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02 December 2016, Volume 20 Issue 50 Previous Issue    Next Issue

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Effects of FAT10 gene silencing on apoptosis of human esophageal cancer cells and the characteristics of cancer stem cells Wang Zhi-qiang, Zhang Jian-jun 2016, 20 (50):  7453-7459.  doi: 10.3969/j.issn.2095-4344.2016.50.001 Abstract ( 285 )   PDF (1152KB) ( 304 )   Save BACKGROUND: Human leukocyte antigen F-associated transcription factor 10 (FAT10) is highly expressed in many tumor cells like colon cancer cells, but its relationship with esophageal cancer is less reported. OBJECTIVE: To investigate the effects of siRNA interference technique on the invasion, apoptosis and the characteristics of EC9706 cells, a human esophageal cancer cell line. METHODS: siRNA sequence was designed and synthesized according to the FAT10 mRNA encoding sequence, and the EC9706 cells were transiently transfected. EC9706 cells were divided into three groups: siRNA FAT10 group, negative control group, and blank control group. The expression levels of bcl-2 and FAT10 were detected by RT-PCR and western blot assay, respectively. Cell counting kit-8 assay was used to measure the proliferation of cells in vitro. Flow cytometry was used to observe the changes of cell cycle, cell apoptosis and the expression of CD44+CD133+. TUNEL staining was used to detect the apoptosis of the cells. Cell invasion in vitro was detected by Transwell invasion assay. RESULTS AND CONCLUSION: RT-PCR and western blot findings showed that compared with the negative control group and blank control group, the expression levels of bcl-2 and FAT10 mRNA and protein were significantly decreased in the siRNA FAT10 group (P < 0.05); the percentage of CD44+CD133+ cells was decreased significantly (P < 0.05); and significantly increased apoptosis rate, and decreased cell proliferation and invasion were also found in the siRNA FAT10 group (P < 0.05). In conclusion, the specific silencing of FAT10 gene can reduce the invasion of esophageal cancer cells, inhibit cell proliferation, reduce bcl-2 expression, and increase the apoptosis rate. Meanwhile, the proportion of CD44+CD133+ cells is decreased.    中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Human umbilical cord mesenchymal stem cells promote the proliferation of HepG-2 cells through interleukin-6/STAT3 signaling pathway Zheng Sheng, Yang Juan, Chen Wen-qin, Liu Jing, Zhang Fan, Wang Yu-bo 2016, 20 (50):  7460-7468.  doi: 10.3969/j.issn.2095-4344.2016.50.002 Abstract ( 323 )   PDF (7747KB) ( 297 )   Save BACKGROUND: Human umbilical cord mesenchymal stem cells (hUC-MSCs) can secrete a variety of factors involved in the regulation of tumor proliferation, metastasis and angiogenesis. Probably,  interleukin-6 (IL-6) is one of the most important inflammatory factors. OBJECTIVE: To explore the effect of hUC-MSCs on the proliferation and migration of HepG-2 hepatocyte carcinoma cells via the IL-6/STAT3 signaling pathway. METHODS: IL-6 expression levels in hUC-MSCs and HepG-2 cells were determined by ELISA. STAT3 and p-STAT3 expression levels were determined by western blot assay. Transcription levels of PCNA, CyclinD1 and STAT3 genes were measured by RT-PCR. HepG-2 cell proliferation was analyzed by flow cytometry and cell counting kit-8 assays. The migration capacity of HepG-2 cells was evaluated through a scratch test and Transwell assays. RESULTS AND CONCLUSION: The IL-6 level in the hUC-MSCs was significantly higher than that in the HepG-2 cells (P < 0.05). Both the hUC-MSC conditioned culture medium and IL-6 could be used for STAT3 activation. The addition of an IL-6 neutralizing antibody significantly weakened the activation of STAT3 in HepG-2 cells by the hUC-MSCs-conditioned culture medium. In the presence of the IL-6 neutralizing antibody or the STAT3 inhibitor, AG490, the mRNA expression levels of HepG-2 proliferation-related genes (PCNA, CyclinD1 and Survivin) were significantly reduced. The proliferation and migration capacity of HepG-2 cells were also significantly decreased by this treatment. Taken together, hUC-MSCs can secrete IL-6 to activate the STAT3 signaling pathway, thereby promoting the proliferation and migration of HepG-2 cells.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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5-Azacytidine induces differentiation of bone marrow mesenchymal stem cells into myocardial cells Jia Xiu-li, Li Chang-ping 2016, 20 (50):  7469-7474.  doi: 10.3969/j.issn.2095-4344.2016.50.003 Abstract ( 244 )   PDF (3956KB) ( 229 )   Save BACKGROUND: Bone marrow mesenchymal stem cells can be differentiated into myocardial cells induced by 5-azacytidine in vitro, which provides an opportunity for cell transplantation in the treatment of heart disease. OBJECTIVE: To study the induction effects of 5-azacytidine at different concentrations on the myocardial differentiation of bone marrow mesenchymal stem cells in rats. METHODS: Passage 3 bone marrow mesechymal stem cells were incubated with DMEM containing 0, 5, 10, 20 μmol/L 5-azacytidine for 24 hours, and then the induced medium was replaced by DMEM containing 5% fetal bovine serum for subsequent 28-day culture. Afterwards, expression of cardiac troponin I was detected by immunocytochemical method. RESULTS AND CONCLUSION: Cell death occurred at 24 hours after 5-azacytidine induction, which was more obvious in the 20 μmol/L group than the 5 and 10 μmol/L groups. The positive expression of cardiac troponin I was significantly lower in the 5 μmol/L group than the 10 and 20 μmol/L groups (P < 0.05), but there was no significant difference between the 10 and 20 μmol/L groups (P > 0.05). These experimental findings indicate that 5-azacytidine can induce the differentiation of rat bone marrow mesenchymal stem cells into myocardial cells in vitro, and its optimal concentration is 10 μmol/L.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Variation of matrix metalloproteinase 2 levels during Kartogenin-induced directional differentiation of human bone marrow mesenchymal stem cells Wang Cheng, Bo Qi-yu, Dai Guo-feng, Yang Wei-wei 2016, 20 (50):  7475-7480.  doi: 10.3969/j.issn.2095-4344.2016.50.004 Abstract ( 343 )   PDF (3788KB) ( 242 )   Save BACKGROUND: Kartogenin can induce chondrogenic differentiation of mesenchymal stem cells as reported in in vitro experiments. The discovery of Kartogenin finds a novel path to cartilage repair, and it is expected to develop into a new drug to treat osteoarthritis. OBJECTIVE: To observe the inductive role of Kartogenin in the process of human bone marrow mesenchymal stem cells differentiating into chondrocytes in vitro. METHODS: In vitro cultured human bone marrow mesenchymal stem cells were grown to the logarithmic phase, and then divided into control group (0 μmol/L Kartogenin), 1 μmol/L Kartogenin group, and 10 μmol/L Kartogenin group. After 72 hours of culture, cell proliferation and differentiation were observed microscopically. Matrix metalloproteinase 2 and type II collagen levels in the cell supernatant were detected by enzyme linked immunosorbent assay and immunofluorescence staining, respectively. RESULTS AND CONCLUSION: Under the microscope, Kartogenin was shown to significantly promote the proliferation and differentiation of human bone marrow mesenchymal stem cells. With the increase of Kartogenin concentrations, the level of type II collagen was increased, while the level of matrix metalloproteinase 2 was decreased. These findings indicate that Kartogenin can induce human bone marrow mesenchymal stem cells to differentiate into chondrocytes, and with the increase of Kartogenin concentration, destruction of the cartilage extracellular matrix may be inhibited.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Biological characteristics of bone marrow mesenchymal stem cells in patients with or whithout coronary heart disease: a comparative study Hou Yong-lan 2016, 20 (50):  7481-7486.  doi: 10.3969/j.issn.2095-4344.2016.50.005 Abstract ( 292 )   PDF (1034KB) ( 193 )   Save BACKGROUND: It is unclear whether the body differences in patients with different types of heart diseases affect the characteristics and performance of stem cells. OBJECTIVE: To explore the biological characteristics of bone marrow mesenchymal stem cells from patients with different types of heart diseases. METHODS: Bone marrow mesenchymal stem cells were extracted using density gradient centrifugation from the bone marrow of 27 patients with coronary heart diseases and 20 patients with other heart diseases. Cell morphology was observed in the two groups. CD13, CD34, CD45, CD54, CD106 and CD44 positive expression was detected by flow cytometry. Cell proliferation was detected by MTT method, and the in vitro cell growth curves of the two groups were described. RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cells of the two groups showed a long spindle shape, and there was no significant difference in the cell morphology between the two groups. In all the patients, the positive rates of CD34 and CD455 were less than 3.0%, while the positive rates of CD13 and CD44 were higher than 95.0%. However, the positive rates of CD54 and CD106 were higher in patients with coronary heart disease as compared with those with other heart diseases (P < 0.05). The in vitro growth curves of cells in the two groups were basically consistent, and the cell proliferation was only a little higher in the patients with other heart diseases compared with those with coronary heart disease. Experimental results show that different types of heart diseases in patients have no influence on morphology and proliferation of bone marrow mesenchymal stem cells, but some function-related proteins may exhibit certain difference in their expressions.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Allogeneic bone marrow mesenchymal stem cell transplantation for ventricular remodeling after myocardial infarction Wang Juan, Jia He-lei, Ji Hong-liang, Ren Dong-dong, Lu Chang-qing 2016, 20 (50):  7487-7493.  doi: 10.3969/j.issn.2095-4344.2016.50.006 Abstract ( 301 )   PDF (1236KB) ( 233 )   Save BACKGROUND: Myocardial infarction leads to ischemic changes in the myocardium, triggering the emergence of ventricular remodeling, which is an important cause of death. Myocardial infarction is a common disease in the middle-aged and elderly population, but autologous bone marrow mesenchymal stem cells from these patients exhibit a weak ability of proliferation and differentiation. Therefore, a positive attempt of allogeneic stem cell transplantation is required in order to obtain better therapeutic outcomes. OBJECTIVE: To explore the effect of allogeneic bone marrow mesenchymal stem cells on ventricular remodeling after myocardial infarction.   METHODS: Bone marrow mesenchymal stem cells from 10 neonatal rats and 10 adult rats were isolated, cultured and identified. Another 40 rats were randomly assigned into four groups (n=10/group): model group, neonatal rat cell transplantation group, adult rat cell transplantation group, or sham group. Animal models of myocardial infarction were made in rats in the all groups except for the sham group in which the rats were given sham operation. Rats in the two cell transplantation groups were given the corresponding cell transplantation. Four weeks postoperatively, heart function of rats was detected in each group, and cardiac tissues were taken to detect changes in collagen formation and blood vessel density in the infarct area. RESULTS AND CONCLUSION: Four weeks after surgery, rats in the model group showed significant changes in cardiac function indexes as compared with the other groups (P < 0.05), while compared with the model group, these cardiac function indexes improved in both two cell transplantation groups, but there was no significant difference between the two cell transplantation groups (P > 0.05). Meanwhile, compared with the model group, significantly decreased collagen formation and increased blood vessel density were found in both two cell transplantation groups (P < 0.05). Additionally, the vascular density of the infarct area was highest in the sham group (P < 0.05). Experimental results show that both neonatal and adult rat bone marrow mesenchymal stem cell transplantation can improve cardiac function of rats, reduce the formation of collagen in the infarct area and delay ventricular remodeling after myocardial infarction.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Immune function regulation by bone marrow mesenchymal stem cells in rats undergoing heart transplantation Zhao Min, Li Xiu-fen 2016, 20 (50):  7494-7499.  doi: 10.3969/j.issn.2095-4344.2016.50.007 Abstract ( 494 )   PDF (3915KB) ( 227 )   Save BACKGROUND: Heart transplantation is an effective method for treatment of end-stage heart failure, but immune rejection that seriously impact therapeutic effacicy is easy to occur after transplantation. OBJECTIVE: To investigate the regulatory effect of bone marrow mesenchymal stem cells on the immune function of rats undergoiong heart transplantation. METHODS: Twenty Lewis rats were enrolled as donors, and 20 Wistar rats as recipients. Heart transplantation models were established in the Wistar rats. These 20 model rats were randomized into cell transplantation and control group with 10 rats in each group. Forty-eight hours after heart transplantation, rats in the cell transplantation group were given bone marrow mesenchymal stem cell suspension (1 mL, 2×108 cells/L) via the tail vein, while rats in the control group were given normal saline in the same dose. Then, the expression levels of serum interleukin-2, interleukin-10 and percentage of CD4+, CD8+, CD4+/CD8+, CD4+CD25high, CD4+CD25high Foxp3+ T cells in the venous blood were detected in the two groups at 7 days after cell transplantation. Additionally, rat myocardial tissues were taken and observed pathologically. RESULTS AND CONCLUSION: The survival time of the cell transplantation group was significantly longer than that of the control group (P < 0.05). The expression level of interleukin-2 showed no significant difference between the two groups (P > 0.05), but the level of interleukin-10 in the cell transplantation group was significantly higher than that in the control group (P < 0.05). Compared with the control group, the percentage of CD4+/CD8+, CD4+CD25high, CD4+CD25high Foxp3+ and CD4+ T cells was significantly higher, and the percentage of CD8+ T cells was significantly lower in the cell transplantation group (P < 0.05). Histopathological findings showed that there were a small amount of infiltrated lymphocytes in the cell transplantation group with the presence of slight bleeding and edema, and these inflammatory reactions were milder than those in the control group. These findings indicate that bone marrow mesenchymal stem cell transplantation can effectively reduce the rejection in rats undergoing heart transplantation.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Changes of insulin and nestin expression during the differentiation of human umbilical cord mesenchymal stem cells into islet-like cells Shen Yi, Wang Yi-zhong, Shi Han, Shan Xia, Wang Lin-lin, Cui Xiao-lan 2016, 20 (50):  7500-7506.  doi: 10.3969/j.issn.2095-4344.2016.50.008 Abstract ( 327 )   PDF (4839KB) ( 302 )   Save BACKGROUND: Domestic and international studies have confirmed that human umbilical cord mesenchymal stem cells could be induced to differentiate into islet-like cells, but little is reported about the changes of insulin and nestin expressions during the differentiation phase. OBJECTIVE: To observe the changes of insulin and nestin expressions during the differentiation of human umbilical cord mesenchymal stem cells into islet-like cells. METHODS: Human umbilical cord mesenchymal stem cells were cultured using UltraCULTURE medium in vitro. Stem cells were cultured for three generations to observe cell morphological changes under an inverted microscope, to test immunophenotype by flow cytometry, and to identify the capacity of osteogenesis and adipogenic differentiation. Induction protocol was divided into two stages. In stage 1, stem cells were induced for 14 days in the UltraCULTURE medium with 4 nmol/L activin A, 25 μg/L epidermal growth factor, 100 μg/L β-nerve growth factor, 10 mmol/L nicotinamide. In stage 2, the cells were cultured in the UltraCULTURE medium with 1% insulin-transferin-selenium, 10 mmol/L nicotinamide, 10 μg/L basic fibroblast growth factor for an additional 14 days. The expressions of nestin and insulin in those differentiated cells were tested by flow cytometry, and zinc ion expression in the islet-like cell clusters was identified by dithizone staining. RESULTS AND CONCLUSION: During the differentiation process, the insulin level was increased gradually in the induction group and reached a higher level on day 28, but the insulin expression showed negative in the control group. In addition, on day 14 of induced differentiation, the nestin expression reached the peak and then gradually reduced along with the prolonged inductive time. On day 28 of induction, islet-like cell clusters formed and were positive for dithizone staining. In this experiment, the umbilical cord mesenchymal stem cells were successfully induced and differentiated into islet-like cells, accompanied with the variation of insulin and nestin expression.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Transplantation of bone marrow mesenchymal stem cells overexpressing Shootin1 for treatment of spinal cord injury Zhang Wei, Zhu Xiao-qi, Zhang De-cai 2016, 20 (50):  7507-7517.  doi: 10.3969/j.issn.2095-4344.2016.50.00 Abstract ( 248 )   PDF (20309KB) ( 219 )   Save BACKGROUND: Genetic modification by Shootin1 aims to effectively improve neural differentiation of bone marrow mesechymal stem cells (BMSCs) in the injured spinal cord, thereby promoting functional recovery from spinal cord injury after cell transplantation. OBJECTIVE: To explore the nerve regeneration ability of transplanted BMSCs overexpressing Shootin1 in rats with spinal cord injury. METHODS: BMSCs were transfected using adenovirus-Shootin1 for 48 hours. Then, immunofluorescence staining was used to detect Nestin and NeuN expression levels in the transfected cells under in vitro neuronal induction and differentiation. Animal models of spinal cord injury were made in rats using modified Allen’s method. Thirty minutes later, Shootin1-transfected BMSCs and non-transfected BMSCs were respectively injected into the subarachnoid space of the rats in the transfection and non-transfection groups, respectively. Rats in the model group were given no treatment. Five weeks after modeling, spinal cord samples were taken from each rat to make frozen sections for detection of nerve related markers RESULTS AND CONCLUSION: After 48-hour adenoviral transfection, Shootin1 expression was successfully detected in BMSCs. After 7-day in vitro induction, the cell morphology in the three groups varied, and there was no significant difference in the expression of Nestin and NeuN between the transfection and non-transfection groups. Basso, Beattie and Bresnahan scores were higher in the two cell transplantation groups than the model group. Increased expression levels of Nestin, NeuN, GFAP, MAP-2, ChAT and SYN were observed in both two cell transplantation groups, indicating a strengthened ability of nerve regeneration. Our experimental findings further confirm that BMSCs transplantation for spinal cord injury has achieved good outcomes, and Shootin1 protein plays a certain role in nerve regeneration and functional recovery after spinal cord injury. However, Shootin1 overexpression shows no obvious additional effects in combination with BMSCs transplantation, and further studies on the optimization of BMSCs transplantation for spinal cord injury are necessary.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cell transplantation alleviates myocardial injury due to systolic hypertension Wang Yong, Zhang Ya-guang 2016, 20 (50):  7518-7523.  doi: 10.3969/j.issn.2095-4344.2016.50.010 Abstract ( 348 )   PDF (4033KB) ( 213 )   Save BACKGROUND: Mesenchymal stem cells can be used to treat a variety of injuries, but little is known about their effect on systolic hypertension. OBJECTIVE: To study the therapeutic effect of bone marrow mesenchymal stem cells on systolic hypertension rats. METHODS: Twenty-four rats were randomized equally into control, model and cell transplantation groups. A rat model of systolic hypertension was made by injection of vitamin K and warfarin sodium in the model and cell transplantation groups. Rats in the cell transplantation group were given injection of bone marrow mesenchymal stem cells into the left ventricle, while those in the other two groups given normal saline. RESULTS AND CONCLUSION: Compared with the model group, bone marrow mesenchymal stem cells could reduce left ventricular mass index, increase cardiac tissue type I collagen volume fraction, collagen volume fraction and perivascular collagen volume fraction, but the ratio of end-diastolic volume/body weight did not change significantly. These findings indicate that bone marrow mesenchymal stem cell transplantation can reduce myocardial hypertrophy and cardiac fibrosis in systolic hypertension rats.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Different doses of human umbilical cord mesenchymal stem cells improve learning and memory ability of dementia rats Zhou Jun, Zhou De-sheng 2016, 20 (50):  7524-7529.  doi: 10.3969/j.issn.2095-4344.2016.50.011 Abstract ( 280 )   PDF (4355KB) ( 270 )   Save BACKGROUND: To delay the onset of Alzheimer’s disease, transplantation of viable and well-differentiated stem cells is expected to repair neural tissue, which has been an issue of concern. OBJECTIVE: To explore the effects of different doses of human umbilical cord mesenchymal stem cells (hUCMSCs) on learning and memory ability of Alzheimer’s disease rats. METHODS: Fifty Sprague-Dawley rats, 7 months of age, were randomized into normal, model, high-, middle- and low-dose hUCMSCs groups (n=10 per group). Rats in model and UCMSCs groups were used to make Alzheimer’s disease animals through intraperitoneal injection of 150 mg/kg D-galactose for 90 days, and rats in the normal group were given intraperitoneal injection of normal saline for 90 days. In the three hUCMSCs group, passage 3 hUCMSCs at doses of 1×105/0.2 mL/20 g, 5×105/0.2 mL/20 g, and 1×106/0.2 mL/20 g were injected via the tail vein, respectively. Forty-five days after cell transplantation, Morris water maze test was used to detect rat’s learning and memory abilities, and hematoxylin-eosin staining was used to observe pathological changes of the rat hippocampal CA1 region. RESULTS AND CONCLUSION: Compared with the normal group, the rats in the model group showed significant reduction in the ability of learning and memory. Compared with the model group, the escape latency was significantly shortened in the middle-dose hUCMSCs group (P < 0.05), while the number of passing times through the platform was increased significantly (P < 0.05). In the model group, the cells in the hippocampal CA1 region were arranged irregularly with unclear nucleoli and a part of cells were concentrated and deeply stained. In the middle-dose hUCMSCs group, the cells in the hippocampal CA1 region were arranged regularly with clear nucleoli, and only individual cells were stained deeply. These findings indicate that middle-dose hUCMSCs transplantation can improve the learning and memory abilities of Alzheimer’s rats.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Biological characteristics of bone marrow mesenchymal stem cells and application in skin defect repair Du Peng, Wang Xiao-chuan, Guan Zhen 2016, 20 (50):  7530-7535.  doi: 10.3969/j.issn.2095-4344.2016.50.012 Abstract ( 281 )   PDF (4123KB) ( 222 )   Save BACKGROUND: An appropriate biomaterial can be directly combined with autologous or allogeneic skin cells to construct tissue-engineered skin, which can accelerate skin repair after transplantation onto the skin wounds. It is a good idea to solve the deficiency in skin sources. OBJECTIVE: To study the biological characteristics of the complex with bone marrow mesenchymal stem cells and to investigate the application of this complex in skin defect repair. METHODS: Twenty-eight Sprague-Dawley rats, SPF grade, were randomized into two groups (n=14 per group). By adjustment of temperature, time, pressure and area of NC perm instrument, scald models were made in rats. Rats in the treatment group were given the repair using composite bone marrow mesenchymal stem cells, while those in the control group were given vaseline gauze repair. Repair effects were compared between two groups. RESULTS AND CONCLUSION: (1) After 72 hours of culture, the composite bone marrow mesenchymal stem cells small with round shape distributed dispersedly. After 5 days of culture, the cells began to stretch and the cell morphology became unstable. After three passages, the cell morphology became stable. The results of antigen identification showed the expression of CD44 and CD29 but the low expression of CD45 and CD34 in bone marrow mesenchymal stem cells. (2) Twenty-eight days after repair, there was no obvious scar on the wound surface of the treatment group, but a little shrinkage and obvious scar still existed in the control group. Moreover, in the control group, the epidermal layer of the skin was relatively thick, and the connection between the basal layer and the dermis was unsatisfactory. In the treatment group, obvious epidermal cell stratification, neat arrangement, and tight connection between the epidermis and dermis were observed. In summary, bone marrow mesenchymal stem cells are a special class of cells that have pluripotent ability and are more readily available. These cells are the preferred target cells for skin defect repair to promote early healing of the skin and improve blood circulation defect site, which are confirmed to have high clinical value.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Human umbilical cord mesenchymal stem cell transplantation for the treatment of acute lung injury Chen Jin-ling, Chen Yan-xia, Zhang Zhi-yong 2016, 20 (50):  7536-7542.  doi: 10.3969/j.issn.2095-4344.2016.50.013 Abstract ( 280 )   PDF (4738KB) ( 495 )   Save BACKGROUND: Studies have shown that human umbilical cord mesenchymal stem cells can improve pulmonary ventilation function by reducing inflammations. OBJECTIVE: To observe the therapeutic effect of human umbilical cord mesenchymal stem cell transplantation on acute lung injury. METHODS: Thirty Sprague-Dawley rats were randomized into normal group, model group and experimental group. Rats in the latter two groups were used to establish animal models of acute lung injury by intratracheal instillation of lipopolysaccharide. One hour after modeling, rats in the experimental group were intratracheally administered human umbilical cord mesenchymal stem cell suspension (0.1 mL, 1×106 cells), and those in the other two groups were given normal saline in the same dose intratracheally. Twenty-four hours after treatment, the pathological changes of lung tissue were observed using hematoxylin-eosin staining; the wet and dry weight ratio of the lung tissue and the levels of serum interleukin-1 and interleukin-8 were detected. RESULTS AND CONCLUSION: Compared with the normal group, the wet and dry weight ratio of the lung tissue and the levels of serum interleukin-1 and interleukin-8 were significantly increased in the model group (P < 0.05), while compared with the model group, these levels were significantly decreased in the experimental group (P < 0.05). Hematoxylin-eosin staining results showed clear alveolar space structure with complete alveolar septum in the normal group. In the model group, the alveolar septum was markedly thickened, and there was visible pulmonary capillary hyperemia, edema, as well as a large amount of inflammatory cell infiltrations in the pulmonary capillaries and alveolar space. Edema fluid rich in proteins was observed in a part of the pulmonary alveoli, and an extensive transparent membrane formed in the alveolar space. In the experimental group, the alveolar structure was clear, but the alveolar septum became thickened, and red blood cells and a small amount of infiltrated inflammatory cells were leaked from the pulmonary interstitial tissue. In conclusion, human umbilical cord mesenchymal stem cell transplantation for treatment of acute lung injury can reduce inflammatory factor levels and alleviate lung injury.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of salinomycin with umbilical cord blood mesenchymal stem cells in breast cancer MCF-7 cells Lv Wen-yue, Wang Zhen-yu, Guo Yang, Liu Yan-xin 2016, 20 (50):  7543-7548.  doi: 10.3969/j.issn.2095-4344.2016.50.014 Abstract ( 268 )   PDF (1020KB) ( 225 )   Save BACKGROUND: Mesenchymal stem cells-released factors can form a microenvironment inhibiting malignant tumor cell proliferation, and moreover, salinomycin also inhibits the growth of tumor cells. OBJECTIVE: To explore the effect of salinomycin with umbilical cord blood mesenchymal stem cells in breast cancer MCF-7 cells and its action mechanism, in order to seek a new target and therapeutic strategy to treat breast cancer. METHODS: Logarithmically growing MCF-7 cells were randomly divided into control, umbilical cord blood mesenchymal stem cell group and combination group. Cells in the control group were given no treatment, while those in the other two groups were given umbilical cord blood mesenchymal stem cell suspension or umbilical cord blood mesenchymal stem cell suspension combined with salinomycin, respectively. RESULTS AND CONCLUSION: Forty-eight hours after the intervention, the proliferation and invasion of MCF-7 cells and expression of POSTN protein in cells were significantly reduced in the combination group compared with the other two groups. These findings indicate that the combination of umbilical cord blood mesenchymal stem cell suspension and salinomycin effectively reduces the proliferation and migration of MCF-7 cells, and this combined use provides a new insight into the treatment of breast cancer.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Adipose-derived mesenchymal stem cell transplantation combined with cordyceps polysaccharide improves renal function and hypercoagulable state of rats with nephrotic syndrome Guo Zhi-bo, Zhang Chen-jie, Ma Li-na, Gao Da-wei, Song Ju-xing, Li Xiao-jing 2016, 20 (50):  7549-7556.  doi: 10.3969/j.issn.2095-4344.2016.50.015 Abstract ( 224 )   PDF (1297KB) ( 262 )   Save BACKGROUND: Cordyceps polysaccharide is a commonly used adjuvant drug for clinical treatment of nephrotic syndrome. As a classic model of nephrotic syndrome induced by adriamycin, the Sprague-Dawley rat model of nephrotic syndrome exhibits similar clinical manifestations and pathological changes to minimal-change nephropathy in humans. OBJECTIVE: To observe the effect of adipose-derived mesenchymal stem cells (ADMSCs) transplantation combined with cordyceps polysaccharide on renal function and hypercoagulable state in rats with nephrotic syndrome. METHODS: ADMSCs suspension was made in vitro and labeled using PKH26. Fifty Sprague-Dawley rats were randomized into normal (no intervention), model, ADMSCs, cordyceps polysaccharide and combined treatment groups (n=10/group). Adriamycin administration was performed to make rat models of nephrotic syndrome in the latter three groups. After modeling, model rats were respectively given no treatment, ADMSCs intravenously for 3 days, cordyceps polysaccharide intragastrically for 12 weeks, or their combined use. Then, 24-hour urinary protein, serum total cholesterol, triglyceride, total protein, albumin, high-density lipoprotein, low-density lipoprotein, creatinine, blood urea nitrogen levels and coagulation changes were detected at 12 weeks. Meanwhile, histopathological changes of renal tissues were observed under light microscope; survival and distribution of PKH26-labeled ADMSCs were observed by fluorescence microscopy; and expression of Hpa gene in renal tissue was detected by RT-PCR. RESULTS AND CONCLUSION: Compared with the model group, the 24-hour urinary protein, serum total cholesterol, triglyceride, low-density lipoprotein, creatinine, and blood urea nitrogen levels were significantly lower, while the serum total protein, albumin and high-density lipoprotein levels were significantly higher in the three treatment groups (P < 0.05). These indicators showed significant differences between the combined group and ADMSCs and cordyceps polysaccharide groups (P < 0.05). Both ADMSCs transplantation and cordyceps polysaccharide significantly relieved the hypercoagulable state of rats with nephrotic syndrome, and their combined effects were stronger (P < 0.05). After treatment, the pathological improvement in the kidney tissues was found in the three treatment groups; moreover, it was better in the combined treatment group than the ADMSCs and cordyceps polysaccharide groups. Better improvement in the number of PKH26-labeled ADMSCs and the expression of Hpa mRNA was observed in the combined treatment group compared with the ADMSCs and cordyceps polysaccharide groups. In conclusion, the combination of ADMSCs transplantation and cordyceps polysaccharide can improve kidney function and hypercoagulable state in rats with nephrotic syndrome, reducing pathological damage to the kidney tissue.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of hyperbaric oxygen on behavior and transplanted neural stem cells in rats with chronic stress depression Mi Kun, Guo Qiang, Sang Wen-hua, Wang Hai-bo, Li Le 2016, 20 (50):  7557-7564.  doi: 10.3969/j.issn.2095-4344.2016.50.016 Abstract ( 284 )   PDF (1217KB) ( 245 )   Save BACKGROUND: Hyperbaric oxygen as a well-recognized therapy for ischemic and hypoxic diseases has to be combined with other treatments. OBJECTIVE: To study the effects of hyperbaric oxygen combined with neural stem cell transplantation via tail vein in chronic stress depression rats. METHODS: Of 60 Sprague-Dawley rats, 15 rats randomly selected were given no treatment as normal group, and the rest 45 ones were used to establish a rat model of depression and randomly divided into three groups: model group (n=15, without giving any treatment), neural stem cell group (n=15, injection of 1 mL neural stem cell suspension (3×106) via the tail vein) and combined group (n=15, hyperbaric oxygen treatment plus neural stem cell injection). Hyperbaric oxygen treatments were carried out four times per day, for 1 week, and all the treatments were given 24 hours after modeling. Rats in each group were subjected to body mass measurement and sugar water consumption test 1 and 2 weeks after treatment, and open field test within 5 minutes after treatment to observe rat behavior changes. The survival and distribution of CM-Dil labeled neural stem cells were observed by fluorescence microscopy. The changes in the size, morphology and number of hippocampus neurons in rat were detected by the method of nylon staining. TUNEL method was used to measure the apoptosis of nerve cells. RESULTS AND CONCLUSION: Compared with the normal group, in the model group, rat’s body weight, sucrose water preference and open field test scores were significantly lower, reduced number of hippocampal neurons with intact structure was found and TUNEL results showed more apoptosis (P < 0.05). Compared with the model group, the body weight, sucrose preference and open field test score increased significantly (P < 0.05), the number of hippocampal neurons was increased, while the number of apoptotic cells was reduced significantly in the neural stem cell and combined groups (P < 0.05). Compared with the neural stem cell group, these indexes were improved more significantly in the combined group (P < 0.05). More CM-Dil positive cells were found in the combined group than the neural stem cell group (P < 0.05). Additionally, the number of hippocampal neurons was higher in the combined group than the neural stem cell group, but lower than the normal group. To conclude, transplantation of neural stem cells combined with hyperbaric oxygen can improve the depression behavior and apoptosis in the hippocampal neurons in chronic stress depression rats.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Enzyme digestion method versus tissue block method in the culture of cardiac stem cells Li Sha, Li Shu-ren, Hao Qing-qing, Yang Ling-ling, Zhang Qian-hui, Ma Yu-long, Zhang Yue-hua, Deng Wen-hao 2016, 20 (50):  7565-7570.  doi: 10.3969/j.issn.2095-4344.2016.50.017 Abstract ( 390 )   PDF (1324KB) ( 439 )   Save BACKGROUND: At present, various methods of extracting cardiac stem cells have been reported in China, but the processes of isolation, culture and purification are not yet standardized. OBJECTIVE: To explore the difference in the culture of rat cardiac stem cells using enzyme digestion method and tissue block method. METHODS: Cardiac stem cells from Sprague-Dawley rats, clean grade, 1 day of age, were extracted and cultured in vitro using the enzyme digestion method (control group) or tissue block method (experimental group). RESULTS AND CONCLUSION: Both of the two methods could produce cardiac stem cells, but the proportion of cultured cells in the experimental group was significantly higher than that in the control group, indicating the tissue block method is superior to the enzyme digestion method in the culture of cardiac stem cells.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Evolutionary mechanisms for stem cell differentiation and stem cell niche distribution Zhang Jian-xin 2016, 20 (50):  7571-7578.  doi: 10.3969/j.issn.2095-4344.2016.50.018 Abstract ( 2805 )   PDF (1557KB) ( 807 )   Save BACKGROUND: More and more stem cells such as hematopoietic stem cells, neural stem cells and induced pluripotent stem cells have been discovered, but there is no unified theory on stem cell classification and stem cell differentiation. Similarly, more and more stem cell niches, such as niches of hematopoietic stem cells, intestinal epithelial stem cells and hair follicle stem cells, have been found, and there is still no unified theory on the formation and distribution of stem cell niches. In the past 20 years, the viewpoint that “the orderly distribution of different types of stem cell niches constitutes the meridian system of Chinese medicine” has not been taken seriously, especially in China. OBJECTIVE: To explore the evolutionary mechanisms for stem cell differentiation and stem cell niche distribution. METHODS: With the reference of the Inner Canon of Yellow Emperor and literature on human anatomy, we further explained the distribution rule of stem cell niches in various tissues from the perspective of meridian theory. We explored the law of stem cell differentiation based on the biogenetic law, species evolutionary tree and the theory of meridians of Chinese medicine. RESULTS AND CONCLUSION: The embryonic development with the formation of three germ layers in higher animals directly transcends the evolutionary history of their invertebrate ancestors in an extremely simple way. The basic objects described in the biogenetic law should first be two stem cell differentiation chains   and    as well as the evolution of the distribution of stem cells in the meridian system. The presently accepted species evolutionary tree assumes that the ancestor of the deuterostomia is a primitive bilaterally symmetric animal, which is completely wrong. It violates the Darwinian principle of evolution from the viewpoint of stem cell differentiation, and the evolutionary tree cannot fork to form two branches, protostomia and deuterostomia, that are unrelated on the their origins. Cockroaches, spearfish and ginkgo cannot be living fossils. The origin of the species is multi-sources, and the species evolution is ongoing in a multi-level wavy manner.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Research progress of stem cells in hair follicle regeneration Ding Yu-wei, Xin Guo-hua, Zeng Yuan-lin 2016, 20 (50):  7579-7585.  doi: 10.3969/j.issn.2095-4344.2016.50.019 Abstract ( 551 )   PDF (1059KB) ( 229 )   Save BACKGROUND: Stem cells have the potential to differentiate into various organs and tissues. In recent years, stem cells have been proved to differentiate into hair follicles under certain conditions. OBJECTIVE: To review the research progress and prospect of stem cells differentiating to hair follicles, thereby providing a reliable basis for clinical treatment of serious hair follicle injury. METHODS: A computer-based search of PubMed, EMBASE, WanFang, CNKI databases was performed for related articles published between 2013 and 2015, using the keywords of “cell hair, follicle stem, medicine regenerative, differentiation” in English and Chinese. A total of 207 articles were retrieved, and finally 34 articles were included in result analysis. RESULTS AND CONCLUSION: Stem cells from different sources all have the ability to differentiate into hair follicles under certain inductions. However, it is important to seek more scientific and rational methods for the differentiation of stem cells into hair follicles based on overcoming their own shortcomings. A great progress has been made in animal experiments and subclinical trials, and even a great breakthrough in some aspects. Further studies on combining the advantages and overcoming the shortcomings of various stem cells during differentiation are required for the clinical treatment of serious hair follicle injury.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Methods of bone marrow mesenchymal stem cells differentiating into hepatocytes and the underlying mechanisms Xie Shu-cai, Zhang Jian-quan, Jiang Xi-li, Zhou Shuai 2016, 20 (50):  7586-7593.  doi: 10.3969/j.issn.2095-4344.2016.50.020 Abstract ( 276 )   PDF (1677KB) ( 248 )   Save BACKGROUND: A number of studies have shown that there are many inducible methods by which bone marrow mesenchymal stem cells can differentiate into hepatocytes, but the specific molecular mechanism is unclear yet. OBJECTIVE: To review the programs and underlying mechanisms by which bone marrow mesenchymal stem cells differentiate into hepatocytes. METHODS: A computer-based online search of CNKI, VIP, WanFang and PubMed databases was performed to retrieve articles about directional differentiation of bone marrow mesenchymal stem cells into hepatocytes published between 2004 and 2015. The key words were “hepatocyte (-like) cells, bone marrow mesenchymal stem cells, differentiation” in Chinese and English, respectively. Finally, 62 articles were included in result analysis. RESULTS AND CONCLUSION: There are many programs for hepatic differentiation of bone marrow mesenchymal stem cells, but the specific molecular mechanism is still unclear. Many studies mainly focus on Notch signaling pathway, Wnt/β-catenin signaling pathway, P38 signal pathway, miR-122 and effect of calcium ions. Bone marrow mesenchymal stem cells that can be induced to differentiate into mature hepatocytes provide an ideal cellular source for hepatocyte transplantation and artificial liver, which is proposed to be a new strategy for clinical treatment of end-stage liver disease.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Icariin promotes the osteogenic differentiation of mesenchymal stem cells: a good candidate for bone defect repair Chen Zhen-dong, Gao Hui, Xu Fang-tian 2016, 20 (50):  7594-7600.  doi: 10.3969/j.issn.2095-4344.2016.50.021 Abstract ( 293 )   PDF (1025KB) ( 231 )   Save BACKGROUND: Recent studies have shown that icariin is a good bone-inducing factor that can promote the osteogenic differentiation of mesenchymal stem cells, providing a new hope for the treatment of bone defects. OBJECTIVE: To review research achievements in pharmacological effects of icariin effects on bone tissue metabolism as well as its effect to promote osteogenic differentiation of mesenchymal stem cells. METHODS: The first author retrieved CNKI and PubMed databases for relevant Chinese and English literatures using keywords of “icariin, stem cell, osteogenesis”, respectively. Articles regarding icariin, stem cells, osteogenesis were included, and repetitive studies were excluded. Totally 754 articles were retrieved initially. In accordance with inclusion and exclusion criteria, 41 articles were included in result analysis. RESULTS AND CONCLUSION: As the main ingredient of Herba epimedii, icariin functions as a good osteogenetic growth factor to promote the osteogenic differentiation of bone marrow mesenchymal stem cells. In recent years, icariin has been shown to promote adipose-, umbilical cord-, and periodontal ligament tissue-derived mesenchymal stem cells to differentiate into osteoblasts. But such studies are less reported. Until now, mesenchymal stem cells still exhibit unsatisfactory osteogenic ability in in vivo experiments. Given this, osteogenetic growth factors contribute to the osteogenic differentiation of mesenchymal stem cells. Therefore, the use of icariin is expected to provide a good strategy for bone defect repair.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics