1.Research progress on the anti-tumor immunity of Th9 cells and its mechanism
Chinese Journal of Clinical Oncology 2017;44(6):294-298
As a newly identified T helper cell subset, Th9 plays an important role in anti-tumor immunity. Th9 can be differentiated from CD4+T cells that have been induced by TGF-beta and IL-4. In addition, other CD4+T helper cell subsets can be developed to Th9 cell in particular situations, thereby showing its plasticity. Results of animal experiments have indicated that Th9 inhibits tumor growth and plays a significant role in anti-tumor immunity by secreting related cytokines such as IL-9. A few cytokines and molecules can regu-late the differentiation and development of Th9 cells in different signaling pathways. This review will focus on the production, anti-tu-mor immunity, related mechanism, and signaling pathways of Th9 cells, thereby providing a new field of vision and idea for anti-tumor therapy in the future.
2.Removal of protein extract from moringa oleifera seed presscake on turbidity water
Jialu XU ; Long WANG ; Kunpeng XIE ; Mingjie XIE
Chinese Journal of Biochemical Pharmaceutics 2017;37(9):7-9
Objective To study the removal effect of Moringa seedprotein on turbidity water. Methods The protein of Moringaoleifera seed was extracted by salting out and salting out methodand the protein concentration of Moringa oleifera seed wasdetermined by Coomassie blue staining;The removal effect of Moringa seed protein on turbidity water was determined by coagulation test.The contents of COD, ammonia nitrogen and nitrate nitrogen in the water were determined to determine the effect of Moringa seed protein on water quality. Results The experimental results showed that Moringa oleifera seed protein has good removal effect on high and medium turbidity water, and its removal effect is in a dose - dependent manner.The removal rate of 7 mg/L of Moringa seed protein to high and medium turbidity water reached 92.25 % and 64.71 % respectively. But the removal efficiency of low turbidity water was less than 7 mg/L, the removal rate of low turbidity water was only 31.91%.The results of determination of COD, ammonia nitrogen and nitrate nitrogen showed that the Moringa seed protein did not increase the content of organic matter in the water while removing turbidity effectively. Conclusion Moringa oleifera seed protein has a certain removal effect on turbidity water, among which the removal effect of high and medium turbidity water is strong, and the removal effect of low turbidity water is poor.Moringa oleifera seed protein had little effect on water quality.
3.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.
4.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.
5.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.
6.Enhancing chondrogenic differentiation in precartilaginous stem cells with 620 nm red light
Kunpeng LI ; Tao XU ; Yu DU ; Chen GONG ; Fei PENG ; Anmin CHEN ; Fengjin GUO
Chinese Journal of Physical Medicine and Rehabilitation 2012;34(3):172-176
Objective To investigate the effect of 620 nm red light on chondrogenic differentiation in rat precartilaginous stem cells (PSCs). Methods Rats' PSCs were isolated and purified using magnetically activated cell sorting and cultured in vitro.The PSCs were exposed once to 620 nm wavelength red light from a light-emitting diode (LED) with an irradiation energy of 0.5 J/cm2,1 J/cm2,2 J/cm2 or 4 J/cm2.Any effect was confirmed by Alcian blue staining,immunohistochemistry and observing histomorphological changes under a light microscope,as well as detection using a reverse transcription polymerase chain reaction (RT-PCR). Results After being induced for 14 d,the PSCs exhibited polygonal and round shapes. Alcian blue and type Ⅱ collagen immunohistoehemistry staining showed positive results,but the control group had no significant change.RT-PCR showed that the mRNA expression of Sox9 and type Ⅱ collagen increased significantly compared with the control group. Conclusion Low energy 620 nm red light can enhance chondrogenic differentiation in PSCs significantly.
7.Influence of head anteflexion on airway sealing pressure during intermittent positive pressure ventilation with ProSeal laryngeal mask airway with an esophageal vent
Chengwen LI ; Fushan XUE ; Kunpeng LIU ; Peng MAO ; Haitao SUN ; Guohua ZHANG ; Yachao XU ; Yi LIU
Chinese Journal of Anesthesiology 2010;30(z1):61-64
Objective To evaluate the influence of head anteflexion on airway sealing pressure during intermittent positive pressure ventilation(IPPV) with ProSeal laryngeal mask airway (PLMA) with an esophageal vent.Methods Fifty ASA Ⅰ or Ⅱ patients (20 males and 30 females), aged 18-51 ye are, weighing 50-70 kg and scheduled for elective plastic surgery under general anesthesia, were enrolled in this study. Anesthesia was induced with fentanyl 2 μg/kg, propofol 2 μg/kg and vecuromium 0.1 mg/kg. PLMA with an esophageal vent was inserted at 2 min after intravenous vecuronium injection.The airway sealing pressure, the anatomic position of the cuff and the efficacy of positive pressure ventilation were checked in the neutral and anteflexed head positions with the cuff deflated and inflated to an intracuff pressure of 60 cm H2 O, respectively.Results The lungs were better ventilated in the head anteflexion position than in the head neutral position whether the cuff was deflated or inflated. There was no significant difference in the volume of air required to achieve an intracuff pressure of 60 cm H2O between the two head positions ( P> 0.05). The airway seating pressure increased from (27 ± 6) cm H2O in the head neutral position to (33 ± 6) cm H2O in the head anteflexion position, with no significant difference between them ( P> 0.05). The expired tidal volume and the peak inspiratory pressure during IPPV were (496 ± 81 ) ml and (14.3 ± 1.9) cm H2O respectively in the head neutral position and (496 ± 81 ) ml and ( 14.5 ± 2.1 )cm H2O respectively in the head anteflexion position.Conclusion Head anteflexion can significantly improve airway sealing but does not affect the anatomic position of the cuff.Appropriate head anteflexion is a simple and effective way to improve IPPV when the airway sealing pressure is inadequate in the head neutral position.
8.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.
9.Role of cyclooxygenase-2 and mitochondrial KATP channels in sufentanil preconditioning-induced delayed cardioprotection against myocardial ischemia-reperfusion injury in rats
Haitao SUN ; Fushan XUE ; Kunpeng LIU ; Li SUN ; Xu LIAO ; Jun XIONG ; Yachao XU ; Yujing YUAN ; Qiang WANG
Chinese Journal of Anesthesiology 2010;30(4):456-460
Objective To investigate the role of cyclooxygenase-2(COX-2)and mitochondrial adenosine tuiphosphate sensitive potassium channels (mito-KATP channels) in sufentanil preconditioning-induced delayed cardiopreteetion against myocardial ischemia-reperfnsion (I/R) injury in rats. Methods Seventy-two adult male Wistsr rats weighing 250-300 g were randomly divided into 6 groups ( n =12 each). Group Ⅰ,Ⅱ,Ⅲ were preconditioned with intraperitoneal (IP) normal saline (NS) 1 ml/kg while group Ⅳ,Ⅴ,Ⅵ with IP sufentanil 20 μg/kg at 24 h before myocardial ischemia. Group Ⅱ and Ⅴ were given IP NS-398 ( COX-2 inhibitor) 5 mg/kg at 30 rain before myocardial ischemla while group Ⅲ and Ⅵ were given intravenous 5-HD (mito-KATP channelblocker) 10 mg/kg at 10 min before ischemia or before being killed. Six animals in each group underwent 45 min myocardial ischemia followed by 120 min reperfusion, while the other six animals in each group were killed immediately before ischemia for determination of myocardial COX-2 expression and myocardial PGF2 and PGF1α content. Myocardial ischemia was induced by occlusion of left anterior descending branch (LAD) of coronary artety for 45 rain followed by 120 min reperfusion. MAP and HR were recorded immediately before ischemia (T0), at 15, 30, 45 rain of ischemia (T1-3) and at 30, 60, 90, 120 vain of reperfusion (T4-7). Heart rate-blood pressure product (RPP) was calculated. Arterial blood samples were obtained at T0.3 and T7 for measurement of plasma CK-MB activity. The animals were killed at the end of 120 nan reperfusion. The hearts were removed for determination of myocardial infarct area (IA) and area at risk (AAR). LA/AAR was calculated. Results There was no significant difference in HR, MAP and RPP at all time points among the 6 groups. Preconditioning with sufentanil significantly decreased plasma CK-MB activity at T3 and T7 and IA/AAR in group Ⅳ as compared with group Ⅰ.Myocardial COX-2 expression was up-regulated and PGE2 and PGF1α, contents were elevated by sufentanil preconditioning in group Ⅳ as eomared with control group (Ⅰ). In group Ⅴ and Ⅳ preconditioning with NS-398/5-HD significantly increased plasma CK-MB concentration and IS/AAB as compared with group Ⅳ, indicating involvement of COX-2 and mito-KATP channels in the sufentanil-induced delayed cardioprotection.The myocardial PGE2 and PGF1α contents were significantly reduced in group Ⅴ as compared with group Ⅳ. There was no significant difference in the myocardial COX-2 expression among group Ⅳ, Ⅴ and Ⅵ. Conclusion Both COX-2 and mito-KATP channels are involved in sufentanil preconditioning-induced delayed cardiopmtection.
10.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.