1.Distribution of resistance genes and virulence genes in multidrug-resistant Salmonella typhimurium strains
Yufeng ZHANG ; He SONG ; Le YAN ; Pengfei XU ; Ruiqing LIU ; Tiantian TANG ; Xiaoyan WANG ; Huiling DENG ; Kairui LEI
Chinese Pediatric Emergency Medicine 2024;31(11):831-835
Objective:To study the distribution of drug resistance genes and virulence genes in multidrug-resistant Salmonella typhimurium strains.Methods:A total of 96 strains of Salmonella typhimurium were collected,and drug sensitivity tests were performed to evaluate the drug resistance and multidrug-resistance of Salmonella typhimurium.Multidrug-resistant Salmonella typhimurium strains were selected to conducted whole genome sequencing,and the distribution of drug resistance genes and virulence genes in the strain were analyzed.Results:Salmonella typhimurium strains had the highest resistance rates to ampicillin and ampicillin/sulbactam,with 89.58% and 76.04%,respectively.Followed by trimethoprim/sulfamethoxazole,ceftriaxone,and aztreonam,with 47.92%,38.54% and 33.33%,respectively,and low resistance rates to ciprofloxacin and levofloxacin,with 8.33% and 4.17%,respectively.Ninety-six strains were all sensitive to carbapenem antibiotics and piperacillin/tazobactam.Fifty-seven strains(59.38%)of Salmonella typhimurium showed multidrug-resistance.Resistance genes were detected in all 57 multidrug-resistant Salmonella typhimurium strains,with higher carrier rates of 98.25%,77.19%,and 59.65% for aac(6')-Iaa,aadA22,and blaTEM-1B,respectively.The multidrug-resistant Salmonella typhimurium strains had the highest carrier rates for invA,sipA,sseL,and sopB.Conclusion:Multidrug-resistant Salmonella typhimurium strains have a high incidence and a high carrier rate for multiple drug resistance genes and virulence genes.The monitoring and prevention of Salmonella typhimurium should be strengthened in the clinic in order to reduce the spreading epidemic of multidrug-resistant strains.
2.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.
3.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.
4.Effect of preoperative mild cognitive impairment on potency of sevoflurane in inhibiting body movement responses during skin incision in aged female patients
Lei WANG ; Zhiqiang NIU ; Sen YANG ; Yufeng GUO ; Min LIU
Chinese Journal of Anesthesiology 2024;44(8):977-980
Objective:To evaluate the effect of preoperative mild cognitive impairment (MCI) on the potency of sevoflurane in inhibiting body movement responses during skin incision in aged female patients.Methods:This was a prospective study. Female American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ patients, aged 60-75 yr, with body mass index of 18.5-23.9 kg/m 2, scheduled to undergo radical mastectomy between January 2022 and March 2023 in our hospital, were selected. The patient′s cognitive function was assessed using Mini-Mental State Examination and Montreal Cognitive Assessment. The patients were divided into 2 groups based on whether they had MCI or not before operation: normal group (group N) and MCI group (group M). General anesthesia was induced by inhaling 8% sevoflurane, and the laryngeal mask airway was inserted after they lost consciousness and their jaws relaxed. According to the Dixon′s up-and-down method, the end-tidal concentration of sevoflurane in the first patient was set at 2%. If the body movement response occurred, the concentration was increased by 2% in the next patient, otherwise the concentration was decreased by 2% in the next patient. The MAC and 95% confidence interval ( CI) of sevoflurane were calculated using the probability regression method. Results:The minimum alveolar concentration of sevoflurane was 1.60% (95% CI 1.48% -1.70%) in group N and 1.38% (95% CI 1.25%-1.49%) in group M, and there was statistically significant difference ( P<0.05). Conclusions:Preoperative MCI can increase the potency of sevoflurane in inhibiting body movement responses during skin incision in aged female patients.
5.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.
6.Effects of weekend admission on total hospitalization expenses for elderly patients with hip fracture under the geriatric orthopedic co-management
Lifang WANG ; Yanwei LYU ; Yufeng GE ; Lei PAN ; Fangfang DUAN ; Shiwen ZHU
Chinese Journal of Orthopaedic Trauma 2023;25(12):1049-1055
Objective:To explore the impact of weekend hospitalization on total hospitalization expenses for elderly patients with hip fracture under the geriatric orthopedic co-management.Methods:A retrospective analysis was conducted to analyze the clinical data of elderly patients with hip fracture who had been hospitalized for surgical treatment at Beijing Jishuitan Hospital from May 2015 to December 2020. They were divided into 2 groups based on their admission date. Group A was admitted from Monday to Thursday while Group B from Friday to Sunday. The general demographic data, diagnostic information, comorbidities, hospitalization expenses of the patients were collected. The differences in total hospitalization expenses, hospitalization time, rate of surgery within 48 hours and rate of hospital mortality between the 2 groups were analyzed by rank sum test, chi square test, correlation analysis, and multiple linear regression.Results:A total of 6,075 patients with hip fracture were included in this study, including 1,675 males and 4,400 females with a median age of 80 (74, 85) years. There were 3,935 ones in group A and 2,140 ones in group B. The total hospitalization expenses for group A was 58,160.52 (49,215.45, 72,748.94) yuan, insignificantly lower than those for Group B [58,412.90 (49,163.58, 72,712.61) yuan] ( P>0.05). The rate of surgery within 48 hours for group A was 75.8% (2,984/3,935), significantly higher than that for group B [49.3% (1,054/2,140)]. The hospitalization time for group A was 5 (4, 7) days, significantly less than that for group B [5 (4, 7) days] ( P<0.05). There was no significant difference in the rate of hospital mortality between the 2 groups ( P>0.05). Multiple linear regression analysis showed that total hospitalization expenses were significantly higher for patients admitted on weekends, hospitalization time was positively correlated with total hospitalization expenses, and total hospitalization expenses were significantly lower for the patients undergoing surgery within 48 hours ( P<0.05). Conclusion:Admission on weekends can increase total hospitalization expenses, prolong hospitalization time, and reduce rate of surgery within 48 hours for elderly patients with hip fracture.
7.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.
8.Expression of pyroptosis related molecules in primary gouty arthritis and its clinical value
Tianyi LEI ; Xiang YU ; Hongyuan XIE ; Jianwei GUO ; Peng WANG ; Zeng ZHANG ; Quanbo ZHANG ; Yufeng QING
Chinese Journal of Rheumatology 2023;27(11):756-765
Objective:To explore the molecular mechanism of cell death of the peripheral blood mononuclear cells (PBMCs) of patients with primary gouty arthritis, and provide new idea for the treatment of gout.Methods:Peripheral blood samples and clinical data were collected from 30 patients with acute gout (AG), 30 patients with intermittent gout (IG) and 40 healthy controls(HC). Real-time fluorescence quantitative detection of cell apoptosis related molecules, including the mRNA expression level of nucleotide binding oligomerization domain like domain like receptor protein 3(NLRP3), cysteine aspartic proteinase-1/4/5 (caspase-1/4/5), Gasdermin A/B/C/E. And NLRP3, precursor caspase-1 (pro-caspase-1), clipped caspase-1 (caspase-1 + p10), Gasdermin D(GSDMD), N segment GSDMD (GSDMD-N), precursor IL-1β (pro IL-1β), mature IL-1β (clevated IL-1β)were detected by western blot. The measurement data of normal or approximate normal distribution were analyzed by independent sample t test or one-way variance analysis (ANOVA), the measurement data of non-normal distribution were analyzed by Mann-Whitney U test or Kruskal-Wallis H test, and the counting data was compared by Chi-square test. Pearson's correlation analysis was used for the continuous variables with normal distribution, and Spearman's correlation analysis was used for the continuous variables with non-normal distribution. The logistic regression analysis was used to assess risk factors. Results:① There were no significant differences in MPR and BMI between AG and IG ( χ2=0.64, P=0.426; t=0.04, P=0.972), and there was significant difference in disease course [25.0 (9.8, 63.0), 54.0 (33.0, 102.0)mouth, Z=2.01, P=0.044]. Comparison of labora-tory parameters: there were statistical significant differences in ESR between AG and IG ( t=5.24, P<0.001), eGFR, GR, LY, RBC, HCT, UA, Creatinin, ALT and AST. ② In the three groups, the expression lev-els of caspase-1, GSDMC, GSDMD, GSDME, NLRP3 mRNA were statistically significantly different. In AG and IG groups, mRNA expression levels of caspase-1 (1.55±0.62), (1.58±0.62), GSDMD (4.7±1.4), (3.5±1.53), NLRP3 [2.63(2.03, 4.10), 2.39(1.57, 3.49)] were higher than those of the HC group [(1.24±0.59), 1.16±0.71, 1.16 (0.50, 2.34)] ( P=0.037, P=0.023, P<0.001, P<0.001, P<0.001, P<0.001). In IG group, mRNA expression levels of GS-DMD (3.53±1.53) were lower than those of AG group (4.68±1.43) ( P<0.001).The mRNA expression levels of GS-DMC and GSDME [0.57(0.33, 0.78), (0.32±0.15)]were lower than those of the HC group [0.80 (0.47, 1.86), (1.06 ± 0.36) ( P=0.004, P<0.001), and the mRNA expression levels of GSDME (0.62±0.29) in the IG group were lower ( P=0.004, P<0.001), However, in the IG group, GSDMC and GSDME [0.87 (0.51, 1.53), (0.62±0.29)] were higher than those in the AG group [0.57 (0.33, 0.78), (0.32±0.15)] ( P=0.003, P<0.001). ③ The expression levels of NLRP3, pro-caspase-1, caspase-1 + p10, GSDMD, GSDMD-N, pro-IL-1β, clevated IL-1β protein were statistically different among the three groups [( F=50.04, P<0.001; F=9.65, P=0.013; F=30.71, P=0.001; F=7.38, P=0.024; F=23.66, P=0.001; F=30.11, P=0.001; F=6.01, P=0.036]. The expression of NLRP3 protein in the AG group (1.14±0.12) was significantly higher than that in the IG and HC group (0.35±0.18), (0.17±0.03) (all P=0.001), the expression levels of Pro caspase-1, caspase-1+p10 protein in the AG (1.11±0.15), (0.93±0.38) and IG (0.98±0.14), (1.14±0.17) group were higher than those in the HC (0.42±0.28), (0.29±0.16) ( P=0.006, P=0.015). The expression levels of GSDMD protein in the AG group (1.04±0.16) were higher than those in the IG and HC group(0.53±0.26), (0.39±0.22) ( P=0.029, P=0.011). The expression level of GSDMD-N protein in the AG and IG group (0.97±0.06), (0.90±0.04) was higher than that in the HC (0.27±0.23) ( P=0.001, P=0.001). The expression level of pro-IL-1β protein in the AG group (1.01±0.06) was significantly higher than that in the IG and HC group (0.32±0.14), (0.64±0.11) ( P<0.001, P=0.006), but lower than that in the HC (0.64±0.11) ( P=0.011). The expression of clevated IL-1β protein was higher in the AG group (1.08±0.20) than in the HC group (0.33±0.24) ( P=0.014). ④ Negative correlation between NLRP3, GSDMD and LY ( r=-0.32, P=0.001; r=-0.24, P=0.017) and positive correlation between GSDMD and WBC, GR ( r=0.43, P<0.001; r=0.23, P=0.019) were found and Logistic regression analysis showed that the GSDMD and NLRP3 were risk factors for AG [ OR ( 95%CI)=11.29 (3.92, 32.48), P<0.001; OR( 95%CI)=2.21(1.00, 4.85), P=0.049]. GSDMD was risk factor for IG [ OR( 95%CI)=6.84(2.52, 18.53), P<0.001]; While GSDMD was the protective factor for IG [ OR( 95%CI)=0.61(0.41, 0.30), P=0.013]. Conclusion:The expression's of NLRP3, Caspase-1 and GSDMD are increased in PBMCs of AG patients, while the expression's of GSDMC and GSDME are decreased. NLRP3/Caspase-1/GSDMD may be associated with the onset of acute gouty arthritis.
9.Clinical characteristics and one-year follow-up outcomes of 6 children with primary nephrotic syndrome infected with SARS-CoV-2 during the Omicron variant epidemic
Rufeng DAI ; Qian SHEN ; Lei YIN ; Yulin KANG ; Yufeng LI ; Jing CHEN ; Mei ZENG ; Hong XU
Chinese Journal of Nephrology 2023;39(10):729-737
Objective:To investigate the clinical characteristics and short-term follow-up outcomes of primary nephrotic syndrome (PNS) children infected with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the Omicron variant outbreak in Shanghai, and to provide a reliable reference for clinicians in the diagnosis and treatment.Methods:It was a case-control study. The clinical data of children with PNS (PNS group) who were diagnosed and followed-up up to 1 year in the nephrology department of four children's medical centers in Shanghai, and the children (control group) who had no underlying diseases and were infected with SARS-CoV-2 in Shanghai Jinshan Public Health Center, including the data when they were infected with SARS-CoV-2, were retrospectively analyzed.Results:(1) From March 30th to April 13th, 2022, 6 PNS children in Shanghai were infected with SARS-CoV-2, including 5 boys and 1 girl. The median age was 4.5 (2.0, 11.0) years old. And 30 children were matched by sex, age and disease type as control group, including 20 males and 10 females. The median age was 4.5 (2.0, 9.0) years. There were no significant differences between the PNS group and the control group in clinical symptoms (including fever duration), treatment regimens, vaccine doses and virus clearance time (all P>0.05). (2) The 6 children with PNS included 3 cases of steroid-sensitive type, 3 cases of steroid-resistant type, 2 cases of minimal change disease, 2 cases of focal segmental glomerulosclerosis and 2 cases with no renal biopsy. Before SARS-CoV-2 infection, their primary disease-PNS were stable, and urine protein was negative, four of them were under maintenance treatment with oral steroids or immunosuppressive drugs. At the time of SARS-CoV-2 infection, the symptoms of all of the 6 cases were mild, no severe, critical or fatal cases, and they were all cured and discharged from hospital through medical isolation observation or symptomatic treatment of infections. (3) Five cases of them still had discomfort symptoms such as cough, anorexia, and fatigue after being discharged from the hospital, which lasted for about 1 week. Within 1 year of follow-up, none of the children have suffered from "recurrent positive PCR results" or "secondary infection" of the SARS-CoV-2. (4) Among them, 4 cases of PNS relapsed after SARS-CoV-2 infection, timely addition of steroids was effective, their urine protein quickly turned negative, and there was no recurrence after 1 year of follow-up. (5) Before infection with SARS-CoV-2, the levels of immunoglobulin IgG were lower than the normal reference value in the 4 cases with PNS recurrence. Conclusions:During the Omicron variant outbreak in Shanghai, the infection of SARS-CoV-2 in children with PNS are resulted in high transmission among household contacts. Most of them have mild symptoms and good prognosis. PNS is prone to relapse after SARS-CoV-2 infection, and steroid therapy is effective and safe for these relapse. IgG may be a potential marker for the prognosis of PNS children infected with SARS-CoV-2.
10.Comparison of characteristic chromatogram and the contents of multiple indicator components of Morus alba decoction powder and decoction at different decoction time
Jinjin JIANG ; Lei WU ; Yufeng ZHU ; Si SUN ; Mingming QIU ; Chao LU
China Pharmacy 2023;34(5):565-568
OBJECTIVE To compare characteristic chromatogram and the contents of multiple indicator components of Morus alba decoction powder and decoction at different decoction time, and to provide experimental basis for the development of M. alba decoction. METHODS Taking decoction powder and decoction at different decoction time as subject, HPLC characteristic chromatogram of 2 kinds of samples were established with Similarity Evaluation Software System of TCM Chromatographic Fingerprint (2012 version), and similarity evaluation was performed. The contents of mulberroside A, geniposide, berberine, baicalin, quercetin and luteolin in decoction powder and decoction were determined by HPLC. The contents of each indicator component and the change of total content were as the evaluation indexes to compare the difference between the two substances during decoction. RESULTS The similarities of characteristic chromatogram of the two substances ranged from 0.943 to 1.000 and 0.975 to 0.998 at different decoction time, respectively. Six indicator components of the decoction powder dissolved faster and had higher contents. The contents of each indicator component in the decoction powder when decocting at 20 minutes was 1.1-1.5 times of the decoction when decocting at 50 min, and the total content in the decoction powder was 1.2 times of the decoction. CONCLUSIONS Compared with decoction, M. alba decoction powder has the advantages of shortening the decoction time and saving traditional Chinese medicine resources. The results of this study lay a research foundation for “Zungu” to develop its preparation.

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