Lipotoxicity is a pivotal factor that initiates and exacerbates liver injury and is involved in the development of metabolic-associated fatty liver disease (MAFLD). However, there are few reported lipotoxicity inhibitors. Here, we identified a natural anti-lipotoxicity candidate, HN-001, from the marine fungus Aspergillus sp. C1. HN-001 dose- and time- dependently reversed palmitic acid (PA)-induced hepatocyte death. This protection was associated with IRE-1α-mediated XBP-1 splicing inhibition, which resulted in suppression of XBP-1s nuclear translocation and transcriptional regulation. Knockdown of XBP-1s attenuated lipotoxicity, but no additional ameliorative effect of HN-001 on lipotoxicity was observed in XBP-1s knockdown hepatocytes. Notably, the ER stress and lipotoxicity amelioration was associated with PLA2. Both HN-001 and the PLA2 inhibitor MAFP inhibited PLA2 activity, reduced lysophosphatidylcholine (LPC) level, subsequently ameliorated lipotoxicity. In contrast, overexpression of PLA2 caused exacerbation of lipotoxicity and weakened the anti-lipotoxic effects of HN-001. Additionally, HN-001 treatment suppressed the downstream pro-apoptotic JNK pathway. In vivo, chronic administration of HN-001 (i.p.) in mice alleviated all manifestations of MAFLD, including hepatic steatosis, liver injury, inflammation, and fibrogenesis. These effects were correlated with PLA2/IRE-1α/XBP-1s axis and JNK signaling suppression. These data indicate that HN-001 has therapeutic potential for MAFLD because it suppresses lipotoxicity, and provide a natural structural basis for developing anti-MAFLD candidates.

Primary intrahepatic stones(PIS)is a refractory disease with a high incidence rate in Southwest China,which greatly affects the life of patients.Metabolites,such as β-glucuronidase produced by chronic biliary tract infection,play an important role in the formation of pigmented stones.In addition to exogenous β-glucuronidase produced by bacteria,endogenous β-glucuronidase produced by intrahepatic bile duct cells also plays an important role in the formation of stones.This article analyzes the research advances in the role of β-glucuronidase in the pathogenesis of PIS,in order to provide a possible method for the prevention and treatment of PIS.

Percutaneous nephrolithotomy(PCNL)is the first-line treatment for complex kidney stones and can be performed in a variety of positions.Semi-supine combined lithotomy position,as a flexible,convenient,safe and effective position for PCNL,can be conducted in a retrograde approach under a single position,so it is suitable for the treatment of complex kidney stones.We will review the development characteristics and advantages of PCNL in semi-supine combined lithotomy position,and its application in the treatment of complex kidney stones.Based on this,we will further propose a minimally invasive treatment strategy for upper urinary tract calculi.

Objective:To evaluate the feasibility and safety of PT Scope (short for intelligent pressure and temperature controlled flexible ureteroscopy)combined with Thulium laser in the intracavitary treatment of upper urinary tract stones.Methods:A retrospective analysis was conducted on the clinical data of 13 patients with upper urinary tract stones who were treated with PT Scope combined with Thulium laser lithotripsy in Sun Yat-Sen Memorial Hospital from February to April 2024. There were 7 males and 6 females. The patients had a mean age of (46±10) years old, with an accumulated stone diameter of (25.8±13.3) mm. There were 7 cases of lower calyx stones (53.8%), and 3 cases of concomitant ureteral stones (23.1%).Four patients (30.8%) had positive preoperative urine cultures, and six patients (46.2%) had leukocyte counts greater than 100 cells/μl in their urine tests. The Thulium laser power was set at 45 W (1.5 J at 30 Hz, 0.3 J at 150 Hz). The renal pelvic pressure threshold was set at 30 mmHg (1 mmHg=0.133 kPa), and the temperature threshold at 43 ℃. Postoperatively, double J stents were placed for 2 to 4 weeks.Results:All 13 patients successfully completed the surgery. The median operative time was 30 (25, 90) minutes. The intraoperative average renal pelvic pressure in these 13 patients ranged from 8 mmHg to 24 mmHg, and the average renal pelvic temperature ranged from 25 ℃ to 34 ℃. Postoperatively, 1 patient experienced a fever (38.0 ℃) and 2 patients required analgesic treatment due to postoperative pain. There were no other intraoperative or postoperative complications. The median postoperative hospital stay was (1.5±0.8) days. The stone-free rate of 1 month was 84.6%(11/13).Conclusions:PT Scope combined with Thulium laser could effectively control renal pelvic pressure and temperature, achieve a high stone-free rate, and have a low complication rate. It is a safe and effective treatment for upper urinary tract stones.

Objective:To investigate the technical key points and clinical effects of laparoscopic surgery using the subgastric approach for infected pancreatic necrosis (IPN).Methods:From October 2020 to October 2021, The clinical data of 6 patients with IPN after severe acute pancreatitis (SAP) undergoing laparoscopic surgery using the subgastric approach at First Hospital and Second Hospital of Hebei Medical University was retrospectively analyzed. Parameters in this report included the operation time, estimated blood loss, and the patient's vital signs, inflammatory marker CRP, and WBC before operation and postoperative 24 h, 3 d, and 1 w, and postoperative complications including pancreatic leakage, organ failure, bleeding, and abdominal infection and incision infection. Follow-up after surgery was completed in outpatient checkups and long-term complications were recorded.Results:There were 4 male and 2 female patients. The median age of the 6 patients was 50 (43.5, 56.5) years. Laparoscopic debridement surgery using the subgastric approach was successfully completed in all the patients and no reoperation was needed. The median operation time was 65 (52.5, 85) min; the median estimated blood loss was 20 (25, 37.5) ml. Median APACHEⅡ score one day before surgery was 11.5 (10.25, 12.75) and the median MCTSI score at initial admission was 8 (7, 8). The inflammatory parameters including CRP, WBC, and neutrophil count on postoperative day 3 and 1w were significantly lower than those before surgery, and all the differences were statistically significant (all P value <0.05). One patient had a postoperative pancreatic fistula and was alleviated after ERCP with pancreatic stent implantation. Another patient had a incision infection after surgery and recovered after complete surgical drainage of the abdominal wall incision. No patients had complications such as heart, lung, and kidney failure, abdominal hemorrhage and infection. During the follow-up, 5 of 6 patients had no newly-occurred diabetes, except one patient who had diabetes before the operation. None of the 6 patients had recurrent IPN. Conclusions:Laparoscopic surgery using the subgastric approach for infected IPN in lesser omental sac is safe and feasible.

Objective:To investigate the correlations between cosmetic and radiographic parameters in patients with congenital scoliosis (CS) with vertebral segmentation failureand the clinical implication of cosmetic parameters.Methods:A total of 27 CS patients were retrospectively reviewed. Anteroposterior and lateral radiographs of the entire spine was taken. Simultaneously, photos were taken from the back in natural standing position and standing forward bending position. Seven cosmetic parameters were measured on the photographs: shoulder area index 1 (SAI1), shoulder area index 2 (SAI2), lumbar area index (LAI), shoulder angle (α 1), axilla angle (α 2), right and left waist angle difference (RLWAD) and hump index. Also, seven radiographic parameterswere measured on the radiographs: radiographic shoulder height difference (RSHD), T 1 tilt, first rib angle (FRA), clavicle angle (CA), clavicle-rib cage intersection (CRCI), clavicle chest cage angle difference (CCAD) and Cobb's angle. The correlation between cosmetic parameters and radiographic parameters was analyzed by Pearson correlation coefficient. Results:Of the 27 patients, 4 were males and 23 were females, with a mean of age 14.0±2.6 years (range 11-18 years). The apical vertebra ranged from T 5 to T 11. LAI was significantly correlatedwith CCAD, but correlation coefficient was only -0.44. The range of correlation coefficientsbetween SAI1 and all radiographic parameters was -0.17 to 0.53, and the range of correlation coefficients between SAI2 and all radiographic parameters was -0.16 to 0.53. However, all correlation coefficients were less than 0.56. Conclusion:Radiographic parameters cannot reflect cosmetic appearances of CS patients with vertebral segmentation failures accurately. More attention should be paid to cosmetic parameters in the evaluation of patients' appearances.

Objective:To investigate the feasibility and effects of modified sequential correction technique combined 3-columns osteotomy for severe kyphoscoliosis.Methods:A retrospective analysis was performed on 18 patients (7 males and 11 females) with severe kyphosis who received modified sequential correction technique combined 3-columns osteotomy in our hospital from June 2019 to April 2020. Preoperative, postoperative and final follow-up clinical and imaging outcomes were evaluated.Results:In this cohort, the average fixed segment was 11.2±3.8. The average operative duration was 401.9±68.9 min and the average intraoperative blood loss was 2 418.8±736.9 ml. The Cobb angle was improved significantly from 65.0°±16.4° pre-operatively to 41.6°±14.1° post-operatively. At final follow-up, it was 41.4°±14.3°, which was not significantly different from that after operation. Global kyphosis (GK) was 65.5°±20.8° pre-operatively and 28.1°±13.8° post-operatively with correction rate of 57.8%±17.8%. However, GK was 29.3°±14 .2° at postoperative 1 year , which was not significantly different from that after operation. There was no significant difference in C 7PL-CSVL ( F=0.449 , P=0.642) or SVA ( F=3.519, P=0.058) among the three time points. There was no alter of SEP and MEP observed during operation. Four patients had temporary lower limb numbness after operation, while the symptoms disappeared at 6 months after operation. There was no instrumental failure during the follow-up. Conclusion:Patients with severe kyphoscoliosis can obtain satisfied local correction by undergoing modified sequential correction technique combined 3-columns osteotomy without significant loss of correction at 1 year after operation. It can effectively avoid instability and dislocation of the osteotomy site and massive bleeding during the operation. As a simplified surgical procedure, it can reduce the difficulty of rod loading without prolonged operation duration. Further, this technique can ensure lower incidence of neurological complications and rod failure.

Objective:To investigate the clinical outcomes and complication of posterior surgery for Scheuermann kyphosis fusing to different distal fusion levels.Methods:From January 2012 to December 2017, a consecutive cohort of 34 patients who were treated with posterior spinal instrumented correction and satisfied the inclusion criteria were retrospectively reviewed, including 29 males and 5 females, aged 17.1±4.3 years (range, 12-30 years). All of the patients had a minimum follow-up of 2 years. According to the distal fusion level, patients were divided into 2 groups. Group sagittal stable vertebra (SSV) (22 cases) included patients whose lowest instrumented vertebra (LIV) was SSV; Group SSV-1 (12 cases) included patients who had a LIV one level above the SSV. Radiographic parameters including global kyphosis (GK), lumbar lordosis (LL), sagittal vertical axis (SVA), pelvic incidence (PI), pelvic tilt (PT), and sacral slope (SS) were measured in the standing radiographs before and after operation and at the latest follow up. Intraoperative and postoperative complications were recorded. The Scoliosis Research Society-22 questionnaire (SRS-22) were conducted at pre-operation and the final follow up to evaluate the clinical outcomes. The sagittal radiographic parameters and the incidence of distal junctional kyphosis (DJK) were compared between the two groups.Results:There were no significant differences in terms of age, sex, radiographic measurements and scores of SRS-22 between two groups preoperatively ( P>0.05). The correction rates of GK in the SSV group and the SSV-1 group were 42.8%±7.6% and 43.2%±8.4% ( t=0.151, P=0.881) respectively. While the correction rates loss were 1.2%±5.2% and 3.9%±7.2% ( t=0.767, P=0.449) at the latest follow up. No significant difference was observed in terms of other radiographic parameters ( P>0.05). During the postoperative follow up period, 3 patients (16.7%) in SSV group and 2 patients (13.6%) in SSV-1 group developed DJK. The incidence of DJK did not show any significant difference between two groups ( χ2=0.057, P=0.812). At the final follow-up, the function scores of SRS-22 in SSV-1 group (4.1±0.6) was significantly higher than SSV group (3.7±0.5) ( t=2.300, P=0.028) and there was no significant difference in the rest of the domain ( P>0.05). Conclusion:Compared with stopping at SSV, fusion to SSV-1 could achieve comparable curve correction with the preservation of more lumbar motility. Moreover, it would not increase the risk of DJK. As a result, we recommend selecting SSV-1 as the ideal LIV for SK patients.

To construct a eukaryotic expression plasmid containing the luciferase reporter gene (Fluc) to quickly detect apoptosis. Four amino acids, Asp-Glu-Val-Asp (DEVD), the recognize motif of Caspase-3, were introduced into the middle of the Fluc-C and N fragment. Meanwhile, four amino acids, Asp-Glu-Val-Gly (DEVG), were selected as a negative control. Subsequently, the recombinant gene was cloned into the N and C terminal end of the split intein, and named as pFluc-DEVD and pFluc-DEVG. Then the plasmids were transfected into cells and renilla luciferase was co-transfected in each sample as an internal control for transfection efficiency. Then the apoptosis level was detected by the double luciferase reporter gene and the Western blotting analysis. The results showed that when apoptosis occurred, the content of firefly luciferase expressed in the pFluc-DEVD plasmid transfected group was about 3 times higher than pFluc-DEVG plasmid transfected group. Furthermore, Western blotting detection indicated that the Fluc level was significantly increased in pFluc-DEVD transfected group when pre-treated by apoptosis stimulants. The activation degree of Caspase-3 was closely related to the expression of Fluc, and had a significant statistical difference. These results confirmed that firefly luciferase protein expressed by pFluc-DEVD plasmid can be cleaved by the intracellular Caspase-3 enzyme, and this plasmid can accurately reflect the cell apoptosis level, which provides a useful method for quantitative detection of apoptosis.
Apoptosis ; Genes, Reporter ; Luciferases, Firefly ; Transfection

Objective To explore the value of ultrasound gray scale ratio (UGSR) in the diagnosis and differential diagnosis of papillary thyroid carcinoma(PTC) with different sizes.Methods A retrospective study was made in 702 patients with 1107 nodules which were confirmed by surgery in the Department of Oncology or fineneedle aspiration of HangZhou First people's Hospital,Zhejiang University of medical school from Jan.2016 to Oct.2017.All the thyroid nodules were divided into three groups:D≤ 1 cm group,1＜D≤2 cm group and ＞2 cm group according to their sizes.The UGSR of the PTC and NG were obtained through the RAD info system.Their differences were analyzed and ROC was established to confirm the optimal threshold in the differential diagnosis between PTC and NG among the groups.Results There were 483 PTC and 624 NG in this study.The UGSR of D≤ 1 cm group,1＜D≤2 cm group and ＞2 cm group of PTC and NG were (0.48±0.12) vs (0.76±0.22)(t=33.21,P=0.00);(0.52±0.17) vs(0.80±0.21)(t=1.30,P=0.00) and (0.63±0.20) vs(0.89±0.24)(t=3.58,P=0.00) respectively.The area under the ROC of UGSR in the differentiation of PTC and NG in the three groups were 0.873,0.840 and 0.811 respectively.The Youden indexes were greatest (0.631,0.536 and 0.535 respectively),when the cut-offs of the UGSR were 0.682,0.652 and 0.831 respectively.The sensitivity and specificity to diagnose PTC were 94.8％ and 68.0％,75.0％ and 78.6％,80.3％ and 73.2％ respectively in the three groups.Conclusions The best UGSR value of PTC was variant in thyroid nodule with different size.Recognition of these differences accurately could improve the pre-operative diagnostic accuracy of PTC.Also the method is simple to operate and easy to apply.