Metabolic dysfunction-associated steatotic liver disease (MASLD), the most prevalent chronic liver condition globally, lacks adequate and effective therapeutic remedies in clinical practice. Recent studies have increasingly highlighted the close connection between the ubiquitin-proteasome system (UPS) and the progression of MASLD. This relationship is crucial for understanding the disease's underlying mechanism. As a sophisticated process, the UPS govern protein stability and function, maintaining protein homeostasis, thus influencing a multitude of elements and biological events of eukaryotic cells. It comprises four enzyme families, namely, ubiquitin-activating enzymes (E1), ubiquitin-conjugating enzymes (E2), ubiquitin-protein ligases (E3), and deubiquitinating enzymes (DUBs). This review aims to delve into the array of pathways and therapeutic targets implicated in the ubiquitination within the pathogenesis of MASLD. Therefore, this review unveils the role of ubiquitination in MASLD while spotlighting potential therapeutic targets within the context of this disease.

[Objective] To analyze the influencing factors of postoperative patency of longitudinal single suture intussusception microsurgical vasoepididymostomy, to provide reference for improving the repetition rate. [Methods] The clinical data of 82 patients with epididymal obstructive azoospermia who underwent longitudinal single suture intussusception microsurgical vasoepididymostomy in our hospital during Sep.2020 and Jan.2023 were retrospectively analyzed.The postoperative patency and spouse pregnancy were followed up by face to face and / or telephone interview.The effects of age, course of disease, body mass index (BMI), previous medical history (epididymitis, operation history, none), preoperative seminal plasma elastase (SPE) level, anastomosis site, unilateral and bilateral lesion, sperm quality, operation time and hospital stay on the postoperative patency rate were analyzed. [Results] All operations were successful, the follow-up rate was 95.12% (78/82), 78 were married, and the postoperative patency was 78.21% (61/78). Of the 61 patients who achieved patency, 56 were married, and the natural pregnancy rate of spouse was 45.21% (33/73). Univariate analysis showed that patients with age <30 years, course of disease <2 years, preoperative SPE level <290 ng/mL, bilateral anastomosis, body or tail anastomosis and motile sperm had higher postoperative patency rate (P>0.05). BMI, previous history, the number of motile sperms examined by epididymal fluid, the length of operation and hospital stay had no significant effects on postoperative patency (P>0.05). The results of multivariate logistic regression analysis showed that preoperative SPE level (OR=0.998, 95%CI: 0.997-1.000, P=0.008) and (OR=10.724, 95%CI: 2.243-51.283, P=0.003) were significantly correlated with postoperative patency. [Conclusion] The preoperative SPE level and anastomosis site are significant influencing factors of postoperative patency, which is higher in patients with age <30 years, course of disease <2 years, preoperative SPE level <290 ng/mL, bilateral anastomosis, body or tail anastomosis and motile sperm.

【Objective】 To analyze the application value of penile vibrating perception threshold measurement in the diagnosis of erectile dysfunction (ED) and provide reference for the seversity of ED patients. 【Methods】 The clinical data, Erectile Hardness Scale (EHS) score, International Index of Erectile Function Questionnaire-5 (IIEF-5) score, and the vibrating perception threshold (VPT) of the penis of 257 patients with decreased erectile function as the main complaint or accompanying symptoms treated during Jan. and Dec.2021 were retrospectively collected and analyzed.Patients with EHS=4 and IIEF-5≥22 were classified into the normal group, and the rest into the ED group.The differences in VPT in patients with different EHS scores were compared, and the correlation between IIEF-5 and VPT was analyzed.The diagnostic value of VPT for ED was evaluated with receiver operating characteristic (ROC) curve. 【Results】 The difference in penile VPT among patients with different EHS scores was statistically significant (P<0.05).With the decrease of EHS score, VPT showed an increasing trend.Glans VPT was negatively correlated with IIEF-5 score (ρ=-0.22, P<0.001), and penile shaft VPT was also negatively correlated with IIEF-5 score (ρ=-0.26, P<0.001).The VPT of glans penis [(4.17±1.37) V vs.(3.47±1.24) V, P=0.009] and the VPT of penis body [(3.73±1.41) V vs.(2.99±1.14)V, P=0.003] in the ED group were both higher than those in the normal group.The area under the ROC curve (AUC) of the glans VPT was 0.642.When the cut-off value was 3.537 V, the sensitivity was 63.4%, and the specificity was 63.6%.The AUC of the penile shaft VPT was 0.659.When the cut-off value was 2.775 V, the sensitivity was 72.3%, with a specificity of 54.5%. 【Conclusion】 The penile VPT of ED patients is higher than that of normal ones, and there is a correlation between VPT and the severity of ED.Severe ED is associated with higher VPT.The measurement of penile VPT is helpful for the clinical diagnosis of ED patients.

【Objective】 To introduce a modified microdot two-layer microsurgical vasovasostomy (MVV) and to analyze its effectiveness in patients with vas deferens obstruction caused by inguinal herniorrhaphy. 【Methods】 Clinical data of patients treated during Mar.2015 and Oct.2020 were retrospectively analyzed. According to different surgical methods, the patients were divided into the modified group and traditional group. The general data, intraoperative conditions, efficacies and complications of the two groups were compared. 【Results】 There were 59 cases in the modified group, 54(91.5%) of whom were successfully followed up, and 41 cases in the traditional group, 38(92.7%) of whom were successfully followed up. There were no significant differences in age, inguinal herniorrhaphy history, and unilateral/bilateral ratio between the two groups (P>0.05). The average operation time for unilateral lesions in the modified group was shorter than that in the traditional group [(89.44±24.86) vs. (112.04±43.40) min, P=0.032]. The postoperative patency rate (83.3% vs.73.7%, P>0.05) and natural pregnancy rate (33.3% vs.28.9%, P>0.05) of the modified group and traditional group were comparable. Incision fat liquefaction occurred in 2 cases (3.70%) in the modified group and in 1 case (2.63%) in the traditional group (P>0.05). 【Conclusion】 The modified microdot two-layer MVV is a safe surgical method with comparable effectiveness as the traditional approach. By adjusting the position of the marking points and the order of suturing, it helps the management of sutures, reduces the difficulty of vasovasostomy, shortens operation time, and can be applied to repair vas deferens obstruction caused by inguinal herniorrhaphy.

Objective:To evaluate the role of secreted protein acidic and rich in cysteine like protein 1 (SPARCL1) in spinal dorsal horns in the development of remifentanil-induced hyperalgesia in mice with incisional pain.Methods:Forty-eight healthy male C57BL/6J mice, aged 8-10 weeks, weighing 18-22 g, were divided into 6 groups ( n=8 each) using a random number table method: control group (group C), incisional pain group (group I), remifentanil group (group R), incisional pain plus remifentanil group (group I+ R), incisional pain plus remifentanil plus negative control group (group I+ R+ N), and incisional pain plus remifentanil plus SPARCL1-siRNA group (group I+ R+ S). In I+ R+ N and I+ R+ S groups, 1×10 8 IFU/ml negative control siRNA and SPARCL1-siRNA 10 μl were intrathecally injected, respectively, once a day for 3 consecutive days.Normal saline 10 μl was intrathecally injected once a day for 3 consecutive days in C, I, R and I+ R groups.After transfection was stable, normal saline 0.1 ml was intravenously injected through the tail vein for 4 consecutive times at 15 min interval in C and I groups, and remifentanil 10 μg/kg (diluted to 0.1 ml in normal saline) was intravenously injected via the tail vein for 4 consecutive times at 15 min interval in R, I+ R, I+ R+ N and I+ R+ S groups.The incisional pain model was established after the first administration via the tail vein in R, I+ R, I+ R+ N and I+ R+ S groups.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 24 h before infusing normal saline or remifentanil (T 0) and 3, 6, 24 and 48 h after stopping infusion (T 1-4). Animals were sacrificed after measuring pain threshold at T 4, and L 4-6 segments of the spinal cord were removed for determination of the expression of SPARCL1 protein and mRNA by Western blot and quantitative real-time polymerase chain reaction, respectively. Results:Compared with group C, MWT was significantly decreased and TWL was shortened at T 1-4 in I+ R and I+ R+ N groups and at T 2-4 in I, R and I+ R+ S groups, and the expression of SPARCL1 protein and mRNA was significantly up-regulated in R, I+ R and I+ R+ C groups ( P<0.05 or 0.01). Compared with group I and group R, MWT was significantly decreased, TWL was shortened, and the expression of SPARCL1 protein and mRNA was up-regulated in group I+ R ( P<0.01). Compared with group I+ R, MWT was significantly increased and TWL was prolonged at T 1-4, and the expression of SPARCL1 protein and mRNA was down-regulated in group I+ R+ S ( P<0.05 or 0.01). Conclusion:Enhanced activity of SPARCL1 in the spinal dorsal horns is involved in the development of remifentanil-induced hyperalgesia in mice with incisional pain.

Objective: To evaluate the role of spinal COX-1 and COX-2 in remifentanil-induced hyperalgesia in mice with incisional pain. Methods: Thirty-two male C57BL/6J mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups (n=8 each) using a random number table method: control group (group C), incisional pain plus remifentanil group (group IR), incisional pain plus remifentanil plus selective COX-1 inhibitor group (group IR+ SC560), and incisional pain plus remifentanil plus selective COX-2 inhibitor group (group IR+ SC236). In IR, IR+ SC560 and IR+ SC236 groups, normal saline 10 μl, SC560 25 μg and SC236 25 μg were intrathecally injected, respectively, 15 min later remifentanil 10 μg/kg was injected via the tail vein for 4 times at 15 min intervals.An incisional pain model was established after the first injection of remifentanil.The mechanical paw withdrawal threshold (MWT) was measured at 24 h before normal saline or remifentanil injection and 3, 6, 24 and 48 h after the last injection (T₀-T₄). The mice were sacrificed after the last measurement of pain threshold, and the L_4-6 segments of the spinal cord were removed for determination of the expression of COX-1 and COX-2 (by Western blot) and expression of COX-1 and COX-2 mRNA (by quantitative real-time polymerase chain reaction). Results: Compared with group C, the MWT was significantly decreased, and the expression of COX-2 protein and mRNA was up-regulated in IR, IR+ SC560 and IR+ SC236 groups (P<0.05). Compared with group IR, the MWT was significantly increased in IR+ SC560 and IR+ SC236 groups (P<0.05). There was no significant difference in the MWT at each time point between IR+ SC560 and IR+ SC236 (P>0.05) .There was no significant difference in the expression of COX-2 protein and mRNA among group IR, group IR+ SC560 and group IR+ SC236 (P>0.05). There was no significant difference in the expression of COX-1 protein and mRNA among the four groups (P>0.05). Conclusion Compared with COX-1, spinal COX-2 plays a major role in the pathophysiological mechanism of remifentanil-induced hyperalgesia in mice with incisional pain.

Objective: To evaluate the role of neuroligin 1 (NL-1) in trafficking of GluR1-containing AMPA receptor to cell membrane in spinal cord dorsal horns during remifentanil-induced hyperalgesia in mice with incisional pain. Methods: Forty SPF healthy male C57BL/6J mice, aged 8-10 weeks, weighing 18-22 g, were divided into 5 groups (n=8 each) using a random number table method: control group (group C), NL1-shRNA plasmid group (group NL), incisional pain plus remifentanil group (group I+ R), incisional pain plus remifentanil plus blank vector group (group I+ R+ B), and incisional pain plus remifentanil plus NL-1-shRNA plasmid group (group I+ R+ NL). Negative lentivirus was intrathecally injected in group I+ R+ B.In NL and I+ R+ NL groups, 10 μl NL-1-shRNA lentivirus at 1×10⁸ IFU/ml was intrathecally injected once a day for 3 consecutive days.Normal saline 10 μl was intrathecally injected at the same time point in C and I+ R groups.After transfection was stable, normal saline 0.1 ml was injected via the caudal vein for 4 consecutive times at 15 min intervals in C and NL groups.In I+ R, I+ R+ B and I+ R+ NL groups, 0.1 ml remifentanil 10 μg/kg was injected via the caudal vein for 4 consecutive times at 15 min intervals, and the model of incisional pain was established after the first administration.The mechanical paw withdrawal threshold (MWT) and tail-flick latency (TFL) were measured at 24 h before normal saline or remifentanil administration (T₀) and at 3, 6, 24 and 48 h after the end of administration (T_1-4). The animals were sacrificed after measurement of pain threshold at T₄, and L_4-6 segments of the spinal dorsal horn were then collected for determination of the expression of NL-1 protein and mRNA and AMPA receptors, and the ratio of AMPA receptor expression in the membrane protein to that in the total protein (m/t ratio) was calculated. Results: Compared with group C, the MWT was significantly decreased, and TFL was shortened at T_1-4, the expression of NL-1 protein and mRNA and GluR1-containing AMPA receptors in membrane and total proteins was up-regulated, and m/t ratio was increased in I+ R and I+ R+ B groups (P<0.05). Compared with I+ R and I+ R+ B groups, the MWT was significantly increased and TFL was prolonged at T_1-4, the expression of NL-1 protein and mRNA and GluR1-containing AMPA receptors in membrane and total proteins was down-regulated, and m/t ratio was decreased in group I+ R+ NL (P<0.05). Conclusion NL-1 in spinal cord dorsal horns can promote the trafficking of GluR1-containing AMPA receptors to cell membrane, which is involved in the development and maintenance of remifentanil-induced hyperalgesia in mice with incisional pain.

Objective To evaluate the effect of oxycodone on intestinal ischemia-reperfusion injury in rats and the role of autophagy.Methods Twenty-four healthy adult male Sprague-Dawley rats,aged 6-9 weeks,weighing 180-220 g,were divided into 4 groups (n =6 each) using a random number table method:sham group (group S),intestinal I/R group (group I/R),oxycodone group (group O) and oxycodone plus autophagy inhibitor 3-methyladenine (3-MA) group (group O+3-MA).Intestinal I/R was induced via clamping the superior mesenteric artery for 1 h,followed by 2-h reperfusion in all the groups except for group S.Oxycodone 0.5 mg/kg was injected via caudal vein at 15 min before ischemia in group O and group O+3-MA.3-MA 30 mg/kg was intraperitoneally injected at 10 min before ischemia in group O+3-MA.Rats were gavaged with fluorescein-isothiocyanate-conjugated dextran (FITC-dextran) immediately before ischemia.Blood samples were collected from the cardiac apex at 2 h of reperfusion to detect the level of serum FITC-dextran.Blood samples were collected from the cardiac apex at 2 h of reperfusion to measure the concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) in serum by enzyme-linked immunosorbent assay.Small intestinal tissues were obtained at 2 h of reperfusion for examination of the pathological changes and for determination of the expression of occludin,Beclin-1 and microtubule-associated protein 1 light chain 3 (LC3) (by Western blot).Intestinal damage was assessed and scored according to Chiu.The ratio of LC3 Ⅱ expression to LC3 Ⅰ expression (LC3 Ⅱ / Ⅰ) was calculated.Results Compared with group S,the Chiu's score,levels of serum FITC-dextran,TNF-α and IL-1β and LC3 Ⅱ/Ⅰ ratio were significantly increased,the expression of Beclin-1 was up-regulated,and the expression of occludin was down-regulated in group I/R (P＜0.05).Compared with group I/R,the Chiu's score and levels of serum FITC-dextran,TNF-α and IL-1β were significantly decreased,LC3 Ⅱ / Ⅰ ratio was increased,and the expression of occludin and Beclin-1 was up-regulated in group O (P＜0.05),and no significant change was found in the parameters mentioned above in group O+ 3-MA (P＞0.05).Compared with group O,the Chiu's score and levels of serum FITC-dextran,TNF-α and IL-1β were significantly increased,LC3 Ⅱ / Ⅰ ratio was decreased,and the expression of occludin and Beclin-1 was down-regulated in group O + 3-MA (P＜ 0.05).Conclusion Oxycodone can ameliorate intestinal I/R injury,and the mechanism may be related to enhancing autophagy in intestinal tissues of rats.

Objective To evaluate the efficacy of preoperative transversus abdominis plane (TAP) block for improving analgesia after radical resection of colorectal cancer in elderly patients.Methods Fiftysix American Society of Anesthesiologists physical status Ⅱ or Ⅲ patients of both sexes,aged 75-86 yr,weighing 52-78 kg,scheduled for elective radical resection of colorectal cancer under general anesthesia,were divided into 2 groups (n=28 each) using a random number table:TAP block group (group T) and control group (group C).After anesthesia induction,ultrasound-guided bilateral TAP block was performed,and 0.40％ ropivacaine 25 ml was injected into each side in group T.Both groups received patientcontrolled intravenous analgesia (PCIA) with fentanyl after surgery.PCIA solution contained fentanyl 600 μg and azasetron 10 mg in 100 ml of normal saline.The PCIA pump was set up with a 2 ml bolus dose,a 15 min lockout interval and background infusion at a rate of 2 ml/h,and visual analogue scale score was maintained≤ 3.When the visual analogue scale score＞3,tramadol 50-100 mg was intravenously injected as rescue analgesic.The number of patients requiring rescue analgesic,consumption of fentanyl during PCIA and the number of successfully delivered doses were recorded within 24 h after surgery.The adverse reactions such as hematoma at the puncture site,nausea and vomiting,respiratory depression,chest tightness and pruritus were recorded.Results Compared with group C,the number of patients requiring rescue analgesic was significantly decreased,and the consumption of fentanyl and the number of successfully delivered doses during PCIA were decreased in group T (P＜0.05).No hematoma was found at the puncture site in group T.No respiratory depression,chest tightness or pruritus was found in the two groups,and there were no significant differences in the incidence of nausea and vomiting between the two groups (P＞0.05).Conclusion Preoperative TAP block can reduce the consumption of fentanyl and enhance the efficacy of analgesia after radical resection of colorectal cancer in elderly patients.

Objective To investigate the effects of hydrogen-rich medium on lipopolysaccharide (LPS)-induced intestinal epithelial barrier dysfunction of human intestinal epithelial (Caco2) cells. Methods Caco2 cells (passages 28-35) were purchased from the Cell Bank of the Shanghai Institute of Cell Biology, Chinese Academy of Sciences in Shanghai, China, and they were cultured in Dulbecco minimum essential medium (DMEM) containing 20% fetal bovine serum. These cells were randomly divided into four groups: control group (group A), hydrogen-rich medium group (group B), LPS group (group C) and LPS + hydrogen-rich medium group (group D). Cells were cultured with normal medium in group A and group C or with hydrogen-rich medium in group B and group D. Meanwhile, 1 g/L LPS was simultaneously added into group C and group D, while an equivalent volume of normal saline was added into group A and group B instead. In vitro intestinal epithelial models were reproduced with monolayer filter-grown Caco2 and intestinal epithelium. The trans-epithelial electrical resistance (TEER) in models of each group was measured at different incubation times (0, 3, 6, 12, 24 and 48 hours). Cell viability and cytotoxicity were assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) release assay, respectively, after incubation for 24 hours. The expression levels of claudin-1 and occludin were respectively determined at 6, 12 and 24 hours of incubation by Western Blot assay. The morphological structure of claudin-1 and occludin was respectively observed after incubation for 24 hours with immunofluorescence staining. Results There was no statistical significance in variables between group A and group B. Compared with group A, it was shown that TEER was time-dependently decreased in groups C and D after 6 hours. Compared with group C, TEER in group D was increased after 6 hours. Compared with group A, the cell viability was significantly reduced in group C [(67.2±7.9)% vs. (100.0±0.0)%, P < 0.05] and cell injury was obvious [LDH release rate: (38.5±2.1)% vs. (1.2±0.3)%, P < 0.05]; the expression levels of claudin-1 and occludin at 6, 12, 24 hours were significantly down-regulated [claudin-1 (gray value): 0.351±0.079, 0.272±0.075, 0.190±0.049 vs. 0.518±0.030; occludin (gray value): 0.416±0.044, 0.290±0.062, 0.226±0.019 vs. 0.602±0.038, all P < 0.05], and the structure of claudin-1 and occludin were profoundly disrupted. Compared with group C, it was shown that the cell viability was significantly increased in group D [(88.8±7.4)% vs. (67.2±7.9)%, P < 0.05] and cell injury was significantly abated [LDH release rate: (16.4±4.3)% vs. (38.5±2.1)%, P < 0.05]; the expression levels of claudin-1 and occludin were significantly up-regulated at 24 hours [claudin-1 (gray value): 0.428±0.046 vs. 0.190±0.049, occludin (gray value): 0.466±0.071 vs. 0.226±0.019, both P < 0.05]; the disrupted structures of claudin-1 and occludin were partially recovered. Conclusion Hydrogen-rich medium can effectively attenuate LPS-induced dysfunction of intestinal epithelial barrier in human Caco2 cells by ameliorating cell viability as well as regulating claudin-1 and occludin expression and structure.