1.Alzheimer's disease diagnosis among dementia patients via blood biomarker measurement based on the AT(N) system.
Tianyi WANG ; Li SHANG ; Chenhui MAO ; Longze SHA ; Liling DONG ; Caiyan LIU ; Dan LEI ; Jie LI ; Jie WANG ; Xinying HUANG ; Shanshan CHU ; Wei JIN ; Zhaohui ZHU ; Huimin SUI ; Bo HOU ; Feng FENG ; Bin PENG ; Liying CUI ; Jianyong WANG ; Qi XU ; Jing GAO
Chinese Medical Journal 2025;138(12):1505-1507
2.Application of Magnetic Resonance Imaging in the Diagnosis and Treatment of Middle Compartment Defect
Shuyu LUO ; Yuchen SUN ; Yuqin LEI ; Tianyi SUN ; Cheng PENG ; Zhiwei ZHAO ; Yali MIAO
Journal of Sichuan University (Medical Sciences) 2025;56(2):577-583
Middle compartment defects,a common subtype of pelvic floor dysfunction(PFD),are primarily characterized by the prolapse of the uterus or vaginal vault.Magnetic resonance imaging(MRI)has emerged as a valuable diagnostic tool for PFD,offering superior soft tissue resolution while eliminating exposure to ionizing radiation.This review comprehensive summarizes current applications of MRI in the diagnosis and treatment of PFD,covering measurement methods,manifestations of three-level structural defects,postoperative efficacy evaluation,vaginal axial assessment,and evaluation of the mesh status.The authors suggest that MRI enables precise preoperative evaluation of three-level defects,thereby facilitating the development of personalized treatment plans.Additionally,MRI provides an accurate postoperative assessment of surgical outcomes and mesh status,offering a new basis for postoperative assessment.MRI demonstrates unique value in the diagnosis and treatment of middle compartment defects.
3.Mid-Term Efficacy Evaluation of Laparoscopic Sacrocolpopexy vs Laparoscopic Pectopexy for Pelvic Organ Prolapse
Yuqin LEI ; Yuchen SUN ; Tianyi SUN ; Xuesong HAN ; Zhiwei ZHAO ; Yali MIAO
Journal of Sichuan University (Medical Sciences) 2025;56(4):1104-1111
Objective To evaluate the perioperative safety and mid-term outcomes of laparoscopic sacrocolpopexy(LSC)and laparoscopic pectopexy(LP)for pelvic organ prolapse(POP).Methods A retrospective analysis was conducted on 274 POP patients,including 178 who underwent LSC and 96 who underwent LP,between August 2017 and January 2023.The extent of prolapse and anatomical restoration were assessed preoperatively and postoperatively using the Pelvic Organ Prolapse Quantification(POP-Q)system.Quality of life outcomes were evaluated with validated questionnaires,including Pelvic Floor Distress Inventory-short form 20(PFDI-20),Pelvic Floor Impact Questionnaire-short form 7(PFIQ-7),and Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire-12(PISQ-12).Postoperative patient satisfaction was assessed during follow-ups.Postoperative anatomical restoration,perioperative status,and postoperative complications,recurrence,and quality of life were compared between the two groups.Multivariate logistic regression was performed to identify postoperative risk factors for recurrence.Results The operative time in the LSC group was significantly shorter than that in the LP group(P<0.05).Intraoperative blood loss was higher in the LSC group compared to that in the LP group(P<0.05).The LSC group also exhibited higher rates of de novo stress urinary incontinence and constipation(P<0.05).The mean follow-up duration was(35.91±16.90)months.The positions of the indicator points(Aa,Ba,C,Ap,and Bp)in the POP-Q classification after the operation were all better than those before the operation.The PFDI-20 score,PFIQ-7 score,and PISQ-12 score all improved compared to those before the operation(P<0.05).Comparison of preoperative and postoperative PFDI-20,PFIQ-7,and PISQ-12 scores showed no intergroup differences.Compared with the LP group,the LSC group had the lower preoperative POP-Q measurements at points Aa and Ba(P<0.05),but superior postoperative measurements for all the indicator points(Aa,Ba,C,Ap,and Bp)(P<0.05).Recurrence occurred in 28 cases in the LP group and 4 cases in the LSC group,with the LP group presenting a significantly higher anatomical recurrence rate than the LSC group did(31.46%[28/89]vs.2.41%[4/166],P<0.05).The subjective cure rate(100%)and objective cure rate(97.59%)in the LSC group were superior to those in the LP group(88.76%and 68.54%,respectively;P<0.05).The results of the multivariate logistic regression analysis showed that,after adjusting for the confounding factors,including age,gravidity,parity,body mass index,and duration of POP,the risk of recurrence after LSC surgery was 0.044 times that after LP(odds ratio[OR],0.044;95%CI,0.015-0.133;P<0.001).Conclusion Mid-term outcomes of LP with partial cervical preservation appear inferior to those of LSC,with LSC demonstrating superior anatomical restoration and lower rates of anatomical recurrence.However,improvements in sexual function and quality of life are comparable between the two procedures.Further evaluation with larger sample sizes and longer follow-up is warranted to better characterize long-term outcomes.
4.Diagnosis,treatment and early outcome evaluation of radicular lumbar disease
Xiaowen LIU ; Tianyi ZHAO ; Lei LIU ; Haoyang SHI ; Yang HOU ; Guodong SHI
Academic Journal of Naval Medical University 2024;45(11):1439-1443
Objective To propose diagnostic criteria for radicular lumbar disease(RLD)and evaluate its surgical outcomes.Methods A retrospective study was conducted on 31 patients with RLD who were admitted to Department of Spinal Surgery of our hospital from Mar.2020 to Oct.2023.The nerve root tension during the operation was recorded,and the surgical complications,visual analog scale(VAS)score,Oswestry disability index(ODI),Japanese Orthopaedic Association(JOA)score,and modified MacNab score were statistically analyzed.Results There were 13 males and 18 females,with an average age of(56.39±9.96)years.All the patients underwent modified transforaminal lumbar interbody fusion(TLIF),including 2 cases of single-level operation,22 cases of two-level operation,4 cases of three-level operation,2 cases of four-level operation,and 1 case of six-level operation.The follow-up duration was(15.29±4.34)months.The nerve root tension during the operation was higher(25.00[22.00,28.25]g),while it decreased immediately after the operation(2.00[0.00,3.00]g),with significant differences(P<0.01).The VAS score,ODI and JOA score after operation were improved compared with those before operation(all P<0.01).At the last follow-up,there were 24 cases of"excellent",5 cases of"good",2 cases of"fair",and no"poor"in the modified MacNab score,and the excellent and good rate was 93.55%(29/31).Surgical complications(including cerebrospinal fluid leakage and incision infection)were occurred in 1 case each.Conclusion The remarkable clinical feature of patients with RLD is high tension of nerve.These patients can be treated by modified TLIF to improve the nerve hypertonia;early and accurate diagnosis and timely intervention are the keys to a good prognosis.
5.APE1 mediates the occurrence and development of colitis-associated colorectal cancer through immunosuppressive tumor microenvironment
Tianyi CHEN ; Chaofan LI ; Lingbo BAO ; Qian CHEN ; Nana HU ; Yuxin YANG ; Lei ZHANG ; Dong WANG
Journal of Army Medical University 2024;46(16):1825-1837
Objective To investigate the regulatory mechanism of apurnic/apyrimidinic endonuclease 1(APE1)in the transformation of chronic intestinal inflammation to colitis-associated colorectal cancer(CAC).Methods C64S mutant(APE1C64S)mice and APE1 wild type(APE1WT)mice were randomly divided into experimental group and control group.In vivo CAC model was established by azoxymethane(AOM)and dextran sulfate sodium salt(DSS)solution.Immunohistochemistry(IHC)and multiple IHC(mIHC)assays were used to observe the expression of APE1 and immune cell infiltration in colon tissues of each group.A mouse colon cancer cell line MC38 with stable knockdown of APE1 was constructed by lentivirus transfection,and subcutaneous tumor bearing experiments were performed in APE1WT and APE1C64S mice to confirm that tumor cell-derived APE1 caused immunosuppressive tumor microenvironment.The expression of APE1 and CXCL1[chemokine(C-X-C motif)ligand 1]and the infiltration of immune cells in tumor-bearing specimens were analyzed by IHC and mIHC assays.The tumor specimens of a 28-year-old female patient with CAC from Army Medical Center of PLA were analyzed for the expression of APE1 and CXCL1 and the infiltration of immune cells in the tumor and adjacent inflammatory tissues by IHC and mIHC assays.Results Compared with the control group and APE1WT erperimental group,APE1C64S erperimental group had significantly reduced disease activity index and tumor formation,polymorphonuclear myeloid-derived suppressor cells(PMN-MDSCs)infiltration,and CD4+and CD8+T cells(P<0.05).No significant differences were observed in tumor growth and immune cells between APE1WT and APE 1C64S mice bearing subcutaneous tumors.However,in the tumor-bearing experiment using tumor cells with knockdown of APE1,the tumor growth was significantly lower and the number of infiltrated PMN-MDSCs was reduced,while those of CD4+and CD8+T cells were significantly increased(P<0.05).Furthermore,high expression of APE1 and increased infiltration of PMN-MDSCs were found in the tumor tissues of the young CAC patient,and CD8+T cells were significantly reduced in the tumor tissues compared with the inflammatory tissues(P<0.05).Conclusion APE1-redox in tumor cells can promote the infiltration of PMN-MDSCs and reduce the number of T cells,thereby forming an immunosuppressive tumor microenvironment and mediating the occurrence and development of CAC.
6.Expression and clinical significance of long non-coding RNA differentially expressed genes, micro RNA-181a-5p and autophagy-related protein 5 in primary gouty arthritis
Jianwei GUO ; Tianyi LEI ; Peng WANG ; Zeng ZHANG ; Guilin JIAN ; Quanbo ZHANG ; Yufeng QING
Chinese Journal of Rheumatology 2024;28(5):303-311
Objective:To explore the expression and clinical significance of long non-coding RNA colorectal neoplasia differentially expressed (lncRNA CRNDE), microRNA-181a-5p (miR-181a-5p) and autophagy related 5 (ATG5) in the peripheral blood of patients with gouty arthritis (GA) patients.Methods:The clinical data, laboratory parameters and peripheral blood samples were collected from 40 patients with acute gout (AG), 40 patients with intermittent gout (IG) and 50 healthy subjects (HC). The expression levels of lncRNA CRNDE, miR-181a-5p and ATG5 mRNA were detected by real-time fluorescence quantification (RT-qPCR) and the expression level of ATG5 protein was detected by Western-blot. The expression levels of lncRNA CRNDE, miR-181a-5p, ATG5 mRNA were compared among the three groups and correlated with clinical indices, and a subject operating characteristic curve (ROC) was constructed to assess the value of lncRNA CRNDE, miR-181a-5p, ATG5 mRNA in the diagnosis of gout. Measurements conforming to normal distribution were analyzed using t test or ANOVA, data with non-normal distribution was analyzed using Mann-Whitney U test or Kruskal-Wallis H test, correlation analysis between variables was analyzed using Spearman's analysis, and the diagnostic value of each indicator was analyzed using ROC curve. Results:① The differences in the expression of lncRNA CRNDE, miR-181a-5p, and ATG5 mRNA between the three groups were statistically significant ( H=32.12, 57.73, 68.32, all P<0.001). Among them, lncRNA CRNDE expression level in the AG group was significantly higher than that in the IG group and healthy control group [61.95(11.39, 108.30)×10 -3, 25.71(15.40, 38.40)×10 -3, 13.80(3.97, 23.99)×10 -3; Z=-3.24, P=0.001; Z=-5.03, P<0.001], and the expression level of IG group was higher than that of healthy control group( Z=-3.56, P<0.001); miR-181a-5p and ATG5 mRNA expression levels in AG group were significantly lower than those in IG group and healthy control group [miR-181a-5p: 39.81(31.22, 69.38)×10 -3, 60.74(44.19, 90.35)×10 -3, 121.30(101.50, 316.90)×10 -3; Z=-3.01, P=0.030; Z=-6.93, P<0.001. ATG5 mRNA: 4.52(2.31, 26.63)×10 -3, 43.63(13.72, 102.70)×10 -3, 153.90(66.62, 365.80)×10 -3; Z=-5.47, -7.36, all P<0.001)], which were expressed at lower levels in the IG group than in the healthy controls ( Z=-5.25, -4.47, all P<0.001). The difference of ATG5 protein expression level among the three groups expressed was statistically significant ( F=6.24, P=0.030), and the AG group was higher than the healthy control group, and the difference was statistically significant [(0.96±0.13) vs.(0.61±0.04), t=4.25, P=0.013], but the difference between the IG group (0.78±0.15) and the AG group and the HC group was not statistically significant ( t=1.51, P=0.206; t=1.85, P=0138). ② Spearman correlation analysis showed that lncRNA CRNDE was negatively correlated with the expression levels of miR-181a-5p and ATG5 mRNA in gout patients ( r=-0.49, P<0.001; r=-0.35, P=0.002); miR-181a-5p was positively correlated with ATG5 mRNA expression levels ( r=0.64, P<0.001); lncRNA CRNDE expression level was positively correlated with ESR and WBC ( r=0.49, P<0.001; r=0.43, P=0.001); miR-181a-5p expression level was negatively correlated with ESR and WBC ( r=-0.29, P=0.009; r=-0.35, P=0.002), and ATG5 mRNA expression levels were negatively correlated with ESR, WBC, and GR ( r=-0.26, P=0.021; r=-0.26, P=0.024; r=-0.27, P=0.021). In the AG group lncRNA CRNDE was positively correlated with ESR and WBC ( r=0.36, P=0.022; r=0.36, P=0.026) and miR-181a-5p was negatively correlated with WBC ( r=-0.34, P=0.038) ③ ROC curve showed that the areas under ROC curve of lncRNA CRNDE, miR-181a-5p and ATG5 mRNA expression levels to predict gout were 0.764, 0.875 and 0.864, respectively. The area under ROC curve of gout predicted by the three combined was 0.928. Conclusion:lncRNA CRNDE, miR-181a-5p, and ATG5 may be involved in the pathoge-nesis of primary gouty arthritis, and are potential biological parameters for studying the pathogenesis of gout.
7.Expression and clinical significance of N6-methyladenosine modification-related genes in peripheral blood mononuclear cells from patients with gouty arthritis
Yanhui LI ; Tianyi LEI ; Yang WANG ; Xin TU ; Mei WANG ; Xiu LI ; Bin MING ; Zeng ZHANG ; Quanbo ZHANG ; Yufeng QING
Chinese Journal of Rheumatology 2024;28(9):640-647
Objective:To investigate the expression of N6-methyladenosine(m6A) modification-related genes and their possible roles in peripheral blood mononuclear cells (PBMCs) of patients with primary gouty arthritis (GA).Methods:Forty-five patients each with acute gout (AG), intermittent gout (IG), and age-and gender-matched healthy controls (HC) were collected from the outpatient clinic of the Department of Rheumatology and Immunology of the Affiliated Hospital of Chuanbei Medical College between October and December of 2023. The expression levels of m6A modification-related genes (METTL3、METTL14、WTAP、FTO、ALKBH5、IGF2BP2、IGF2BP3、YTHDF1、YTHDC2) in PBMCs among the 3 groups were detected by RT-qPCR and correlation analysis with clinical indicators was performed. Measurements conforming to normal distribution were analyzed using ANOVA or t-tests, and data were analyzed using the Kruskal-Wallis H-test and Mann-Whitney U-test for data that is not-normaly distributed. The value of m6A modification-related genes for the diagnosis of GA was evaluated using subject characterization curve ROC. Results:①There were statistically significant differences in the expression of IGF2BP2 ( Z=-3.59, P<0.001)、WTAP ( Z=-5.25, P<0.001)、METTL14 ( Z=-3.62, P<0.001)、YTHDF1 ( Z=-2.12, P=0.034)and YTHDC2 ( Z=-2.00, P=0.045) in the disease group and the normal control group. Among them, the expression of IGF2BP2 in the GA group [28.08 (17.99, 47.06)×10 -4] was significantly higher than that in the HC group [19.23 (12.90, 25.78)×10 -4], and the expressions of WTAP、METTL14、YTHDF1 and YTHDC2 in the GA group [298.61 (213.61, 377.80)×10 -4, 9.94 (6.43, 13.46)×10 -4, 52.63 (28.22, 72.77)×10 -4, 40.24 (20.74, 73.32)×10 -4] were significantly lower than those in the HC group [398.45(339.88, 454.89)×10 -4, 13.27(11.07, 15.85)×10 -4, 64.43(43.61, 87.10)×10 -4, 53.11(36.37, 79.28)×10 -4]. Further subgroup analysis revealed statistically significant differences in the expression of IGF2BP2、WTAP、METTL14、YTHDF1 and YTHDC2 among the 3 groups ( H=19.62、31.73、13.14、16.64、28.90, all P≤0.001). The expressions of WTAP and METTL14 in the AG group [311.13(234.96, 426.67)×10 -4, Z=-3.27, P=0.001; 9.64 (5.21, 15.21)×10 -4, Z=-2.71, P=0.008] and IG group [272.27 (203.29, 347.95)×10 -4, Z=-5.78, P<0.001; 10.40(6.88, 12.88)×10 -4, Z=-3.54, P=0.003] were lower than those in the HC group [398.45 (339.88, 454.89)×10 -4, 13.27(11.07, 15.85)×10 -4]. However, there was no significant difference between AG and IG group ( P>0.05). Both YTHDF1 and YTHDC2 were significantly lower in the AG group [38.10(16.19, 56.78)×10 -4, 24.31 (14.35, 42.77)×10 -4] than those in the IG group [64.13 (48.28, 74.40)×10 -4(Z=-3.54, P<0.001, 65.49 (39.89, 91.23)×10 -4(Z=-4.96, P<0.001)] and HC group [64.43 (43.61, 86.92)×10 -4(Z=-3.51, P<0.001), 53.11 (36.37, 79.28)×10 -4(Z=-4.25, P<0.001)]. But there was no statistically significant difference between IG and HC groups ( P>0.05); IGF2BP2 was significantly lower in the AG group [25.32(16.40, 40.43)×10 -4, Z=-2.46, P=0.014] and HC group [19.23 (12.90, 25.78)×10 -4, Z=-4.54, P<0.001] than in the IG group [31.10(22.60, 49.58)×10 -4], but the comparison between AG and HC showed no statistically significant difference( P>0.05). ②Spearman correlation analysis showed that in GA patients, the expression of IGF2BP2、METTL14 and YTHDF1 was positively correlated with plasma glucose、blood uric acid(sUA) and total cholesterol level respectively ( r=0.22, P=0.037; r=0.38, P=0.003; r=0.23, P=0.034), and WTAP was negatively correlated with GLU ( r=-0.25, P=0.020). ③The ROC curve for the joint prediction of the five differential genes showed that the 95% CI for area under the curve in GA was 0.90 (0.84, 0.95). Conclusion:The m6A modification-related genes are abnormally expressed in GA and are correlated with clinical indicators such as GLU and UA, which are hypothesized to be involved in the pathogenesis of GA and have a certain reference value for the evaluation of metabolism in GA patients.
8.Expression of decoy receptor 3 and its signaling pathway in ankylosing spondylitis and its clinical significance
Yi JIANG ; Xia LIAO ; Shunbing WANG ; Yixi HE ; Tianyi LEI ; Zeng ZHANG ; Jianwei GUO ; Yufeng QING
Chinese Journal of Rheumatology 2023;27(1):28-33,C1-4
Objective:To investigate the expression and clinical significance of decoy receptor 3 (DcR3) and its signal pathway-related molecules in PBMCs of patients with ankylosing spondylitis (AS).Methods:Peripheral blood samples, clinical data and laboratory test results were collected from 100 patients with ankylosing spondylitis [50 patients with AS activity (ASA), 50 patients with AS stability (ASS)], 30 patients with osteoarthritis and 30 patients with gouty arthritis (as disease control group), and 60 healthy controls (HC). The mRNA expression levels of DcR3 and its signal pathway related genes (DR3, TL1A, Fas, FasL, LIGHT, LIGHTR, LTβR) were measured by real-time fluorescence quantitative polymerase chain reaction. Measurement data among the three groups in normal distribution were analyzed by t test or one-way analysis of variance, pairwise comparisons using LSD- t test, non-normal distribution data were analyzed by Mann-Whitney test or Kruskal-Wallis H test, χ2 test was used for correlation analysis of categorical variables. Correlation analysis between variables were analyzed using Spearman correlation analysis. Results:① By comparing the AS group, disease control group and HC group, the expression levels of DcR3 mRNA and DR3 mRNA in the AS group were lower than those in disease control group and HC group, and DcR3 mRNA and DR3 mRNA in disease control group were lower than those in the HC group {DcR3mRNA: [6.21 (3.89, 10.70)]×10 -4vs [9.51 (5.89, 16.65)]×10 -4vs [17.81 (11.27, 24.20)]×10 -4, H=55.28, P<0.001; DR3 mRNA: [41.05 (24.09, 66.95)]×10 -4vs [58.28 (28.41, 94.38)]×10 -4vs [94.79 (54.07, 144.51)]×10 -4, H=37.10, P<0.001}. The expression level of TL1A mRNA in the AS group was higher than that in disease control group {[14.71(4.91, 42.22)]×10 -4vs [4.00(1.07, 16.60)]×10 -4vs [7.70 (3.52, 27.83)]×10 -4, H=17.71, P<0.001}; The expression level of Fas mRNA in AS group and disease control group was lower than that in HC group {[20.99(4.63, 62.89)]×10 -4vs [23.97(15.82, 38.99)]×10 -4vs [78.45 (27.32, 146.46)]×10 -4, H=31.17, P<0.001}. The expression level of FasL mRNA in AS group was higher than that in disease control group and HC group {[42.87(6.57, 91.21)]×10 -4vs [5.45(2.83, 10.32)]×10 -4vs [6.88 (4.57, 23.79)]×10 -4, H=46.42, P<0.001}. The expression level of LIGHTR mRNA in AS group was lower than that in disease control group {[52.66 (7.20, 143.21)]×10 -4vs [98.80 (53.11, 166.24)]×10 -4vs [63.47(40.85, 138.07)]×10 -4, H=11.96, P<0.001}. There were no significant differences in LIGHT mRNA and LTβR mRNA among all groups ( H=0.86, P>0.05; H=3.18, P>0.05). ②The expression levels of DcR3 mRNA, DR3 mRNA and Fas mRNA in ASA group and ASS group were lower than those in HC group. DcR3 mRNA in ASA group was higher than that in ASS group, and DR3 mRNA in ASA group was lower than that in ASS group {DcR3 mRNA: [7.28 (4.92, 16.56)]×10 -4vs [4.59 (2.49, 7.03)]×10 -4vs [17.81 (11.27, 24.20)]×10 -4, H=62.63, P<0.001; DR3 mRNA: [30.93(16.18, 66.66)]×10 -4vs [47.17(29.91, 67.40)]×10 -4vs [94.79(54.07, 144.51)]×10 -4, H=41.48, P<0.001; Fas mRNA: [20.04(3.29, 62.30)]×10 -4vs [22.49(5.63, 64.79)]×10 -4vs [78.45(27.32, 146.46)]×10 -4, H=23.54, P<0.001}. The expression levels of TL1A mRNA and LTβR mRNA in the ASA group were higher than those in the ASS group and the HC group {TL1A mRNA: [32.36(10.09, 97.84)]×10 -4vs [9.98(1.29, 21.63)]×10 -4vs [7.70(3.52,27.83)]×10 -4, H=21.14, P<0.001; LTβR mRNA: [6.13(2.16,20.06)×10 -4vs [2.13(0.53,8.04)]×10 -4vs [2.72 (1.24,5.73)]×10 -4, H=12.86, P<0.001}. The expression level of FasL mRNA in the ASA group and the ASS group was higher than that in the HC group {[60.70 (8.16, 106.16)]×10 -4vs [30.14 (5.37, 78.40)]×10 -4vs [6.88 (4.57, 23.79)]×10 -4, H=18.99, P<0.001}. The expression level of LIGHTR mRNA in ASS group was lower than that in HC group {[49.79(10.75, 168.48)]×10 -4vs [15.92(3.27, 105.91)]×10 -4vs [63.47(40.85, 138.07)]×10 -4, H=11.80, P<0.001]. There was no significant difference in LIGHT mRNA among all groups ( H=4.15, P>0.05). ③Spearman correlation analysis showed that DcR3 level was positively correlated with BASDAI score and hsCRP in AS patients ( r=0.52, P<0.001; r=0.35, P<0.01), and DR3 level was negatively correlated with BASDAI score, ESR and hsCRP level ( r=-0.28, P<0.001; r=-0.25, P<0.001; r=-0.31, P<0.001). TL1A was positively correlated with BASDAI score, ESR and hsCRP level ( r=0.23, P=0.046; r=0.26, P=0.015; r=0.25, P=0.017). Conclusion:DcR3 and its signal pathway-related molecules are differentially expressed in PBMCs of patients with AS, suggesting that they may participate in the occurrence and development of AS.
9.Isolation and cultivation of HIV strains circulating in China and the screening and identification of the standard strains
Siqi LI ; Guoxin ZHU ; Lin LI ; Jingyun LI ; Hanping LI ; Yongjian LIU ; Lei JIA ; Xiaolin WANG ; Tianyi LI ; Bohan ZHANG ; Jingwan HAN ; Chuntao MA
Chinese Journal of Experimental and Clinical Virology 2023;37(3):286-295
Objective:To screen the HIV standard strains with typical biological characteristics of HIV strains circulating in China through the isolation, culture, genotype and phenotype identification of HIV from the whole blood samples of HIV-infected persons, confirm genetic characteristics, traceability, and in line with the Standard Strains of Pathogenic Microorganism-technical Specifications for Establishment of HIV Strains (T/CPMA 027—2023).Methods:Whole blood samples were collected from 48 HIV infected patients. Peripheral blood mononuclear cells (PBMCs) were isolated from the samples and co-cultured with PBMCs isolated from healthy persons′ whole blood samples to isolate and culture HIV from infected persons. We determined concentration of p24 antigen and the virus titer in the culture supernatant. The viral RNA was extracted from the successfully isolated strains, and the gag, pol genes and env C2V3 fragments of the viral genome were amplified and sequenced. The genotype, gene recombination and drug resistance sites were determined according to the viral gene sequences. Virus infection and replication were monitored by inoculating the virus culture supernatant into Ghost cells expressing CCR5 or CXCR4 to determine the viral tropism.The formation of syncytium was observed by inoculating the virus culture supernatant into MT-2 cells to determine whether was a syncytium-induced phenotype. Results:Fourteen strains with p24 antigen concentration > 1 ng/ml in culture supernatant were isolated and cultured from 48 fresh EDTA anticoagulated whole blood samples of HIV infected persons. Of the 14 strains, only one strain with a titer≥10 5 TCID 50/ml, 8 strains with titers ≥10 4 TCID 50/ml, and the other 5 strains with titers≥10 3 TCID 50/ml. Phylogenetic analysis showed that the genotypes of the strains were 9 strains of subtype B, 3 strains of CRF01_AE and 2 strains of CRF07_BC recombinant. Genotypic resistance analysis showed that 11 strains contained drug resistance sites. Ghost cells were used to verify the tropism of the strains, and it was found that 8 strains were CCR5 tropism, 6 strains were CXCR4 & CCR5 dual tropism. Only 2 of the 14 strains could induce MT-2 cytopathic effect, which was syncytium-inducing phenotype. Conclusions:Fourteen HIV strains with typical biological and genetic characteristics were isolated to screen the standard HIV strains. Among which, 1 strain was evaluated as a standard HIV strain that meets the Standard Strains of Pathogenic Microorganism-technical Specifications for Establishment of HIV Strains (T/CPMA 027—2023). This study can also provide technical guidance for the screening of the HIV standard strains. Next step is to complete the application and reserve database construction according to the sharing mechanism of the HIV standard strains, to provide resources for the researches of HIV vaccines and drugs.
10.Changes and clinical significance of microRNA-146b and signal transducer and transcriptional activator 1/3 in male primary gouty arthritis
Zeng ZHANG ; Peng WANG ; Jianwei GUO ; Tianyi LEI ; Quanbo ZHANG ; Shaowei NIU ; Xia LIAO ; Yufeng QING
Chinese Journal of Rheumatology 2023;27(6):385-392
Objective:To This study was to investigate the expression and possible clinical significance of microRNA-146b (miR-146b) and signal transducer and transcriptional activator 1 and 3 (STAT1/3) in peripheral blood mononuclear cells (PBMCs) of patients with primary gouty arthritis (GA).Methods:The peripheral blood samples, clinical data and laboratory indexes of 120 male cases of GA [including 57 cases of acute (AG group) and 63 cases of intermittent (IG group)]and 66 healthy subjects (HC group) were collected. The expression levels of miR-146b and STAT1/3 in PBMCs were detected by real-time fluorescence quantitative PCR (RT-qPCR). The differences among the three groups were compared and the correlation between them and clinical indexes was analyzed. The receiver operating characteristic curve (ROC) was constructed to evaluate its diagnostic value in GA. After the PBMCs of 18 healthy subjects were stimulated by 100 μg/ml MSU for 3 hours to simulate acute gout inflammatory environment, the transcriptional changes of IL-1β, miR-146b and STAT1/3 were detected by RT-qPCR, and the expressions of IL-1β, STAT1/3 protein and phosphorylated protein were detected by Western blotting. T test or one-way ANOVA and LSD- t test were used in accordance with the normal measurement data, Kruskal-Wallis H and Mann-Whitney U test were used in the non-normal data, Spearman correlation analysis was used in the correlation between variables, and the diagnostic value was evaluated by the receiver working characteristic curve ROC. Results:①There were statistical differences in the expression of miR-146b, STAT1 and STAT3 among the three groups ( F=7.02、19.52、17.07, all P<0.001). The expression of miR-146b in gout group [(0.32±0.28)] was significantly higher than that in HC group (0.19±0.18)( t=2.96, P=0.003), while STAT1(0.019±0.012) and STAT3(0.014±0.010) were significantly lower than those in HC group (0.038±0.029),(0.025±0.016)( t=6.26, 5.56, both P<0.001). Further subgroup analysis showed that the expression of miR-146b in AG and IG groups was higher than that in HC group[(0.27±0.17), (0.38±0.35), (0.19±0.18), t=2.09, 3.30, both P<0.05], but that in AG group was lower than that in IG group ( t=2.02, P<0.05). The expression of STAT1 mRNA in AG and IG groups was lower than that in HC group [(0.020±0.012), (0.019±0.012), (0.038±0.029), t=4.89, 4.56, both P<0.001], but there was no statistical significance between AG and IG groups ( t=0.24, P>0.05). The expression of STAT3 mRNA in AG and IG groups was lower than that in HC group [(0.016±0.012), (0.012±0.008), (0.025±0.016), t=5.64, 3.33, both P<0.01], and the expression of STAT3 mRNA in AG group was higher than that in IG group ( t=2.12, P<0.05). ② Spearman correlation analysis showed that the expression of miR-146b in GA was negatively correlated with HCY ( r=-0.37, P=0.014), STAT1 was negatively correlated with Crea ( r=-0.29, P=0.019), positively correlated with eGFR ( r=0.25, P=0.047), and STAT3 was negatively correlated with HDL-C ( r=-0.27, P=0.033). ③ROC curve showed that the AUC (95% CI) of miR-146b, STAT1 and STAT3 were 0.679(0.582, 0.776), 0.710(0.629, 0.791) and 0.705(0.626, 0.783), and the combined AUC(95% CI) of the three was 0.836 (0.765, 0.907). ④Compared with blank control group and negative control group, the expression of miR-146b in PBMCs of 18 cases of HC was significantly decreased ( H=14.44, P=0.003), while the expression of IL-1β, STAT1 and STAT3 mRNA was significantly increased after 3 h of MSU stimulation ( H=26.44、27.26、15.90, all P<0.001). The expression of IL-1β, STAT1 and STAT3 protein and phosphorylated protein in the model group were significantly higher than those in the blank control group, and the differences were statistically significant ( t=9.97、6.63、7.48、11.25、6.28, all P<0.01). Conclusion:The abnormal expression of miR-146b and STAT1/3 in GA is related to some clinical indicators, suggesting that it may be involved in the regulation of gout immune inflammatory response and metabolism, and the specific mechanism is worth further study.

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