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Our previous study demonstrated that human KIAA0100 gene is a novel acute monocytic leukemia-associated antigen (MLAA) gene. But the functional characterization of human KIAA0100 gene has remained unknown to date. Here, firstly, bioinformatic prediction of human KIAA0100 gene was carried out using online software;Secondly, human KIAA0100 gene expression was downregulated by the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9 system in U937 cells. Cell proliferation and apoptosis were next evaluated in KIAA0100-knockdown U937 cells. The bioinformatic prediction showed that human KIAA0100 gene was located on 17q11.2, and human KIAA0100 protein was located in the secretory pathway. Besides, human KIAA0100 protein contained a signal peptide, a transmembrane region, three types of secondary structures (alpha helix, extended strand, and random coil) , and four domains from mitochondrial protein 27 (FMP27). The observation on functional characterization of human KIAA0100 gene revealed that its downregulation inhibited cell proliferation, and promoted cell apoptosis in U937 cells. To summarize, these results suggest human KIAA0100 gene possibly comes within mitochondrial genome; moreover, it is a novel anti-apoptotic factor related to carcinogenesis or progression in acute monocytic leukemia, and may be a potential target for immunotherapy against acute monocytic leukemia.
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BACKGROUND:There is a certain cel subset in esophageal cancer tissues, with certain invasive and metastatic properties, which is closely related to the clinical therapeutic effect on tumors. OBJECTIVE:To isolate tumor stem cel spheres in human esophageal carcinoma cel lines KYSE-150 and TE-1 and to analyze their proliferation and invasion ability. METHODS:KYSE-150 and TE-1 cels were cultured in serum-free medium to observe the formation of cel spheres. Cel proliferation and invasion were detected using MTT and Transwel chamber culture. Surface markers of cels were detected using flow cytometry. RESULTS AND CONCLUSION: Cel spheres that were stably subcultured were obtained from KYSE-150 and TE-1 cels cultured in serum-free medium. The proliferation and invasion abilities of cel spheres were significantly stronger than those of parent cels (P < 0.05). The number of CD44+, CD271+ and CD44+CD271+ cels in TE-1 and KYSE-150 cel spheres was significantly higher than that in the TE-1 and KYSE-150 parent cels (P < 0.05). These experimental results show that cel spheres isolated from human esophageal carcinoma cel lines TE-1 and KYSE-150 have tumor stem cel properties as wel as strong proliferation and invasion abilities. And moreover, CD44 and CD271 can be used as important surface markers of esophageal carcinoma stem cels. Cite this article:Wang YL, Wang ZM, Wang Y, Tao YP, Han GY.Culture, differentiation, proliferation and invasion of tumor stem cels in human esophageal carcinoma cel lines KYSE-150 and TE-1. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):55-59.
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BACKGROUND:Stromal cel derived factor-1 is a smal molecular protein with a wide range of biological activity that can cause immune cel chemotaxis, and it also has a chemotactic effect on bone marrow stem cels and periodontal ligament cels. OBJECTIVE:To investigate the effect of stromal cel derived factor-1 with different concentrations on the proliferation of bone marrow stem cels and to probe the best concentration. METHODS:Bone marrow stem cels from beagle dogs were culturedin vitro and stimulated by different concentrations of stromal cel derived factor-1 (100, 200, 300 μg/L). MTT was used to detect the influence of stromal cel derived factor-1 on the proliferation of bone marrow stem cels so as to screen the best concentration of stromal cel derived factor-1. Then, stromal cel derived factor-1 at the best concentrations was used to intervene the bone marrow stem cels, and MTT was used again to detect the proliferation of bone marrow stem cels. RESULTS AND CONCLUSION:Stromal cel derived factor-1 at concentrations of 100, 200, 300 μg/L could promote the proliferation of bone marrow stem cels, and the effect was more notable at 200 and 300 μg/Lbut withno significant difference. Therefore, 200 μg/L was considered to be the best concentration of stromal cel derived factor-1 for intervention of bone marrow stem cels. Compared with the blank control group, 200 μg/L stromal cel derived factor-1 could significantly promote the proliferation of bone marrow stem cels. Taken together, stromal cel derived factor-1 can promote the proliferation of bone marrow stem cels, and its best concentration is 200 μg/L.
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BACKGROUND:Cel replacement therapy as an effective strategy for reconstruction of the central nervous system has very broad application prospects. OBJECTIVE:To investigate the effect of stereotactic transplantation of neural stem cels into the brain on the neuromotor function of craniocerebral trauma rats. METHODS:Twenty male Sprague-Dawley rats were equivalently randomized into study and control groups. Animal models of craniocerebral trauma were made using the improved free-fal method in the rats. Then, model rats in the study and control groups were given parenchymal transplantation of embryonic neural stem cels and the same volume of culture medium with no stem cels at 1 day after injury, respectively. Neuromotor function of rats was assessed based on the neurological severity scores. At 2 weeks after transplantation, brain tissues were taken for hematoxylin-eosin staining, anti-BrdU, glial fibrilary acidic protein, β-tubulin III and tyrosine hydroxylase immunohistochemistry staining. RESULTS AND CONCLUSION:The neurological severity scores in the study group were significantly lower than those in the control group at 1 and 2 weeks after injury (P< 0.05). In the study group, there were many BrdU-positive neural stem cels in the brain tissues, some of which were positive for glial fibrilary acidic protein, β-tubulin III and tyrosine hydroxylase; while in the control group, there was no BrdU-positive cel in the brain tissues. Experimental findings show that neural stem cels stereotacticaly transplanted into the brain can proliferate and differentiate in the brain lesion, and thereby notably improve the neuromotor function of rats with craniocerebral trauma.
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BACKGROUND:Isoflurane cannot only induce a wide range of large neuronal apoptosis, but also inhibit hippocampal neurogenesis in neonatal rats, thereby resulting in hippocampus-dependent learning and memory defects. OBJECTIVE:To investigate the isoflurane effect on proliferation and differentiation of the hippocampal neural stem cels. METHODS:Twenty-six Sprague-Dawley rats were randomly divided into air group and isoflurane group (n=13 per group). Rats in the isoflurane group were subjected to 2.5% isoflurane inhalation for 3 minutes folowed by 1.5% isoflurane inhalation for 4 hours. Rats in the air group only breathed in air. After the intervention, blood glucose and arterial blood gas changes were detected in the two groups. Additionaly, rats in the two groups were given intraperitoneal injection of 5-bromodeoxyuridine before and after intervention. At 24 hours after the last injection of 5-bromodeoxyuridine, brain tissues were taken to make frozen sections for immunofluorescence staining. RESULTS AND CONCLUSION:There were no significant difference in pH, PaO2, PaCO2, HCO3, BE and SaO2 levels between the two groups (P> 0.05). Compared with the air group, the number of BrdU+ cels was significantly less in the isoflurane group (P < 0.05), while the number of NeuroD+/BrdU+ cels was significantly higher in the isoflurane group (P < 0.05). The incidence of adverse reactions was 23% in the isoflurane group, which was significantly higher than that in the air group (7.7%;P < 0.05). These findings indicate that isoflurane can inhibit the proliferation of neural stem cels in the hippocampal dentate gyrus, and promote their differentiation into neurons.
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BACKGROUND:To establish a rapid and effective method to obtain sufficient spermatogonial stem cels that can meet the clinical need is urgent to be solved in the spermatogonial stem cel transplantation. OBJECTIVE:To study the effect of rhodiola polysaccharide on the proliferation of spermatogonial stem celsin vitro. METHODS:Under sterile conditions, spermatogonial stem cels and Sertoli cels were isolated from the testis of mice, and spermatogonial stem cels were seeded onto the feed layer of Sertoli cels. Then, the co-cultured cels were assigned into experimental group 1 (simple cel culture medium), experimental group 2 (cel culture medium containing 150 mg/L rhodiola polysaccharide) and experimental group 3 (cel culture medium containing 150 mg/L rhodiola polysaccharide, 1 U/L leukemia inhibitory factor and 10 μg/L glial cel line-derived neurotrophic factor). After 7 days of co-culture, flow cytometry was used to detect cel proliferation in vitro, and cel viability and positive expression of GFRa-1, Thy-1 and C-kit were calculated. RESULTS AND CONCLUSION:After 7 days of co-culture, the cels grew rapidly and presented with colony and thyrsiform growth, and the number of cel masses increased significantly, al of which were in line with the proliferative features of spermatogonial stem cels. The GFRa-1, Thy-1 and C-kit proteins were expressed in the cel membrane and cytoplasm, mainly in the cel membrane. The viability of spermatogonial stem cels and positive expression of GFRa-1 and Thy-1 were ranked as folows: experimental group 3 > experimental group 2 > experimental group 1, and there were significant differences between groups (P < 0.05). The positive expression of C-kit had no difference between experimental groups 1 and 2, but it was significantly higher in the experimental group 3 than the other two groups (P < 0.05). These findings indicate that rhodiola polysaccharide used alone or combined with leukemia inhibitory factor and glial cel line-derived neurotrophic factor can enhance the proliferative ability of spermatogonial stem celsin vitro.
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Abstract BACKGROUND:Previous studies have found that cryptotanshinone represses multiple tumors, but little is reported on its effect on renal carcinoma. OBJECTIVE:To explore the effect of cryptotanshinone on the proliferation and apoptosis of the renal carcinoma stem cels. METHODS:CD133+ renal carcinoma stem cels were separated from OS-RC-2 cels by immunomagnetic bead separation. Effects of 0, 0.2, 1, 5 mg/L cryptotanshinone on the proliferation and apoptosis of CD133+ renal carcinoma stem cels were detected by MTT and flow cytometry, respectively. Expression levels of Ki67, Bcl-2, Caspase-3 and p-Caspase-3 protein were detected by western blot assay. RESULTS AND CONCLUSION:After magnetic cel sorting, the percentage of CD133+ cels was increased from 6.32% to 82.73%, and there was a significant difference before and after cel sorting (P < 0.001). Cryptotanshinone could repress the proliferation of CD133+ renal carcinoma stem cels and promote cel apoptosis in a dose-dependent manner. The protein expression levels of Ki67 and Bcl-2 in the 5 mg/L cryptotanshinone group were significantly decreased compared with the control group, while the protein expression level of p-Caspase-3 protein was significantly increased. In addition, there was no difference in the protein expression of Caspase-3 between cryptotanshinone and control group. These findings indicate that cryptotanshinone may be a potent anticancer drug for the treatment of renal carcinoma by inhibiting expression of Ki67 and Bcl-2 and promoting protein expression of p-Caspase-3. Cite this article:Feng M, Jia MH. Cryptotanshinone effects on the proliferation and apoptosis of renal carcinoma stem cels. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):49-54.
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BACKGROUND:Studies have found that sodium arsenite can cause the malignant transformation and tumorigenicity of HaCaT cels, but whether low concentrations of sodium arsenite can cause the malignant transformation is rarely reported. OBJECTIVE:To study the effect of sodium arsenite on the malignant transformation of human immortalized keratinocyte cel lines. METHODS:HaCaT cels were treated with different concentrations of sodium arsenite. MTT assay was used to determine the effect of sodium arsenite on HaCaT cel morphology and proliferation, flow cytometry used to detect the effect of sodium arsenite on HaCaT cel cycle, and soft agar colony formation experiments assay used to determine the effect of sodium arsenite on HaCaT cel colony formation capacity. RESULTS AND CONCLUSION: HaCaT cels grew wel when the concentration of sodium arsenite was 5 mol/L, but the cel growth was inhibited under intervention with 10 and 50 mol/L sodium arsenite. HaCaT cels treated with 0.1 mol/L sodium arsenite were passaged to the 20th generation, and cel morphology had no notable changes; cels at passage 25 exhibited enlarged size and multiple nucleoli, which had a continued proliferation trend. Compared with the primarily cultured cels, 0.1 mol/L sodium arsenite-treated HaCaT cels at passages 15 and 25 had an increased proportion at S phase and G2/M phase, with strengthened proliferation ability and increased colony-forming efficiency, and moreover, the proliferation ability and colony-forming efficiency of passage 25 cels were higher than those of passage 15 cels. These experimental data show that high concentrations of sodium arsenite reduce HaCaT cel viability, and low concentrations of sodium sulfite have a certain influence on the morphology, cel cycle, proliferation ability and colony-forming efficiency of HaCaT cels, and moreover, the proliferation ability and colony-forming efficiency of human immortalized keratinocytes wil be strengthened with the increase of passage.
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BACKGROUND:Stem cels from the apical papila are a new kind of mesenchymal stem cels, and whether it can be used in root regeneration is the key to the present study. OBJECTIVE:To culture rat stem cels from the apical papila and periodontal ligament stem celsin vitro, and to compare the biology behaviors of these two kinds of cels, thereby providing experimental basis for the application of stem cels from the apical papila in root regeneration. METHODS:The apical papila, as wel as the periodontal ligament tissues from the healthy mandibular teeth of young rats were digested and cultured. Immunophenotypes of stem cels from the apical papila and periodontal ligament stem cels were detected by immunofluorescence technique. Then, cel growth curves were determined by MTT method and mineralized nodule formation was observed by alizarin red staining. RESULTS AND CONCLUSION:Stem cels from the apical papila and periodontal ligament stem cels were both positive for STRO-1. Stem cels from the apical papila were positive for CD90 and weakly positive for CD146. Periodontal ligament stem cels were positive for CD146 and weakly positive for CD90. The absorbance values of stem cels from the apical papila and periodontal ligament stem cels increased with the increasing of time and became stable at 8 days. Since the 4th day, the proliferation capacity of stem cels from the apical papila was significantly stronger than that of periodontal ligament stem cels (P < 0.05). Both of stem cels are visible to have mineralized nodule formation. Compared with the periodontal ligament stem cels, stem cels from the apical papila were stained obviously deeper and had more mineralized nodules. These results show that stem cels from the apical papila have stronger proliferation capacity and mineralization ability than periodontal ligament stem cels. Cite this article:Zhao L, Yu L, Yuan P, Zhou CM, Wu PL.Stem cels from the apical papila versus periodontal ligament stem cels: biological behaviors. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):113-117.
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BACKGROUND:Isoflurane is an anesthesia drug that has a certain effect on the nervous system. It possibly causes neurologic disorders through impacting nerve stem cel function or morphology. OBJECTIVE:To investigate the effects of isoflurane on the proliferation and differentiation of neural stem cels in the hippocampus of rats. METHODS:Neural stem cels from the hippocampus of neonatal Sprague-Dawley rats, aged 7 days, were induced and differentiated. Passage 3 cels were obtained and divided into two groups: isoflurane group (a mixture gas of 2.8% isoflurane, 5% CO2 and 95% O2) and control group (a mixture of 5% CO2 and 95% O2). After intervention of 6 hours folowed by 2 hours of routine culture, anti-BrdU monoclonal antibody immunofluorescent staining was used to detect cel proliferation, and western blot assay to detect the expression of β3-tubulin and glial fibrilary acidic protein. RESULTS AND CONCLUSION:Compared with the control group, the number of BrdU positive cels in the isoflurane group reduced significantly, indicating that isoflurane inhibits the proliferation of neural stem cels. Compared with the control group, the expression of glial fibrilary acidic protein in the isoflurane group up-regulated, but the expression of β3-tubulin had no changes, indicating isoflurane promotes the differentiation of neural stem cels into astrocytes. Cite this article:Min N, Hu QF, Li XP, Nie XH, Yang LL.Isoflurane effects on the proliferation and differentiation of neural stem cels in the hippocampus of neonatal rats. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):118-122.
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BACKGROUND:Studies have shown that cytokine inhibitor pirfenidone can inhibit biological activity of fibroblasts by regulating a variety of cytokines. It has made good progress in the research and application of anti-fibrosis of internal organs, but the effect and mechanism for hypertrophic scars and skin fibroblasts are unclear. OBJECTIVE:To investigate the effect of pirfenidone on human hypertrophic scar fibroblasts. METHODS:Human hypertrophic scar fibroblasts were cultured using tissue culture method. Passages 3-6 cel s grew wel in the logarithmic growth phase were col ected. Cel s were divided into the control group (0 g/L pirfenidone), 0.15, 0.3 and 1 g/L pirfenidone groups according to different mass concentrations. Cel s were intervened for 12, 36 and 48 hours. RESULTS AND CONCLUSION:MTT, reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay results demonstrated that compared with the control group, cel proliferation, transforming growth factorβ1 mRNA expression, types I and III col agen secretion were decreased in the 0.15, 0.3 and 1 g/L pirfenidone groups (P<0.05), and the decrease was most significant in the 1 g/L pirfenidone group (P<0.05). At 24, 48 and 72 hours after intervention, significant differences in inhibitory rate of cel proliferation and the secretion of types I and III col agen were detected among 0.15, 0.3 and 1 g/L pirfenidone groups (P<0.05). Results confirmed that pirfenidone apparently inhibited the secretion of col agen of hypertrophic scar fibroblasts cultured in vitro, transforming growth factorβ1 expression and cel proliferation and viability.
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BACKGROUND:In recent years, many reports have focused on inflammatory cytokines, growth factors and mechanical loads affecting the cartilage and subchondral regeneration, but there is a lack of comprehensive understanding about the mechanism of osteoarthritis. OBJECTIVE: To explore the correlation between function status of bone marrow mesenchymal stem cels and disease progression in patients with osteoarthritis. METHODS:Femoral bone marrow was extracted from patients with femoral neck fractures (control group), mild (mild group) and severe (severe group) osteoarthritis to isolate and culture bone marrow mesenchymal stem cels. Cel counting kit-8 was used to detect the proliferative ability of bone marrow mesenchymal stem cels from different patient groups, and passage 3 bone marrow mesenchymal stem cels were subject to 2-week chondrogenic induction folowed by toluidine blue staining. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cels were isolated and cultured from the femoral bone marrow of different groups. The proliferative ability of cels in the control group was significantly higher than that in the mild and severe groups. After chondrogenic induction, bone marrow mesenchymal stem cels varied obviously in the morphology that was from fusiform to qusi-circular or polygon, the percentage of nucleoplasm became smaler, and cels were positive for toluidine blue staining. The number of chondrocytes generated in the severe group was less than that in the control group, but there was no great difference in cel morphology. These findings indicate that the occurrence of osteoarthritis is negatively correlated with the functional status of autologous bone marrow mesenchymal stem cels.
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BACKGROUND:Studies have shown that stem cels transfected with human telomerase reverse transcriptase (hTERT) gene can stably express high-level telomerase activity and strengthen cel proliferation, which lays the foundation to establish geneticaly engineered immortalized stem cel lines. OBJECTIVE:To explore the effects of hTERT transfection on proliferation and cel cycle of rat fetal liver stem cels in vitro. METHODS:Rat fetal liver stem cels culturedin vitro were transfected by recombinant adeno-associated virus carrying hTERT genes. RT-PCR and western blot assay were used to detect the expression of hTERT mRNA and protein, respectively. Cel Counting Kit-8 method and cel growth curve were used to detect cel growth and proliferation. Changes in cel cycle distribution were determined by flow cytometry. RESULTS AND CONCLUSION:Compared with the control group and empty virus group, in the hTERT infection group, hTERT expressed at gene and protein levels, the growth rate of the cels increased, and the number of cels at G0/G1 phase and S phase was decreased and increased, respectively. The results show that recombinant adeno-associated virus as a vector of hTERT gene used for gene transfection can promote the in vitro proliferation of rat fetal liver stem cels and play an optimal role in cel culture.
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BACKGROUND:There is no clear understanding about the effect of let-7f and interleukin-6 (IL-6) on the proliferation of bone marrow mesenchymal stem cels and their relationship. OBJECTIVE: To explore the effects of expression levels of let-7f and IL-6 on the proliferation of bone marrow mesenchymal stem cels and their relationship. METHODS:(1) LV-rno-let-7f-up and LV-rno-let-7f-down were constructed and transfected into bone marrow mesenchymal stem cels of Sprague-Dawley rats, respectively. Then, there were four groups in the study: transfection upregulation group transfected with LV-rno-let-7f-up), transfection inhibition group (transfected with LV-rno-let-7f-down), negative control group (transfected with FU-RNAi-NC-LV), and untransfected group. The expression level of let-7f in each group was detected by qRT-PCR. The proliferation ability of cels and expression levels of IL-6 when let-7f expression was at different levels were detected by MTT, flow cytometry and ELISA. The expression of Cyclin D1 at mRNA and protein levels was detected by qRT-PCR and western blot, respectively. (2) To predict the potential target gene of let-7f, the wild-type/mutant IL-6 3’UTR reporter gene vectors were constructed, and cotransfected with let-7f/let-7f inhibitor respectively into the 293T cels to measure the luciferase. RESULTS AND CONCLUSION: Compared with the negative control group, the proliferative and cloning capacities of cels in the transfection upregulation group were higher; the number of cels was significantly decreased at G1 stage and increased at S stage, and the apoptotic cels were reduced in number (P 0.05). Luciferase activity of cels transfected with wide-type IL-6 3’UTR and let-7f was significantly reduced (P < 0.05). These findings indicate that up-regulation of let-7f can promote the proliferative and cloning capacities of bone marrow mesenchymal stem cels and reduce cel apoptosis, but downrelation of let-7f exhibits an inhibitory effect. Overexpression of IL-6 can suppress the proliferation of bone marrow mesenchymal stem cels, which is considered to be a target gene of let-7f, and let-7f may suppress the expression of IL-6 to promote the cel proliferation.
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BACKGROUND:Different local anesthetic solutions for tumescent liposuction are necessary and have a certain effect on the biological characteristics of human adipose stem cels. OBJECTIVE:To explore the effect of different tumescent local anesthesia on the proliferation and differentiation of human adipose stem cels culturedin vitro. METHODS: Passage 3 human adipose stem cels were intervened by different tumescent local anesthesia (bupivacaine, ropivacaine, lidocaine). Cels cultured in low-glucose DMEM served as controls. Proliferation ability, lactate dehydrogenase activity in the supernatant and percentage of cels under adipogenic differentiation were detected. RESULTS AND CONCLUSION:After intervention for 1, 2, 3, 4, 5 days, the supernatant lactate dehydrogenase activity in the bupivacaine, ropivacaine, lidocaine groups was significantly higher than that in the control group (P 0.05). Overal, these findings indicate that different tumescent local anesthesia (bupivacaine, ropivacaine, lidocaine) can al inhibit the proliferation of human adipose stem cels, but have no certain effects on the adipogenic differentiation.
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BACKGROUND:Studies have shown that microRNAs (miRNAs) have moderating effect on the renewal and differentiation of cancer stem cels. However, there is no complete understanding on the effect of microRNA-17-92 gene on gastric cancer stem cel renewal and proliferation. OBJECTIVE:To explore the effect of miRNA-17-92 in promoting self-renewal and proliferation of gastric cancer stem cels. METHODS:(1) The gradualy reduced miRNAs during gastric cancer stem cel self-renewal were investigated using miRNA array based on RNAs from differentiated and adherent cels. (2) The miRNA-17-92 was constructed and transfected to gastric cancer stem cels. (3) The effects of miRNA-17-92 on the self-renewal of gastric cancer stem cels were studied by tumor sphere assayin vitro. (4) The effects of miRNA-17-92 on the proliferation of gastric cancer stem cels were investigated by MTT assay and colony formation assay. RESULTS AND CONCLUSION:(1) miR-19b/20a/92a expression gradualy reduced in the adhesion and differentiation of gastric cancer stem cels. (2) The expression of lentivirus carrying miRNA-17-19 gene in MKN28 cels and CD44-/EpCAM- cels were significantly increased; transient transfection of pre-miR-19b/20a/92a increased the expression of CD44-/EpCAM- and MKN28 miRNA, transient transfection of pre-miR-19b/20a/92a antagonists reduced the expression of SGC7901 and CD44+/EpCAM+ miRNA; overexpression of lenti-miR-19b/20a/92a significantly increased the ability of gastric cels to form tumor spheres; chemotherapy drugs prolonged the survival time of lenti-miR-19b/20a/92a-infected cels; transient transfection of pre-miR-19b/20a/92a significantly increased the number of CD44+/EpCAM+ cels, but transfection of pre-miR-19b/20a/92a antagonist reduced the number of CD44+/EpCAM+ cels. (3) MTT proliferation assay showed that gastric cancer cel proliferation rate in miR-19b/20a/92a stably expressing group was faster than that in the control group. Transient transfection of miR-19b/20a/92a precursor accelerated the growth rate of gastric cancer cels, and transient transfection of its antagonist slowed down the growth rate of gastric cancer cels. Colony formation assay showed that transient transfection of miR-19b/20a/92a precursor significantly increased the colony formation number as compared with the control group; transient transfection of miR-19b/20a/92a antagonist reduced the colony formation as compared with the control group. These findings indicate that miR-19b/20a/92a gene presents with continuous deletion in gastric cancer stem cel differentiation process, and miRNA-17-92 gene can promote the renewal and proliferation of gastric cancer stem cels.
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BACKGROUND:Previous studies have shown that the survival state of oral mucosal cels directly is directly affected by the nutritional status of the body, therefore, it may be a good indicator of the surgical nutritional status. But whether this index also influences surgical trauma stimulus is stil unclear. OBJECTIVE: To observe the effect of breast augmentation with silicone implants on the survival status of oral mucosal cels. METHODS: Thirty-two female patients scheduled for breast augmentation with silicone implants, including 13 cases of reproductive history and 19 cases of non-reproductive history, aged 23-45 years old. Sub-G1 peak method and Ki-67 were used to detect the proliferation and apoptosis rates of oral mucosa cels before and after breast augmentation with silicone implants for statistical comparisons. RESULTS AND CONCLUSION:Al the 32 cases successfuly completed breast augmentation with silicone implants, and no serious adverse events occurred perioperatively. The oral mucosa cel apoptosis and proliferation rates were (27.3±6.2)% and (27.8±5.8)% before breast augmentation, and (27.8±5.8)% and (20.2±7.7)% after breast augmentation. There was no difference before and after breast augmentation (P > 0.05). These findings indicate that the proliferation and apoptosis of human oral mucosa cels cannot be significantly by breast augmentation-related trauma, and the survival state of oral mucosa cels has some value to assess the nutritional state of breast augmentation recipients.
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BACKGROUND:Studies have reported that puerarin can reduce bone resorption and promote bone formation. OBJECTIVE: To further explore the effects of puerarin at different doses on the proliferation of osteoblasts culturedin vitro. METHODS: Osteoblasts isolated from the cranium of newborn rats were culturedin vitro, and then passage 3 cels were cultured in DMEM medium with the presence of 0, 20, 40, 80 and 100 μmol/L puerarin+10% fetal bovine serum for 48 hours. The viability of osteoblasts, alkaline phosphatase activity and mineral node formation were determined using MTT, alkaline phosphatase kit and alizarin red staining, respectively. RESULTS AND CONCLUSION:The viability of osteoblasts treated with puerarin at either 40 or 80 μmol/L was significantly higher than that of the control group. Alkaline phosphatase activity and the number of mineral modules were significantly increased in osteoblasts cultured with puerarin at 40 or 80 μmol/L when compared with the untreated cels. These findings demonstrate that puerarin is able to promote osteoblast proliferation in vitro.
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BACKGROUND:Jiegu Qili Tablet is a compound Chinese medicine which can promote fracture healing, but its mechanism is stil unclear. OBJECTIVE: To observe the effects ofJiegu Qili Tablet on the proliferation and mineralization of MC-3T3 cels. METHODS:MC-3T3 cels cultured in vitro were selected. (1) After 24 hours of routine culture, cels were cultured in serum-free DMEM for another 24 hours to synchronize cel cycles. Then, the culture solution was removed, and cels were cultured in DMEM complete medium with (experimental group) or without (control group)Jiegu Qili Tablet solution. At 24 and 48 hours of culture, MTS assay was used to detect the proliferative ability of cels. (2) In the experimental group, MC3T3-E1 cels at a density of 1×105 were inoculated into 96-wel plates, and after 4 hours of culture, cels were cultured in an osteogenic induction medium. After 21 days of osteogenic induction, the mineralization of MC3T3-E1 cels was examined using alizarin red S staining. Cels in the control groups were treated without osteogenic induction. RESULTS AND CONCLUSION:Compared with the control group, the cel proliferation rate was up-regulated in the experimental group at 24 and 48 hours after culture (P < 0.05), and the number of calcium nodules was also larger in the experimental group at 21 days after osteogenic induction (P < 0.01). These findings indicate that Jiegu QiliTablet solution can up-regulate the proliferation and mineralization of MC-3T3 cels.
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BACKGROUND:Fetal liver stem cel s have the potential to differentiate into hepatocytes and bile duct cel s, and participate in the repair and reconstruction of the liver, which is an important source of hepatocytes. But there are a little amount of fetal liver stem cel s in human body, and how to obtain a certain number of high-purity fetal liver stem cel s is currently a hot research. OBJECTIVE:To construct a siRNA carrier that can effectively inhibit the expression of connexin 43 (Cx43) in rat fetal liver stem cel s, and to investigate the effect of Cx43 inhibition on the proliferation and cel cycle of fetal liver stem cel s cultured in vitro. METHODS:Fetal liver stem cel s were cultured by the suspension culture in vitro, siRNA sequences targeting Cx43 (Cx43-siRNA) and negative control sequence (NC-siRNA) were designed and synthesized. Then, rat fetal liver stem cel s were transferred electrophoretical y and divided into three groups:blank group, NC-siRNA group, Cx43-siRNA group. Real-time PCR and western blot were used to assess the knockdown efficiency. Cel ular proliferation was determined by cel growth curve and cel counting kit-8 assay. The cel cycle was analyzed by flow cytometry. RESULTS AND CONCLUSION:After transfection, the Cx43 gene and protein expression levels were declined dramatical y in the Cx43-siRNA, NC-siRNA and blank groups, and the cel s grew faster. The number of cel s at G0/G1 phase decrease, but the number of cel s in S phase increased. There were significant differences between the groups (P<0.05). Electrophoretic transfer of Cx43-siRNA can promote the proliferation of cultured fetal liver stem cel s and optimize the cel culture.