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  • 2024 Volume 34 Issue 04
    Published: 23 July 2024
      
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  • Eicosapentaenoic acid inhibited lipopolysaccharideinduced mesangial cell pyroptosis by regulating Nrf2 pathway
    LI Jie, FENG Ye, LU Qingxue, et al.
    2024, 34(04): 277-282.
    Abstract ( ) PDF ( )   Knowledge map   Save
    Objective： To explore the regulatory effect of eicosapentaenoic acid (EPA) on lipopolysaccharide (LPS)induced HBZY1 cell pyroptosis based on the Nrf2/NLRP3/GSDMD signaling pathway. Additionally, nuclear factor E2 related factor 2 (Nrf2) agonist (4OI) and nucleotidebound oligomerized domainlike receptors (NLRP3) receptor inhibitor (MCC950) were chosen to explore the regulating mechanism of EPA on Nrf2. Methods： According to different drug treatments strategy, HBZY1 cells were divided into five groups: control group, LPS group, LPS+EPA group, LPS+EPA+4OI group, and LPS+EPA+MCC950 group, in which 10 μg/mL LPS was applied to induce the cell pyroptosis in vitro, and the concentration of EPA, 4OI, MCC950 was all 200 μmol/L, the drugs were added to the culture plate and treated for 48 h. In addition, CCK8 assay, LDH release test, and TUNEL staining were performed to detect cell damage. The release of inflammatory factors IL1β and IL18 in the cell supernatant were detected by ELISA kits, protein levels of Nrf2, NLRP3, Caspase1 and GSDMD were detected by Western Blotting. Results： Cell viability assay showed that 10 μg/mL LPS stimulation for 48 hours significantly reduced the cell viability (P<005), but 20-200 μmol/L EPA had significantly protective effects (P<005). Moreover, 200 μmol/L EPA significantly attenuated the cell damage caused by LPS (P<001), reduced the release of inflammatory factors (P<001), LDH release (P<001) and DNA fragmentation, inhibited the activation of the pyroptosis, and promoted the expression of Nrf2 protein (P<001). In addition, cotreatment with 4OI and MCC950 found that 4OI can promote the inhibitory effect of EPA on pyroptosis (P<005), reduce the expression of related proteins and the release of IL1β and IL18 (P<005), and enhance the protection effect of EPA (P<001), while MCC950 has no significant on the effect of EPA. Conclusion： EPA inhibits the expression of various molecules in the pyroptosis of HBZY1 cells induced by LPS through Nrf2, and exerts a significant cytoprotective effect.
  • Expression of pyroptosis related proteins in fetal membranes of pregnant women with premature rupture of membranes and its relationship with pregnancy outcome
    QIAN Yunying, QIAN Guiying, CAI Ximei, et al.
    2024, 34(04): 283.
    Abstract ( ) PDF ( )   Knowledge map   Save
    Objective： To explore the expression of pyroptosis related protein cysteinyl aspartate specific proteinase1 (Caspase-1) and nucleotidebinding oligomerization domainlike receptor protein-3 (NLRP3) in fetal membranes of pregnant women with premature rupture of membranes (PROM) and their relationship with pregnancy outcomes. Methods： A total of 135 pregnant women with PROM who received medical treatment and delivered at the Obstetrics and Gynecology Department of Changshu Traditional Chinese Medicine Hospital from January 2020 to January 2023 were selected as the observation group, and other 135 healthy pregnant women who received prenatal examination and delivered during the same period were selected as the control group. The expression of NLRP3 mRNA and Caspase-1 mRNA in the fetal membrane tissue of the two groups of pregnant women was compared. One hundred and thirtyfive pregnant women with PROM were divided into a poor pregnancy group (n=50) and a good pregnancy group (n=85) based on pregnancy outcomes. Independent risk factors for poor pregnancy in pregnant women with PROM were analyzed using univariate and multivariate Logistic analysis. Logistic regression model and restricted cubic splines (RCS) were used to analyze the doseresponse relationship between the expression of pyroptosis related proteins in fetal membranes of pregnant women with PROM and adverse pregnancy. A nomogram prediction model was constructed by using independent influencing factors and the model was also verified. Results： The expression of NLRP3 mRNA and Caspase-1 mRNA in the fetal membrane tissue of the observation group was significantly higher than that of the control group (P<0.001). Visit time, reproductive tract infections, and the mRNA expression levels of NLRP3 and Caspase-1 are independent risk factors for poor pregnancy outcomes in pregnant women with PROM (P<0.05). The RCS results showed that the expression levels of NLRP3 and Caspase-1 in the fetal membrane tissue of pregnant women with PROM showed a non-linear doseresponse relationship with adverse pregnancy outcomes. The highest risk of adverse pregnancy outcomes occurred at NLRP3 mRNA expression levels of 1.20 (OR=1.818, 95%CI: 1.673-1.932) and Caspase-1 mRNA expression levels of 1.25 (OR=2.735, 95%CI: 1.132-3.821). The constructed nomogram prediction model has good discrimination, accuracy, and clinical applicability. Conclusion： Caspase-1 mRNA and NLRP3 mRNA are highly expressed in the fetal membrane tissue of pregnant women with PROM, and they are independent risk factors for adverse pregnancy outcomes in pregnant women with PROM. With the increase of expression levels, the risk of adverse pregnancy in pregnant women with PROM also increases.
  • Clinical efficacy of flexible ureteroscopic lithotripsy with flexible vacuum suction ureteral access sheath and tubeless discharge in the treatment of >2 cm upper urinary tract calculi
    IANG Xiaofeng, LI Yunzhi, JIN Ming, et al.
    2024, 34(04): 301.
    Abstract ( ) PDF ( )   Knowledge map   Save
    Objective： To investigate the clinical efficacy of flexible ureteroscopic lithotripsy（FURL） using flexible vacuum suction ureteral access sheath (FVSUAS) in the treatment of upper urinary tract calculi >2 cm, through the combination of crushing and suction technology, without indwelling double J tube before discharge. Methods： Clinical data of 43 patients with >2 cm upper urinary tract stones treated with FVSUAS during FURL from January 2022 to October 2023 were collected and compared with 40 randomly selected patients treated with conventional FURL during the same period. The trial group chose to undergo FURL with FVSUAS and was discharged from the hospital with removal of the ureteral stent tubes 3 days after the operation. In the control group, conventional ureteral access sheath (UAS) was used, and ureteral stent tubes were routinely placed in place postoperatively, and the ureteral stent tubes were removed under second-stage cystoscopy/ureteroscopy at 4 weeks; general data and perioperative indices of patients of the two groups were recorded, and the postoperative stonefree rate, postoperative complications, and patient satisfaction were obtained. Results： The stone-free rate on the 3rd postoperative day in the trial group was 93.02% (40/43), which was significantly higher than that in the control group of 65.00% (26/40), and the difference was statistically significant (P=0.002). When haematuria, pain, and fever between the trial group and the control group on the 3rd postoperative day was compared, the difference was not statistically significant (P>0.05); the proportion of haematuria, pain, and fever in the control group at 4 weeks after the surgery was significantly higher than that of the trial group, the difference was statistically significant (all P<0.05, the satisfaction of the trial group at 4 weeks postoperatively was significantly higher than that of the control group (P<0.01). Conclusion： FURL in combination with FVSUAS for the treatment of >2 cm upper urinary tract stones and tubeless discharge with ureteral stent tubes removed 3 days postoperatively achieves high stonefree rate and significantly reduces the incidence of fever, haematuria and pain complications, and it can be concluded that it is a safe and effective treatment modality.
  • Flexible ureteroscopic holmium laser lithotripsy and rigid ureteroscopic holmium laser lithotripsy with NTrap in the treatment of upper ureteral calculi: a randomized controlled clinical trial
    HAN Xiaozhou, ZHAO Cheng, ZHOU Mingmei, et al.
    2024, 34(04): 307.
    Abstract ( ) PDF ( )   Knowledge map   Save
    Objective： To analyze the clinical efficacy of flexible ureteroscopic holmium laser lithotripsy (FURL) and rigid ureteroscopic holmium laser lithotripsy (RURL) with N-Trap in the treatment of upper ureteral calculi. Methods： A total of 92 cases of upper ureteral calculi with a diameter greater than 10 mm admitted to Shanghai TCM-Integrated Hospital, Shanghai University of Traditional Chinese Medicine from October 2021 to April 2023 were selected as research subjects. The cases were randomly divided into FURL group of 46 cases and RURL group of 46 cases. The FURL group received the operation of FURL, while the RURL group received the operation of RURL with N-Trap. The operation success rate between FURL group and RURL group were compared. The surgical time, severe complications, 1-month stone clearance rate and 3-month stone clearance rate of the successful operation cases between the FURL group and the RURL group were compared. After dividing the two groups of calculi cases into upper FURL group, lower FURL group, and upper RURL group, lower RURL group based on whether the calculus was above the lower pole of the kidney or not, the indexes mentioned above were compared between the two groups in subgroups. Results： The operation success rate of the FURL group (95.7%) was significantly higher than that of the RURL group (80.4%). The operation success rate of the upper FURL group (96.0%) was significantly higher than that of the upper RURL group (70.8%), while there was no significant difference between the lower FURL group (95.2%) and the lower RURL group (90.9%, P>0.05). The indexes of the successful operation cases of the two groups were analyzed as follows. The operation duration of the FURL group, the upper FURL group, and the lower FURL group were longer than those of the RURL group, the upper RURL group, and the lower RURL group respectively (P<0.05). There were no significant differences in the incidence of severe complications between the FURL group, the upper FURL group, the lower FURL group and the RURL group, the upper RURL group, the lower RURL group respectively (P>005). There was no significant difference in 1-month stone clearance rate between the FURL group (84.1%) and the RURL group (67.6%, P>0.05). The 1-month stone clearance rate of the upper FURL group (83.3%) was higher than that of the upper RURL group (52.9%), while there was no significant difference in 1-month stone clearance rate between the lower FURL group (850%) and the lower RURL group (80.0%, P>0.05). There were no significant differences in 3-month stone clearance rate between the FURL group, the upper FURL group, the lower FURL group and the RURL group, the upper RURL group, the lower RURL group respectively (P>0.05). Conclusion： Both FURL and RURL are safe operations in the treatment of upper ureteral calculi. For the upper ureteral calculi above the lower renal pole, the success rate and 1month stone clearance rate are higher with the treatment of FURL than those with the treatment of RURL, it may be implicated that FURL is a better choice than RURL in the treatment of calculi in this segment of upper ureter. For the upper ureteral calculi below the lower renal pole, the success rate and stone clearance rate are similar between FURL and RURL, while the operation time of RURL is shorter than that of FURL.
  • Detection of melamine in human kidney stones by infrared spectroscopy combined with chemometrics
    DAI Jiangming, LONG Qilai, XU Zhibing, et al.
    2024, 34(04): 312.
    Abstract ( ) PDF ( )   Knowledge map   Save
    Objective： To establish a method for identifying melamine in stones by fourier transform infrared spectrometer (FTIR) combined with chemometrics. By preparing in vitro simulated calcium oxalate (CaOx) stones containing melamine, the characteristic group information of melamine was analyzed, and the characteristic response bands of 1 535~1 548 cm-1, 983~997 cm-1 and 704~716 cm-1 were selected for stoichiometric analysis. The training set was established with simulated CaOx stones and human kidney stone samples, and the validation set was established with human kidney stones, and the IR spectra of obtained stone samples were preprocessed by baseline correction, smoothing, vector normalization, secondorder derivation, and standard normal variable transformation. OPUS software and Unscrambler software were used for cluster analysis and principal component analysis of stone samples to identify melamine in human kidney stones. Results： In the training set, the identification rate of melaminecontaining stones was 90.9% by cluster analysis, and 91.8% was identified by principal component analysis. In the validation set, 95.0% of the stones containing melamine were successfully identified. Conclusion： FTIR combined with chemometrics can effectively identify melamine in stones.
  • Curcumin enhances cisplatin sensitivity of OVCAR-3 cells by decreasing the expression of miR-135b-5p and FEN1
    MA Rong, LI Juan, WANG Fang
    2024, 34(04): 324.
    Abstract ( ) PDF ( )   Knowledge map   Save
    Objective： To investigate the effect of curcumin on cisplatin (DDP) sensitivity of ovarian cancer OVCAR-3 cells and its potential mechanism. Methods： (1) Ovarian cancer parental cell line OVCAR-3 was routinely cultured in vitro and cisplatin-resistant cell line OVCAR-3/DDP was constructed. OVCAR-3 cells were divided into Control (conventional culture), DDP-1 (1 μmol/L cisplatin), DDP-5 (5 μmol/L cisplatin), DDP-10 (10 μmol/L cisplatin), DDP-15 (15 μmol/L cisplatin), DDP-20 (20 μmol/L cisplatin), DDP-30 (30 μmol/L cisplatin), curcumin+DDP-1 (20 μmol/L curcumin+1 μmol/L cisplatin), curcumin+DDP-5 (20 μmol/L curcumin+5 μmol/L cisplatin), curcumin+DDP-10 (20 μmol/L curcumin+10 μmol/L cisplatin), curcumin+DDP-15 (20 μmol/L curcumin+15 μmol/L cisplatin), curcumin+DDP-20 (20 μmol/L curcumin+20 μmol/L cisplatin) and curcumin+DDP30 groups (20 μmol/L curcumin+30 μmol/L cisplatin). OVCAR-3/DDP cells were divided into Control, DDP-10 (10 μmol/L cisplatin), DDP-20 (20 μmol/L cisplatin), DDP-30 (30 μmol/L cisplatin), DDP-40 (40 μmol/L cisplatin), DDP-50 (50 μmol/L cisplatin), DDP-60 (60 μmol/L cisplatin), curcumin+DDP-10 (20 μmol/L curcumin+10 μmol/L cisplatin), curcumin+DDP-20 (20 μmol/L curcumin+20 μmol/L cisplatin), curcumin+DDP-30 (20 μmol/L curcumin+30 μmol/L cisplatin), curcumin+DDP-40 (20 μmol/L curcumin+40 μmol/L cisplatin), curcumin+DDP-50 (20 μmol/L curcumin+50 μmol/L cisplatin), curcumin+DDP-60 (20 μmol/L curcumin+60 μmol/L cisplatin) groups. CCK-8 method was used to detect the cell viability, the median inhibitory concentration of cisplatin (IC50) was calculated, and the appropriate concentration of cisplatin and curcumin was screened. (2) OVCAR-3 cells were divided into Control group, DDP group (20.5 μg/mL cisplatin), curcumin group (20 μmol/L curcumin) and curcumin+DDP group (20 μmol/L curcumin+20.5 μg/mL cisplatin). OVCAR-3/DDP cells were divided into Control group, DDP group (42.1 μg/mL cisplatin), curcumin group (20 μmol/L curcumin) and curcumin+DDP group (20 μmol/L curcumin+42.1 μg/mL cisplatin). The apoptosis rate was detected by flow cytometry, the expression of miR-135b-5p was detected by quantitative real-time PCR (qRT-PCR), and the mRNA and protein expressions of flap endonuclease 1 (FEN1) were detected by qRT-PCR and Western blotting, respectively. Results： Compared with DDP group, the activity of OVCAR-3 and OVCAR-3/DDP cells in curcumin+DDP group was significantly decreased at the same cisplatin concentration (P<0.001). The IC50 of DDP on OVCAR-3 and OVCAR-3/DDP cells were 20.5 μg/mL and 42.1 μg/mL, respectively. Compared with OVCAR group or OVCAR/DDP group, curcumin or cisplatin significantly increased the apoptosis rate of OVCAR-3 and OVCAR-3/DDP cells (all P<0.001), and the combined effect of curcumin and cisplatin was more significant. In addition, curcumin or cisplatin treatment significantly decreased the expression of miR-135b-5p and FEN1 expression in OVCAR-3 and OVCAR-3/DDP cells (P<0.01 or <0.001), and the combined effect was more significant. Conclusion： Curcumin could enhance cisplatin sensitivity in ovarian cancer OVCAR-3 cells, which may be attributed to the reducing expression of miR-135b-5p and FEN1.
  • Effect of the expression of transducer protein β-like 1 X linked receptor-1 on proliferation and migration of ovarian cancer A2780 cells
    CHU Xiu, JIN Wei.
    2024, 34(04): 331.
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    Objective： To investigate the expression of transducer protein β-like 1 X linked receptor 1 (TBL1XR1) in the tissues of ovarian cancer patients and its effect on the proliferation and migration of ovarian cancer A2780 cells. Methods： Realtime quantitative fluorescent PCR (qRT-PCR) was used to detect the expression of TBL1XR1 mRNA in 10 pairs of ovarian cancer tissues and adjacent tissues, and IOSE80, A2780, CP70 and SKOV-3 human ovarian cancer cell lines, the latter was used to screen the cell lines with high expression of TBL1XR1 mRNA. BALB/C nude mice aged 4-6 weeks were selected to establish patient derived tumor xenografts (PDTX) model. Ten PDTX model mice were divided into siR-NC group and si-TBL1XR1 group, with 5 mice in each group; siR-NC and si-TBL1XR1 were given via local injection, respectively, 10 mg/kg, once every 3 days. After 18 days, cancer tissues were harvested from each group, and their volume and weight were calculated. Ovarian cancer A2780 cells were taken and divided into siRNC group, si-TBL1XR1 group, pcDNA3.1 group and PCDNA3.1-TBL1XR1 group; and they were treated with siR-NC, si-TBL1XR1, empty vector plasmid pcDNA3.1 and PCDNA3.1-TBL1XR1 plasmid, respectively. The expression of cyclin in ovarian cancer cells in each group was detected by Western blotting, the cell viability was detected by MTT, the cell cycle and the proportion of apoptotic cells were detected by flow cytometry, and the cell migration ability was detected by Transwell assay. Results： The expression of TBL1XR1 mRNA in ovarian cancer tissues was significantly higher than that in adjacent tissues (P<0.05), and the expression of TBL1XR1 mRNA in human ovarian cancer A2780 cell line was significantly higher than that in IOSE80, CP70 and SKOV-3 cell lines (P<0.05). Compared with siRNC group, the tumor volume and weight of si-TBL1XR1 group were significantly decreased on the 18th day (both P<0.05). Compared with the siR-NC group, the expression of cyclin in si-TBL1XR1 group was significantly decreased (P<0.05), while the expression in PCDNA3.1-TBL1XR1 group was significantly increased compared with pcDNA3.1 group (P<0.05). Compared with siR-NC group, the number of ovarian cancer cell migration in si-TBL1XR1 group was greatly decreased (P<0.05), and the proportion of early and late apoptotic cells was markedly increased (P<0.05); and compared with pcDNA3.1 group, the migration number of ovarian cancer cells in pcDNA3.1-TBL1XR1 group was significantly increased (P<0.05), and the proportion of early apoptotic and late apoptotic cells was greatly decreased (P<0.05). Conclusion： TBL1XR1 is highly expressed in ovarian cancer tissues, and reducing TBL1XR1 mRNA expression could significantly inhibit the proliferation and migration of ovarian cancer A2780 cells.
  • Clinical efficacy of metformin combined with orlistat in the treatment of obese polycystic ovary syndrome with insulin resistance
    KANG Yinlin, LI Yi, TANG Yunlin, et al.
    2024, 34(04): 338.
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    Objective： To investigate the clinical efficacy of metformin combined with orlistat in obese patients with polycystic ovary syndrome (PCOS) and insulin resistance. Methods： A total of 150 obese PCOS patients with insulin resistance were randomly divided into control group and research group, with 75 cases in each group. The control group was given basic treatment such as metformin and ethinylestradiol cycloproterone, and the research group was treated with basic treatment combined with orlistat. After 3 months of treatment, the differences in anthropometric indicators, glycolipid metabolism indicators and sex hormone levels between the two groups were compared. Results： After 3 months of treatment, compared with the control group, the body weight, body mass index (BMI), waist-to-hip ratio (WHR), body fat percentage, fasting plasma glucose (FPG), 2 h postprandial plasma glucose (2hPG), fasting insulin (FINS), glycosylated hemoglobin (HbA1c), and homeostasis model assessment-IR (HOMA-IR) of the research group were significantly decreased (P<0.05); and the triglyceride, total cholesterol, low density lipoprotein cholesterol in the research group were also significantly decreased (P<0.05), while high density lipoprotein cholesterol was significantly increased (P<0.05); the levels of luteinizing hormone (LH), estradiol, testosterone, prolactin and LH/follicle stimulating hormone （FSH） were greatly decreased (P<0.05), while there was no significant difference in FSH (P>0.05). Conclusion： Metformin combined with orlistat in the treatment of obese PCOS with insulin resistance could significantly improve the glucose and lipid metabolism, reduce weight and insulin resistance and improve sexual hormone disorder.