1.Compare the effect of hepatic oval cells and bone marrow mesenchymal stem cells transplanted by spleen to treat liver fibresis in mice
Bo LIU ; Nan LIN ; Kunpeng HU
Journal of Chinese Physician 2009;11(9):1191-1194
Objective To compare the effect of hepatic oval cells and bone marrow mesenchymal stem cells to treat liver fibrosis in mice. Methods The C57BL/6 mice were fed with 3,5-diethoxycarbonyl-1, 4-dihydro-collidine (DDC) in a standard food at a concentra-tion of 0.1% to activate the oval cells proliferation, then the nonparenchymal cells were separated by in situ perfusion and density gradient centrifugation, and finally Sca-1 antibody was used in conjunction with magnetic activated cell sorting (MACS) to separate the Sca-1 +cells. Other mice were injected subcutaneously with CCl4 to establish experimental liver fibrosis model. Finally, the fibrosis mice were trans-planted with HOC or MSC or saline by spleen injection according to the group they belonged to. Four weeks later, the liver function index, the hydroxyproline and the histopathological change among the groups were used to evaluate the effect of the treatment. Result The liver function index was improved after transplantation. HOC and MSE treatment could reduce both the hydroxyproline level and alleviate the fibro-sis, but HOC seems more effective in the group with continuing CCl4 injection(P < 0.05). When stopping injecting CCl4, the change of fi-brosis was not visible(P >0.05). Conclusion Both hepatic oval cells and bone marrow mesenchymal stem cells can improve the liver function, and alleviate the fibrosis level by transpleen transplantation, but hepatic oval cells were more effective.
2.Experimental study on treatment of liver fibrosis by hepatic oval cell transplantation via spleen injection
Bo LIU ; Nan LIN ; Kunpeng HU
Journal of Chinese Physician 2009;11(8):1040-1044
especially in the group of ceasing CCl4 injection. Conclusion After transplanted into the liver by spleen injection, hepatic oval cells could plant in the portal area, which improve liver function and alleviate the cirrhosis level.
3.The expression status of ZIC2 is an independent prognostic marker of hepatocellular carcinoma
Kong QINGLEI ; Li WENCHAO ; Hu PAN ; Zeng HUA ; Pan YUHANG ; Zhou TAICHENG ; Hu KUNPENG
Liver Research 2020;4(1):40-46
Background:Zinc finger protein of cerebellum 2(ZIC2)is a transcriptional activator or repressor that is important for the organogenesis of the central nervous system.Previous studies have reported that ZIC2 is widely upregulated in a variety of tumors. Methods:Oncomine database was used to evaluate the expression levels of ZIC2 in hepatocellular car-cinoma(HCC)and normal liver tissues.Quantitative real-time polymerase chain reaction and immu-nohistochemistry were conducted to validate the results from the database.Cox regression analysis and survival curves were performed to assess the survival effect of ZIC2 in HCC. Results:Increased expression of ZIC2 was detected in HCC tissues compared with normal liver tissues.In addition,patients with high ZIC2 levels had a poor prognosis.Multivariate analysis showed that clinical stage(T or M classification)and ZIC2 levels were independent prognostic factors for overall survival.Moreover,a subgroup analysis revealed that patients with moderate or poor grade tumors,T1-2 tumors,N0 tumors,early American Joint Committee on Cancer(AJCC)stage,and old age were more likely to present with overexpression of ZIC2.To conclude,ZIC2 is upregulated in HCC and associated with the histology and survival of HCC patients. Conclusions:The expression status of ZIC2 may serve as an independent prognostic marker of HCC.
4.The effect of thermal ablation combined TACE on the prognosis of small liver carcinoma
Jizong LIN ; Chenhu WANG ; Nan LIN ; Kunpeng HU ; Shuxian CHEN ; Heping FANG ; Ruiyun XU
Chinese Journal of General Surgery 2013;28(11):874-877
Objective To analyze the short-term and long-term effectiveness of radiofrequency ablation combined with transcatheter arterial chemoembolization (TACE) for the treatment of hepatocellular carcinoma (HCC).Methods The clinical data of 70 HCC patients who had received thermal ablation (group A) done or in combination with TACE (group B) were retrospectively analyzed.Results The rate of intrahepatic distant recurrence in group B (25 cases) was lower than that in group A (45 cases) (X2 =3.845,P =0.046) and the tumor-free survival rate was higher than group A (X2 =5.020,P =O.030).There were no differences in the local tumor progression rate (X2 =0.853,P =0.374) and overall survival (x2 =2.316,P =0.154) between two groups.Incidence of bone marrow suppression in group B was higher than that of group A (X2 =5.642,P =0.042).Major complications didn't occur in any group(X2 =2.016,P =0.183).The costs was higher(t =7.738,P <0.001) and the hospital stay was longer (t =5.921,P =0.003) in group B than group A.Conclusions Compared with ablation alone,combined therapy is able to reduce short-term recurrence,and improve tumor-free survival.Combine therapy is safe and effective method for small liver carcinoma.
5.The construction of bioartificial liver by BMSC and alginate scaffold
Jizong LIN ; Zhaofeng TANG ; Heping FANG ; Nan LIN ; Kunpeng HU ; Jun YANG ; Peng XIANG ; Ruiyun XU
Chinese Journal of General Surgery 2009;24(3):234-237
Objective To construct the bioartificial liver by bone mesenchymal stem cell (BMSC) and alginate scaffold. Method Alginate scaffold was used as the cell carrier for the cultivation of BMSC and the differentiation from BMSC into hepatic like cells was induced by the cell factors of HGF, EGF and FGF-4 in the scaffold in vitro. The compatibility of the cells and the scaffold was observed by microscopy and the function of the differentiatd cells was tested. The gene of AFP and ALB was detected by RT-PCR. The secretion of ALB and the urea synthesis of the cells were tested by ALB kit and urea kit respectively. The glycogen synthesis and the CK-18 was tested by the glycogen stanning method and the immunofluorescence test. Results BMSC was able to attach, grow and proliferate well in the alginate scaffold, the well compatibility was observed by microscopy. ALB and urea were detected in the cultivating medium, the gene of ALB and AFP was identified by RT-PCR. The glycogen synthesis ability and the expression of CK-18 were induced during the differentiation. Conclusion The three dimensional atginate scaffold exhibited well compatibility with BMSC, BMSC could be differentiated into the hepatic like cell in the scaffold. BMSC and the alginate scaffold could be used to construct the bioartificial liver for the hepatic tissue engineering.
6.Rat bone marrow mesenchymal stem cells induce hepatic stellate cells apoptosis in vivo
Nan LIN ; Shujie XIE ; Weidong PAN ; Kunpeng HU ; Si CHEN ; Yutian CHONG ; Peng XIANG ; Ruiyun XU
Chinese Journal of Tissue Engineering Research 2010;14(10):1769-1774
BACKGROUND:It is reported that bone marrow mesenchymal stem cell(BMSC)transplantation might be a promising treatment for liver fibrosis.But the mechanism is still unclear.OBJECTIVE:To observe the hepatic stellate cells apoptosis induced by BMSC transplantation,and to study the mechanism of BMSC in treating hepatic fibrosis in vivo.METHODS:CCl_4 subcutaneous injection was performed to induce rat liver fibrosis.After 8 weeks of CCU injection,20 rats which underwent successful model establishment were randomly divided into experimental group and control group,10 in each group.The experimental group received MSC transplantation via tail vein injection,and the control group were given DMEM instead.The rats were killed and the livers were harvested at three time point,the day of MSC transplantation,3 days after transplantation,and 7 days after transplantation.The hydroxyproline content was detected by HE and Masson staining,and the expression changes of α-smooth muscle actin(α-SMA)proteins were determined using immunohistochemistry.The apoptosis of hepatic stellate cells were determined by α-SMA and TUNEL(terminal dUTP nick-end labeling)dual-staining.RESULTS AND CONCLUSION:After 8 weeks of CCU injection,the hydroxyproline content increased and histology indicated progress of liver fibrosis.At 7 days after MSC transplantation,the hydroxyproline in the liver was decreased,and the liver fibrosis was alleviated in the experimental group but aggravated in the control group.Immunohistochemistry indicated that α-SMA positive cells were increased at 8 weeks after CCU injection.At day 7 after transplantation,α-SMA positive cells in the experimental group were significantly less than control group(P < 0.05).At 3 days after transplantation,the hepatic stellate cells apoptosis in the experimental group was significantly aggravated compared with control group(P < 0.05).This suggested that MSC transplant was an effective treatment for liver fibrosis.MSC inducing hepatic stellate cells apoptosis may be one of the mechanisms.
7.In vitro regulation effect of human bone marrow mesenchymal stem cells on hepatic stellate cells
Kunpeng HU ; Nan LIN ; Jizong LIN ; Meihai DENG ; Zhaofeng TANG ; Peng XIANG ; Ruiyun XU
Chinese Journal of Tissue Engineering Research 2009;13(27):5257-5260
BACKGROUND: There is no accepted treatment for liver fibrosis recently. Bone marrow meaenchymal stern cells (BMSCs) used in the treatment of liver fibrosis has been reported as an effectively treatment, but the mechanism is unclear.OBJECTIVE: To study the regulation of hepatic stellate cells mediated by human BMSCs in vitro.DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Center for Stem Cells and Tissue Engineering of Sun Yat-sen University and the Central Laboratory of Third Affiliated Hospital of Sun Yat-sen University from June to December 2008.MATERIALS: Human bone marrow masenchymal stem cells were collected from normal youth volunteers; Human hepatic stellate cells and normal liver call line L-O2 were supplied by the Animal Experimental Center of Sun Yat-sen University.METHODS: The purified human BMSCs and hepatic stellate calls were set up in Transwell co-culture system. The incubation density was 2×104cells/well. L-O2 was set up instead of human BMSCs as negative control. Hepatic stellate cells cultured alone served as blank control group. The culture was performed for 72 hours.MAIN OUTCOME MEASURES: Morphology of hepatic stellate cells and results of immunocytochemical staining. Apoptosis of hepatic stellte calls was determined by flow cytometry. Western blot were used to assay the expression of α-actin.RESULTS: Activated hepatic stellate cells presented fiat and thin shape under an inverted microscope. Fat drop was lack in cytoplasm, a -actin located in hepatic stellate calls, with the presence of high tension fibers. Compared with the L-O2 + hepatic stellate cell and hepatic stellate call groups, the apoptotic rate of hepatic stellate cells was significantly increased in the BMSC + hepatic stellate cell group (P < 0.05). α -actin expression was significantly down-regulated.CONCLUSION: Human BMSCs can inhibit activation of hepatic stellate ceils and promote them apoptosis, which may be the anti-hepatic fibrosis mechanism of BMSCs.
8.Clinical study on Tuina plus physical agents for lateral collateral ligament injury of ankle in gymnasts
Lijun HU ; Junliang WANG ; Gang LIU ; Kunpeng LIU ; Fei GU ; Kaiwei ZHANG
Journal of Acupuncture and Tuina Science 2022;20(4):309-315
Objective: To compare the efficacy of Tuina (Chinese therapeutic massage) plus physical agents and physical agents alone for lateral collateral ligament injury of ankle in gymnasts, and to explore the feasibility of Tuina for injury intervention in competitive athletes.Methods: A total of 64 gymnasts with types Ⅰ-Ⅱ lateral collateral ligament injury of ankle were selected and divided into a control group and an observation group according to a full analysis set based on the intention-to-treat principle, with 32 cases in each group. Patients in the control group received ultrasound and microwave treatment, while those in the observation group received additional Tuina manipulations. The efficacy was evaluated by total effective rate, visual analog scale (VAS) score, and American Orthopedic Foot and Ankle Society ankle-hindfoot scale (AOFAS-AHS) score. Results: The total effective rate was 96.9% in the observation group and 90.6% in the control group. There was no statistical difference in the total effective rate between the two groups (P>0.05). The markedly effective rate was 75.0% in the observation group and 46.9% in the control group. The markedly effective rate in the observation group was higher than that in the control group (P<0.05). After treatment, the VAS scores of both groups showed a downward trend, and there were statistical differences between different treatment time points in the same group (P<0.05). After one and three months of treatment, the VAS scores of the observation group were lower than those of the control group (P<0.05). There were statistical differences in the AOFAS-AHS score before and after treatment within the same group (P<0.05). After one month of treatment, there was no statistical difference in the AOFAS-AHS score between the two groups (P>0.05). After three months of treatment, the AOFAS-AHS score in the observation group was higher than that in the control group, indicating statistical significance (P<0.05). There was an interaction between time and group (P<0.05). Conclusion: Tuina plus physical agents can improve the symptoms of lateral collateral ligament injury of ankle in gymnasts. This combined treatment is superior to physical agents alone in relieving pain and improving joint functions. Therefore, Tuina plus physical agents can be used as a treatment for lateral collateral ligament injury of ankle in gymnasts.
9.Changes in apoptosis-related genes in bone marrow mesenchymal stem cells after cocultured with hepatic stellate cells
Kunpeng HU ; Bo LIU ; Zhicheng YAO ; Jizong LIN ; Meihai DENG ; Weidong PAN ; Nan LIN ; Cheng CHEN ; Ruiyun XU
Chinese Journal of Tissue Engineering Research 2014;(28):4444-4449
BACKGROUND:Previous studies have confirmed that bone marrow mesenchymal stem cells in vitro can promote hepatic stel ate cellapoptosis and inhibit its activity, in which the mechanism of action remains unknown. OBJECTIVE:To screen out apoptosis-related genes during hepatic stel ate cellapoptosis regulated by bone marrow mesenchymal stem cells using gene chip technology. METHODS:Purified human bone marrow mesenchymal stem cells were seeded in 6-wel Transwel plate and cocultured with hepatic stel ate cells. Cultured human bone marrow mesenchymal stem cells alone served as control group, and cultured for 72 hours. The alterations in apoptosis-related genes were analyzed between culture alone group and coculture group using gene chip technology. The genes strongly associated with regulation of hepatic stel ate cells were selected. RESULTS AND CONCLUSION:By the functional classification of second-generation SABiosciences Gene chips, apoptotic gene screening found that after coculture, significantly upregulated genes in bone marrow mesenchymal stem cells contained:AKT1, PIK3R2, DAPK1, DHCR24, NOTCH2 and BDNF. Combined with previous findings, we hypothesized that NOTCH may play a key role in the regulation of hepatic stel ate cells by bone marrow mesenchymal stem cells.
10.Association of age, gender and carotid artery atherosclerosis with carotid artery tortuosity
Kunpeng CHEN ; Junjie WANG ; Lijun WANG ; Jun LU ; Peng QI ; Shen HU ; Ximeng YANG ; Haifeng WANG ; Daming WANG
Chinese Journal of Geriatrics 2017;36(5):528-532
Objective To investigate the relationship of age,gender,carotid artery atherosclerosis with carotid artery tortuosity.Methods The data of patients with carotid computed tomography angiography (CTA)at Beijing Hospital from January 2014 to June 2016 were retrospectively reviewed.A total of 100 patients aged ≥50 years with normal carotid arteries and 100 patients with atherosclerotic carotid artery diseases were selected with age-and gender-stratified and random sampling method.Three-dimensional images of bilateral carotid arteries were reconstructed by using Mimics software.The common carotid artery(CCA)tortuosity index,extracranial internal carotid artery(EICA) tortuosity index,and the CCA-ICA bifurcation angle were measured.Association of laterality,age and gender with carotid artery tortuosity was analyzed and the differences in parameter values of carotid artery tortuosity were compared between normal carotid artery subjects and patients with atherosclerotic carotid artery.Results In normal carotid artery group,the EICA tortuosity index and ICA angle were larger in left-side than in right side(P<0.001),and CCA tortuosity index was less in left side than in right side(all P<0.001).The bilateral EICA and CCA tortuosity index were positively correlated with age(the left ICA:rs=0.284,P =0.004;the right ICA:rs =0.308,P =0.002;the left CCA..rs=0.371,P<0.001;the right CCA:rs=0.243,P=0.015).The EICA and CCA were more tortuous in women than in men[the left EICA:1.180(0.200)vs.1.105(0.140),Z=-2.996,P=0.003;the right EICA:1.150(0.128)vs.1.105(0.123),Z=-2.189,P=0.029;the left CCA:1.060(0.073)vs.1.045(0.060),Z=-2.145,P=0.032;the right CCA:1.145(0.158)vs.1.080(0.083),Z=2.324,P=0.020].The ICA angle was not related with advanced age or gender(P>0.05).Manne-Whitney tests showed no statistically significant differences in EICA tortuosity,ICA angle or CCA tortuosity between patients with versus without atherosclerotic carotid artery stenosis(P >0.05).Conclusions The tortuosity in both EICA and CCA increases with age,and the EICA and CCA are more tortuous in female.There is no significant correlation between carotid artery tortuosity and atherosclerotic carotid artery disease.