Objective To investigate the effect of blockading OX40-OX40L co-stimulatory signaling on the survival time of liver allograft in rat.Methods siRNA-expression vectors were constructed to targeting OX40.3～5 minutes before DA to Lewis orthotopic liver transplantation was performed,5×109 pfu of targeting OX40 siRNA plasmid DNA were diluted in 5 ml of phosphate buffered saline(PBS)and inlected intravenously into recipient Lewis rat over a period of 10 seconds.Serum IL-2 and IFN-γ levels were assayed by ELISA,and mix lymphocyte response(MLR)were tested by 3H-thymidine.Results The survival time of recipients in siRNA treatment group(74.0±9.3)was significantly longer than that in control group[(7.3±0.5)days].In experiment group,the inflammatory cell infihration and liver tissue structure destruction were very slight.The concentration of serum IL-2 was much lower in siRNA treatment group[(46±8.4)pg/ml]than that in control group[(286.5±14.6)pg/ml].Meanwhile,the concentration of serum IFN-γ was much lower in siRNA treatment group [(202.7±14.6)pg/ml]than that in control group[(1682.7±87.9)pg/ml].Conclusion Administration of OX40-siRNA can blockade OX40-OX40L co-stimulatory signaling pathway.hence inhibit the rejection of liver allograft.

Psammosilene Radix is a famous miao national herb and it is rare and endangered nowadays. However, it is often confused with the root of Silene viscidula. In this study, the ITS2 regions was used to identify Psam-mosilene Radix and its adulterants. All DNA samples of Psammosilene Radix and its adulterants were extracted. The ITS2 regions were amplified and sequenced, and the final sequences were assembled using the CondonCode Aligner. The genetic distances were calculated by kimura 2-parameter (K2P) model and the Neighbor-Joining(NJ) phylogenetic trees were constructed using MEGA5.1.BLAST 1, the nearest distance and phylogenetic tree (NJ-tree)methods were used to assess the identification efficiency of the ITS2 region. Results indicated that the length of ITS2 sequences of Psammosilene Radix w 229 bp, the identification effficiency of ITS2 region using BLAST 1 was 100%;and the maximum intra-specific K2P distance were lower than the minimum inter-specific K2P distance. Additionally, the NJ tree based on ITS2 sequence indicated that Psammosilene Radix and its adul-terants could be distinguished clearly . In conclusion , the ITS2 region as DNA barcodes could identify Psammosi-lene Radix and its adulterants stably and accurately. Furthermore, the application of ITS2 barcode in the identifi-cation of TCM has a good prospective .

Objective:To explore the clinical effectiveness of arthroscopic treatment of tibial eminence avulsion fracture by four-point fixation with suture anchors.Methods:A retrospective analysis was performed of the 58 patients with tibial eminence avulsion fracture who had been treated by the same group of surgeons using four-point fixation technique with suture anchors under arthroscopy at Department of Sports Medicine, The Affiliated Hospital to Qingdao University from January 2015 to December 2018. They were 33 males and 25 females, with an average age of 18.4 years (from 14 to 32 years). By the modified Meyers-McKeever classification, 15 fractures were type Ⅱ, 19 type Ⅲ and 24 type Ⅳ. Recorded and compared were knee Lysholm scores, International Knee Documentation Committee (IKDC) scores and tibial eminence height between preoperation and one year postoperation; recorded at the last follow-up were range of knee motion and results of Lachman and pivot-shift tests.Results:The 58 patients were followed up for a mean of 20.7 months (from 12 to 33 months). Bony union was achieved in all patients within 12 weeks after operation. In this cohort, the Lysholm score (85.2±4.9) and IKDC score (86.2±4.3) at one year postoperation were significantly higher than the preoperative values (43.2±5.2 and 51.2±4.9), and the post-operative tibial eminence height [(9.1±1.2) mm] was significantly lower than the preoperative value [(12.6±1.2) mm] (all P<0.05). The correlation coefficients between the tibial eminence height and the Lysholm & IKDC scores at one year postoperation were -0.16 and -0.17, respectively. The last follow-up showed a 132°±5° range of knee motion for all patients, a positive result of pivot-shift test (grade Ⅱ) for 3 and a positive result of Lachman test (grade Ⅰ) for 2. Conclusion:Arthroscopic treatment of tibial eminence avulsion fracture by four-point fixation with suture anchors can lead to satisfactory effectiveness, showing advantages of minimal invasion, anatomic reduction, reliable fixation, and little impact on the epiphysis plate.

Objective:To study the effect of microglia depletion combined with bone marrow mesenchymal stem cells (BMSC) transplantation for spinal cord injury (SCI) repair.Methods:GFP-BMSCs were cultured, identified and detected for expression levels of growth factors. The effects of BMSCs ondorsal root ganglion (DRG) axon outgrowth were observed by the co-culture of BMSCs with DRGs. Mice were depleted of microglia by administrating the colony stimulating factor 1 receptor (CSF1R) inhibitor PLX3397. The spinal cords of these microglia-depleted mice were subjected to crush injury. BMSCs were transplanted into SCI area after microglia depletion. Mice were randomly divided into control group (SCI+BMSCs) and experimental group (PLX3397+SCI+BMSCs). Mice were sacrificed at corresponding time points after transplantation for observing the survival of transplanted BMSCs and the repair of spinal cord. BMS score was used for evaluation of motor function recovery.Results:BMSCs secreted a large number of neurotrophic factors and promoted the growth of DRG axons when co-cultured with DRGs. Depletion of microglia significantly improved the survival of transplanted BMSCs. Compared with BMSCs transplantation alone, the combined treatments slightly but non-significantly reduced the area of the lesion ( t=2.141, P=0.065). Immunofluorescence staining showed that both BMSC transplantation alone and the combined treatments did not cause the corticospinalaxons across the lesion and into distal spinal cord. BMS scores were (1.20±0.45), (3.20±0.45), (3.80±0.45), (4.20±0.45), and (4.60±0.55) points in control group at 1, 7, 14, 21 and 28 d after injury. The experimental groups were(0.60±0.55), (3.00±0.71), (3.80±0.84), (4.20±0.84), and (4.40±0.89) points, respectively. Conclusion:Depletion of microglia improves the survival of transplanted cells, depletion of microglia combined with BMSC transplantation did not result in a significant reduction in lesion area. At the same time, the damaged CST axons were notregenerated. Thus, combining cell transplantation with axon-promoting strategy may be necessary for SCI repair.

Objective:To explore the key pathways and genes involved in microglia inflammation through transcriptome sequencing and bioinformatics analysis.Methods:BV2 cells were stimulated by lipopolysaccharide to establish microglia inflammation model. The levels of IL-6 and TNF-α were detected by ELISA and RT-qPCR. The established microglia inflammation model was sequenced by transcriptome sequencing, and the differentially expressed genes were screened by bioinformatics method. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of differentially expressed genes were performed. The protein-protein interaction network of differentially expressed genes was constructed by using string database, and the protein-protein interaction network was visualized by using Cytoscape software. The protein interaction network module was extracted by using MCODE app. The hub gene was extracted by using cytohubba app and was verified through RT-qPCR. We conducted enrichment analysis of hub genes, predicted their targeted miRNAs and interacting drugs.Results:The microglia inflammation model was successfully established and verified by ELISA and RT-qPCR. We screened 434 differentially expressed genes by bioinformatics analysis of transcriptome sequencing results. GO analysis showed that these differentially expressed genes were mainly concentrated in cellular response to cytokine stimulus, inflammatory response, regulation of response to external stimulation. KEGG analysis showed that these differentially expressed genes were mainly concentrated in Chemokine signaling pathway, TNF signaling pathway, IL-17 signaling pathway. We constructed the protein interaction network of these differentially expressed genes, and carried out module analysis and extraction of hub genes. Most of hub genes are located in module 1, and the seed gene of module 1 is S1pr1. Hub genes include S1pr1, Cxcr4, Cx3cl1, Cx3cr1, Cxcl10, Cxcl2, Ccl4, Ccl5, Ccl9, Fpr1. RT-qPCR results showed that compared with the culture medium group, the mRNA expressions of S1pr1, Cxcr4, Cx3cl1 and Cx3cr1 were down-regulated, and the mRNA expressions of Cxcl10, Cxcl2, Ccl4, Ccl5, Ccl9 and Fpr1 were up-regulated in the LPS group. The enrichment analysis of hub genes mainly focused on chemokine-mediated signaling pathway, Class A/1 (Rhodopsin-like receptors), cell chemotaxis and so on. Drugs and miRNAs that may interact with hub genes were predicted. Conclusion:Through transcriptome sequencing and bioinformatics analysis of microglia inflammation model, differentially expressed genes were screened, hub genes and seed genes were extracted, which will help us further understand the molecular mechanism of microglia inflammation and provide potential targets for the treatment of related diseases.

Objective To investigate the abnormal region of gray matter and its' structural changes closely related to mild cognitive impairment in patients with Parkinson's disease (PD) using voxel-based morphometry(VBM) method.Methods Thirty-seven clinically defined PD patients and 20 normal controls (NC),collected in our hospital from March 2011 to February 2013,were examined using T1WI three-dimensional brain volume sequence (3D-fast spoiled gradient echo,3D-FSPGR).We classified PD patients into 2 subgroups according to the extent of cognitive impairment:20 patients with mild cognitive impairment (MCI) and 17 patients with none mild cognitive impairment (nMCI).The data of three groups were analyzed using VBM based on SPM5 to generate gray matter map.Results As compared with NC group,PD patients showed extensively decreased gray matter volume,involving bilateral frontal,temporal,occipital and parietal lobes,insular,parahippocampal gyrus,amygdale and right uncus.As compared with PD-nMCI patients,decreased gray matter volume in PD-MCI patients was observed in bilateral midfrontal gyrus,inferior frontal gyrus,bilateral insular,left precentral gyrus,left superior temporal gyrus and right midtemporal gyrus.Conclusions Areas of decreased gray volume in PD patients locate in widespread brain regions involving limbic system and neocortex.Gray matter atrophy in bilateral midfrontal gyrus,inferior frontal gyrus,insular and left precentral gyrus is related to the mild cognitive impairment.

ObjectiveTo observe the effect of Feiyanning prescription （FYN） on cisplatin （DDP） resistance in non-small cell lung cancer （NSCLC） and explore the underlying mechanism. MethodCell counting kit-8 （CCK-8） assay was used to detect the proliferation of A549 and A549/DDP （DDP-resistant） cells treated by DDP （0， 2.0， 4.0， 6.0， 8.0， 10.0 mg⋅L^-1） and the proliferation of A549/DDP cells treated by FYN （0， 100， 200， 300， 400， 500， 600 mg⋅L^-1）. Based on immunofluorescence staining and Western blot （WB）， the expression of epithelial mesenchymal transition （EMT）-related proteins in A549 and A549/DDP groups was observed. A549/DDP cells were classified into control group， FYN group （200 mg⋅L^-1）， DDP group （6.0 mg⋅L^-1）， and combination group ［FYN （200 mg⋅L^-1） + DDP （6.0 mg⋅L^-1）］ and respectively treated with corresponding drugs. Then， invasion ability of each group was examined by transwell assay， and the expression of EMT-related proteins in each group by WB. Moreover， real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） and immunofluorescence staining were separately applied to detect the mRNA and protein expression of drug resistance-related factors in each group， respectively. ResultCompared with A549 group， A549/DDP group showed high resistance to DDP （P<0.01）， low expression of E-cadherin， and high protein expression of Vimentin， N-cadherin， and Snail （P<0.05， P<0.01）. As compared with the control group， FYN inhibited the proliferation of A549/DDP cells in a concentration-dependent manner （P<0.01）， and the FYN group， DDP group， and combination group demonstrated low invasion ability （P<0.01）. In addition， the invasion ability in the combination group was particularly lower than that in the DDP group （P<0.01）. The expression of E-cadherin protein was higher and the protein expression of N-cadherin， Vimentin， and Snail was lower in the in FYN group than in the control group （P<0.01）. The protein expression of E-cadherin， N-cadherin， and Vimentin was lower and the expression of Snail was higher in the DDP group than in the control group （P<0.05，P<0.01）. The protein expression of E-cadherin， N-cadherin， Vimentin， and Snail in the combination group decreased as compared with that in the control group （P<0.01）. Compared with the DDP alone， the combination raised the expression of E-cadherin and lowered the protein expression of N-cadherin， Vimentin， and Snail （P<0.01）. The protein and mRNA expression of lung resistance-related protein （LRP） and multidrug resistance 1 （MDR1） was lower and the protein and mRNA expression of topoisomerase Ⅱα （TOPO Ⅱα） was higher in the FYN group than in the control group （P<0.01）. The protein and mRNA expression of LRP， MDR1， and TOPO Ⅱα was higher in the DDP group than in the control group （P<0.01）. The expression of LRP protein and mRNA showed no significant variation， but the protein and mRNA expression of MDR1 and TOPO Ⅱα increased in the combination group compared with those in the control group （P<0.01）. Compared with the DDP group， FYN group and combination group showed low protein and mRNA expression of LRP and MDR1 and high protein and mRNA expression of TOPO Ⅱα （P<0.01）. Compared with FYN， the combination elevated the protein and mRNA expression of LRP， MDR1， and TOPO Ⅱα （P<0.01）. ConclusionFYN prescription can reverse the DDP resistance of NSCLC by modulating EMT.

Hip fractures are among the most common fractures in the elderly, presenting to be a leading cause of disability and mortality. Surgical treatment is currently the main treatment method for hip fractures. The incidence of perioperative malnutrition is increased after hip fractures in the elderly due to the comorbidities, decreased basal metabolic rate, accelerated protein breakdown, weakened anabolism and surgical stress. However, malnutrition not only increases the incidence of postoperative complications, but also leads to increased mortality, indicating an important role of perioperative nursing management of nutrition for the elderly patients with hip fractures. At present, there still lacks scientific guidance and application standards on perioperative nursing management of nutrition for the elderly patients with hip fractures. Therefore, the Orthopedic Nursing Committee of Chinese Nursing Association and the Editorial Board of Chinese Journal of Trauma organized relevant experts to formulate the Expert consensus on perioperative nursing management of nutrition for elderly patients with hip fractures ( version 2023) according to evidence-based medical evidences and their clinical experiences. Fourteen recommendations were made from aspects of nutrition screening, nutrition assessment, nutrition diagnosis, nutrition intervention and nutrition monitoring to provide guidance for perioperative nursing management of nutrition in elderly patients with hip fractures.