Objective:To investigate variation and correlation of type 2 intrinsic lymphocyte(ILC2)and related factors in acute and chronic brucellosis,to identify immune role of ILC2 in chronic brucellosis.Methods:Forty-three patients with acute brucel-losis and 45 patients with chronic brucellosis were compared with 49 healthy controls.ILC2 level in each group was detected by flow cytometry.mRNA level of GATA3 in PBMC was detected by fluorescence quantitative PCR.Serum IL-33,ST2,IL-4 and IL-13 levels were detected by ELISA.ALT and AST levels were measured by fully automatic biochemical analyzer.Results:ILC2 level,GATA3 mRNA in PBMC,serum IL-33,IL-4 and IL-13 levels in chronic brucellosis group were significantly higher than healthy control group and acute brucellosis group(P<0.01).Correlation analysis showed that ILC2 level was positively correlated with GATA3 mRNA,IL-33,IL-4 and IL-13 levels,while negatively correlated with ST2 level.ROC curve suggested that ILC2,GATA3,IL-33 and ST2 had good predictive ability for severity of brucellosis patients(AUC>0.7).Conclusion:Increase of ILC2 and its related factors are closely related to chronic brucella infection,which may play an important immune role in pathogenesis of brucella infection.

Objective:To investigate variation and correlation of type 2 intrinsic lymphocyte(ILC2)and related factors in acute and chronic brucellosis,to identify immune role of ILC2 in chronic brucellosis.Methods:Forty-three patients with acute brucel-losis and 45 patients with chronic brucellosis were compared with 49 healthy controls.ILC2 level in each group was detected by flow cytometry.mRNA level of GATA3 in PBMC was detected by fluorescence quantitative PCR.Serum IL-33,ST2,IL-4 and IL-13 levels were detected by ELISA.ALT and AST levels were measured by fully automatic biochemical analyzer.Results:ILC2 level,GATA3 mRNA in PBMC,serum IL-33,IL-4 and IL-13 levels in chronic brucellosis group were significantly higher than healthy control group and acute brucellosis group(P<0.01).Correlation analysis showed that ILC2 level was positively correlated with GATA3 mRNA,IL-33,IL-4 and IL-13 levels,while negatively correlated with ST2 level.ROC curve suggested that ILC2,GATA3,IL-33 and ST2 had good predictive ability for severity of brucellosis patients(AUC>0.7).Conclusion:Increase of ILC2 and its related factors are closely related to chronic brucella infection,which may play an important immune role in pathogenesis of brucella infection.

This study was performed to investigate the variation characteristics and functions of Siglec-9 immune checkpoint in pulmonary tuberculosis.R language was used for bioinformatics analysis of GSE83456 chip data.Total of 48 patients with confirmed pulmonary tuberculosis(PTB)and 46 healthy controls were included.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the mRNA expression of Siglec-9 in peripheral blood;flow cytometry was used to detect the proportion of Siglec-9+CD4+T cells.The levels of TNF-α,IL-6,IFN-γ and IL-4 were detected by Cytometric Bead Array(CBA).Furthermore,the correlation of the proportion of Siglec-9+CD4+T cells with TNF-α and other cytokines were analyzed.Bioinformatics analysis showed that Siglec-9 was significantly increased in patients with PTB(P＜0.001),and was related to TNF-α/NF-κB,IL-6/JAK/STATA3,PI3K/AKT/mTOR signal pathways.Experimental data showed that the proportion of Siglec-9+CD4+T cells was significantly increased in patients with PTB(P＜0.001)and negatively correlated with the levels of TNF-α and other cytokines.In conclusion,the higher levels of Siglec-9 mRNA and Siglec-9+CD4+T cells in PTB may inhibit the function of CD4+T cells and participate in the occurrence and development of PTB.

This study was performed to investigate the variation characteristics and functions of Siglec-9 immune checkpoint in pulmonary tuberculosis.R language was used for bioinformatics analysis of GSE83456 chip data.Total of 48 patients with confirmed pulmonary tuberculosis(PTB)and 46 healthy controls were included.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the mRNA expression of Siglec-9 in peripheral blood;flow cytometry was used to detect the proportion of Siglec-9+CD4+T cells.The levels of TNF-α,IL-6,IFN-γ and IL-4 were detected by Cytometric Bead Array(CBA).Furthermore,the correlation of the proportion of Siglec-9+CD4+T cells with TNF-α and other cytokines were analyzed.Bioinformatics analysis showed that Siglec-9 was significantly increased in patients with PTB(P＜0.001),and was related to TNF-α/NF-κB,IL-6/JAK/STATA3,PI3K/AKT/mTOR signal pathways.Experimental data showed that the proportion of Siglec-9+CD4+T cells was significantly increased in patients with PTB(P＜0.001)and negatively correlated with the levels of TNF-α and other cytokines.In conclusion,the higher levels of Siglec-9 mRNA and Siglec-9+CD4+T cells in PTB may inhibit the function of CD4+T cells and participate in the occurrence and development of PTB.

Objective:To study the clinical efficacy of laparoscopic radical cholecystectomy in the treatment of stage Ⅲ gallbladder cancer.Methods:The clinical characteristics and postoperative follow-up data of 184 patients (male 66, and female 118) who underwent radical cholecystectomy for stage Ⅲ gallbladder cancer at Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, from May 2015 to May 2022, were retrospectively analyzed. The age was (67.0±8.6) years old (range 38 to 85 years old). There were 71 patients in the laparoscopic group and 113 in the open group. The general medical data, surgery-related indicators and complications were analyzed. Follow-up was completed by outpatient visits and by telephone.Results:The laparoscopic group showed better postoperative alanine aminotransferase [67.5 (40.0, 138.5) vs. 104.0 (45.0, 252.2) U/L] and aspartate aminotransferase [41.5 (26.0, 71.2) vs. 53.0 (30.2, 153.5) U/L] recovery, higher albumin levels [32.05 (30.18, 35.20) vs. 30.50 (27.70, 33.50) g/L], earlier abdominal drainage tube removal [8.00(6.00, 10.25) vs. 10.00(6.00, 13.00)d], shorter hospital stay [10.00(8.00, 15.25) vs. 14.00(9.00, 19.00) d] and lower incidences of complications [(14.1%(10/71) vs. 31.9%(36/113)] when compared with the open group (all P<0.05). The 1 year (49.1% vs 61.0%), 2 years (24.0% vs. 28.5%), 3 years (16.0% vs. 14.5%) overall survival ( P=0.640), and the 3 years progression-free survival (18.3% vs. 15.0%, P=0.463) showed no significant difference between the 2 groups. Conclusion:Laparoscopic surgery for AJCC TNM stage Ⅲ gallbladder cancer showed comparable results with open surgery. When compared with open surgery, laparoscopic radical resection of gallbladder cancer had the advantages of earlier removal of abdominal drainage tube, lower incidence of postoperative complications, and shorter hospital stay.

Objective:To study the effect of preoperative transcatheter arterial chemoembolization (TACE) on postoperative complications after hepatectomy in patients with hepatocellular carcinoma (HCC) by propensity score matching analysis.Methods:Of 1 666 patients with HCC undergoing hepatectomy in Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology and Tianyou Hospital of Wuhan University of Science and Technology from March 2015 to March 2021 were retrospectively screened. Of 262 patients were enrolled, including 236 males and 26 females, aged (50.3±11.8) years. Of 131 patients were enrolled in both the single surgery group and the combined group (preoperative TACE + surgical resection). Factors affecting the complications after hepatectomy in patients with HCC were analyzed using univariate and multivariate logistic regression method.Results:After matching the propensity score, the incidence of postoperative complications in the single surgery group was 22.1% (29/131), lower than that in the combined group [41.2% (54/131), χ 2=11.02, P<0.001]. The incidence of bile leakage in the single surgery group [2.3% (3/131)] was also lower than that in the combined group [(9.2% (12/131), χ 2=5.73, P=0.017]. Multivariate logistic regression analysis showed that the combined group ( OR=2.43, 95% CI: 1.28-4.61, P=0.007) had an increased incidence of postoperative complications, so did patients with a preoperative alpha-fetoprotein > 400 μg/L, anatomic hepatectomy, long operation time, and hilar occlusion. Conclusion:Preoperative TACE could be a risk factor for postoperative complications in patients with HCC, especially for the postoperative biliary leakage.

Objective:To investigate the physicochemical and biological properties of different magnesium modified calcium phosphate bone cements.Methods:The different magnesium modified calcium phosphate bone cements were divided into magnesium citrate, magnesium lactate, magnesium malate, magnesium phosphate and magnesium glycinate groups, each of which was added with different magnesium agents in the proportion of 0%, 1%, 3% and 5% of the total weight of calcium phosphate bone cements. The initial and final setting time, injectability, anti-collapse performance and compressive strength of different magnesium modified calcium phosphate bone cements were tested. Furthermore, the screened bone cement extracts were used to culture with third generation osteoblasts. Bioactivity assays were performed using the Cell Proliferation and Toxicity Assay Kit (CCK-8). Alkaline phosphatase (ALP) staining and Alizarin Red S (ARS) staining were performed on osteoblasts to observe the osteogenic activity of magnesium malate modified calcium phosphate bone cements.Results:The addition of different proportions of different magnesium agents led to the shortening of the initial and final setting time of modified calcium phosphate bone cements. Moreover, the final setting time of 5% magnesium malate modified calcium phosphate bone cements was the shortest (<40 minutes), which was significantly shorter compared with other magnesium agents in the same proportion (all P<0.05). With the addition of different magnesium agents in different proportions, the injectability of bone cements was gradually increased, and the injectability of 5% magnesium malate calcium phosphate bone cements reached the highest for (87.3±1.9)%, which was significantly increased compared with other magnesium agents in the same proportion (all P<0.05). The anti-collapse performance of bone cements was decreased with the addition of different magnesium agents in different proportions. Magnesium citrate, magnesium phosphate and magnesium glycinate modified calcium phosphate bone cements could not resist the flushing of deionized water. In particular, magnesium malate modified calcium phosphate bone cements had the best anti-collapse performance, with the maximum weight loss rate for only (9.8±2.3)% after 30 minutes of deionized water flushing, which was better than the rest of the groups (all P<0.05). The compressive strength of magnesium lactate and magnesium phosphate modified calcium phosphate bone cements showed a decrease compared with original calcium phosphate bone cements, while the compressive strength of magnesium citrate and magnesium malate modified calcium phosphate bone cements was significantly increased compared with original calcium phosphate bone cements, of which 3% magnesium malate modified calcium phosphate bone cements had the greatest compressive strength of (6.2±0.2)MPa, significantly higher than the rest of the groups (all P<0.05). The sieve test yielded magnesium malate modified calcium phosphate bone cement, which had a weight loss of (27.0±0.9)% at 35 days in vitro. The release of magnesium ions was increased with increasing magnesium malate dose in the in vitro environment of magnesium malate modified calcium phosphate bone cements in different ratios. A stable magnesium ion release was achieved within 35 days.Also, the pro-proliferative and osteogenic effects of modified calcium phosphate bone cements on osteoblasts were more obvious with increase of magnesium malate dose. For 5% magnesium malate modified calcium phosphate bone cements, the cell number, ALP staining area ratio and calcium nodule area ratio were significantly increased compared with the groups in the proportion of 0% and 1% magnesium malate (all P<0.05). Conclusions:Among magnesium citrate, magnesium lactate, magnesium malate, magnesium phosphate and magnesium glycinate modified calcium phosphate bone cements, magnesium malate modified calcium phosphate bone cements have relatively suitable setting time, excellent anti-collapse performance and mechanical strength. Meanwhile, 5% magnesium malate modified calcium phosphate bone cements have better biological activity among different ratios of magnesium malate modified calcium phosphate bone cements, suggesting a potential value for clinical application.

When nanoparticles were introduced into the biological media, the protein corona would be formed, which endowed the nanoparticles with new bio-identities. Thus, controlling protein corona formation is critical to

Objective:To compare the clinical effect of combined anterior and posterior approach and posterior median approach to treat O'Driscoll type III b fracture of ulnar coronoid process.Methods:A retrospective case control study was made on 67 patients with O'Driscoll type III b fracture of ulnar coronoid process treated in Honghui Hospital, Xi'an Jiaotong University from January 2015 to January 2019, including 35 males and 32 females, aged from 21 to 61 years [(38.0±9.4)years]. Among them, 31 patients were treated with combined anterior and posterior approach for reduction and internal fixation (combined approach group), and 36 patients with median posterior elbow approach group for reduction and internal fixation (posterior elbow approach group). The operation time, amount of intraoperative blood loss and fracture healing time were compared between groups. The visual analogue score (VAS), elbow joint range of motion and Mayo elbow performance score (MEPS) were assessed for pain and function evaluation at postoperative 1, 3, 6 months and at the last follow-up. The occurrence of complications were observed as well.Results:All patients were followed up for 12 to 28 months [(20.1±4.2)months]. There was no significant difference in operation time and VAS between the two groups ( P>0.05). The intraoperative blood loss [(133.6±20.3)ml] and fracture healing time [(12.3±1.7)months] in combined approach group were less or shorter than those in posterior elbow approach group [(144.4±22.1)ml, (13.2±2.0)months] ( P<0.05). The range of flexion and extension of elbow joint in combined approach group [(88.7±10.8)°, (111.1±13.9)°, (121.3±14.1)°, (127.1±13.3)°] was higher than that in posterior elbow approach group [(74.5±11.8)°, (97.6±12.6)°, (111.3±13.0)°, (115.2±12.7)°] at postoperative 1, 3, 6 months and at the last follow-up ( P<0.05). The MEPS in combined approach group [(31.7±8.6)points, (55.6±9.3)points, (84.6±10.5)points, (85.0±10.3)points] was higher than that in posterior elbow approach group [(27.2±8.2)points, (50.7±8.7)points, (77.4±11.2)points, (80.1±9.4)points] at postoperative 1, 3, 6 months and last follow-up ( P<0.05). The incidence of complications in combined approach group [10%(3/31)] was lower than that in posterior elbow approach group [31%(11/36)]( P<0.05). Conclusion:Compared with the simple posterior elbow median approach, the combined anterior and posterior elbow approach for treatment of O'Driscoll type IIIb fracture of ulnar coronoid process has lower intraoperative blood loss, faster fracture healing, lower incidence of complications and better elbow function.

Objective To investigate the effects and mechanisms of shRNA interfered with expression of leucine-rich repeat containing G-protein coupled receptor 5 (Lgr5) on the malignant behaviors of colorectal cancer stem cells (CSCs).Methods The experimental study was conducted.The CSCs expressing Lgr5+ were sorted by fluorescence activated cell sorting.Lgr5+ cells that were transfected with Lgr5-shRNA lentiviral vector and nontarget shRNA lentiviral vector were respectively allocated into the experimental group and control group.The percentage of Lgr5+ cells was analyzed by flow cytometery.The relative expression of Lgr5 mRNA was detected by fluorescence quantitative real-time polymerase chain reaction (qRT-PCR).The capacity of self-renewal was detected by sphere forming assay.The tumorigenesis in vitro and in vivo were respectively measured by colony formation assay and xenografting experiment.The mRNA expressions of stem cells related genes (Oct4,Sox2,Nanog,KLF4),CSCs genes (CD133,CD44,ALDH) and Wnt/β-catenin pathway key genes (Axin2,Wnt5a,Wnt3a,Fzd3,c-myc,VEGF,Ascl2,claudin-1) were detected by qRT-PCR.Measurement data with normal distribution were represented as-x±s.Comparison between groups was analyzed using the t test.Results (1)Transfection efficiency of shRNA lentiviral vector induced Lgr5 by flow cytometery was respectively 6.8％± 1.0％ in the experimental group and 92.7％±3.3％ in the control group,with a statistically significant difference (t =43.148,P＜0.05).The relative expression of Lgr5 mRNA measured by qPT-PCR was respectively 0.168±0.057 in the experimental group and 1.148±0.004 in the control group,with a statistically significant difference (t=28.778,P＜0.05).(2) The capacity of self-renewal was detected by sphere forming assay.The results of sphere forming assay:the number of spheres was 29±6 in the experimental group and 410± 10 in the control group,with a statistically significant difference (t =41.070,P＜0.05).The results of colony formation assay:the numbers of colonies in the experimental group and control group were respectively 72±4 and 412± 19,showing a statistically significant difference (t =31.433,P＜ 0.05).The results of tumorigenesis:the volumes of tumors in the experimental group and control group were respectively (81± 15)mm3 and (328±24)mm3,with a statistically significant difference (t=11.304,P＜0.05).(3) The effects of Lgr5 down-regulation on related genes,results of qRT-PCR detection:① The mRNA relative expressions of Oct4,Sox2,Nanog and KLF4 (stem cells related genes) were 0.377±0.093,0.662±0.104,3.591±0.300,0.425±0.091 in the experimental group and 1.957± 0.026,2.137±0.015,5.831±0.165,1.536±0.014 in the control group,with statistically significant differences (t=23.079,22.261,8.446,19.186,P＜0.05).② The mRNA relative expressions of CD133,CD44 and ALDH (CSCs genes) were 1.490±0.155,5.535±0.487,1.640±0.039 in the experimental group and 2.488± 0.061,9.908±0.332,5.718±0.292 in the control group,with statistically significant differences (t =8.170,9.667,27.849,P＜0.05).③The mRNA relative expressions of Axin2,Wnt5a,Wnt3a,Fzd3,c-myc,VEGF,Ascl2 and claudin-1 genes in the Wnt/β-catenin pathway were respectively 1.592±0.267,0.528±0.138,2.153±0.078,1.480±0.064,0.248±0.128,1.492±0.025,0.658±0.095,1.647±0.087 in the experimental group and 3.651±0.224,2.570±0.093,2.301±0.157,1.636±0.058,1.415±0.080,2.610±0.159,2.480±0.123,3.432±0.273 in the control group.There were statistically significant differences in the mRNA relative expressions of Axin2,Wnt5a,c-myc,VEGF,Ascl2 and claudin-1 genes between the 2 groups (t =7.316,15.332,12.649,12.320,14.831,9.063,P＜0.05),and no statistically significant difference in the mRNA relative expressions of Wnt3a and Fzd3 between the 2 groups (t =2.887,2.242,P＞0.05).Conclusion The malignant behaviors of colorectal CSCs are suppressed after shRNA lentivirus interfered with expression of Lrg5,and its mechanism is related to inhibiting activity of Wnt/β-catenin pathway.