explore the process of 3D printing bone tissue engineering scaffold containing pearl composite material by low-temperature condensation deposition method,and further test the physicochemical properties and in vitro biological functions of the composite scaffold. METHODS:Pearl powder was prepared by grinding and sieving.The pearl powder of different qualities was added into the poly-L-lactic acid ink,so that the mass ratio of pearl powder to poly-L-lactic acid was 0,0.1,0.2,0.3,and 0.5,respectively.The 3D-printed poly-L-lactic acid/pearl powder scaffolds were prepared using the low-temperature condensation deposition method.The microstructure,compressive properties,water contact angle,cytocompatibility,and in vitro bone differentiation ability of the printed poly-L-lactic acid/pearl powder composite scaffolds were detected. RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that the five groups of scaffolds all had micropores with a diameter of 2 μm or even smaller,irregular shapes and interconnectivity.(2)All the five groups had good compressive properties.The compressive strength of the pearl powder 0.5 group was higher than that of the other four groups(P<0.05).The water contact angle of the pearl powder 0.2 group and the pearl powder 0.5 group was smaller than that of the pearl powder 0 group(P<0.01,P<0.001).(3)Bone marrow mesenchymal stem cells were co-cultured with five groups of scaffolds for 1,3,and 5 days,respectively.The cell proliferation in pearl powder 0.1,0.2,0.3,and 0.5 groups cultured for 3 and 5 days was faster than that in pearl powder 0 group(P<0.05).After 1 day of culture,live-dead staining exhibited that the number of cells on the scaffold was small,but all of them were living cells.(4)Bone marrow mesenchymal stem cells were inoculated on the scaffold surface of the pearl powder 0 group and pearl powder 0.1 group respectively for osteogenic differentiation.The alkaline phosphatase activity induced for 4 and 6 days in the pearl powder 0.1 group was higher than that in the pearl powder 0 group(P<0.05).(5)The results showed that the poly-L-lactic acid/pearl powder composite scaffold had good compressive strength,hydrophilicity,cytocompatibility,and osteogenic properties.
9.Risk genes for the development of chronic hepatitis B cirrhosis assessed by prediction analysis of microarrays
Chunyang XIAO ; Qinkai LI ; Jingxi YU
Journal of Clinical Hepatology 2016;32(7):1323-1329
ObjectiveTo investigate the risk genes for predicting the development of chronic hepatitis B (CHB) cirrhosis using gene chip technology. MethodsA total of 40 CHB patients who visited Shanghai First People′s Hospital from April 2008 to December 2010 were enrolled as a clinical cohort and were divided into S0, S1, S2, S3, and S4 groups, with 8 patients in each group. Liver biopsy was performed to determine fibrosis stage with the Scheuer pathological score as the criteria, and clinical data and liver tissue samples were reserved. The Human Affymetrix GeneChip was used to establish the gene expression profiles of liver tissues in CHB patients, and the significance analysis of microarrays (SAM) and prediction analysis of microarrays (PAM) were used to screen out the risk genomes for predicting the development of CHB cirrhosis. Quantitative real-time PCR was used to measure the mRNA expression of risk genes in liver tissue. The chi-square test was used for comparison of categorical data. The t-test and a one-way analysis of variance were used for comparison of normally distributed continuous data, and SNK-q test was used for further comparison between any two groups; the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data. ResultsA total of 1674 differentially expressed genes were screened out by Affymetrix GeneChip. A cluster analysis of these genes showed that gene expression showed differences between groups with different fibrosis stages, which suggested that the gene expression profile was well consistent with fibrosis stage. Four different classification methods were used for analysis, and 87 significant genes were screened out by SAM and 14 “high-risk” genes were screened out by PAM. The quantitative real-time PCR showed the expression of 6 risk genes (CD24, CXCL6, EHF, ITGBL1, LUM, and SOX9) differed significantly between groups S0, S1-3, and S4 (P<0.05), and the S1-3 and S4 groups showed significantly upregulated expression of these genes compared with the S0 group (all P<0.05). ConclusionThe 6 high-risk factors screened out and verified by gene chip technology help to predict the probability of developing liver cirrhosis in CHB patients and can be used as the diagnostic genes for predicting hepatitis B cirrhosis.
10.Three-dimensional bioprinting of osteochondral composite tissue with innovative bio-ink and PCL to repair articular cartilage defects.
Qiang ZUO ; Xiao ZHANG ; Yang LIU ; Chunyang LUO ; Weimin FAN ; Feng LIU
Chinese Journal of Orthopaedics 2021;41(16):1073-1080
Objective:A new type of bio-ink and polycaprolactone (PCL) were used to construct an integrated osteochon-dral composite tissue block by multi-nozzle 3D bioprinter. And the repair results to osteochondral defects were evaluated.Methods:In freeze-drying group: Freeze-dried composite scaffold made by silk fibroin (SF) and β-tricalcium phosphate was used to repair osteochondral defects, as control. In the 3D printing group: PCL was used to printed a hollow multi-layer cylinder frame by 3D biological printer. Extracellular matrix, SF and bone marrow mesenchymal stem cells were used as chondral bio-ink. Then, chon-dral bio-ink was used to print tissue-engineered cartilage on top of PCL frame. Before implantation of cartilage defect, autogenous cancellous bone was filled in PCL frame, then the tissue-engineered osteochondral composite was used to repair osteochondral defects. In mosaic group: Autologous osteochondral transplantation was performed. The repair results of the above three groups were compared by histological score, biochemical analysis and biomechanical test to evaluate the effect of repairing rabbit cartilage defects.Results:The compression modulus of neo-cartilage in the 3D print group 2.56±0.30 MPa was close to that of the mosaic group 2.51±0.13 MPa ( P>0.05), and significantly higher than that of freeze-dried group 1.37±0.14 MPa ( F=11.058, P<0.05). The sGAG contents in the 3D print group 14.49±0.7 μg/mg was close to that of the mosaic group 14.98±0.81 μg/mg ( P>0.05), and significantly higher than that of freeze-dried group 8.72±0.73 μg/mg ( F=20.973, P<0.05). However, there was no significant difference in collagen content between the three groups ( P>0.05). The results of ICRS cartilage repair histology score showed that the scores of the 3D print group were close to those of the mosaic group in the matrix, cell distribution, cell viability and subchondral bone ( P>0.05), and were significantly higher than those of freeze-dried group in the surface and cartilage mineralization scores ( F=19.544, P<0.05). Conclusion:Using the new bio-ink to make bone cartilage composite scaffold by 3D bio printing can simplify the construction of tissue-engineered bone cartilage composite tissue in vitro, and can repair cartilage defects in vivo.