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    01 July 2016, Volume 20 Issue 28 Previous Issue    Next Issue
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    miR-106a regulates the osteogenic differentiation of human bone marrow mesenchymal stem cells by targeting bone morphogenetic protein receptor 2
    Gu Su
    2016, 20 (28):  4109-4116.  doi: 10.3969/j.issn.2095-4344.2016.28.001
    Abstract ( 297 )   PDF (1479KB) ( 442 )   Save

    BACKGROUND: Under normal physiological conditions, there is a homeostasis between the osteogenic and adipogenic differentiation of human bone marrow mesenchymal stem cells. Osteogenic differentiation is an important process in the formation of skeleton in which differentiated and mature osteoblasts are indispensable.
    OBJECTIVE: To explore the role of miR-106a and its target gene, bone morphogenetic protein receptor 2 (BMPR2) in the differentiation of human bone marrow mesenchymal stem cells into osteoblasts.
    METHODS: Human bone marrow mesenchymal stem cells were induced to differentiate into osteoblasts by osteoblast-specific induction medium, and this process was detected by western blot and alkaline phosphatase staining. Overexpression of miR-106a was elicited by transfecting miR-106a mimics and the BMPR2 knockdown achieved by RNA interference. The expression levels of miR-106a and BMPR2 were detected by real-time PCR and western blot, respectively. The interaction of miR-106a and BMPR2 was verified by TargetScan software and dual luciferase report experiment assay.
    RESULTS AND CONCLUSION: The expression of miR-106a was decreased whereas the expression of BMPR2 increased with the progress of osteogenesis differentiation. When miR-106a was overexpressed, alkaline phosphatase activity was declined and the expressions of runt-related transcription factor 2 and osteocalcin, markers of osteogenesis differentiation, both decreased. The expression of BMPR2 was decreased as well. BMPR2 was predicted to be the target gene of miR-106a by TargetScan software and this prediction proved by dual luciferase report experiments assay. Additionally, osteogenesis differentiation was inhibited by knocking down the expression of BMPR2. These results indicate that miR-106a regulates the differentiation of bone marrow mesenchymal stem cells into osteoblasts by targeting BMPR2.

     

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    Correlation between bone marrow stromal stem cells and apoptosis in epilepsy
    Wang Hao, Ren Xiang-yang, Ma Cong-min, Huang Chao, Zhou Hai-tao
    2016, 20 (28):  4117-4122.  doi: 10.3969/j.issn.2095-4344.2016.28.002
    Abstract ( 300 )   PDF (3731KB) ( 273 )   Save

    BACKGROUND: There is a close relationship between epilepsy and apoptosis. The appearance of epilepsy can lead to the loss of neurons in the hippocampus, triggering a series of programmed cell death.
    OBJECTIVE: To investigate the effect of bone marrow stromal stem cell transplantation on apoptosis in epilepsy.
    METHODS: After modeled to be of epilepsy 45, Sprague-Dawley model rats were randomly divided into three groups, followed by given no intervention (moldel group), normal saline (normal saline group) or bone marrow stromal stem cell transplantation (transplantation group). At 1, 2 and 4 weeks after modeling, the number of Bax-positive cells, Bcl-2-positive cells and Bax/Bcl-2 were detected by immunohistochemistry.
    RESULTS AND CONCLUSION: The number of Bax-positive cells, Bcl-2-positive cells and Bax/Bcl-2 presented no obvious changes in the normal saline group at different time points. However, the number of Bax-positive cells and Bax/Bcl-2 in the transplantation group was significantly decreased, while the number of Bcl-2-positive cells significantly increased compared with the other two groups at 1, 2 and 4 weeks after modeling (P < 0.05). Moreover, the above indicators varied significantly in the transplantation group at different time points after modeling (P < 0.05). These results show that bone marrow stromal stem cell transplantation can affect the apoptosis and effectively reduce the apoptosis in rats with epilepsy by up-regulating the number of Bax-positive cells and down-regulating the number of Bcl-2-positive cells.

     

     

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    Effects of human umbilical cord mesenchymal stem cells via intramuscular injection on the myocardial micrangium and collagen expression in normal rats
    Guo Yu-xiu, Wang Si-ping, Zhang Wen-xiang, Mao Cheng-gang, Gao Hong, Li Zi-pu
    2016, 20 (28):  4123-4129.  doi: 10.3969/j.issn.2095-4344.2016.28.003
    Abstract ( 249 )   PDF (4136KB) ( 204 )   Save

    BACKGROUND: To date, it is still unclear whether the intramuscular injection of heterogeneous umbilical cord mesenchymal stem cells (UC-MSC) can cause cardiac ectopic pathological angiogenesis as well as increase collagen synthesis to promote myocardial fibrosis.
    OBJECTIVE: To explore the effects of intramuscular injection of human UC-MSCs on myocardial micrangium and collagen expression in normal Wistar rats.
    METHODS: After 2 weeks of feeding, 60 male SPF Wistar rats were randomly assigned to receive intramuscular injection of PBS (normal group), DMEM (culture medium group), human UC-MSCs supernatant (supernatant group), 0.25×105, 1.0×105, 4.0×105 human UC-MSCs (low-, moderate- and high-dose groups), respectively (n=10 per group). All the rats were subjected to second injection (same dose) at 4 weeks after first intramuscular injection. Then, the rats were killed under anesthesia at 4 weeks after second injection, to take heart tissues from the left ventricle for pathological observation, immunohistochemical examination and Masson staining.
    RESULTS AND CONCLUSION: No alteration of the response, activity, victualage, faeces, weight growth, and fur was found, and there was no death in rats during the experiment. All the rats had no symptoms of molt, inflammation, skin ulcer, scleroma. Strong positive expression of CD34 for the micrangium in the myocardial tissue was observed, and positive expression of the collagen in the myocardial tissue observed by Masson staining. There were no significant differences in the microvessel density and collagen expression in the myocardium among the groups (F=0.110 and 0.585, P > 0.05). To conclude, hUC-MSCs or its supernatant via intramuscular injection has no effect on the micrangium and collagen expression in normal rats.

     

     

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    Characterization of UGT1a1, UGT1a6 and mGST1 at different stages of differentiation of mouse embryonic stem cells
    Xu Ling-li
    2016, 20 (28):  4130-4135.  doi: 10.3969/j.issn.2095-4344.2016.28.004
    Abstract ( 308 )   PDF (3865KB) ( 265 )   Save

    BACKGROUND: Until now, limited information has been neported on the characterization of udines 5-diphosphateglucuronosyl transferase (UGT) 1a1, UGT1a6, and microsomal glutathione S-transferase 1 (mGST1) during the differentiation of mouse embryonic stem cells.
    OBJECTIVE: To observe the expression characteristics of UGT1a1, UGT1a6 and mGST1 at different stages of differentiation of mouse embryonic stem cells.
    METHODS: Embryonic fibroblasts from Wistar rats were isolated and cultured as feeder cells. Mouse embryonic stem cells seeded onto these feeder cells were induced to differentiate into hepatocytes. Subsequently, expression of UGT1a1, UGT1a6 and mGST1 was detected using western blot assay, and catalytic activity of mGST1 was determined by high performance liquid chromatography.
    RESULTS AND CONCLUSION: Increasing UGT1a1 expression was visible during the whole cell differentiation, while UGT1a6 exhibited no expression initially but a higher level at 18 days of differentiation. mGST1 expression was visible at high level throughout the differentiation, but its expressive abundance was still lower than that in the adult mouse stem cells. At 18 days after the beginning of differentiation, the catalytic activity of mGST1 in microsomal liver tissue was 7.65 μmol/(min•g). Taken together, stabilized UGT1a1, increased UGT1a6 and little mGST1 expression are confirmed in the differentiation of embryonic stem cells into hepatocytes.

     

     

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    Different culture methods for human umbilical cord mesenchymal stem cells
    Liu Feng, Jiang Xiang-lin, Liu Ming-ming
    2016, 20 (28):  4136-4141.  doi: 10.3969/j.issn.2095-4344.2016.28.005
    Abstract ( 362 )   PDF (962KB) ( 241 )   Save

    BACKGROUND: Umbilical cord stem cells mainly derive from full-term infants, and common culture methods include tissue-attached method and trypsin-digestion mehod. However, effects of different culture methods on the separation of umbilical cord mesenchymal stem cells remain many disputes.
    OBJECTIVE: To observe the effects of different culture methods on umbilical cord mesenchymal stem cells.
    METHODS: Umbilical cords of 30 healthy full-term and caesarean delivery infants were selected, and cultured using tissue-attached method or trypsin-digestion method to isloate and culture human umbilical cord mesenchymal stem cells. Meanwhile, cell growth was measured by flow cytometry.
    RESULTS AND CONCLUSION: The fusiform-shaped cells began to separate from the umbilical cord tissue that was primary cultured using tissue-attached method, and 10 days later, the cell fusion reached 80%; after the umbilical cord was cultured using collagenase-trypsin digestion for 5 days, a small amount of adherent cells with different shapes appeared, and the fiber-like cells reached 80% of confluence until 2-week culture. There was no significant difference in the growth of umbilical cord mesenchymal stem cells cultured by different culture methods (P > 0.05). Moreover, cells cultured by two methods were all positive for CD13, CD29, CD44 and CD105. These results demonstrate that human umbilical cord mesenchymal stem cells exhibit a high success rate in primary culture using tissue-attached method, which is superior to the trypsin-digestion method.

     

     

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    Effects of fatty acid binding protein-5 silencing on proliferation, apoptosis, invasion of human glioma cells and expressions of CD44+ and CD133+
    Liu Hai-peng, Wang Ji-wei, Zheng Ke-bin
    2016, 20 (28):  4142-4148.  doi: 10.3969/j.issn.2095-4344.2016.28.006
    Abstract ( 296 )   PDF (1292KB) ( 251 )   Save

    BACKGROUND: It has been demonstrated that there are different expressions of fatty acid binding proteins (FABPs) in most malignant tumors such as breast cancer, prostate cancer, liver cancer, lung carcinoma and bladder carcinoma; therefore, FABPs are closely related to the occurrence, metastasis, invasion and drug resistance of malignant tumors.
    OBJECTIVE: To investigate the effects of fatty acid binding protein-5 (FABP-5) silencing on the proliferation, invasion and apoptosis of U251 cells (human glioma cells).
    METHODS: siRNA molecules targeting the mRNA of FABP-5 was designed and chenically synthesized, which was transiently transfected into U251 cells. U251 cells were divided into three groups: Lv-shRNA-FABP-5 was added into FABP-5-shRNA group, LV-shRNA-NC added into negative control group, and blank control group underwent normal culture. mRNA and protein expressions of FABP-5 were detected by RT-PCR and western blot assay, respectively. The cell proliferation in vitro was determined by cell counting kit-8 assay, the cell cycle, apoptosis and expressions of CD44+ and CD133+ were detected by flow cytometry, and the apoptosis was observed using TUNEL staining.
    RESULTS AND COUNCLUSION: mRNA and protein expressions of FABP-5 in the FABP-5-shRNA group were significantly lower than those in the negative control and blank control groups. Compared with the negative control and blank control groups, the cell growth rate was significantly decreased, the cell cycle arrested in the G0/G1 phase, and the cell number in the S phase was decreased, and moreover, the proportion of CD44+/CD133+ expression was significantly decreased in the FABP-5-shRNA group (P < 0.05). Besides, compared with the negative control and blank control groups, the apoptosis rate was significantly increased, and the cell proliferation and invasiveness were significantly decreased in the FABP-5-shRNA group (P < 0.05). In conclusion, it is possible that FABP-5 directly or indirectly regulates the cell cycle and apoptosis of glioma cells, and its expression changes share a close relationship with the invasiveness of tumor cells.

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    ABCE1 silencing enhances sensitivity to beta-eiemene of electrotransferred ovarian cancer stem cells
    Wang Jun, Wang Jian-zhong
    2016, 20 (28):  4149-4154.  doi: 10.3969/j.issn.2095-4344.2016.28.007
    Abstract ( 265 )   PDF (1107KB) ( 278 )   Save

    BACKGROUND: It is likely to find that ABCE1 plays an important part in the occurrence and progression of cancers through in-depth study.
    OBJECTIVE: To investigate the effects of ABCE1 silencing on the proliferation and sensitivity to β-eiemene of ovarian cancer stem cells via electrotransfection.
    METHODS: siRNA sequence of ABCE1 was designed and synthesized, and then was electrotransfection into ovarian cancer stem cells. Subsequently, expressions of ABCE1 mRNA and protein in ovarian cancer stem cells after ABCE1 silencing were detected by RT-PCR and western blot assay; the cell cycle was detected using flow cytometry; the sensitivity of ovarian cancer stem cells to β-eiemene after ABCE1 silencing was analyzed by MTT assay and colony-formation assay.
    RESULTS AND CONCLUSION: After ABCE1 silencing, expressions of ABCE1 mRNA and protein in ovarian cancer stem cells were significantly reduced. And the cell cycle was arrested in G0/G1 phase, and the number of cells in S phase significantly decreased. Furthermore, the sensitivity of ovarian cancer stem cells to β-eiemene after ABCE1 silencing was significantly enhanced. In conclusion, ABCE1 silencing cannot only significantly inhibit the proliferation of ovarian cancer stem cells, but also enhance the sensitivity of cancer stem cells to β-eiemene.

     

     

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    Osteogenic efficiency of induced adipose-derived stem cells under Transwell co-cultured condition  
    Sun Shi-chen, Dong Teng-zhe, Huang Xin, Zhang Yun-long, Chen Gang
    2016, 20 (28):  4155-4161.  doi: 10.3969/j.issn.2095-4344.2016.28.008
    Abstract ( 268 )   PDF (1417KB) ( 574 )   Save

    BACKGROUND: Under co-culture conditions, mesenchymal stem cells could regulate osteogenic differentiation and osteogenesis of osteoblasts.
    OBJECTIVE: To observe the osteogenic efficiency of osteoblastic precursor cells co-cultured with undifferentiated bone marrow-derived mesenchymal stem cells, umbilical cord-derived mesenchymal stem cells, or placenta-derived mesenchymal stem cells in mineralization medium.
    METHODS: Adipose-derived stem cells were induced in osteogenic differentiation medium for 7 days before being indirectly co-cultured with undifferentiated mesenchymal stem cells isolated from different tissues (bone marrow group, umbilical cord group and placenta group) in Transwell plates. Induced adipose-derived stem cells cultured alone served as control group. At different experimental intervals, quantitative analysis of alkaline phosphatase activity and calcified matrix was preformed to observe the effects of mesenchymal stem cells from different sources on the osteogenic efficiency of induced adipose-derived stem cells.
    RESULTS AND CONCLUSION: Expression of alkaline phosphatase was significantly higher in different experimental groups than the control group (P < 0.05), and it was also higher in the bone marrow group than the umbilical cord and placenta groups (P < 0.05). Quantitative analysis of calcified matrix revealed that the experimental groups were significantly higher than the control group (P < 0.05); and in experimental groups, the umbilical cord group was higher than bone marrow group and placenta group(P < 0.05). These findings indicate that the osteogenic efficiency of induced adipose-derived stem cells is improved dramatically under co-culture conditions.

     

     

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    Expression of serum inflammatory cytokines in rats with cerebral infarction undergoing bone marrow mesenchymal stem cell transplantation combined with edaravone
    Song Nai-guang, Sun Jing-jing, Zhang Yao-long, Meng Ling-hai, Gao Shu-huan, Xue Jian, Sun Cai-yue
    2016, 20 (28):  4162-4168.  doi: 10.3969/j.issn.2095-4344.2016.28.009
    Abstract ( 216 )   PDF (1212KB) ( 260 )   Save

    BACKGROUND: To inhibit the expressions of prothrombin activator inhibitor 1 and tissue plasminogen activator is one potential target for the treatment of cerebral infarction.
    OBJECTIVE: To investigate the expressions of serum inflammatory cytokines in rats with cerebral infarction undergoing bone marrow mesenchymal stem cell transplantation combined with edaravone.
    METHODS: Sixty Sprague-Dawley rats were enrolled to prepare models of focal cerebral infarction by middle cerebral artery occlusion, and were randomly divided into four groups. Rats were given intravenous injection of PBS via tail veins for 5 consecutive days as model group, rats were subjected to intravenous injection of 2.0×109 /L bone marrow mesenchymal stem cell suspension (1 mL) via tail veins, twice daily for 5 days as stem cell transplantation group, and those were given intravenous injection of 30 mg edaravone combined with intravenous injection of 2.0×109/L bone marrow mesenchymal stem cell suspension (1 mL) via tail veins for 5 days, twice daily, as combined group.
    RESULTS AND CONCLUSION: Compared with the model group, modified neurologic severity scores were lower, expressions of serum prothrombin activator inhibitor 1 and tumor necrosis factor-α mRNA in the brain decreased, and the infarct area reduced in the stem cell transplantation and combined groups. And the changed levels of above indicators in the combined group were significantly larger than those of the stem cell transplantation group. In conclusion, combination of bone marrow mesenchymal stem cell transplantation with edaravone can promote neural function recovery after cerebral infarction.

     

     

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    Basic fibroblast growth factor-transfected bone marrow mesenchymal stem cell transplantation for acute kidney injury
    Cui Li-de, Li Tie-min
    2016, 20 (28):  4169-4175.  doi: 10.3969/j.issn.2095-4344.2016.28.010
    Abstract ( 220 )   PDF (1202KB) ( 235 )   Save

    BACKGROUND: It has been confirmed that basic fibroblast growth factor (bFGF) can promote the growth, proliferation, differentiation and functional expression of most cells derived from neuroderm and mesoderm.
    OBJECTIVE: To investigate the effect of bFGF-transfected bone marrow mesenchymal stem cell transplantation in rats with acute kidney injury.
    METHODS: bFGF genes were transfected into bone marrow mesenchymal stem cells via an adenovirus vector, and then expression of bFGF in transfected cells was identified using RT-PCR technology. Rat models of acute kidney injury were prepared by clipping bilateral renal pedicles, and then randomized into three groups (n=20): rats were given injection of bone marrow mesenchymal stem cell suspensions via tail vein as negative transfected group, those given injection of bFGF-transfected bone marrow mesenchymal stem cell suspensions via tail vein as bFGF-transfected group, and the others given injection of DMEM via tail vein as model group. Four weeks later, levels of serum creatinine and urea nitrogen were detected, expressions of connective tissue growth factor and growth factor in renal tissues were detected by Western blot assay, and morphology of renal tissues was observed using hematoxylin-eosin staining.
    RESULTS AND CONCLUSION: bFGF genes were successfully transfected into bone marrow mesenchymal stem cells. Compared with the model group, the levels of serum creatinine and urea nitrogen were significantly reduced in bFGF-transfected and negative transfected groups, especially in the bFGF-transfected group (P < 0.05), while expressions of connective tissue growth factor and transforming growth factor in renal tissues in bFGF-transfected and negative transfected groups were significantly weakened in these two groups (P < 0.05), but there were no significant differences between the bFGF-transfected group and negative transfected group (P > 0.05). Besides, renal tissues necrosis and inflammatory reactions were mitigated in the negative transfected group; renal tubules with normal outlines and no overt necrotic cells could be found in the bFGF-transfected group. These findings show that bFGF-transfected bone marrow mesenchymal stem cell transplantation plays a better role in acute kidney injury repair in rats.

     

     

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    Stem cell transplantation protects the kidney in diabetic rat models
    Gan Li-ming
    2016, 20 (28):  4176-4181.  doi: 10.3969/j.issn.2095-4344.2016.28.011
    Abstract ( 259 )   PDF (1129KB) ( 234 )   Save

    BACKGROUND: So far numerous theoretical studies have shown the treatment effect of stem cell transplantation for chronic complications of diabetes, while its treatment effects on diabetic nephropathy have not yet been confirmed in animal models.
    OBJECTIVE: To investigate the protective effect of bone marrow mesenchymal stem cell transplantation on the kidney in rat models of diabetes.
    METHODS: Rats were fed with high-sugar and high-fat diet for 4 weeks, and then were given injection of streptozotocin to establish type 2 diabetic rat models. At 2 days after modeling, bone marrow mesenchymal stem cells were injected via the tail vein (stem cell transplantation group). In the meanwhile, control and diabetes groups were established. At 21 days after cell transplantation, levels of glucose, triglyceride and insulin in the tail vein were detected. Additionally, morphological observations of kidney and pancreatic tissues were performed.
    RESULTS AND CONCLUSION: The levels of blood sugar, insulin and triglycerides in the diabetes group were significantly higher than those of the control group (P < 0.05). Blood glucose and insulin levels in the stem cell transplantation group were significantly lower than those of the diabetes group (P < 0.05). In addition, mesangial area and glomerular volume in the stem cell transplantation group were significantly lower compared with the diabetes group (P < 0.05). These results confirm that bone marrow mesenchymal stem cell transplantation can reduce levels of blood glucose and serum insulin, contributing to the repair of damaged pancreas and kidney.

     

     

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    Bone marrow mesenchymal stem cell transplantation for rat cerebral infarction: recovery of neurological function and expression of synaptophysin
    Cheng Lv-fang
    2016, 20 (28):  4182-4188.  doi: 10.3969/j.issn.2095-4344.2016.28.012
    Abstract ( 292 )   PDF (4746KB) ( 225 )   Save

    BACKGROUND: Synaptophysin plays an important role in the recovery of neural function after cerebral ischemia.
    OBJECTIVE: To investigate the effects of bone marrow mesenchymal stem cell transplantation on nervous function and expression of synaptophysin after cerebral infarction.
    METHODS: Totally 60 rats were equivalently randomized into four groups, including sham operation, control, model and stem cell treatment groups. Rats in the control, model and stem cell treatment groups were used for preparing cerebral infarction models, and the remaining underwent the sham operation. After 1 day of modeling, bone marrow mesenchymal stem cells were transplanted into the rat lateral ventricle in the stem cell treatment group, and rats in the control group was given the injection of the same amount of PBS. After 1, 7 and 14 days of treatment, rat’s neurological function was scored on beam-walking test, rotarod test and screen test, and expression of synaptophysin was detected by RT-PCR and immunohistochemical assay.
    RESULTS AND CONCLUSION: At 7 and 14 days after treatment, the beam-walking test, rotarod test and screen test scores in the stem cell treatment group were significantly lower than those in the control and model groups (P < 0.05), and the above scores showed no significant differences between the control group and model group (P > 0.05). At 1 day after treatment, the mRNA expression of synaptophysin and the number of synaptophysin-positive cells in the sham operation group were significantly higher than those in the other three groups (P < 0.05); at 7 and 14 days after treatment, the mRNA expression of synaptophysin and the number of synaptophysin-positive cells in the stem cell treatment group were significantly increased compared with the other three groups (P < 0.05), and additionally, the mRNA expression of synaptophysin and the number of synaptophysin-positive cells in the sham operation group were significantly lower than those in the model and control groups (P < 0.05). These findings suggest that bone marrow mesenchymal stem cell transplantation can effectively promote the recovery of neurological function in cerebral infarction rats, and partially promote the formation of synaptophysin.

     

     

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    Protective effect of bone marrow stromal stem cells against acute lung injury after hip fracture in aged rats with chronic obstructive pulmonary disease
    Zhang Gui-tong, Feng Jie-yang, Liu Jia, Zhang Yan-jin, Xiang Zi-min, Meng Fan-xing,Sun Tian-sheng
    2016, 20 (28):  4189-4195.  doi: 10.3969/j.issn.2095-4344.2016.28.013
    Abstract ( 282 )   PDF (4347KB) ( 198 )   Save

    BACKGROUND: The elderly hip fracture patients with chronic obstructive pulmonary disease (COPD) have a higher postoperative mortality than those only with hip fractures. In recent years, it has become an issue of concerns. Because the mechanism is unknown, however, there are no effective clinical interventions for these patients.
    OBJECTIVE: To observe the effect of bone marrow stromal stem cells (BMSCs) on the level of pulmonary inflammation in aged rats with chronic obstructive pulmonary disease (COPD) after hip fractures.
    METHODS: Thirty male Sprague-Dawley rats, 12 months old, were exposed to smoking for 4 months and randomized into three groups, smoking, smoking+hip fracture+normal saline, smoking+hip fracture+ BMSCs groups. Animal models of hip fracture were made in the latter two groups. Twenty-four hours after hip fracture, the lower lobe and peripheral blood samples were taken from all rats in the three groups, to evaluate the pathological changes of lung tissue and detect levels of inflammatory factors in the lung tissue and peripheral blood.
    RESULTS AND CONCLUSION: After closed hip fracture, aged COPD rats exhibited inflammatory cell infiltration, mucus secretion, airway stenosis or occlusion; the levels of interleukin-6, tumor necrosis factor-α and interleukin-10 in the lung tissue and peripheral blood were increased. After intravenous injection of BMSCs, the pathological changes of the lung tissue were reduced, and the levels of pro-inflammatory factors, tumor necrosis factor-α and interleukin-6, decreased, but the level of anti-inflammatory factor interleukin-10 further increased, which were significantly different from those in the normal saline group (P < 0.05). These findings indicate that BMSCs can relieve acute lung injury in aged COPD rats with hip fractures．

     

     

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    Influences of CD133+ cells on human umbilical cord blood mononuclear cell transplantation for treating heart failure
    Ma Li, Xie Yi-xu, Chang Yu, Yao Lei
    2016, 20 (28):  4196-4202.  doi: 10.3969/j.issn.2095-4344.2016.28.014
    Abstract ( 260 )   PDF (4304KB) ( 198 )   Save

    BACKGROUND: Cell purification can eliminate the biological variability of cells, providing new insight into cell regeneration therapy.
    OBJECTIVE: To study the Influence of CD133+ cells on human umbilical cord blood mononuclear cell transplantation for treatment of heart failure.
    METHODS: Human cord blood mononuclear cells were isolated using lymphocyte separation medium method, and CD133+ and CD133- cells were sorted using immunomagnetic beads at a cell density of 1×108/L. Forty Sprague-Dawley rats were randomized into five groups: sham group, model group, CD133+ cell group, CD133- cell group and mononuclear cell group. Animal models of heart failure were made using intraperitoneal injection of isoproterenol in all the groups except for the sham group. Rats in the CD133+ cell group and CD133- cell group were given 1 mL CD133+ cells plus 1 mL PBS and 1 mL CD133- cells plus 1 mL PBS via the tail vein, respectively. Rats in the mononuclear cell group were given 1 mL CD133+ cells plus 1 mL CD133- cells via the tail vein, and those in the sham and model groups given 2 mL PBS via the tail vein. After 4 weeks, cardiac pathology, degree of myocardial fibrosis and colonization of CD133+ cells in myocardial tissues were observed.
    RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that myocardial tissues arranged disorderly in the model group, but regularly in the sham group; myocardial disorders were mildest in the CD133+ cell group, successively followed by the mononuclear cell group, and severest in the CD133- cell group and model group. Masson staining showed that in the model group, collagen fibers were proliferated, arranged irregularly and even broken, while in the sham group, the collagen fibers were less in number and arranged in order. Additionally, there was less reduction in collagen fibers and milder myocardial disorders in the CD133+ cell group compared with the other groups. Area of collagen fibers was increased significantly in all the groups except for the sham group (P < 0.05), but this increment was the minimal in the CD133+ cell group. Findings from immunohistochemistry and immunofluorescence staining showed that there were no CD133+ cells in the myocardial tissues of rats. Therefore, our data indicate that compared with the mononuclear cell transplantation, CD133+ cell transplantation exerts superiorities in relieving myocardial damage and reducing myocardial fibrosis. However, CD133+ cells are not colonized in the myocardial tissue after transplantation.

     

     

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    Protective effect of retinal stem cell transplantation on retinal ganglion cells in glaucoma
    Gu Zhi-min, Zhou Li-xiao, Qi Ruo
    2016, 20 (28):  4203-4209.  doi: 10.3969/j.issn.2095-4344.2016.28.015
    Abstract ( 362 )   PDF (4265KB) ( 246 )   Save

    BACKGROUND: Stem cell transplantation is a new method for blinding eye disease. But there is a lack of research about the protective effect of retinal stem cell transplantation on retinal ganglion cells in glaucoma.
    OBJECTIVE: To explore the protective effect of retinal stem cell transplantation on retinal ganglion cells of rats with glaucoma.
    METHODS: Forty-five Sprague-Dawley rats were randomly divided into three groups (n=15 per group) including control, model and retinal stem cell transplantation groups. Rat models of glaucoma were prepared in the latter two groups, and at 7 days after modeling, rats in the three groups were given intravitreal injection of 1 mL retinal stem cells (5x106 cells), the same amount of PBS, and no treatment, respectively. Subsequently, relative indicators were detected at 2 weeks after transplantation.
    RESULTS AND CONCLUSION: The expressions of brain-derived neurotrophic factor and insulin-like growth factor I protein as well as the number of retinal ganglion cells were the highest in the control group, followed by the retinal stem cell transplantation group model group, and the lowest in the model group (P < 0.05). The number of apoptotic retinal ganglion cells in model group was significantly higher than that of control group (P < 0.05), and which in the retinal stem cell transplantation group was significantly lower than that in the model group (P < 0.05), but higher than that in the control group (P < 0.05). These results suggest that retinal stem cell transplantation for rat glaucoma can exert a protective effect on retinal ganglion cells.

     

     

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    Insulin-secreting cells from induced pluripotent stem cells regulate blood glucose levels in vitro
    Lei Lei, Liang Ying-zi, Su Ying-jun, Ma Xian-jie, Cui Xin, Guo Shu-zhong
    2016, 20 (28):  4210-4217.  doi: 10.3969/j.issn.2095-4344.2016.28.016
    Abstract ( 450 )   PDF (5296KB) ( 292 )   Save

    BACKGROUND: Mouse pluripotent stem cells are induced to differentiate into insulin-secreting cells that can effectively improve blood glucose levels in diabetic mice.
    OBJECTIVE: To detect mRNA and protein levels of insulin-like cell clusters from induced pluripotent stem cells and to investigate the function of insulin-secreting cells in vitro and in vivo.
    METHODS: Mouse induced pluripotent stem cells cultured in vitro were induced to differentiate into insulin-secreting cells using combined inducers through three stages. The morphology of endodermal cells, islet-derived progenitor cells and mature islet cells in each stage was observed and relative gene expression levels were detected by PCR. Mature insulin-like cell clusters underwent dithizone staining and functions of insulin released in vitro were observed by ELISA assay. Finally, the insulin-secreting cells were transplanted into the subrenal capsule of diabetic mice, and then blood glucose levels were observed.
    RESULTS AND CONCLUSION: The mature spherical insulin-like cell clusters were successfully obtained in vitro, which were in iron red by dithizone staining, and expression of insulin mRNA was determined by PCR. The insulin-like cell clusters could secrete insulin in response to various blood glucose levels by ELISA assay. In addition, after the cells clusters were transplanted into the subrenal capsule of mice with type 1 diabetes, the blood glucose levels were marbedly improved.

     

     

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    Haploidentical hematopoietic stem cell transplantation for leukemia in 23 cases
    Jiang Qu, Zhang Hong-bin, Yang Li, Luo Hong, Miao Qiao, Deng Xue-mei
    2016, 20 (28):  4218-4225.  doi: 10.3969/j.issn.2095-4344.2016.28.017
    Abstract ( 276 )   PDF (4192KB) ( 225 )   Save

    BACKGROUND: Allogeneic hematopoietic stem cell transplantation (HSCT) is one of the effective methods in the treatment of leukemia. The haploidentical HSCT is an option for the patients who need a HSCT without a human leukocyte antigen (HLA)-matched donor.
    OBJECTIVE: To study the clinical efficacy of HLA-haploidentical HSCT on leukemia and its complications.
    METHODS: A total of 23 patients (4 cases of acute lymphoblastic leukemia, 12 of acute myelogenous leukemia, and 7 of chronic granulocytic leukemia) who had been treated with HLA-haploidentical HSCT from November 2007 to March 2015 were enrolled. Conditioning regimen I was set as cyclohexyl nitrosourea+cytarabine+busulfan+cyclophosphamide; regimen II as cyclophosphamide+total body irradiation; regimen III as fludarabine+cytarabine+busulfan+cyclophosphamide; and regimen IV as busulfan+cyclophosphamide. Cyclosporin A, mycophenlate mofetil, antithymocyte globulin and methotrexate were used to prevent graft-versus-host disease (GVHD). Hematopoietic remodeling, complications and prognosis were observed in all patients undergoing HLA-haploidentical HSCT.
    RESULTS AND CONCLUSION: Of the 23 patients, 22 achieved reconstitution of the granulocyte series, and 19 achieved reconstruction of the megakaryocyte series. Additionally, there were 7 cases of acute GVHD and 4 of chronic GVHD. Transplant-related mortality was 22% (5/23) within 100 days post transplantation including graft failure, acute GVHD, intracranial hemorrhage and disseminated infections. There were 14 cases of disease-free survival from 100 days to 24 months post transplantation, 2 cases of death due to GVHD and fungal infection, or recurrence and chronic GVHD, and 2 cases of recurrence under treatment. These findings indicate that HLA-haploidentical HSCT is an effective approach for the treatment of patients with leukemia, which is worth further investigation in clinical practice.

     

     

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    Cardiac stem cells improve the electrophysiological stability and ventricular fibrillation threshold via ANGII/AT1R/TGF-beta1/SMAD/CX43 signaling pathway
    Yan Ping, Hou Jing-ying, Zheng Shao-xin, Long Hui-bao, Zhong Ting-ting, Zhou Chang-qing, Guo Tian-zhu, Wu Quan-hua, Wang Tong
    2016, 20 (28):  4226-4233.  doi: 10.3969/j.issn.2095-4344.2016.28.018
    Abstract ( 234 )   PDF (1416KB) ( 282 )   Save

    BACKGROUND: Previous studies have demonstrated that the electrophysiological stability and ventricular fibrillation threshold after myocardial infarction in rats are significantly improved in the mid-term of cardiac stem cell transplantation, but relative regulatory mechanism and pathway remain unclear.
    OBJECTIVE: To explore the relative molecular regulatory mechanism of cardiac stem cells improving the electrophysiological stability and ventricular fibrillation threshold after myocardial infarction in rats. 
    METHODS: Myocardial infarction was induced in 20 Sprague-Dawley rats by ligation of the left anterior descending coronary, which were then randomized into two groups (n=10 per group) and were subjected to the injection of cardiac stem cells labeled with PKH26 in phosphate buffer solution (cardiac stem cell group) or the same amount of phosphate buffer solution (PBS) alone (PBS group) into the local infarct zone at 2 weeks after modeling, respectively. Six weeks later, relevant signaling molecules involved in the ANGII/AT1R/TGF-β1/SMAD/Cx43 pathway were all examined in myocardial tissues of the left ventricle and harvested blood samples.
    RESULTS AND CONCLUSION: Compared with the PBS group, expressions of connexin 43 in different zones of the left ventricle were significantly increased in the cardiac stem cell group (P < 0.01); there was a significant reduction of the angiotensin II level in plasma and different regions of the left ventricular (P < 0.05; P < 0.01). Furthermore, in the cardiac stem cell group, expressions of angiotensin II type I receptor, transforming growth factor-β1, SMAD2 and SMAD3 were significantly decreased (P < 0.01). Whereas SMAD7 was significantly elevated (P < 0.05) in different areas of the left ventricle compared with the phosphate buffer solution group. These findings suggest that the cardiac stem cell transplantation can improve the electrophysiological stability and ventricular fibrillation threshold after myocardial infarction by enhancing the expression of connexin 43 via ANGII/AT1R/TGF-beta1/SMAD/CX43 signaling pathway.

     

     

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    Preconditioning strategies for promoting mesenchymal stem cell homing
    Wang Guo-ren, Bai Zhi-ming
    2016, 20 (28):  4234-4242.  doi: 10.3969/j.issn.2095-4344.2016.28.019
    Abstract ( 311 )   PDF (1107KB) ( 292 )   Save

    BACKGROUND: Homing is the initial and key procedure of stem cells-based tissue restoration. Current studies have shown that the inability to recruit bone marrow mesenchymal stem cells to target tissue with high efficiency remains a significant barrier to tissue restoration. Preconditioning strategy provides a new insight to promote stem cell homing.
    OBJECTIVE: To review preconditioning strategies for promoting the homing of stem cells.
    METHODS: In PubMed database, different combinations of terms from “stem cell, mesenchymal stem cells, preconditioning, homing, migration” served as search terms to retrieve articles referring preconditioning strategies for promoting mesenchymal stem cell homing published from January 2000 to September 2015. According to the inclusion criteria, 72 articles were selected for final review.
    RESULTS AND CONCLUSION: Pretreating target tissue or mesenchymal stem cells ahead of cell transplantation, known as tissue preconditioning or cell preconditioning, prominently promotes the homing of mesenchymal stem cells, therefore enhancing tissue restoration effect. Tissue preconditioning is designed to up-regulate expression of chemokines by varying the local microenvironment, thereby increasing homing ability of mesechymal stem cells. Mesenchymal stem cell preconditioning strategies, for example, gene modification and cytokine induction, are mainly to up-regulate expression of chemokine receptors on the surface of mesenchymal stem cells as effectors, and thus promote targeted cell homing. Overall, preconditioning strategy will bring great hope to apply stem cell therapy into the clinic.

     

     

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    Traditional Chinese medicine in stem cell proliferation and tissue engineering
    Zhang Ye, Cui Yuan-lu
    2016, 20 (28):  4243-4249.  doi: 10.3969/j.issn.2095-4344.2016.28.020
    Abstract ( 244 )   PDF (1123KB) ( 248 )   Save

    BACKGROUND: Some Chinese herbal ingredients produce the analogical and pharmacological effects equivalent to growth factors, which can promote cell proliferation as well as tissue and organ recovery and regeneration. Additionally, they possess low price and relative stable physicochemical property. Thus, the applications of traditional Chinese medicine in tissue engineering has attracted more attentions of researchers all over the world.
    OBJECTIVE: To review the application of the traditional Chinese medicine in stem cell proliferation and tissue engineering.
    METHODS: A computer-based retrieval of PubMed and CNKI databases was performed for articles concerning the application of traditional Chinese medicine in stem cell proliferation and tissue engineering published from 1978 to 2015. The keywords were “the traditional Chinese medicine, tissue engineering, application, stem cell, anti-inflammatory, cross linking, surface modification, osteoblast, blood vessel” in English and Chinese, respectively.
    RESULTS AND CONCLUSION: Natural medicine and the traditional Chinese medicine are expected to be applied in tissue engineering as a surrogate of growth factor and antibiotics, which exert anti-inflammatory, angiogenesis, surface modification, anticoagulation and cross-linking roles as well as promote proliferation and differentiation of stem cells in tissue engineering.

     

     

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    Endometriosis and stem cell theory
    Yang Yun
    2016, 20 (28):  4250-4256.  doi: 10.3969/j.issn.2095-4344.2016.28.021
    Abstract ( 431 )   PDF (1048KB) ( 258 )   Save

    BACKGROUND: Pathogenesis of endometriosis involves many factors that ultimately lead to the emergence of endometrial glands and stroma outside the uterine cavity.
    OBJECTIVE: To review the research progress in the theory of endometrial stem cells, and to provide suggestions for the pathogenesis and treatment of endometriosis.
    METHODS: VIP database and PubMed database were retrieved for articles addressing endometriosis stem cell theory, including original research, experimental analysis, case analysis and review, published from January 2005 to October 2015. Keywords were "endometriosis, stem cells" in Chinese and English, respectively. After removal of repetitive articles, 48 articles were included for final analysis.
    RESULTS AND CONCLUSION: Endometrial stem/progenitor cells may exist in the endometrium, and these cells and bone marrow stem cells are speculated to be involved in the occurrence of endometriosis. Therefore, treatment of endometriosis is targeted to endometrial stem/progenitor cells and abnormal regulation on signal transduction pathway, which provides new insight into the mechanism of action and treatment of endometriosis.

     

     

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