1.Effect of ABCE1 gene silencing by electroporation on proliferation and migration of human glioma cells line U87MG
Journal of Chinese Physician 2014;16(11):1508-1512
Objective To investigate the effect of ATP-binding cassette protein E 1 (ABCE1) gene silencing by electroporation on the survival,cell cycle and invasion of human glioma cells line U87MG.Methods The siRNA against ABCE1 was constructed and transfected into U87MG cells by electroporation.The expression of ABCE1 was detected by real time-polymerase chain reaction (RT-PCR) and Western blot.Flow cytometry was used to detect the cell cycle distribution and apoptosis.The effects of ABCE1 gene silencing by electroporation on proliferation,migration and invasion of U87MG cell line were evaluated by cell counting kit-8 (CCK-8) assay,wound closure assay,chemotactic migration,and cell invasive experiments,respectively.Results Compared to the control and blank groups,the mRNA and protein levels were significantly decreased in the experimental group when ABCE1 gene silencing by electroporation.The cell cycle was arrested at G0/G1 phase,and cell number in S phase was decreased in U87MG cell line (P < 0.05).The cell growth inhibition ratio in the experimental group was significantly higher than that in the control and blank groups (P <0.01).Compared to the control and blank groups,the experimental group U87MG cell proliferation was inhibited significantly (P < 0.05).Scratch healing experiments showed the experimental group migration ability was decreased significantly (P < 0.05).Transwell chamber method showed the experimental group U87MG cell invasion ability was decreased significantly (P < 0.05).Conclusions ABCE1 is involved in the progression of human glioma cells,and inhibiting the expression of ABCE1 by electroporation can decrease migration,invasion,and proliferation ability of tumor cells in vitro.
2.Propofol intervention affects spinal cord edema and hindlimb electrophysiology in a model of spinal cord injury
Chinese Journal of Tissue Engineering Research 2016;20(5):640-645
BACKGROUND:A large number of studies have verified that propofol could effectively reduce secondary nerve injury by improving microenvironment of spinal cord injury. OBJECTIVE: To study the effects of propofol on spinal cord edema and electrophysiology of the hind limb in rats with spinal cord injury. METHODS: Rat models of acute spinal cord injury were established by using the modified Alen method. A total of 40 rat models were randomly divided into spinal cord injury group and propofol group (n=20). Rats in the propofol group were injected with propofol through the caudal vein. The spinal cords of an additional 20 rats were exposed in the sham surgery group. Motor function was evaluated using BBB score and inclined plate test before modeling, 1, 3 days, 1-4 weeks after modeling. Neuronal apoptosis was detected after spinal cord injury using TUNEL assay at 72 hours after modeling. AQP4/9, matrix metaloproteinases 9/2 mRNA and protein expressions were measured using RT-PCR and western blot assay. At 4 weeks after modeling, pathological changes of the spinal cord were observed using immunohistochemistry and hematoxylin-eosin staining. Neurophysiological recovery was analyzed using motor evoked potentials and somatosensory evoked potentials. RESULTS AND CONCLUSION: (1) At 1-4 weeks after modeling, BBB score and inclined plate test score were higher in the propofol group than in the spinal cord injury group (P < 0.05), but lower than in the sham surgery group (P < 0.05). (2) The number of apoptotic cels was significantly more in the spinal cord injury group than in the propofol group (P < 0.05). No apoptotic cels were found in the sham surgery group. (3) At 72 hours after spinal cord injury, AQP4/9 and matrix metaloproteinases 9/2 mRNA and protein expression was higher in the propofol group than in the sham surgery group (P < 0.05). AQP4/9 and matrix metaloproteinases 9/2 mRNA and protein expression was significantly reduced in the propofol group (P < 0.05). (4) At 4 weeks after modeling, the spinal cord was loose, and the cavity was smal. Partial neuronal necrosis could be seen. The degree of nerve fiber density in the propofol group was between the sham surgery group and spinal cord injury group. (5) Motor evoked potentials and somatosensory evoked potentials were obviously recovered, the latency was short, amplitude was increased in the propofol group, which showed significant differences as compared with the sham surgery group and the spinal cord injury group (P< 0.05). Results suggested that propofol can reduce apoptosis in rat neurons after spinal cord injury, reduce spinal cord edema-related gene expression, and improve electrophysiological function and limb motor function.
3.Propofol with bone marrow mesenchymal stem cell transplantation improves the hind limb function and electrophysiological changes in rats with spinal cord injury
Chinese Journal of Tissue Engineering Research 2015;(41):6659-6664
BACKGROUND:Bone marrow mesenchymal stem cels can be used to repair neurons, but have no ideal outcomes on nervous system injuries. OBJECTIVE:To investigate the effects of bone marrow mesenchymal stem cel transplantation combined with propofol on the hind limb function and electrophysiological changes of rats with spinal cord injury. METHODS:Eighty adult Wistar rats were selected to make animal models of spinal cord injury, and then randomized into four groups (n=20): bone marrow mesenchymal stem cel group, control group, combination group, propofol group. At 6 hours after modeling, rats in these four groups were injectedvia the tail vein with bone marrow mesenchymal stem cel suspension, cel culture medium, bone marrow mesenchymal stem cel suspension+propofol solution, and propofol solution using a 1 mL syringe, respectively. Rat motor function was assessed by Basso Beattie Bresnahan score, modified Tarlov score and inclined plane test before and at 1 day, 3 days, 1-4 weeks after modeling. Under fluorescence microscope, the survival and distribution of PKH-26-labeled bone marrow mesenchymal stem cels were observed at 4 weeks after modeling, and meanwhile, hematoxylin-eosin staining was used for pathological observation. Horseradish peroxidase tracer analysis was performed to analyze regeneration of nerve fibers, and motor and somatosensory evoked potentials were used to analyze the neurophysiological recovery of rats. RESULTS AND CONCLUSION:(1) The motor function of the rat hind limb recovered best in the combination group, better in the bone marrow mesenchymal stem cel group and propofol group, but worse in the control group. (2) There were a smal amount of nerve axon-like structures and smal syringomyelia in the bone marrow mesenchymal stem cel group and propofol group, but the combination group had more axon-like structures and no syringomyelia. In the control group, no axons but spinal cord defects and syringomyelia formed. (3) The amount of horseradish peroxidase-positive nerve fibers and the number of PKH-26 positive cels were ranked as folows: control group < propofol group and bone marrow mesenchymal stem cel group < combination group. There were significant differences between the groups (P < 0.05). (4) The latencies of motor and somatosensory evoked potentials were ranked as folows: control group> propofol group and bone marrow mesenchymal stem cel group > combination group, and there were significant differences between the groups (P < 0.05). (5) Amplitudes of motor and somatosensory evoked potentials were arranged as folows: control group < propofol group and bone marrow mesenchymal stem cel group < combination group, and the differences were statisticaly significant (P < 0.05). These findings indicate that both propofol and bone marrow mesenchymal stem cels can promote synaptic regeneration and improve the electrophysiological function and motor function of rats with spinal cord injury. Their combination has a better role than propofol and bone marrow mesenchymal stem cels used alone.
4.CT manifestation and clinical analysis of appendiceal mucinous tumor
Dafeng XU ; Zhoupeng MA ; Yiping XIE ; Tianke WANG
Chinese Journal of Postgraduates of Medicine 2016;39(9):846-849,854
Objective To analyze the CT manifestation and clinical characteristics of appendiceal mucinous tumor for improving the diagnostic and therapeutic level. Methods The CT and clinical data of 7 patients with appendiceal mucinous tumor verified by histopathology were retrospectively analyzed. Results Among the 7 cases, mucinous cystadenomas was in 6 cases, and mucinous cystadenocarcinoma was in 1 case. One case had no obvious discomfort;3 cases visited because of pain on right hypogastrium and fever;3 cases had the medical history ofchronic appendicitis, among whom 2 cases had the mass on right hypogastrium which had existed for 1 day or 2 years. The unenhanced CT showed that all of 7 cases had the cystic tumors on right hypogastrium near the cecum, and the maximum traverse diameter was 25-208 (67 ± 27) mm. The cyst walls of mucinous cystadenoma in 6 cases were flimsy, symmetrical, 2.3-3.5 mm thickness and smooth. Three cases had cyst walls calcification; the cyst wall of mucinous cystadenocarcinoma was thick and asymmetrical, and the thickness of cyst wall was 3.5-5.7 mm. Small nodes could be found inside the walls. 7 cases had much mucilage, with CT value 14.0-33.5 HU. Four cases had slight septa. The enhanced CT showed that the cyst walls of mucinous cystadenoma in 6 cases were mild to moderate continuous enhancement during venous phase; the cyst wall and nodes mucinous cystadenocarcinoma was obvious and continuous enhancement. Four cases showed clear boundary, while 3 cases accompanied with acute inflammation showed dim edge. The enlargement of lymphatic nodes could be seen near mesentery in 1 case. All the 7 cases were treated by surgical treatment. One patient who survived after 26 months showed the metastasis of peritoneal pseudomyxoma after 20 months. The 6 patients with mucinous cystadenoma were followed up for 18 - 36 months, they did not had metastasis or recurrence by CT review. Conclusions Appendiceal mucinous tumor is often short of characteristic in clinical symptom and physical sign, but has favourable prognosis. CT is a vital tool for its diagnosis and identification.
5.Genetic mutation and clinical features of osteogenesis imperfecta type V.
Shizhen GUAN ; Xue BAI ; Yi WANG ; Zhigang LIU ; Xiuzhi REN ; Tianke ZHANG ; Mingyan JU ; Keqiu LI ; Guang LI
Chinese Journal of Medical Genetics 2017;34(6):797-801
OBJECTIVETo explore genetic mutations and clinical features of osteogenesis imperfecta type V.
METHODSClinical record of five patients (including one familial case) with osteogenesis imperfecta type V were retrospectively analyzed. Peripheral blood samples of the patients, one family member, as well as healthy controls were collected. Mutation of IFITM5 gene was identified by PCR amplification and Sanger sequencing.
RESULTSA heterozygous mutation (c.-14C>T) in the 5-UTR of the IFITM5 gene was identified in all of the patients and one mother. The clinical findings included frequent fractures and spine and/or extremities deformities, absence of dentinogenesis imperfecta, absence of hearing impairment, and blue sclera in 1 case. Radiographic findings revealed calcification of the interosseous membrane between the radius-ulna in all cases. Hyperplastic callus formation was found in 3 cases. Four had radial-head dislocation.
CONCLUSIONA single heterozygous mutation c.-14C>T was found in the 5-UTR of the IFITM5 gene in 5 patients with osteogensis imperfecta type V. The patients showed specific radiological features including calcification of interosseous membrane, hyperplastic callus formation, and radial-head dislocation.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Humans ; Male ; Mutation ; Osteogenesis Imperfecta ; diagnostic imaging ; genetics ; Young Adult
6.Advance of in vitro oral static biofilm model.
Huabing ZHAO ; Tingting SHI ; Fengzhu LI ; Rui WANG ; Fuping LU ; Zhengmei HUANG
Chinese Journal of Biotechnology 2022;38(9):3267-3278
Oral static biofilm model is an important tool for in vitro simulation of oral microecological environment, which has become an important method for studying the pathogenesis of various oral diseases and testing the efficacy of various drugs, oral care products and foods due to its low cost, high throughput, good reliability and easy operation. The establishment of oral static biofilm models allows the selection of different devices, inoculum sources, media, substrates and culture conditions according to the purpose of the study, and the evaluation of biofilm growth by various methods such as measuring biomass, metabolic activity, community structure and performing visualization analysis. This paper summarizes the methodological elements reported in recent years for the establishment and evaluation of oral static biofilm models, and analyzes and discusses the applicability of various methods in the hope of contributing to the research and production practice in related fields.
Biofilms
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Reproducibility of Results