OBJECTIVES: Uridine diphospho-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE) myopathy is a progressive neurodegenerative disease associated with homozygous or compound heterozygous missense mutations in the GNE gene. This study aims to explore the impact of the homozygous p.V543M mutation in on cell phenotype and to gain preliminary insights into the underlying mechanisms. METHODS: Human embryonic kidney 293T (HEK 293T) cells were used to construct wild-type (WT-GNE) and mutant (MUT-GNE) GNE overexpression models. Western blotting and immunofluorescence were used to assess GNE protein expression levels and subcellular localization. Cell adhesion, proliferation, apoptosis, and mitochondrial membrane potential were evaluated using the cell counting kit-8 (CCK-8) assay, crystal violet staining, flow cytometry, Hoechst 33342/propidium iodide (PI) staining, and tetramethylrhodamine ethyl ester (TMRE) staining. Sialic acid synthesis levels and GNE enzymatic activity were measured, and the mRNA expression of sialic acid biosynthesis-related enzymes was quantified by real-time PCR. RESULTS: Western blotting confirmed successful establishment of GNE overexpression models. Immunofluorescence showed significantly reduced co-localization of GNE protein with Golgin-97 in the MUT-GNE group compared to WT-GNE (Pearson's correlation coefficient: 0.65±0.08 vs 0.83±0.06, P<0.05). Compared with WT-GNE, cells in the MUT-GNE group exhibited increased adhesion, decreased proliferation, and reduced mitochondrial membrane potential (P<0.05). No significant differences in apoptosis were observed between groups. The MUT-GNE group showed reduced sialic acid production, significantly decreased kinase activity, and downregulated transcription of sialic acid biosynthesis-related enzymes compared to WT-GNE (P<0.001). CONCLUSIONS The p.V543M mutation in the GNE gene alters cellular phenotype by reducing GNE enzymatic activity and the transcription of sialic acid biosynthesis enzymes, ultimately impairing sialic acid production.
Humans ; Mutation, Missense ; HEK293 Cells ; Apoptosis/genetics* ; Phenotype ; Multienzyme Complexes/metabolism* ; Cell Proliferation ; Homozygote ; Cell Adhesion/genetics* ; Distal Myopathies/genetics*

OBJECTIVES: To explore the therapeutic mechanism of compound Centella asiatica formula (CCA) for alleviating Schistosoma japonicum (Sj)-induced liver fibrosis in mice. METHODS: The active components and targets of CCA were identified using the TCMSP database with cross-analysis of Sj-related liver fibrosis targets. A "drug-component-target-pathway-disease" network was constructed using Cytoscape 3.9.1. Functional enrichment analysis (GO/KEGG) was performed using DAVID. Molecular docking study was carried out to validate interactions between the core targets and the key compounds. For experimental validation of the results, 36 mice were divided into control group, Sj-infected model group, and CCA-treated groups. In the latter two groups, liver fibrosis was induced via abdominal infection with Sj cercariae for 8 weeks, followed by 8 weeks of daily treatment with CCA decoction or saline. Hepatic pathology of the mice was assessedwith HE and Masson staining, and hepatic expressions of collagen-I and collagen-III were detected using immunohistochemistry; serum IL-6 and TNF-α levels were determined with ELISA. Hepatic expressions of TLR4 and MyD88 proteins were analyzed with Western blotting. RESULTS: We identified a total of 107 bioactive CCA components and 791 targets, including 37 intersection targets linked to Sj-induced fibrosis. The core targets included TNF, TP53, JUN, MMP9, and CXCL8, involving the IL-17 signaling, lipid metabolism, TLR4/MyD88 axis, and cancer pathways. Molecular docking study confirmed strong binding affinity between quercetin (a primary CCA component) and TNF/TP53/JUN/MMP9. In Sj-infected mouse models, CCA treatment significantly attenuated hepatic inflammatory cell infiltration, reduced collagen-I and collagen-III deposition, improved tissue architecture, reduced serum IL-6 and TNF-α levels, and downregulated TLR4 and MyD88 expressions in the liver. CONCLUSIONS CCA mitigates Sj-induced liver fibrosis by targeting TNF, TP53, JUN, and MMP9 to modulate the TLR4/MyD88 pathway, thereby suppressing pro-inflammatory cytokine release, inhibiting hepatic stellate cell activation, reducing collagen deposition, and preventing granuloma formation in the liver.
Animals ; Toll-Like Receptor 4/metabolism* ; Mice ; Myeloid Differentiation Factor 88/metabolism* ; Schistosoma japonicum ; Liver Cirrhosis/parasitology* ; Schistosomiasis japonica ; Signal Transduction ; Molecular Docking Simulation ; Inflammation ; Centella/chemistry* ; Drugs, Chinese Herbal/pharmacology* ; Tumor Necrosis Factor-alpha/metabolism*

Objective To investigate the effect of transcutaneous electrical acupoint stimulation(TEAS)on emergence agitation(EA)in children undergoing selective posterior rhizotomy(SPR).Methods Forty-two children with cerebral palsy undergoing SPR were selected,20 males and 22 females,aged 6-12 years,BMI 13-24 kg/m2,ASA physical status Ⅰ or Ⅱ,randomly divided into two groups:TEAS group(group T)and control group(group C),21 children in each group.Children in group T re-ceived TEAS at the bilateral acupoints of Neiguan and Hegu from 30 minutes before anesthesia induction to the end of surgery.In group C,electrodes were placed on the same acupoints without electrical stimulation.All children in both groups received total intravenous anesthesia.HR and MAP were recorded at the time of entry,extubation,5,15,30 minutes after extubation.The consumption of remifentanil and propofol during the procedure were recorded.The time of operation and extubation were recorded.The Wong-Baker faces pain scale-revised(FPS-R)and the pediatric anesthesia emergence delirium(PAED)were recorded at 15 minutes after extubation,and the incidence of EA was assessed by PAED.While,the occurrence of post-operative nausea and vomiting(PONV)was recorded.Results Compared with that at the time of entry,HR at the time of extubation,5,15 minutes after extubation and MAP at the time of extubation,5,15,30 minutes after extubation in group C were significantly increased(P＜0.05),HR and MAP at the time of extubation,5,15 minutes after extubationin group T were significantly increased(P＜0.05).Compared with group C,HR was significantly slower and MAP was significantly lower in group T at the time of extuba-tion,5,15,30 minutes after extubation(P＜0.05).Compared with group C,the consumption of remifen-tanil during operation was reduced,the time of extubation was significantly shortened,the FPS-R and PAED were significantly decreased,and the incidence of EA were significantly reduced(P＜0.05).There were no significant differences in the time of operation,the consumption of propofol during operation and the inci-dence of PONV.Conclusion TEAS is beneficial to prevent the occurrence of EA in pediatric patients un-dergoing SPR,stabilize hemodynamics,reduce the dosage of opioids during surgery,reduce the postopera-tive pain,andaccelerate the time of anesthesia resuscitation.

In recent years,the rapid development of fascia plane block technique has played a cru-cial role in the development of multi-modal analgesic strategies in spinal surgery.Compared with traditional intraspinal anesthesia,fascia plane block has the advantages of simple operation,safety,good analgesic effect,no interference to intraoperative nerve monitoring and early postoperative inspection.In this artical,we reviewed the clinical application advances of fascia plane blocks commonly used in spinal surgery,such as transversal abdominis plane block(TAPB),thoracolumbar interfascial plane block(TLIPB),erector spinae plane block(ESPB),mid-point transverse process to pleura block(MTPB),multifidus plane block(MPB),and inter semispinal plane block(ISPB),in order to provide reference for multi-mode analgesia in spinal surgery.

Objective:To evaluate the relationship between preoperative plasma trimethylamine oxide (TMAO) concentrations and postoperative delirium (POD) in elderly patients.Methods:One hundred and eighty patients, aged ≥65 yr, of American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ, with body mass index of 17-30 kg/m 2, scheduled for elective hip replacement, were enrolled in this study. Venous blood samples were collected at 1 day before surgery, and the plasma TMAO concentrations were measured by high throughput liquid chromatography-tandem mass spectrometry. The patients were divided into 3 groups according to the concentrations of TMAO: low-concentration TMAO group (group L, ≤1.2 μmol/L), moderate-concentration TMAO group (group M, 1.3-1.9 μmol/L), and high-concentration TMAO group (group H, ≥2.0 μmol/L). All patients received superior inguinal fascia iliaca compartment block combined with intravenous general anesthesia. POD was identified by the Confusion Assessment Method-Chinese Reversion scale at 1, 2, 3 and 7 days after operation. The patients were divided into POD group and non-POD group according to whether POD occurred. Binary logistic regression analysis was used to evaluate the relationship between different plasma TMAO concentrations and POD. The receiver operating characteristic curve was used to evaluate the efficacy of plasma TMAO concentrations in predicting the occurrence of POD. Results:There were 60 cases in L group, 67 cases in M group and 53 cases in H group, and the incidence of POD was 12%, 22% and 32% in L, M and H groups, respectively. The incidence of POD was significantly higher in group H than in group L ( P<0.05). The plasma TMAO concentration was significantly higher in POD group than in non-POD group ( P<0.05). Logistic regression analysis showed that the risk of POD was 3.91 times higher in group H than in group L ( P<0.05). The area under the receiver operating characteristic curve of preoperative plasma TMAO concentrations in predicting POD was 0.754 (95% confidence interval 0.671-0.838, P<0.05). When the Youden index was 0.426, the optimal cut-off value of plasma TMAO concentrations was 1.625 μmol/L, and the specificity and sensitivity were 0.795 and 0.631, respectively. Conclusions:Elevated preoperative plasma TMAO concentration is associated with an increased risk of POD in elderly patients, and it has a good predictive value for POD.

Objective:To explore the correlation between preoperative echocardiography indicators and surgical prognosis of children with ventricular septal defect (VSD) and conduct verification based on significant indicators and indicator ratios.Methods:A total of 1 357 children with VSD who were admitted to the Children′s Hospital, Zhejiang University School of Medicine from June 2016 to June 2021 were selected. Various measurements including the size of the VSD, left ventricular ejection fraction (LVEF), left atrial (LA) diameter, the aortic (AO) flow rate, the tricuspid regurgitation velocity and pressure gradient were extracted from preoperative echocardiography reports. This paper explored the correlation between echocardiography reports indicators, indicator ratios and postoperative auxiliary ventilation time, respectively. The patients were divided into two groups according to whether there were complications, and the differences of echocardiography reports indicators between the two groups were compared. A linear regression model was established to predict the postoperative auxiliary ventilation time using these indicators, and the least absolute shrinkage and selection operator (LASSO) regression model was used for variable selection.Results:The VSD size and AO flow velocity were weakly correlated with the postoperative auxiliary ventilation time ( r=0.32, 0.25; all P<0.01). There was no significant correlation between VSD flow velocity and postoperative auxiliary ventilation time. The AO flow velocity/VSD flow velocity and LVEF/VSD flow velocity were strongly correlated with the postoperative auxiliary ventilation time ( r=0.67, 0.51; all P<0.01). In the significance test, there were no significant differences in tricuspid regurgitation flow velocity, tricuspid regurgitation pressure gradient, LA diameter, and LVEF between the complication group and the non-complication group(all P>0.01). However, the ratio of LVEF/tricuspid regurgitation velocity in the complication group was significantly lower than that in the non-complication group, and the ratio of tricuspid regurgitation pressure gradient/LA diameter was significantly higher than that in the non-complication group (all P<0.01). The postoperative auxiliary ventilation time of VSD patients was predicted on an independent test set, with an R2 of 0.51. Conclusions:Echocardiography report indicator ratios of AO flow velocity/VSD flow velocity and LVEF/VSD flow velocity have strong correlations with postoperative auxiliary ventilation time in children with VSD, and the ratios of LVEF/tricuspid regurgitation velocity and tricuspid regurgitation pressure gradient/LA diameter are significantly different between groups with and without postoperative complications. The ratios of indicators can significantly improve this correlation and difference, which can be used to predict the prognosis of VSD operation.

Objective To evaluate the clinical efficacy of adult kidney transplantation from unilateral pediatric donor kidney. Methods Clinical data of pediatric donors (n=10) and adult recipients (n=19) undergoing kidney transplantation were retrospectively analyzed. The changes of renal function, liver function and the maximal diameters of the kidney allografts were compared at 1, 7, 14, 28, 60 d after operation. The short-term survival and incidence of postoperative complications of the recipients were analyzed. Results Ten donors included 6 males and 4 females, aged (7±3) years old, with a body mass index (BMI) of (16.3±3.8) kg/m². All donors were donation after brain death followed by cardiac death. Among 19 recipients, 12 were males and 7 were females, aged (34±12) years old, with a BMI of (20.3±1.3) kg/m².An oblique incision was created in the lower right abdomen of the recipients. The arteries and veins of donor kidney were anastomosed with the external iliac arteries and veins of the recipients. The ureter of donor kidney was anastomosed with the bladder of the recipients. After anastomosis, the kidney was placed and fixed in the right iliac fossa. The serum creatinine and blood urea nitrogen levels of the recipients were decreased at 1 week after kidney transplantation, and restored to normal range at postoperative 2 weeks. All parameters related to liver function were normal after operation. At postoperative 1 month, the maximal diameters of the kidney allografts were (9.5±0.3) cm on average, which basically reached those of normal adults. The 1-year survival rate of 19 recipients was 95%. One recipient died from pulmonary infection after ineffective treatment. Two recipients developed rejection, and 1 recipient experienced urinary system infection, who were healed after corresponding treatment. Conclusions Adult kidney transplantation from unilateral pediatric donor kidney is safe, feasible and effective, which can be utilized to enlarge the source of donor kidneys.

Objective:To evaluate the effect of regional oxygen saturation (rSO 2)-guided low-dose norepinephrine on postoperative cognitive dysfunction (POCD) in elderly patients undergoing hip replacement under general anesthesia. Methods:One hundred and twenty patients of both sexes, aged 65-80 yr, with body mass index of 18-24 kg/m 2, of American Society of Anesthesiologists physical statusⅠ-Ⅲ, scheduled for hip replacement under general anesthesia, were divided into 2 groups ( n=60 each) using a random number table method: control group (group C) and low-dose norepinephrine guided by rSO 2 group (group RN). The patients in both groups received superior inguinal fascial space block combined with general anesthesia under laryngeal mask placement.In group C, the fluctuation range of mean arterial pressure (MAP) was not more than 20% of the baseline, vasoactive agents were administered according to the changes in blood pressure, rSO 2 was monitored continuously, but the change rate of rSO 2 was not used as the regulating index.In group RN, norepinephrine was infused continuously via the central vein at 0.01-0.10 μg·kg -1·min -1 after anesthesia induction, the dose was adjusted according to rSO 2, the rSO 2 change rate was maintained≤10%, the fluctuation range of mean arterial pressure was not more than 20% of the baseline, and vasoactive agents were administered when necessary.MAP, end-tidal pressure of carbon dioxide (P ETCO 2) and rSO 2 were recorded after inhalation of oxygen (T 0), at 5 min after anesthesia induction (T 1), at 30 min after skin incision (T 2), at the end of surgery (T 3) and after recovery and extubation (T 4), and the change rate of rSO 2 was calculated.The occurrence of adverse events and amount of vasoactive drugs used were recorded.The cognitive function was assessed using Montreal Scale at 1 day before surgery and 7 days after surgery, and the development of postoperative cognitive dysfunction (POCD) was calculated using Z score.The postoperative hospital stay time was recorded. Results:Compared with group C, MAP and rSO 2 were significantly increased, and the change rate of rSO 2 was decreased at T 1, 2 in group RN ( P<0.05). Compared with group C, the requirement for intraoperative vasoactive drugs was significantly decreased, the consumption of norepinephrine was increased, MoCA total score, attention and delayed recall sub-score were increased at 7 days after surgery, the incidence of POCD was decreased, and the postoperative hospital stay time was shortened in group RN ( P<0.05). Conclusion:Low-dose norepinephrine guided by rSO 2 can decrease the development of POCD in elderly patients undergoing hip replacement under general anesthesia.

Objective To compare the efficacy and safety of turning holmium laser enucleation of the prostate (THoLEP) and transurethral holmium laser enucleation of the prostate (HoLEP) in the treatment of benign prostatic hyperplasia (BPH).Methods A retrospective analysis of 532 patients with BPH surgery from January 2016 to December 2017 was performed,including 289 cases of transurethral resection of holmium laser enucleation (THoLEP) and 43 cases of transurethral holmium laser enucleation (HoLEP).Cases with incomplete data were excluded.Finally,100 patients were enrolled and divided into THoLEP group and HoLEP.The mean ages of patients in the THoLEP group and the HoLEP group were (74.6 ± 8.2) years and (75.3 ± 7.7) years,respectively.The prostate weights were (52.3 ± 23.1) g and (52.6 ± 22.7) g,respectively.International prostate symptom scores (IPSS) were (22.8 ± 1.3) and (23.1 ± 1.6),and the quality of life score (QOL) scores were (4.4 ± 0.5) and (4.3 ± 0.8),respectively.The rate of mximum flow rate (Qmax) was (5.9 ± 2.7) ml/s and (6.3 ± 2.8) mL/s,respectively.The residual urine volume was (149.3 ± 8.7) ml and (145.3 ± 9.7) ml,respectively.There was no significant difference between the two groups (P ＞ 0.05).Results Compared with HoLEP group,THoLEP group showed shorter operation time [(44 ± 13) min vs.(61 ± 15) min,P ＜ 0.05] and less intraoperative bleeding [(101.6 ± 13.2) ml vs.(119.9 ± 18.0) ml,P ＜ 0.05].The weight of the resected tissue in the THoLEP group was slightly larger than that in the HoLEP group,however the difference was not statistically significant [(30.5 ± 9.5) g vs.(28.7 ± 8.5) g] (P ＞ 0.05).The incidence of transient urinary incontinence in THoLEP group was less than that in HoLEP group [2 and 4 cases,P ＜ 0.05].There were no cases of urinary incontinence in both groups after 3 months,and no permanent urinary incontinence in both groups.THoLEP was slightly better than HoLEP group,but the difference was not statistically significant (P ＞0.05).Conclusions Compared with HoLEP,THoLEP has similar short-term results,and has more advantages in shortening the operation time,reducing intraoperative bleeding and transient urinary incontinence.It is a safer and effective method for the treatment of benign prostatic hyperplasia.

Objective To establish a pediatric echocardiographic normal reference system based on clinical BigData and overcome limitations such as insufficient sample size and diverse in methods of normalization . Methods Measurements were extracted from total 71 831 pediatric echocardiography reports in the past 5 years by using the Natural Language Processing ( NLP) technology . Among them ,a total of 12 732 reports were labeled as normal and were used to establish the normal reference system . A local regression ( LOESS ) approach was used to optimize both the reference value and variance across 5 grow th variables ( aortic diameter ,left atrium diameter ,left ventricle end‐diastolic endocardial diameter ,left main coronary artery diameter ,and right main coronary artery diameter) . T wo Z scores adjusted for age/sex and body surface area ( BSA ) were established respectively . In addition , 4 459 echocardiography reports with BSA information were used to evaluate these two Z scores . Results T wo Z scores generated from 4 459 reports showed pretty good normal distribution . T here were close strong correlations among two Z scores with Z scores generated based on the Pediatric Heart Network ( PHN ) . T he average correlation coefficient between BSA‐adjust Z scores and PHN Z scores was 0 .954 . T he average correlation coefficient between age/sex‐adjust Z scores and PHN Z scores was 0 .895 . T he results of this project were available as Z score calculator using the following link :http ://hdb .nbscn .org/zscore . Conclusions BigData provides a more efficient and better approach to establish normal reference systems in pediatric echocardiography .