Effects of biological clock gene Bmal1 on the expression of cell cycle-associated genes in chondrocytes / 基础医学与临床

Objective To explore the intrinsic relationship between circadian clock and cell cycle in osteoarthritis(OA)chondrocytes,especially the regulation of cell cycle-related genes by the clock gene Bmal1.Methods The chondroid ATDC5 cells induced by insulin-transfering-selenium(ITS)were divided into control group,OA group and LV-Bmal1 group.The cell viability of each group was detected by CCK8 method.The expression of Bmal1,Per1,Wee1,Cdk1,Ccnb1 and Mmp13 mRNA in each group was detected by RT-qPCR.The expression of BMAL1,PER1,WEE1,CDK1,CCNB1 and MMP13 protein in each group was detected by Western blot.The effects of Bmal1 on different stages of cell cycle and apoptosis was analyzed by flow cytometry.The regulation of Bmal1 on Per1,Wee1,Cdk1,Ccnb1 and Mmp13 and their roles in OA were analyzed.Results Compared with the normal group,the cell viability of the OA group was increased,the relative mRNA expression of Bmal1 and Wee1 in the OA group decreased,and the relative mRNA expression of Per1,Cdk1,Ccnb1 and Mmp13 increased signif-icantly.The cell viability of LV-Bmal1 group decreased,the relative expression of Bmal1 and Wee1 mRNA in-creased,and the relative expression of Per1,Cdk1,Ccnb1 and Mmp13 mRNA decreased(P＜0.05).Correlation analysis showed that Bmal1 was positively correlated with Wee1 and they were negatively correlated with Per1,Cdk1,Ccnb1 or Mmp13.The results of Western blot showed that protein expression in different groups were con-sistent with the trend of PCR.The results of cell cycle and apoptosis showed that compared with the normal group,the S phase and G2/M phase of the OA group were shortened,the proportion of cells decreased significantly,and the proportion of early and late apoptosis increased.The S phase and G2/M phase of the LV-Bmal1 group were prolonged,the proportion of cells was increased,and the proportion of early and late apoptosis was decreased.Conclusions Circadian clock gene Bmal1 in inflammatory chondrocytes might regulate the expression of cell cycle-related genes.

Diffusion tensor imaging analysis along the perivascular space derived intracerebral glymphatic system circulatory function on neuromyelitis optica spectrum disorder: a follow-up study for curative effect / 中华放射学杂志

Objective:To evaluate the value of curative effect in neuromyelitis spectrum disease (NMOSD) based on circulatory function evaluation of intracerebral glymphatic system by using diffusion tensor imaging analysis along the perivascular space.Methods:The clinical and imaging data of 23 patients diagnosed with NMOSD at Tianjin Medical University General Hospital from March 2018 to December 2019 were retrospectively analyzed in this study. The clinical data included expanded disability status scale (EDSS), average relapse rate (ARR) and retinal nerve fiber layer (RNFL) thickness at baseline and 1 year follow-up after treatment. Among the 23 NMOSD patients, there were 22 females and 1 male, aged from 21 to 71 (45±13) years old. All the patients underwent MR scans at both baseline and 1 year after treatment, and the scanning sequences included cerebral 3D-T 1WI, T 2WI, diffusion tensor imaging and cervical spinal sagittal 3D-T 2WI, and the cervical spinal cord volume and bilateral diffusion tensor imaging analysis along the perivascular space index (ALPS index) were calculated. The partial correlation test was used to analyze the correlations between ALPS index and the clinical indicators such as EDSS, ARR, and bilateral RNFL, with the control variables as gender, age, years of education and course of disease. The multiple linear regression model was used to analyze the independent predictors for ALPS index and EDSS after treatment. Receiver operating characteristic curve (ROC) and area under the curve (AUC) were used to evaluate the diagnostic value of NMOSD treatment outcome by using ALPS index. Results:When controlling for gender, age, years of education and course of disease, there were significant negative correlations between right ALPS index and EDSS ( r=-0.50, P=0.048), bilateral average ALPS index and EDSS ( r=-0.53, P=0.034), left ALPS index and ARR ( r=-0.58, P=0.018), while there was significant positive correlations between right ALPS index and RNFL ( r=0.88, P=0.008) at 1 year follow-up after treatment. Multiple linear regression analysis showed that cervical spinal cord volume was an independent impact factor of bilateral average ALPS indexes (β=0.24, 95%CI 0.10-0.38, P=0.002), and bilateral average ALPS indexes (β=-3.22, 95%CI -5.97--0.48, P=0.024) and right RNFL (β=-0.05, 95%CI -0.08--0.02, P=0.002) at baseline were the independent impact factors of EDSS after treatment. ROC curve analysis showed that the bilateral average ALPS index at baseline had the best efficacy in predicting the curative effect of NMOSD patients with AUC=0.92. Conclusions:After treatment, NMOSD patients with severe clinical disability, high frequency of disease attack, poor visual performance, and severe cervical spinal cord atrophy have more serious impairment of intracerebral glymphatic system circulatory function. The ALPS index could help in predicting the clinical curative effect of NMOSD patients.

Application of radiomics captured from CT to predict the EGFR mutation status and TKIs therapeutic sensitivity of advanced lung adenocarcinoma / 中华肿瘤杂志

Objective@#To explore the ability of computed-tomography (CT) radiomic features to predict the Epidermal growth factor receptor (EGFR) mutation status and the therapeutic response of advanced lung adenocarcinoma to EGFR- Tyrosine kinase inhibitors (TKIs) treatment.@*Methods@#A retrospective analysis was performed on 253 patients diagnosed as advanced lung adenocarcinoma, who underwent EGFR mutation detection, and those with EGFR sensitive mutation were treated with TKIs. Using the Lasso regression model and the 10 fold cross-validation method, the radiomic features of predicted EGFR mutation status and the screening of TKIs for sensitive populations were obtained. 715 radiomic features were extracted from unenhanced, arterial phase and venous phase, respectively.@*Results@#The area under curve (AUC) values of the multi-phases including unenhanced, arterial phase and venous phase of the EGFR mutation status validation group were 0.763, 0.807 and 0.808, respectively. The number of radiomic features extracted from the multi-phases were 5, 18 and 23, respectively, which could distinguish the EGFR mutation status. The AUC values of the multi-phases of the EGFR-TKIs sensitive validation group were 0.730, 0.833 and 0.895, respectively. The number of radiomic features extracted from the multi-phases were 3, 7 and 22, respectively, which can be used to screen the superior population for TKIs treatment. The efficiency of radiomic features extracted from venous phase in predicting EGFR mutant status and EGFR-TKIs sensitivity was significantly superior than those of unenhanced and arterial phase.@*Conclusions@#The radiomic features of CT scanning can be used as the radiomics biomarker to predict the EGFR mutation status of lung adenocarcinoma and to further screen the dominant population in TKIs therapy, which provides the basis for targeted therapy.

Progress of immunotherapy in neuromyelitis optica spectrum disorders / 中华全科医师杂志

Neuromyelitis optica spectrum disorder (NMOSD) is an autoimmune demyelinating disease in central nervous system,mainly involving the optic nerves and spinal cord.NMOSD is a relapsing disease with severe residual disability,therefore,prevention of attacks is critical for the preservation of visual and neurologic function.The conventional immunosuppressants,including prednisone,azathioprine,mycophenolate mofetil and methotrexate,can be used to prevent relapse of NMOSD.And immunotherapeutics such as rituximab,tocilizumab and eculizumab are increasingly used in recent years.This review summarizes the current status and the future strategies in treatment of NMOSD based on the progress of NMOSD pathogenesis.

Co-culture with vascular endothelial progenitor cells:effects on proliferation and apoptosis of neural stem cells and vascular remodeling in rats with ischemia reperfusion injury / 中国组织工程研究

BACKGROUND:Neural stem cel (NSC) transplantation is a common method for various ischemicencephalopathies, but inability to survive in the transplantation region limits its further use in clinical practice. OBJECTIVE:To explore the effect of vascular endothelial progenitor cel s (VEPCs) on the proliferation and apoptosis of co-cultured NSCs as wel as vascular remodeling in rats with ischemia reperfusion injury. METHODS:125 Sprague-Dawley rats were randomly divided into five groups, 25 rats in each group, including sham operation, ischemia, NSCs, co-culture, and VEPCs groups. Rat models of ischemia reperfusion injury were made in al groups except for the sham operation group, fol owed by corresponding interventions. The proliferation and apoptosis of neural stem cel s were detected, and vascular remolding in the ischemic region was observed in each group. RESULTS AND CONCLUSION:At different time points after transplantation, BrdU positive cel s were not observed in VEPCs, ischemia and sham operation groups;the number of BrdU positive cel s in the co-culture group was significantly higher than that in the NSCs group (P<0.05);BrdU+/Caspase-3+cel were observed in both co-culture and NSCs groups, and the apoptosis rate of the co-culture group was significantly lower than that in the NSCs group (P<0.05);there were new blood vessels in al the groups except for the sham operation group, and the number of new bone vessels was highest in the co-culture group. To conclude, our experimental results show that VEPCs promotes the proliferation of co-cultured NSCs, inhibits cel apoptosis and and promote angiogenesis in the ischemic penumbra of rats with ischemia reperfusion injury.

Application progress of disease modifying drugs in neuromyelitis optica spectrum disorder / 天津医药

Neuromyelitis optica spectrum disorder (NMOSD) is an autoimmune disease of the central nervous system that is characterized by inflammatory demyelinating lesions and axonal degeneration in optic nerve and spinal cord. It has high relapse rate and high disability rate. In recent years, as the studies on mechanisms of NMOSD, related immune therapy strategies have been developed rapidly. These immunologic interventions are collectively called the disease modifying therapy, which are helpful in improving the course and outcome of the disease. In this review, the latest treatment progress in NMOSD is summarized to guide clinical choice and application of the medication.

Clinical observation of baixuan xiatare capsules combined with ebastine tablets and orsalin ointment therapy in elderly patients with chronic eczema / 中国基层医药

Objective To observe clinical effects of Baixuan xiatare capsules combined with Baskin and compound Flumetasone on elderly patients with chronic eczema .Methods 160 elderly patients with chronic eczema were randomly divided into observation group and control group ,80 cases in each group , the observation group with XTR combined with Baskin , compound Flumetasone treatment , control group using Baskin combined with compound Flumetasone treatment for 4 weeks,the clinical efficacy was compared between the two groups .Results In the obser-vation group,the effective rate was 94.7%,the effective rate of control group was 82.1%,two groups have statistical significance with the efficiency difference (χ2 =5.862,P<0.05).Conclusion The XTR combined with Baskin and compound Flumetasone have better clinical effects than Baskin combined with compound flumetasone treatment with elderly patients with chronic eczema .

Body weight supported treadmill training on nerve cells differentiation after stem cells transplantation in rats of spinal cord injury / 中华物理医学与康复杂志

Objective To investigate the effect of bone marrow stem cells (BMSCs) transplantation combined with body weight supported treadmill training on neural functional recovery of rats with spinal cord injury.Methods T11 complete spinal cord injury (SCI) was introduced into 40 Sprague-Dawley male rats using an improved simple device,imitating the Allen method.The rats were randomly divided into a stem cell transplantation group,a body weight support treadmill training group,a combined treatment group and a control group,each of 10 assigned according to a random number table.One week after the SCI operation,stem cell transplantation was performed on the rats in the stem cell transplantation group and the combined treatment group.One day before their transplantation,the third passage BMSCs were labeled with 5-ethynyl-2'-deoxyuridine (Edu).The rats in body weight support treadmill training group and combined treatment group were received body weight support treadmill training,while the other two groups were not given any training.At the 1 st,2nd,3rd,4th and 5th week after SCI modeling,Basso-Beattie-Bresnahan (BBB) movement function score was used to evaluate the motor function recovery of all rats.At the 5 th week after SCI,immunohistochemical staining and immunofluorescence staining was applied to detect neural specific markers including the neuron specific enolase (NSE),microtubule associated protein (MAP-1 β) and vimentin (VIM).The survival and differentiation of the transplanted cells,as well as the nerve fiber recovery in the lesion were also observed.Results The average BBB score of the combined treatment group was (6.60 ± 0.97) at the 2nd week after SCI operation,significantly higher than the other 3 groups,while that of the stem cell transplantation group and the body weight support treadmill training group was (5.00 ± 0.67) and (4.80 ± 0.63) respectively,both higher than that of the control group but without significant differences (P ＞ 0.05).In the third week after modeling,however,the value of the stem cell transplantation group rose to (8.00 ± 0.67),significantly higher than that of the body weight support treadmill training group (6.80 ± 0.79).The immuohistochemical staining showed that a variable degree of neural specific markers (NSE,MAP-1 β,VIM) positive cells were observed in the SCI tissues of all groups,with those in the combined treatment group significantly higher than the other 3 groups.The immunofluorescence also found significantly higher fluorescence expression of neural specific protein markers including NSE,MAP-1 βand VIM in the combined treatment group than the rest group,with obvious proliferation and differentiation of nerve fibers.Conclusions Stem cell transplantation combined with partial body weight supported treadmill training can effectively promote the recovery of neural function of rats with spinal cord injury,and the effect is better than solely stem cell transplantation or the weight loss training.

Significance of aquaporin-4 antibodies detection in diagnosis and treatment of neuromyelitis optica spectrum disorders / 中华神经医学杂志

Objective To test the level of aquaporin-4 (AQP4) antibody and assess the frequency and specificity of AQP4 antibodies in patients with neuromyelitis optica spectrum disorders (NMOSDs).Methods One hundred and fifty-three patients with demyelinating disease of the central nervous system,admitted to our hospital from May 2007 to May 2012,were chosen in our study; of them,102 had NMOSDs; AQP4 antibodies in the sera of these 102 patients were detected by cell-based immunofluorescence assay (CBA); the distinctive features of NMOSDs by autoantibodies status (AQP4 antibody-positive or antibody-negative) were analyzed retrospectively.Results The sensitivity and specificity of CBA assay for detecting AQP4 antibodies were 83.3％ and 100％,respectively.In patients with positive AQP4 antibodies,the female one took 78.8％,while in patients with negative AQP4 antibodies,the female one only took 28.6％; significant difference was noted between the two (P=0.007); the patients with both AQP4 antibody-positive and other systemic autoimmune diseases showed higher expanded disability status scale scores at the disease onset (5.9±2.0) as compared with the patients only with AQP4 antibody-positive (4.2±1.9,t=3.806,P=0.033); AQP4 antibody deduced even lost during the remission stage,and immunosuppressant medicine could remove the AQP4 antibody.Conclusion Testing for AQP4 antibodies is helpful for the differential diagnosis between NMOSDs and multiple sclerosis,and also useful for predicting the relapse and treating of NMOSDs.

Investigation on bioactive protective function of amino acids to insulin by molecular simulation / 生物医学工程学杂志

Heat sensitive protein medicines are increasingly exhibiting their critical importance on treatment of various diseases at present. But their popularization and application meet a great challenge because of their heat instability. In the present study, insulin was taken as a heat sensitive protein medicine and amino acid as bio-protective agent in order to investigate if these amino acids can protect the insulin from losing its bioactivity due to desiccation. The experiment was performed by using replica exchange molecular simulation (REMD) method and Gromacs software with Gromos96 (53a6) force field. The REMD results indicated that these amino acids could protect the bioactive structure of insulin during desiccation. The configurations of the protected insulin were preserved very well. Those results proved that amino acid is a kind of good bioactive protective agent for the heat sensitive protein medicines.

Investigation on bioactive protection of LEA protein for insulin by molecular simulation in the low-temperature drying process / 生物医学工程学杂志

Nowadays various protein medicines are increasingly playing significant roles in the treatment of many diseases, but the bioactive structures of such kinds of protein medicines are unstable because they are heat sensitive. Therefore, it is very important to explore a protective method and to explain the protective mechanism of protein medicines. In the present research, insulin was chosen as a heat-sensitive protein medicine, and a Group 3 late embryogenesis abundant (LEA) protein was chosen as its bioactive protectant during desiccation. The results of replica exchange molecular dynamics simulation suggest that comparing with insulin without any protection, the bioactive 3D structure and secondary structure of the insulin protected by LEA protein were preserved very well. All analyzing results proved that the LEA protein was a good bioactive protectant for heat sensitive protein medicines.

Myasthenia gravis with muscle specific receptor tyrosine kinase antibodies / 中华神经科杂志

Objectives To compare the characteristics of myasthenia gravis (MG) with different antibodies.Methods The muscle specific receptor tyrosine kinase (MuSk) and acetylcholine receptor (AChR) antibodies were detected in the sera of 119 MG patients,and fluoroimmunoprecipitation assay and cell based assay were applied. The clinical features of AChR-Ab positive,MuSK-Ab positive and serum negative MG patients were compared.Results There were 90 AChR-Ab positive sera tested out from the 119 MG sera,and 5 sera found with MuSK-Ab in the 29 AChR-Ab negative sera.There was no significant difference among the three groups regarding sex and age at onset.There were 3/5 of MuSK-Ab positive patients with predominantly bulbar paralysis,2/5 of MuSK-Ab positive patients were classified as MGFA Ⅲ to Ⅴ,and 79.2% (19/24) of serum negative patients were classified as MGFA Ⅰ.There was significantly positive relation between the levels of MuSK antibodies and disease severity (r=0.941,P=0.014).Neither thymic hyperplasia nor hymoma were found in MuSK-Ab positive patients.Conclusions MuSK antibodies are only detected out in the sera without AChR-Ab.The MuSK-Ab positive patients are mainly involved bulbar muscles,and without thymus abnormality.MuSK-MG is different with the AChR MG.