Objective:To investigate the inhibitory effect of antisense human telomerase RNA(hTR) gene on implanted hepatocellular carcinoma in nude mice.Methods: HepG2 cells were subcutaneously inoculated into BALB/c nude mice at the axilla to establish implanted hepatocellular carcinoma model.The retrovirus plasmid containing antisense telomerse RNA(PLXSN-hTR-BamHⅠ) was injected into the tumor(0.2 ml every time,5 times).Retrovirus plasmid containing sense telomerase RNA(PLXSN-hTR-EcoRⅠ) and normal saline were inoculated as control groups.Tumor volume was determined and the inhibitory rate was calculated.Tumor necrosis was observed by histological analysis and cell apoptosis was analyzed by terminal transferase dUTP nick end labeling(TUNEL).Results: Tumor growth in antisense hTR group was significantly inhibited compared with the two control groups.The tumor inhibitory rate(26.78%) of antisense hTR group was significantly higher than that of sense hTR group(1.93%,P

Objective To observe the influence of preoperative regional chemotherpy on colon cancer cell cycle.Methods30 colorectal cancer patients received intraarterial chemotherapy 10 days before the radical resection.The expression of p16 and Rb protein was examined by immunohistochemistry.Result was compared with the control group.Results Labeling index (LI) of p16 protein was (35?19)% in treatment group, while in control group L1 was (16?8)%,( P

Objective To explore the motion features of cervical spine based on continuous X-ray images.Methods The cervical spontaneous continuous multi-frame sagittal images from flexion to extention positions were selected from cervical spondylosis patients (patients group) and healthy adult (normal group).After preprocessing and registration,the trajectory of single vertebral body were recorded,and the feature points of each vertebral body were extracted.Meanwhile,the rel ative geometry parameters and movement rate were calculated.Results The motion trajectory of patients' cervical spine C4-6 were different from healthy people.The angles between the left edge curve of the vertebral body (C4 and C5 vertebra) and the baseline of patients group were smaller than those of normal group in flexion position (all P＜0.05).There were instability in the movement of C4 vertebral body in patient group,and the volatility of the angle change,the rate of change and the frequency were larger.The relative position change of the adjacent single vertebral body in the patient group are smaller.Conclusion Through the preprocessing,registration,parameter extraction and result analysis,the changes of functional features in cervical spondylosis patients are truly reflected.And it also provides a new idea for dynamic analysis of cervical vertebrae based on X-ray images.

BACKGROUND: Some studies have showed that after indirect co-culture, bone marrow mesenchymal stem cells can differentiate into myocardial cells and hepatocypte-like cells. OBJECTIVE: To investigate the possibility of human umbilical cord blood mesenchymal stem cells (UCB-MSCs) to differentiate into hepatocytes by co-culture with human hepatocyte line LO2 in vitro.METHODS: Full-term umbilical cord blood samples were obtained sterilely. The UCB-MSCs were isolated by density gradient centrifugation and directly adherence growth, then passaged with trypsin digestion at 80% cell fusion. By utilizing cell culture plate insets with microporous membrane combined with 6-well plate, the LO2-/UCB-MSCs co-culture system was established. UCB-MSCs were plated into the wells of 6-well plate at a density of 1×10~7/L. LO2 cells were plated into the cell culture plate insert at a density of 1×10~5/L. UCB-MSCs were plated in both layers in the control group. Surface markers of adhered cells were detected by flow cytometry. Morphological changes of UCB-MSCs were observed by inverted phase contrast microscopy. mRNA expression was detected by reverse transcriptase PCR. RESULTS AND CONCLUSION: HUCB MSCs expressed CD44 and CD29 strongly, but CD34 and CD45 were expressed negatively. After 5 days, fusiform-shaped cells were reduced in the co-culture group; while, the time passing by, cells shaped irregular round or polygonal were increased, which were similar to hepatocytes. At 4 weeks after culture, UCB-MSCs were still fusiform-shaped in the control group. At day 5 after culture, alpha fetoprotein mRNA expressed positively, but other expressed negatively in the co-culture group; at day 14 after culture, cytokeratin-19 mRNA and albumin mRNA expressions were observed; moreover, with the time passing by, the expression of albumin mRNA was increased, but the expression of alpha fetoprotein-19 mRNA was decreased. Antigenic expressions in the control group were negative. This suggested that UCB-MSCs could differentiate into hepatocypte-like cells by co-culture with human hepatocyte line LO2 in vitro.

Objective To explore the feasibility of labeling in vitro rabbit osteoblasts with Qtracker and the features of Qtracker-labeled rabbit osteoblasts. Methods A healthy male rabbit, 3 months old, weighing 2 kg, was used in this study. After bone marrow was aspirated, bone marrow stromal cells (BMSCs) were isolated and cultured using the adherence method in vitro. The third passage of BMSCs was induced into osteablasts before incubation with Qtracker at concentrations of 1, 2, 4, 8, 16, 32 nmol/10~6 cells (Groups A, B, C, D, E, F respectively). Cells not labeled by Qtracker served as negative control (Group G). The following parameters were measured: induction, differentiation and determination of rabbit osteoblasts; the optimal mass concentration of Qtracker labeling by fluorescence microscopy and flow cytometry; the cell sur-vival rates at various concentrations of Qtraeker labeling by trypan-blue exclusion; Qtracker-labeled cell pro-liferation by MTr. Results The primary and the passage rabbit BMSCs were chiefly of fusiform shape. Rabbit BMSCs differentiated into osteoblasts following induction. The osteoblasts cytoplasm showed green fluorescence under fluorescence microscopy after being labeled by Qtracker. The mean labeling rate increased with the increased concentration of Qtracker, reaching up to (93.58±2.08) % after incubation at 8 nmol/ 10~6 cells by fluorescence microscopy, and (95.24±1.31) % by flow cytometry. There were no significant differences between Groups D, E, F(P>0.05), but significant differences were found between Groups A, B, C and Groups D, E, F (P<0.05). The labeling rate for Group G was 0. The cell survival rates were all above 96% (P>0.05) . No significant differences were found in the cell proliferation among various con-centrations (P>0.05). Conclusions Qtraeker can be used as a labeling marker for rabbit osteoblasts. When the concentration is at 8 nmol/10~6 cells, optimal labeling effect can be achieved. Rabbit osteoblasts labeled with Qtracker are of high efficiency and safety.

Objective To explore the proliferation and differentiation of osteoblasts from 2 sources co-cultured with SD rat Schwann cells(SCs) . Methods Bone marrow stromal cells (BMSCs) were obtained by washing the femoral and tibial bone marrow cavities in SD rats. Osteoblast differentiation of the third passage of BMSCs was induced by incubation in osteogenic medium. Primary rat calvarial osteoblasts were obtained by digestion of the calvarial bone in one day old SD rats. The cells were cultured in DMEM supplemented with 10% fetal bovine serum(FBS) . SCs of passage 2 were obtained by digestion of sciatic nerve. The SCs were identified by S-100. The proliferation of 2 kinds of osteoblasts co-cultured with SCs was tested using 96 co-culture plate by methyl thiazdyl tetrazolium(MTF). Real-time PCR was used to test the osteoblast differentiation through co-culturing with SCs in 3 d and 7 d. The osteoblasts were implanted in the subtus chamber. The SCs were implanted in the superior chamber. Results SCs enhanced significantly the proliferation of calvarial osteoblasts at 7 time points. The expression levels of OPN mRNA, OCN mRNA, ALP mRNA, and BMP-2 mRNA of the osteoblasts were significantly lower in the experiment group than in the control group in 3 d and 7 d. SCs also enhanced significantly the proliferation of the induced osteoblasts in 5 d, 7 d and 9 d. The expression levels of OPN mRNA, OCN mRNA, ALP mRNA, and BMP-2 mRNA of the induced osteoblasts were significantly higher in the experiment group than in the control group in 3 d and 7 d, except the level of ALP mRNA in 7 d.Conclusions The BMSCs-induced osteoblasts cocultured with SCs may be used as seed cells to construct neurotized tissue engineered bone.

In order to remove the endotoxin from the blood of endotoxemia patients, we prepared a new adsorbent with heparin space arm and polymyxin B (PMB) ligand. The carrier of chloromethyl polystyrene resin was activated and heparin space arm was grafted, and then PMB ligand was immobilized onto adsorbent with glutaraldehyde. We employed in vitro FITC-lipopolysaccharide (FITC-LPS) static adsorption to characterize the adsorption properties on the adsorbent, and conducted in vitro lipopolysaccharide (LPS) static adsorption to measure quantitavely the adsorption capacity and rate, and then evaluated the blood compatibility. The in vitro static adsorption indicated that the adsorbent had the removal rate of LPS above 70% with the adsorption equilibrium time for 2 hours. Blood compatibility experiment showed that the adsorbent had little negative effects on blood cells and plasma protein, and their adsorption rates were less than 10% for hemocytes and 20% for plasma protein respectively. This adsorbent exhibited high selectivity, high adsorption capacity and good biocompatibility, and presented a promising clinical application in the treatment of endotoxemia.
Adsorption ; Endotoxemia ; therapy ; Endotoxins ; isolation & purification ; Hemofiltration ; instrumentation ; methods ; Heparin ; chemistry ; Humans ; Ion Exchange Resins ; chemistry ; Ligands ; Polymyxin B ; chemistry ; Sorption Detoxification ; methods

Objective:To investigate the relationship between manganese superoxide dismutase (MnSOD) gene Val16Ala polymorphism and the susceptibility to prostate cancer.Methods:All literatures related to MnSOD gene polymorphism and susceptibility to prostate cancer were retrieved from PubMed, Embase, China Knowledge Network and Wanfang databases. The retrieval time was from the database establishment to September 19, 2018. Literature screening and data extraction were independently done by 2 investigators, and Meta-analysis was performed by using Stata 12.0 software.Results:A total of 17 studies were included in the final analysis, including 7 101 prostate cancer cases and 9 146 healthy controls (people working at hospital and the ordinary people). The results of Meta-analysis showed that MnSOD gene Val16Ala polymorphism was not associated with susceptibility to prostate cancer under the homozygous model ( OR = 1.12, 95% CI 1.02-1.23, P = 0.435), heterozygous model ( OR = 1.14, 95% CI 1.05-1.24, P = 0.765), dominant model ( OR = 1.14, 95% CI 1.05-1.23, P = 0.552) and allele comparison model ( OR = 1.07, 95% CI 1.02-1.12, P = 0.106). Conclusion:There may not be the relationship between MnSOD gene Val16Ala polymorphism and the susceptibility to prostate cancer.

Traumatic brain injury (TBI) contributes to the key causative elements of neurological deficits. However, no effective therapeutics have been developed yet. In our previous work, extracellular vesicles (EVs) secreted by stem cells from human exfoliated deciduous teeth (SHED) offered new insights as potential strategies for functional recovery of TBI. The current study aims to elucidate the mechanism of action, providing novel therapeutic targets for future clinical interventions. With the miRNA array performed and Real-time PCR validated, we revealed the crucial function of miR-330-5p transferred by SHED-derived EVs (SHED-EVs) in regulating microglia, the critical immune modulator in central nervous system. MiR-330-5p targeted Ehmt2 and mediated the transcription of CXCL14 to promote M2 microglia polarization and inhibit M1 polarization. Identified in our in vivo data, SHED-EVs and their effector miR-330-5p alleviated the secretion of inflammatory cytokines and resumed the motor functional recovery of TBI rats. In summary, by transferring miR-330-5p, SHED-EVs favored anti-inflammatory microglia polarization through Ehmt2 mediated CXCL14 transcription in treating traumatic brain injury.
Animals ; Brain Injuries, Traumatic/therapy* ; Chemokines, CXC/metabolism* ; Extracellular Vesicles/metabolism* ; Histocompatibility Antigens/metabolism* ; Histone-Lysine N-Methyltransferase/metabolism* ; Humans ; MicroRNAs/metabolism* ; Microglia/metabolism* ; Rats ; Stem Cells/metabolism*

Objective: To investigate the morphology and position changes of displaced disk with reduction after treatment by modified Twin-block. Methods : 19 patients were diagnosed as displaced disk with reduction and they were treated with modified Twin-block from July 2015 to June 2016. 28 temporomandibular joints (TMJ) of these patients were included in the analysis. The disk length, disk-condylar distance, and disk-condylar angle were measured with MRI before and after treatment. Paired t-test was used. Disk morphology before and after treatment was also documented and analyzed by Wilcoxon signed rank test. The statistical significance was set at P<0.01. Results : 28 TMJ disks were anteriorly displaced with reduction. 24 of them were repositioned while the other 4 were still anteriorly positioned after treatment. The disk length was increased significantly (P<0.01) while the disk-condylar distance and disk-condylar angle were decreased significantly after treatment (P<0.01). The disk morphology as hemiconvex (16) and bi planar (9) were the majority before treatment, while biconcave (16) and biplanar (10) were changed to the larger part after treatment. There was improvement on the disk deformation with a statistical significance (P<0.01). Larger disk-condylar distance, disk-condylar angle and severer deformation of disks were observed in the 4 disks without reposition. Conclusion Modified twin-block is an effective appliance for disk displacement with reduction by repositioning the disk and modifying the disk deformation. However the effect is not good for disks with severer deformation and displacement. Further studies with larger sample and stratified group are still needed.