To investigate the mechanism of cAMP/Ca2+ signaling pathway inducing bone marrow mesenchymal stem cells to differentiate into neuronal cells, we cultured the bone marrow mesenchymal stem cells D1 cells in the present study. D1 cells were divided into two groups: control group and salidroside inducing groups. Control group was cultured with complete culture solution D/F12, while salidroside inducing groups were induced with 100 mg·L–1 salidroside for different time periods (24, 48 and 72 hours). PCR-array assay was used to detect expression of 84 calcium related mRNA, and significantly different genes were chosen to analyse. As a result, there were 4 significantly upregulated mRNAs inclu-ding DNA damage-inducible transcript 3 (Ddit3), heat shock protein 5 (Hspa5), protein phosphatase 1 regulatory subunit (Ppp1r15a) and prostaglandin-endoperoxide synthase 2 (Ptgs-2), and 4 significantly downregulated mRNAs including glucagon (Gcg), interleukin 2 (Il2), tumor necrosis factor (Tnf) and somatostatin (Sst) in the cAMP/Ca2+ signaling pathway. They probably had an effect on the process of salidroside induced D1 cells differentiating into neuronal cells.
Objective To analyze the changes of gene expression profiles during the process that human bonemarrow mesenchymal stem cells (hBMSCs) are induced to differentiate into cardiomyogenic cells with 5-azacytidine (5-aza). Methods hBMSCs were isolated from marrow of obsolete ribs and induced with 5-aza. Then immunocytochemicalstaining was used to detect the expressions of α-actin, cardiac troponin T (cTnT), and connexin 43, and the percentage ofcTnT positive cells was tested with flow cytometry. In the process of differentiation, variation of gene expression was screenedwith Genechi ps Operating System of human gene expression profiles. And the differentially expressed genes were functionallyanalyzed and hierarchical clustered. Results When BMSCs were induced in vitro with 5-aza, part of the cells turnedinto myogenic cells morphologically. Before induction, immunocytochemical staining for α-actin and cTnT showed sl ightpositive and for connexin 43 showed negative. While after 3 weeks of induction, immunocytochemical staining for α-actin,cTnT, and connexin 43 showed all positive. With flow cytometry, the percentage of cTnT positive cells was 7.43% ± 0.02%before induction, but it was 49.64% ± 0.05% after induction. During differentiation, 1 814 differentially expressed geneswere reported by gene chi ps. Of them, 647 genes were divided into 5 groups with hierarchical clustering. They had variousbiological functions, involving signal transduction, cell metabol ism, prol iferation, differentiation, development, andtopogenesis. Conclusion hBMSCs can differentiate into cardiomyogenic cells with the induction of 5-aza in vitro. Multi plegenes related with signal transduction, transcri ption, and growth factors are involved during this process.
Objective To evaluate the early outcome of anterior cervical discectomy and fusion (ACDF) using a Zero-profile implant system (Zero-P) for interbody fusion in the treatment of cervical spondylosis. Methods Between March 2010 and June 2011, 25 patients with cervical spondylosis underwent ACDF with Zero-P. There were 13 males and 12 females with an average age of 44.2 years (range, 26-67 years), including 14 cases of nerve root cervical spondylosis, 6 cases of spinal cervical spondylosis, and 5 cases of mixed cervical spondylosis. The disease duration was 3-120 months (median, 25 months). Single segment was involved in 20 cases, 2 segments in 4 cases, and 3 segments in 1 case. A total of 31 Zero-P were implanted (3 at C3, 4, 8 at C4, 5, 12 at C5, 6, and 8 at C6, 7). Primary cervical operation was performed in 23 cases and re-operation in 2 cases. Before and after operation, the height of intervertebral space and the cervical Cobb angle were measured; clinical outcome was evaluated using visual analogue scale (VAS) score for pain in the neck and upper limb, and Japanese Orthopaedic Association (JOA) score for myelopathy; and intervertebral fusion and the incidence of dysphagia were also observed. Results All incisions healed by first intention. All the patients were followed up 12-16 months (mean, 13.9 months). Interbody bone fusion was obtained, and the fusion time was 2.7-6.0 months (mean, 3.8 months). Three patients had dysphagia after operation; symptom disappeared at 1 week and 3 months after operation in 2 cases and 1 case, respectively. No fixation loosening, subsidence, or breakage occurred. The height of intervertebral space was significantly improved (P lt; 0.05) from (4.5 ± 0.5) mm at preoperation to (6.0 ± 0.7) mm at 1 week and (5.7 ± 0.6) mm at 12 months after operation; the cervical Cobb angle was significantly improved (P lt; 0.05) from (11.9 ± 6.1)° at preoperation to (21.2 ± 4.1)° at 1 week and (20.2 ± 3.7)° at 12 months after operation; and there was no significant difference between at 1 week and 12 months after operation (P gt; 0.05). The VAS score was significantly reduced (P lt; 0.05) from 7.1 ± 0.8 at preoperation to 1.9 ± 0.8 at 3 months and 1.0 ± 0.5 at 12 months after operation; the JOA score was significantly increased (P lt; 0.05) from 9.6 ± 1.3 at preoperation to 13.5 ± 1.0 at 3 months and 14.9 ± 1.0 at 12 months after operation; and there was significant difference between at 3 months and at 12 months after operation (P lt; 0.05). Conclusion The early outcome of ACDF using a Zero-P in the treatment of cervical spondylosis is satisfactory and reliable, and it can restore and maintain the cervical alignment and disc height, and disc has low incidence of postoperative dysphagia.
Objective To evaluate the safety and effectiveness of applying self-stabilizing zero-profile three-dimensional (3D) printed artificial vertebral bodies in anterior cervical corpectomy and fusion (ACCF) for cervical spondylotic myelopathy. Methods A retrospective analysis was conducted on 37 patients diagnosed with cervical spondylotic myelopathy who underwent single-level ACCF using either self-stabilizing zero-profile 3D-printed artificial vertebral bodies (n=15, treatment group) or conventional 3D-printed artificial vertebral bodies with titanium plates (n=22, control group) between January 2022 and February 2023. There was no significant difference in age, gender, lesion segment, disease duration, and preoperative Japanese Orthopedic Association (JOA) score between the two groups (P>0.05). Operation time, intraoperative bleeding volume, hospitalization costs, JOA score and improvement rate, incidence of postoperative prosthesis subsidence, and interbody fusion were recorded and compared between the two groups. Results Compared with the control group, the treatment group had significantly shorter operation time and lower hospitalization costs (P<0.05); there was no significant difference in intraoperative bleeding volume between the two groups (P>0.05). All patients were followed up, with a follow-up period of 6-21 months in the treatment group (mean, 13.7 months) and 6-19 months in the control group (mean, 12.7 months). No dysphagia occurred in the treatment group, while 5 cases occurred in the control group, with a significant difference in the incidence of dysphagia between the two groups (P<0.05). At 12 months after operation, both groups showed improvement in JOA scores compared to preoperative scores, with significant differences (P<0.05); however, there was no significant difference in the JOA scores and improvement rate between the two groups (P>0.05). Radiographic examinations showed the interbody fusion in both groups, and the difference in the time of interbody fusion was not significant (P>0.05). At last follow-up, 2 cases in the treatment group and 3 cases in the control group experienced prosthesis subsidence, with no significant difference in the incidence of prosthesis subsidence (P>0.05). There was no implant displacement or plate-screw fracture during follow-up.Conclusion The use of self-stabilizing zero-profile 3D-printed artificial vertebral bodies in the treatment of cervical spondylotic myelopathy not only achieves similar effectiveness to 3D-printed artificial vertebral bodies, but also reduces operation time and the incidence of postoperative dysphagia.
There is a shared problem in current optical imaging technologies of how to obtain the optical parameters of biological tissues with complex profiles. In this work, an imaging system for obtaining the optical parameters of biological tissues with complex profile was presented. Firstly, Fourier transformation profilometry was used for obtaining the profile information of biological tissues, and then the difference of incident light intensity at different positions on biological tissue surface was corrected with the laws of illumination, and lastly the optical parameters of biological tissues were achieved with the spatial frequency domain imaging technique. Experimental results indicated the proposed imaging system could obtain the profile information and the optical parameters of biological tissues accurately and quickly. For the slab phantoms with height variation less than 30 mm and angle variation less than 40o, the maximum relative errors of the profile uncorrected optical parameters were 46.27% and 72.18%, while the maximum relative errors of the profile corrected optical parameters were 6.89% and 10.26%. Imaging experiments of a face-like phantom and a human’s prefrontal lobe were performed respectively, which demonstrated the proposed imaging system possesses clinical application value for the achievement of the optical parameters of biological tissues with complex profiles. Besides, the proposed profile corrected method can be used to combine with the current optical imaging technologies to reduce the influence of the profile information of biological tissues on imaging quality.
ObjectiveTo discuss the risk of abortion related to lamotrigine (LTG) and its safety profile during pregnancy. MethodsRetrospectively studied pregnant women in our epilepsy clinics who took LTG from 2011 to 2015 as monotherapy and experienced embryo damage or abortion. Here, we present an extensive review of related literatures regarding possible mechanisms, clinical features and safty of LTG during pregnancy. ResultsIn our study, fourty-five pregnancies were administered monotherapy LTG, and three of these patients suffered embryo damage. ConclusionsAlthough LTG is considered safe for pregnant women and the embryo or fetus,it also has risk of embryo damage or abortion, which should be carefully considered before prescription. Using monotherapy and the lowest effective drug dose, monitoring LTG serum concentrations during pregnancy, supplementing folate administration before and after conception and conducting regular prenatal diagnostic tests might reduce the risk of abortion.
ObjectiveTo analyze the clinical outcome of anterior cervical discectomy and fusion using a Zero-profile interbody fusion and fixation device (Zero-P) for cervical spondylotic myelopathy. MethodsBetween April 2011 and September 2013, 26 cases of cervical spondylotic myelopathy underwent anterior cervical discectomy and fusion with the Zero-P. Of 26 cases, 12 were male and 14 were female, aged 43-82 years (mean, 58.3 years). The disease duration was from 3 months to 10 years (mean, 5.9 years). The involved segments included C3,4 in 5 cases, C4,5 in 3 cases, C5,6 in 6 cases, and C6,7 in 12 cases. The clinical outcome was evaluated using visual analogue scale (VAS) score, Japanese Orthopaedic Association (JOA) score, and Neck Disability Index (NDI) score before operation and after operation. ResultsThe operations were successful and the operation time was 75-140 minutes (mean, 105 minutes); and blood loss was 20-150 mL (mean, 45 mL). There was no complications of infection, neural injury, esophageal fistula, prevertebral hematoma, or leakage of cerebrospinal. Dysphagia occurred in 1 case within 1 week after operation,and disappeared after 1 month. All patients were followed up for an average of 15.3 months (range, 12-18 months). The clinical symptoms were relieved after operation. During follow-up, no implant displacement or subsidence, screw breakage, and cervical instability were observed. At 3 and 12 months after operation, the VAS score and NDI reduced significantly (P<0.05); the JOA score increased significantly (P<0.05); and the intervertebral space height and the cervical Cobb angle improved significantly (P<0.05). But there was no significantly difference between at 3 and 12 months (P>0.05). According to JOA evaluation, the results were excellent in 14 cases, good in 10 cases, and fair in 2 cases, with an excellent and good rate of 92.3% at last follow-up. ConclusionThe clinical outcome of anterior cervical discectomy and fusion using a Zero-P is satisfactory and reliable in the treatment of cervical spondylotic myelopathy. It can restore the cervical physiological curve and the intervertebral space height and decrease the incidence of postoperative dysphagia.
ObjectiveTo analyze the expression profile changes of osteogenic-related genes during spontaneous calcification of rat bone marrow mesenchymal stem cells (BMSCs). MethodsBMSCs were isolated from 3-day-old healthy Sprague Dawley rats;cells at the 4th generation were used to establish the spontaneous calcification model in vitro. Spontaneous calcification process was recorded by inverted phase contrast microscope observation and alizarin red staining after 7 and 14 days of culture. For gene microarray analysis, cell samples were collected at 0, 7, and 14 days after culture; the differentially expressed genes were analyzed by bioinformatics methods and validated by real-time quantitative PCR (RT-qPCR) assay. ResultsRat BMSCs calcified spontaneously in vitro. When cultured for 7 days, the cells began to aggregate and were weakly positive for alizarin red staining. After 14 days of culture, obvious cellular aggregation and typical mineralized nodules were observed, the mineralized nodules were brightly positive for alizarin red staining. A total of 576 gene probe-sets expressed differentially during spontaneous calcification, corresponding 378 rat genes. Among them, 359 gene probe-sets expressed differentially between at 0 and 7 days, while only 13 gene probe-sets expressed differentially between at 7 and 14 days. The 378 differentially expressed genes were divided into 6 modes according to their expression profiles. Moreover, according to their biological functions, differentially expressed genes related to bone cell biology could be classified into 7 major groups:angiogenesis, apoptosis, bone-related genes, cell cycle, development, cell communication, and signal pathways related to osteogenic differentiation. In cell cycle group, 12 down-regulated genes were linked with each other functionally. Matrix metalloproteinase 13 (Mmp13), secreted phosphoprotein 1 (Spp1), Cxcl12, Mmp2, Mmp3, Apoe, and Itga7 had more functional connections with other genes. The results of genes Spp1, Mgp, Mmp13, Wnt inhibitory factor 1, Cxcl12, and cyclin A2 by RT-qPCR were consistent with that of gene microarray. ConclusionThe first 7 days after rat BMSCs were seeded are a key phase determining the fate of spontaneous calcification. Multiple genes related with cell communication, bone-related genes, cell cycle, transforming growth factor-β signaling pathway, mitogen-activated protein kinase signaling pathway, and Wnt signaling pathway are involved during spontaneous calcification.
The microarray technology used in biological and medical research provides a new idea for the diagnosis and treatment of cancer. To find different types of cancer and to classify the cancer samples accurately, we propose a new cluster ensemble framework Dual Neural Gas Cluster Ensemble (DNGCE), which is based on neural gas algorithm, to discover the underlying structure of noisy cancer gene expression profiles. This framework DNGCE applies the neural gas algorithm to perform clustering not only on the sample dimension, but also on the attribute dimension. It also adopts the normalized cut algorithm to partition off the consensus matrix constructed from multiple clustering solutions. We obtained the final accurate results. Experiments on cancer gene expression profiles illustrated that the proposed approach could achieve good performance, as it outperforms the single clustering algorithms and most of the existing approaches in the process of clustering gene expression profiles.
ObjectiveTo evaluate the effectiveness of very low profile/variable angle locking plate (VLP) internal fixation for posterior Pilon fractures extending to the medial malleolus by posteromedial approach. MethodsA retrospective analysis was made on the clinical data from 13 patients with posterior Pilon fractures extending to the medial malleolus between December 2011 and August 2012. There were 4 males and 9 females with an average age of 48.9 years (range, 23-68 years). Fractures were caused by falling in 9 cases and by traffic accident in 4 cases. The locations were the left ankle in 6 cases and the right ankle in 7 cases. According to the Orthopedic Trauma Association (OTA) classification, 6 cases were rated as type 43B1, 4 cases as type 43B2, and 3 cases as type 43B3. Twelve cases had fibular fractures, including 11 cases of Denis-Weber type B, 1 case of Denis-Weber type C. The interval of injury and operation was 7-14 days (mean, 11.4 days). Open reduction was performed and VLP internal fixation was used for posterior malleolar fracture by posteromedial approach. ResultsPrimary healing of incision was obtained in all patients. Tibial nerve palsy was observed in 2 cases, and was cured after oral administration of mecobalamin. Twelve cases were followed up 12-18 months (mean, 14.5 months). According to the Burwell-Charnley's radiological evaluation system, 11 cases achieved anatomical reduction, 1 case achieved fair reduction. The fracture union time was 3-6 months (mean, 3.7 months). No loosening or breakage of internal fixation occurred during follow-up. According to the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot scale, the results were excellent in 7 cases, good in 4, and fair in 1; the excellent and good rate was 91.7%. ConclusionVLP internal fixation for posterior Pilon fractures extending to the medial malleolus by posteromedial approach can achieve good short-term effectiveness. The high quality of reduction is acquired under direct vision with low rate of soft tissue complications.