【Abstract】ObjectiveTo investigate the effect of genetic modulation of the hepatic graft with IL-10 during liver preservation in rat liver transplantation. MethodsEleven cases of orthotopic liver transplantation were performed in Lewis to BN rats according to the cuff’s technique. All rats were divided into 3 groups,which were control group(n=3), Lipo group(n=4) and Lipo-rIL-10 group(n=4). Lipofectamine 2000-pCR3.1 complex and Lipofectamine 2000-pCR3.1 rIL10 complex were respectively injected into portal vein and kept for 45 minutes to transfect grafts during cold preservation in vitro. All rats were killed on postoperative day 6. Serum samples were collected for decting IL-10 by means ELISA. Transgene expression of rIL-10 was assessed by means of RT-PCR and immunohistochemistry. ResultsIn Lipo-rIL-10 group, levels of IL-10 from suprahepatic vena cava were significant higher than those from infrahepatic vena cava (P=0.024), transgene expression of rIL-10 in Lipo-rIL-10 were higher than those of control group and Lipo group assessed by means of RT-PCR and immunohistochemistry. ConclusionDuring cold preservation in vitro through portal vein injection to donor liver, liposome mediated gene transfection can successfully achieve local gene expression.
Objective To investigate the expression of SAPCD2 in the lung adenocarcinoma cells, and to study the effect of SAPCD2 regulating Hippo signaling pathway on the proliferation, invasion, migration and apoptosis of the lung adenocarcinoma cells and its mechanism. Methods Quantitative real-time PCR (qRT-PCR) and Western blot were used to detect the expression levels of SAPCD2 mRNA and protein in four types of lung cancer cells (HCC827, H1650, SK-MES-1, A549) and human normal lung epithelial cells (BESA-2B), respectively. Then, lung cancer cells with relatively high levels of SAPCD2 expression were selected for subsequent experiments. The experiment cells were divided into a normal control group (NC group), a si-SAPCD2 group, and a pathway inhibitor group (si-SAPCD2+XMU-MP-1 group). Firstly, SAPCD2 mRNA was silenced using small interfering RNA (siRNA) technology, and then qRT-PCR was used to detect the expression of SAPCD2 in transfected lung cancer cells; using clone plate assay to detect the proliferation of lung cancer cells after silencing; using flow cytometry to detect the apoptosis of lung cancer cells after silencing; observe the number of lung cancer cells at different stages through cell cycle experiments; then Transwell experiment was used to analyze the effect of silencing SAPCD2 on the migration and invasion of lung cancer cell migration. Finally, Western blot was used to detect the expression of ki-67, Bcl-2, Caspase-3, NF2, P-MST1, P-LATS1, P-YAP, YAP, and TAZ proteins.Results SAPCD2 had the highest expression level in lung adenocarcinoma A549 cells (P<0.01). Silencing SAPCD2 significantly decreased the proliferation ability of A549 cells (P<0.01), inhibited their migration (P<0.05) and invasion (P<0.01), and promoted A549 cell apoptosis (P<0.01); more than half of the cells remained in the G0/G1 phase. Compared with the NC group, A549 cells showed a significant increase in G0/G1 phase cells (P<0.01), a significant decrease in G2/M and S phase cells (P<0.01), and a significant increase in the proportion of early apoptotic cells (P<0.01). Western blot results showed that silencing SAPCD2 down-regulated the expression of ki-67, Bcl-2, YAP, and TAZ proteins compared to the NC group (P<0.01), and up-regulated the expression of Caspase-3, NF2, P-MST1, P-LATS1, and P-YAP proteins (P<0.01). Conclusions The expression of SAPCD2 in lung adenocarcinoma A549 cells is significantly higher than that in normal lung epithelial cells (BESA-2B), which promotes the proliferation, migration and invasion of A549 cells and inhibits apoptosis. The mechanism may be related to the inhibition of Hippo signaling pathway.
Objective To investigate feasibility and curative effect of ultrasound-guided percutaneous transhepatic cholangioscopy in treatment of complicated hepatolithiasis. Methods The data of 42 patients with complicated hepatolithiasis from June 2012 to June 2017 in the Hepatobiliary Surgery, the First Affiliated Hospital of Xi’an Jiaotong University were retrospectively analyzed. All the patients were treated with ultrasound-guided percutaneous transhepatic cholangioscopy, including the first stage of dilation and drainage and the second stage choledochoscopy. Results The operations of the 42 patients were successfully performed. No case was converted to the conventional laparotomy. The puncture sites of 10 cases were at the right intrahepatic bile duct, 25 cases were at the left intrahepatic bile duct, and 7 cases were at the bilateral intrahepatic bile duct. The residual stones were removed by two stage choledochoscopy in the 31 patients, 11 patients had the residual stones. After the first stage, there were 4 cases of the bile duct hemorrhage, 8 cases of the cholangitis, 1 case of the pleural effusion and 1 case of the infection, 2 cases of the postoperative drainage tube shedding. After the second stage, there were 3 cases of the cholangitis and 3 cases of the postoperative drainage tube shedding. The stones of the 10/31 patients with stone removal occurred and the diseases of 9/11 patients with stone residual were stable during the following-up of (18.6±7.8) months. Conclusion Ultrasound-guided percutaneous transhepatic cholangioscopy including the first stage of dilation and drainage plus the second stage choledochoscopy is safe and effective in treatment of complex intrahepatic bile duct stones, it is an effective supplement to traditional surgery.
ObjectiveTo detect the difference in the osteogenesis ability of biphasic calcium phosphate (BCP) ceramic granular materials with different mesoporous diameters prepared at different sintering temperatures through in vivo and in vitro experiments, so as to provide evidence for screening BCP materials with better clinical application parameters.MethodsThree kinds of BCP (materials 1, 2, 3) were prepared by mixing hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP) at a ratio of 8∶2 and sintered at 1 050, 1 150, and 1 250℃ for 3 hours, respectively. The internal porosity and the diameter, volume, and area of the mesopore were measured by Brunauer-Emmett-Teller test (BET); the composition of the material was evaluated by X-ray diffraction (XRD); the microscopic surface morphology of the material was observed by scanning electron microscopy (SEM). The 3rd generation bone marrow mesenchymal stem cells (BMSCs) from Sprague-Dawley rats were co-cultured with the materials 1, 2, and 3 for 7 days in vitro respectively (groups A, B, and C), and the cells adhesion on the materials was observed by SEM and phalloidine staining, respectively. Cell proliferation activity was measured by cell counting kit 8 method. In vivo, 9 muscle bags were made in dorsal muscles of 9 beagles, respectively. The muscle bags were randomly divided into 3 groups (3 per beagle in each group) and materials 1, 2, and 3 were placed into the muscle bags of groups A, B, and C, respectively. After 1, 2, and 3 months of operation, 3 beagles were anesthetized and the samples were stained with HE, Masson, and Safranin, and the bone formation area ratio in the BCP gap was calculated. Real-time fluorescence quantitative PCR (qRT-PCR) was performed to detect the expressions of bone-related genes [including alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OC)].ResultsThe BET test showed that with the increase of sintering temperature, the internal porosity of the particles did not change significantly, but the diameter, volume, and area of the mesopores gradually decreased. The XRD detection showed that the XRD waves of HA and β-TCP could be seen in all 3 kinds of materials; SEM showed that there were widely distributed macropores on the surface of 3 kinds of BCPs, and the interpores connected with the others. In vitro, BMSCs adhered and proliferated on the surfaces of 3 kinds of BCPs, and the cell biocompatibility of the materials in groups B and C was better than that in group A. In vivo, obvious osteoid tissue deposition could be observed in the intergranular space of 3 kinds of BCPs from 2 months after implantation. The bone formation area ratio of each group increased with time. The bone formation area ratio in group A was significantly higher than that in groups B and C at 2 and 3 months after implantation, and in group A than in group B at 1 month (P<0.05). qRT-PCR showed that the expressions of osteogenic related genes peaked at 2 months in group A, and gradually increased with time in groups B and C. The relative expressions of ALP and OPN mRNAs in group A were significantly higher than those in groups B and C at 1 month after implantation, the relative expression of OC mRNA in group A was significantly higher than that in groups B and C at 2 months after operation, the relative expression of ALP mRNA in groups B and C and the relative expression of OPN mRNA in group B were significantly higher than those in group A, all showing significant differences (P<0.05); there was no significant difference in the relative expression of each gene among the other groups at each time point (P>0.05).ConclusionThe mesoporous diameter of BCP decreases with the increase of sintering temperature. Different mesoporous diameters lead to different ectopic osteogenesis of BCP materials. BCP material with mesoporous diameter of 12.57 nm has better osteogenic ability which can activate the osteogenic gene earlier. The mesoporous diameter is expected to be an adjustable index for optimizing the osteogenic capacity of BCP materials.
Stroke is an acute cerebrovascular disease in which sudden interruption of blood supply to the brain or rupture of cerebral blood vessels cause damage to brain cells and consequently impair the patient's motor and cognitive abilities. A novel rehabilitation training model integrating brain-computer interface (BCI) and virtual reality (VR) not only promotes the functional activation of brain networks, but also provides immersive and interesting contextual feedback for patients. In this paper, we designed a hand rehabilitation training system integrating multi-sensory stimulation feedback, BCI and VR, which guides patients' motor imaginations through the tasks of the virtual scene, acquires patients' motor intentions, and then carries out human-computer interactions under the virtual scene. At the same time, haptic feedback is incorporated to further increase the patients' proprioceptive sensations, so as to realize the hand function rehabilitation training based on the multi-sensory stimulation feedback of vision, hearing, and haptic senses. In this study, we compared and analyzed the differences in power spectral density of different frequency bands within the EEG signal data before and after the incorporation of haptic feedback, and found that the motor brain area was significantly activated after the incorporation of haptic feedback, and the power spectral density of the motor brain area was significantly increased in the high gamma frequency band. The results of this study indicate that the rehabilitation training of patients with the VR-BCI hand function enhancement rehabilitation system incorporating multi-sensory stimulation can accelerate the two-way facilitation of sensory and motor conduction pathways, thus accelerating the rehabilitation process.