Objective To investigate the dynamic changes of interleukin-1 receptor-associated kinases (IRAK-4) in hypoxic neurons and explore their role in regulation of inflammatory reaction. Methods The B35 cells exposed to hypoxia of 3% O2,5% CO2 and 92% N2 were cultured for 1,3,6, 12,24,48,72 and 96 hours respectively. Then, mRNA and protein expressions of IRAK-4 were detected by RT-PCR and Western blot, the expression of IRAK-4 in the cells were observed by laser scanning con-focal microscope (LSCM), and the concentration of IL-6 was measured by ELISA method. Results After hypoxia, the mRNA and protein expressions of IRAK-4 were increased at one hour, reached the peak at six hours (P<0.05), kept at a high level at 12 hours (P<0.05) , but decreased gradually to the normal oxygen level at 24 hours (P < 0.05) and to below the normal oxygen level at 48 hours (P < 0.05). Immunofluorescence results showed that the fluorescence intensity of IRAK-4 was gradually increased with time. The changes of IL-6 in the supernatant were positively correlated with protein expression of IRAK-4 (r =0.84, P < 0.05 ). Conclusions Hypoxia can increase the expression of IRAK-4 at transcription and translation levels in a certain period of time, which may participate in down-stream inflammatory reaction and lead to increase of IL-6 expression.

Objective To investigate the impact of graft position shift on anterior cruciate ligament reconstruction induced by femoral fixation of interference screw. Methods Nineteen fresh cadaveric knees were used and assigned to three groups. 1) Study of graft position shift: 5 knees were randomly selected, interference screws of 7 mm, 8 mm and 9 mm were used in autologous tendon fixation, then the graft position shift were measured. 2) Study of isometry: 7 knees were randomly divided into the isometric reconstruction group (D group). In the D group, Retrobutton, interference screw and interference screw in location-corrected bone tunnel were used respectively as fixation. The isometry of grafts was evaluated. 3) Study of tibia anterior translation: 7 knees were randomly divided into the anatomic reconstruction group (J group). In the J group,the tibia anterior translation was measured in four different conditions in the same joint: intact knee joint,knee joint without ACL, ACL anatomic reconstruction by interference screw fixation, and ACL anatomic reconstruction by interference screw fixation with corrected bone tunnel location. Results 1) With 7 mm, 8mm and 9 mm interference screw fixation, graft position shift were (2.36±0.11) mm, (2.72±0.06) mm and (3.00±0.06) mm respectively. 2) Graft length change: graft length change in Retrobutton group and corrected bone tunnel group were less than 3 mm, while graft length change in those fixed with interference screw were stretched in more than 3 mm. 3) Study of tibia anterior translation: there was no difference among the intact group, the anatomic group and the corrected group at 0° and 15°. However, the difference was found between the intact group and other groups at 30°、60° and 90° of flexion, as well as between these two reconstructed methods at 20° joint flexion (P＜0.05). Conclusion In both isometric and anatomic ACL reconstruction with interference screw, the graft is pushed tightly toward the femoral tunnel wall, which shifts the graft away from the desired position. In our study we find out that the corrected location of the femoral bone tunnel significantly improves the isometry of ACL reconstruction and anatomic reconstruction.

Objective:To analyze the clinical effect of different surgical methods on giant-cell tumor of bone (GCTB). Methods:Between January 2007 and June 2012, 37 patients receiving treatment for GCTB were enrolled in this study. These patients included 20 males and 17 females whose ages ranged from 20 to 56 (average=31.6). The prosthesis replacement and nonprosthetic replacement op-eration on the postoperative recurrence, limb function, comprehensive evaluation of the curative effect, as well as morbidity and recur-rence rate among different nations and in the upper and lower limbs of GCTB patients were analyzed. Results:The predilection age of the cases ranged from 20 to 50 years old. The incidence was higher in the Uighur population of Xinjiang, China. However, the recur-rence rate in ethnic difference was not statistically significant (P>0.05). Conclusion:The predilection age of GCTB cases was 20~50 years old, with higher morbidity in Uighur than in Han and other ethnic groups. The morbidity and recurrence rate were higher in the lower limbs than in the upper limbs of patients. The scraping bone graft and bone cement filling, the internal fixation of the scraping bone graft, and the artificial joint replacement of tumor resection were all reliable modes of treatment.

A microwave digestion system was preparation for the digestion of the sediment samples of 16 stations in the Southe mid_Atlantic ridge by using HNO3_H2 O2_HF as the digestion reagent. The rare earth elements ( RE ) in sediments were determined by inductively coupled plasma mass spectrometry, and distribution characteristics of rare earth elements were studied. The microwave digestion_ICP_MS method was used for the determination of rare earth elements with a good linear relationship ( r=0 . 9997-1 . 0000 ) for each element. The detection limit reached ng/L level, the relative standard deviation ( RSD, n=3 ) was less than 3% and the relative error was 6%. The total amount of rare earth elements (ΣRE) in sediment samples from 16 stations varied in the range of 37. 25-134. 77 μg/g, the ratio range of light RE/heavy RE ( LRE/HRE) was 0 . 61-1 . 70 , the average value was 1 . 27 , and the enrichment of light rare earth elements in sediments was slightly obvious. The RE distribution patterns were basically the same in each station with obvious fractionation between LRE and HRE. The RE distribution patterns were also similar in sediments from different sources with slightly difference between terrestrial and marine sediments. The δEu and δCe in the sediments had negative anomaly which showed that the rare earth elements in sediments came from the seawater. This study first analyzed the content and distribution of rare earth elements in the southern Atlantic, providing data and technical support for further study of the distribution of rare earth elements in the Atlantic.

BACKGROUND:In mass spectrometry analysis, the same protein in different samples labeled with isobaric tags for relative and absolute quantitation presents the same mass-to-charge ratio, while in the tandem mass spectrometry analysis, the ion signal shows different mass-to-charge ratio (114-121). Thus the quantitative information of the related proteins can be obtained. OBJECTIVE:To establish the protein spectrum of spinal cord tissue differences proteins after acute spinal cord injury, and to explore the spinal differential protein expression on the molecular level using isobaric tags for relative and absolute quantitation combined with LC-MS/MS mass spectrometry technique. METHODS:Eight Sprague Dawley rats were selected to establish the acute spinal cord injury models using Al en’s method. The rats were randomly divided into 0 hour spinal cord injury group and 8 hours spinal cord injury group, four rats in each group. The spinal cord tissues were col ected after injury, and the spinal cord tissue differences proteins were analyzed with isobaric tags for relative and absolute quantitation technique after acute spinal cord injury. RESULTS AND CONCLUSION:Total y 220 differential y expressed proteins were identified in this research, the number of up-regulation proteins was 116 and the number of down-regulation proteins was 104. There were 12 differential proteins related to neural regeneration, and among the 12 proteins, there were seven up-regulation proteins and five down-regulation proteins. The various identified differential proteins and significantly expressed nerve growth factors in this experiment can be used as the biomarkers of acute spinal cord injury or used as the strong evidence for the clinical management and monitoring of the injury process and target therapy of acute spinal cord injury, as wel as the effect evaluation.

Objective To construct recombinant retroviral vector containing human hepatocellular carcinoma-related gene ANGPTL4 ( angiopoietin-like 4) cDNA and to evaluate antitumor effect of recombinant retroviral vector-mediated human ANGPTL4 gene transfer. Methods ANGPTL4 cDNA was cloned in vitro from human liver cell lines HL-7702 and subcloned into plasmid vector pMSCV and sequenced. High-tiler recombinant retrovirus pMSCV-ANGPTLA and blank retrovirus pMSCV packaged under mediation of lipofectamine infected HepG2 cells in vitro, respectively. Flow cytometry and fluorescence microscopy detected expression of GFP (green fluorescence protein) in HepG2 cells. The expression of ANGPTL4 mRNA in HepG2 cells was determined. Results Recombinant retroviral vector pMSCV-ANGPTL4 was constructed successfully. Titer of recombinant retrovirus pMSCV-ANGPTL4 packaged is 1. 4 ? 106 infective viral grains /ml. Titer of blank retrovirus pMSCV packaged was 1. 5 ? 106 infective viral grains /ml. Positive cell rate of HepG2-ANGPTL4 cells group expressing GFP was 68.45% , and average intensity of fluorescence of HepG2-ANGPTL4 cells group was 31.67 -fold as that of HepG2 cells group. Positive cell rate of HepG2-pMSCV cells group expressing GFP was 77.72%, and average intensity of fluorescence of HepG2-pMSCV cells group was 64. 87 -fold as that of HepG2 cells group. The expression of ANGPTL4 mRNA in HepG2-ANGPTL4 cells group was higher than that in HepG2-pMSCV cells group (154%) and HepG2 cells group( 161%). The proliferation rate of HepG2-ANGPTL4 cells group in vitro was lower than HepG2-pMSCV cells group and HepG2 cells group (P

We studied the effect of the Mycobacterium tuberculosis prokaryotic ubiquitin-like protein-proteasome system on mono-resistant to rifampin resistance to M.tuberculosis.A resazurin-based assay was employed to evaluate minimum inhibitory concentration (MIC) and comparative research on mono-resistant to rifampin MTB with Pup,Dop,PafA,Mpa genes expression and deletion of the difference.Above testing strains,respectively,carbonyl cyanide chlorobenzene hydrazone (CCCP),reserpine (RP),verapamil (VP)and chlorpromazine (CPZ) were tested.We compared and analyzed the change of rifampicin MICs on the various strains.Compared with rifampin resistant MTB,overexpression of Pup,Dop,PafA and Mpa genes were able to make monorifampicin of M.tuberculosis to enhance resistance to rifampin.Deletion of Pup gene,Mpa gene,Dop gene,PafA gene significantly decreased the resistance to rifampicin alone MTB,and the P value was ＜0.05.Results indicated that 4 kinds of efflux pump inhibitors can reduce the degree of rifampin MIC in different strains.Through the factorial analysis,there were some interactions between MTB and PPS efflux pump inhibitors,and the P value was ＜0.05.MTB PPS has influence on mono-rifampin resistance to MTB and it may regulate the efflux pathway related protein to influence its resistance.

Objective To study the molecular mechanism of oridonin-induced apoptosis of ghoma SHG44 cells.Methods A growth curve was plotted using CCK-8 colorimetric method with different concentrations of oridonin (0,1.25,2.5,5,10,20,and 40 μmol/L)to observe its effect on the growth of SHG44 cells.Hoechst33258 and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining were used to examine the changes in cell morphology and flow cytometry was used to detect cell apoptosis.Western blotting was used to analyze the expression of apoptosis-related proteins (Caspase-3,cleaved Caspase-3,Bax,and Bcl-2)in SHG44 cells.Results SHG44 cell proliferation was significantly suppressed after 24 and 48 h Oridonin treatment,with a half-maximal inhibitory concentration of 7.865 and 4.74 μmol/L,respectively.Hoechst33258 and TUNEL staining showed changes in cell morphology such as shrinkage and nucleus fragmentation and morphogenesis,which are indicative of apoptosis.Western blotting analysis showed that oridonin inhibited the expression of Bcl-2 and activated the expression of Caspase-3 and Bax.Conclusion Oridonin can inhibit the proliferation and induce the apoptosis of SHG44 cells by regulating the expression of apoptosis-related proteins.

Objective To explore the correlation of baseline CCL19+dendritic cell(CCL19+DC)infiltration in lung adenocarcinoma microenvironment with immunotherapy efficacy and CD8+T cell infiltration.Methods We retrospectively analyzed the data of patients with lung adenocarcinoma hospitalized at First Affiliated Hospital of Henan University of Science and Technology from January,2020 to December,2023,and collected tissue samples from 96 patients undergoing immunotherapy for assessing CCL19+DC and CD8+T cell infiltration using immunofluorescence assay.We evaluated the predictive value of baseline CCL19+DCs for patient responses to immunotherapy using receiver-operating characteristics(ROC)curves and analyzed the correlations of baseline CCL19+DC expression with immunotherapy efficacy and CD8+T cell and cytotoxic T lymphocyte(CTL)infiltrations.In co-culture systems of lung adenocarcinoma PC9 cells,CD8+T cells and DCs(overexpressing CCL19 with or without anti PD-1 antibody treatment),the expressions of granzyme B,perforin,IFN-γ,and Ki-67 in T cells were analyzed using flow cytometry.Results The patients with partial or complete remission following immunotherapy had a significantly higher baseline CCL19+DC infiltration level in lung adenocarcinoma tissues than those with poor responses.CCL19+DC infiltration had an area under ROC curve of 0.785,a sensitivity of 75.6%,and a specificity of 62.8%for predicting immunotherapy efficacy.The expression of CD8+T cell surface molecules Granzyme B(P<0.01),Perforin(P<0.01),IFN-γ(P<0.01)and Ki-67(P<0.001)in patients with high expression of CCL19+DC were higher than those in patients with low expression of CCL19+DC.The baseline CCL19+DC infiltration level was positively correlated with immunotherapy efficacy(P=0.003),CTL infiltration of(r=0.6657,P<0.001)and CD8+T cell infiltration(P=0.007).In the co-cultured cells,CCL19 overexpression combined with anti-PD1 treatment of the DCs more strongly enhanced cytotoxicity and proliferation of CD8+T lymphocytes than either of the single treatments(P<0.01 or 0.001).Conclusion The baseline CCL19+DC infiltration level in lung adenocarcinoma microenvironment is positively correlated with immunotherapy efficacy and CTL infiltration and can thus predict the response to immunotherapy.

Objective To explore the correlation of baseline CCL19+dendritic cell(CCL19+DC)infiltration in lung adenocarcinoma microenvironment with immunotherapy efficacy and CD8+T cell infiltration.Methods We retrospectively analyzed the data of patients with lung adenocarcinoma hospitalized at First Affiliated Hospital of Henan University of Science and Technology from January,2020 to December,2023,and collected tissue samples from 96 patients undergoing immunotherapy for assessing CCL19+DC and CD8+T cell infiltration using immunofluorescence assay.We evaluated the predictive value of baseline CCL19+DCs for patient responses to immunotherapy using receiver-operating characteristics(ROC)curves and analyzed the correlations of baseline CCL19+DC expression with immunotherapy efficacy and CD8+T cell and cytotoxic T lymphocyte(CTL)infiltrations.In co-culture systems of lung adenocarcinoma PC9 cells,CD8+T cells and DCs(overexpressing CCL19 with or without anti PD-1 antibody treatment),the expressions of granzyme B,perforin,IFN-γ,and Ki-67 in T cells were analyzed using flow cytometry.Results The patients with partial or complete remission following immunotherapy had a significantly higher baseline CCL19+DC infiltration level in lung adenocarcinoma tissues than those with poor responses.CCL19+DC infiltration had an area under ROC curve of 0.785,a sensitivity of 75.6%,and a specificity of 62.8%for predicting immunotherapy efficacy.The expression of CD8+T cell surface molecules Granzyme B(P<0.01),Perforin(P<0.01),IFN-γ(P<0.01)and Ki-67(P<0.001)in patients with high expression of CCL19+DC were higher than those in patients with low expression of CCL19+DC.The baseline CCL19+DC infiltration level was positively correlated with immunotherapy efficacy(P=0.003),CTL infiltration of(r=0.6657,P<0.001)and CD8+T cell infiltration(P=0.007).In the co-cultured cells,CCL19 overexpression combined with anti-PD1 treatment of the DCs more strongly enhanced cytotoxicity and proliferation of CD8+T lymphocytes than either of the single treatments(P<0.01 or 0.001).Conclusion The baseline CCL19+DC infiltration level in lung adenocarcinoma microenvironment is positively correlated with immunotherapy efficacy and CTL infiltration and can thus predict the response to immunotherapy.