Objective To investigate the expression and mechanistic role of the M2-type pyruvate kinase(PKM2)/signal transducer and activator of transcription 3(STAT3)signaling pathway in the development of experimental autoim-mune uveitis(EAU).Methods Eighteen 4-to 5-week-old C57BL/6J mice were selected and randomly divided into a control group(normal breeding,set as the 0-day state post-modeling),Experimental Group A,and Experimental Group B(both established as stable EAU mouse models using the interphotoreceptor retinoid-binding protein peptide 651-670,com-plete Freund's adjuvant,and pertussis toxin,designated as the 14-day and 21-day states post-modeling,respectively),with 6 mice in each group.The anterior segment of the mice was observed using a slit-lamp microscope,fundus findings were collected using a fundus camera,and clinical and histopathological scores were evaluated after hematoxylin-eosin stai-ning.The protein expression level of serum interleukin(IL)-17A was detected by ELISA.The expression level of PKM2 protein in retinal tissue was detected by immunohistochemistry.The protein expression levels of PKM2,phosphorylated STAT3(p-STAT3),and STAT3 in mouse retinal tissue were detected by Western blot.Results The clinical scores of the anterior segment and fundus,as well as the retinal histopathological scores in Experimental Group A and Experimental Group B,were all significantly higher than those in the control group(all P＜0.05).The serum IL-17A protein expression levels in the control group,Experimental Group A,and Experimental Group B were(69.05±0.45)ng·L-1,(75.06±0.46)ng·L-1,and(72.04±0.82)ng·L-1,respectively.The optical density values of PKM2 protein expression in retinal tissue were(18.51±2.59)％,(37.35±4.67)％,and(29.75±2.17)％,respectively.The expression levels of serum IL-17A protein,retinal PKM2 protein,and retinal STAT3 and p-STAT3 proteins in Experimental Group A and Experimental Group B were all significantly higher than those in the control group(all P＜0.05).Conclusion The expression levels of key factors in the PKM2/STAT3 signaling pathway are positively correlated with the severity of EAU,indicating that this sig-naling pathway,as a positive regulator of the immune response,is involved in the pathological process of EAU.

BACKGROUND:Currently,there are various rat models of lumbar disc herniation used in experiments,each with its own advantages and disadvantages.The most common modeling methods include autologous nucleus pulposus transplantation and annulus fibrosus puncture models.OBJECTIVE:To establish two autologous nucleus pulposus transplantation models(with spinous process and mastoid process excision and with transverse process and mastoid process excision)as well as an annulus fibrosus puncture model,and to compare and evaluate the characteristics of the three models.METHODS:Forty male adult Sprague-Dawley rats were randomly divided into four groups(n=10 per group):sham surgery group,spinous process group,transverse process group,and annulus fibrosus puncture group.In the sham surgery group,surgical preparation was performed,the skin was incised,and the spinous process was exposed,and then sutured.In the spinous process group,L5 spinous process and transverse process were excised,and two pieces of tail nucleus pulposus were placed in the intervertebral foramen.In the transverse process group,L5 transverse process and transverse process were excised,and two pieces of tail nucleus pulposus were placed in the intervertebral foramen.In the annulus fibrosus puncture group,the transverse process was excised and annulus fibrosus puncture and intervertebral disc injection of interleukin 1β were then performed.Thermal paw withdrawal latencies were tested before and after modeling.Lumbar spine MRI was performed 2 weeks after modeling.Pathological changes in the intervertebral discs were observed using hematoxylin-eosin staining and safranin-O-fast green staining.Immunofluorescence was used to observe CD68+positive expression.RESULTS AND CONCLUSION:(1)Thermal withdrawal threshold testing results showed that compared with the sham surgery group,pain sensitivity and tolerance time of rats decreased significantly after modeling(P<0.05).(2)Lumbar spine MRI images showed that the spinous process and transverse process groups had obvious protrusion of nucleus pulposus tissue,which more closely resembled MRI images of patients with common lumbar disc herniation.(3)Hematoxylin-eosin staining revealed that compared with the sham surgery group,nucleus pulposus tissues in the model groups showed varying degrees of degeneration,inflammatory cell infiltration,and degradation of spinal cord cells,with the appearance of cystic changes,among which the annulus fibrosus puncture group had the most severe pathological changes.(4)Safranin-O-fast green staining showed that compared with the sham surgery group,the boundaries of nucleus pulposus tissues in the three model groups were blurred,with extensive inflammatory reactions and varying degrees of degeneration in the annulus fibrosus.(5)CD68+immunofluorescence staining results showed that compared with the sham surgery group,the expression of CD68+in the model groups was higher and more widespread,with the annulus fibrosus puncture model showing the highest expression.All the three methods could be used to effectively establish rat models of lumbar disc herniation,with the annulus fibrosus puncture model established after excision of the transverse process being superior to the autologous nucleus pulposus transplantation model(spinous process+mastoid process),and the first two models being superior to the autologous nucleus pulposus transplantation model(transverse process+mastoid process).

Objective To investigate the effect of sodium glucose cotransporter 2 inhibitor(SGLT2i)on cardiac function in T2DM rats with heart failure(HF)by regulating the silencing signaling protein 1/mitochondrial uncoupling protein 2(SIRT1/UCP2)signaling pathway.Methods Forty SD rats were randomly divided into normal control(NC)group,T2DM combined with HF(T2DM-HF)group,SGLT2i group,and SGLT2i+SIRT1 inhibitor(EX527)(SGLT2i+EX527)group.Fasting blood glucose(FBG),cardiac function,and oxidative stress indicators were tested in each group.The pathological morphological changes of myocardial tissue were evaluated by HE and Masson staining,and the expression of SIRT1 and UCP2 proteins in myocardial tissue was evaluated by Western blot.Results Compared with NC group,the T2DM-HF,SGLT2i,and SGLT2i+EX527 groups showed a decrease in body weight(P<0.05)and an increase in cardiac mass index(P<0.05).Compared with T2DM-HF group,the SGLT2i group showed decreased protein expression of FBG,LVESD,LVEDD,LVEDP,MDA,and UCP2(P<0.05),while increased protein expression of LVFS,LVEF,LVSP,±dp/dtmax,SOD,and SIRT1(P<0.05).Compared with SGLT2i group,the SGLT2i+EX527 group showed an increase in FBG,LVESD,LVEDD,LVEDP,MDA,and UCP2 opal levels,while LVFS,LVEF,LVSP,±dp/dtmax,SOD,and SIRT1 opal levels decreased(P<0.05).Conclusions SGLT2i can improve the cardiac function in T2DM rats combined with HF,enhance antioxidant capacity,and exert a protective effect on cardiac function.Its mechanism of action may be related to the regulation of SIRT1/UCP2 signaling pathway.

Objective To compare the ability of single-phase,dual-phase,and triphasic models in forecasting postoperative pancreatic fistula(POPF)after pancreaticoduodenectomy(PD)using radiomics based on triphasic enhanced CT.Methods A total of 181 patients who underwent multi-phase enhanced CT prior to PD were retrospectively selected,and the collection phase included non-contrast,arterial phase(AP),and equilibrium phase(EP).3D Slicer software was utilized to segment the region of interest(ROI)for the postoperative pancreatic remnant on each phase.Radiomics feature extraction was performed using R software,followed by feature selection through least absolute shrinkage and selection operator(LASSO)regression with five-fold cross-validation to prevent model overfitting.The effective features selected were combined in a weighted linear manner to obtain a Radiomics score(Radscore).The patients were divided into training set and test set in a 7︰3 ratio.Logistic regression was employed to construct seven POPF prediction models(three single-phase,three dual-phase,and one triphasic models)based on different phase combinations.The diagnostic performance of the models was evaluated using the area under the curve(AUC)of receiver operating characteristic(ROC)curve,accuracy(ACC),sensitivity(SEN),and specificity(SPE).The DeLong test was applied to compare the differences in AUC among different models.Results After LASSO regression,24 effective features associated with POPF were selected from different phases.In the test set,the triphasic model exhibited the highest AUC and ACC(AUC=0.76,ACC=0.808).The calibration curve demonstrated the strongest agreement between the estimated probabilities and observed probabilities for the triphasic model.The decision curve analysis(DCA)curve indicated that the triphasic model had the largest threshold range with a higher net benefit.Conclusion Compared with single-phase and dual-phase models,the triphasic model based on enhanced CT provides better prediction of POPF after PD,aiding clinical decision-making and improve prognosis.

BACKGROUND:Currently,there are various rat models of lumbar disc herniation used in experiments,each with its own advantages and disadvantages.The most common modeling methods include autologous nucleus pulposus transplantation and annulus fibrosus puncture models.OBJECTIVE:To establish two autologous nucleus pulposus transplantation models(with spinous process and mastoid process excision and with transverse process and mastoid process excision)as well as an annulus fibrosus puncture model,and to compare and evaluate the characteristics of the three models.METHODS:Forty male adult Sprague-Dawley rats were randomly divided into four groups(n=10 per group):sham surgery group,spinous process group,transverse process group,and annulus fibrosus puncture group.In the sham surgery group,surgical preparation was performed,the skin was incised,and the spinous process was exposed,and then sutured.In the spinous process group,L5 spinous process and transverse process were excised,and two pieces of tail nucleus pulposus were placed in the intervertebral foramen.In the transverse process group,L5 transverse process and transverse process were excised,and two pieces of tail nucleus pulposus were placed in the intervertebral foramen.In the annulus fibrosus puncture group,the transverse process was excised and annulus fibrosus puncture and intervertebral disc injection of interleukin 1β were then performed.Thermal paw withdrawal latencies were tested before and after modeling.Lumbar spine MRI was performed 2 weeks after modeling.Pathological changes in the intervertebral discs were observed using hematoxylin-eosin staining and safranin-O-fast green staining.Immunofluorescence was used to observe CD68+positive expression.RESULTS AND CONCLUSION:(1)Thermal withdrawal threshold testing results showed that compared with the sham surgery group,pain sensitivity and tolerance time of rats decreased significantly after modeling(P<0.05).(2)Lumbar spine MRI images showed that the spinous process and transverse process groups had obvious protrusion of nucleus pulposus tissue,which more closely resembled MRI images of patients with common lumbar disc herniation.(3)Hematoxylin-eosin staining revealed that compared with the sham surgery group,nucleus pulposus tissues in the model groups showed varying degrees of degeneration,inflammatory cell infiltration,and degradation of spinal cord cells,with the appearance of cystic changes,among which the annulus fibrosus puncture group had the most severe pathological changes.(4)Safranin-O-fast green staining showed that compared with the sham surgery group,the boundaries of nucleus pulposus tissues in the three model groups were blurred,with extensive inflammatory reactions and varying degrees of degeneration in the annulus fibrosus.(5)CD68+immunofluorescence staining results showed that compared with the sham surgery group,the expression of CD68+in the model groups was higher and more widespread,with the annulus fibrosus puncture model showing the highest expression.All the three methods could be used to effectively establish rat models of lumbar disc herniation,with the annulus fibrosus puncture model established after excision of the transverse process being superior to the autologous nucleus pulposus transplantation model(spinous process+mastoid process),and the first two models being superior to the autologous nucleus pulposus transplantation model(transverse process+mastoid process).

Objective To investigate the expression and role of T cell immunoglobulin and mucin domain-containing protein 3(Tim-3)in the pathogenesis of experimental autoimmune uveitis(EAU).Methods A total of 12 male C57BL/6J mice,aged 4 to 5 weeks,were selected and divided into the control group(n=3)and the experimental group(n=9)using a random number table.The control group(modeling time point:0 days after modeling)received no treatment,while the experimental group was induced to establish an EAU model(divided into three subgroups according to the modeling time points:7 days,14 days,and 21 days after modeling,with 3 mice in each subgroup).Firstly,the interphotoreceptor retinoid-binding protein 651-670 and complete Freund's adjuvant were fully mixed and emulsified.Then,the emulsion was subcutaneously injected into the two thighs,tail base,and neck of mice in the experimental group(each mouse received 200 μL of immune emulsion containing 500 pg of interphotoreceptor retinoid-binding protein 651-670).Subsequently,each mouse in the experimental group was also intraperitoneally injected with 1 μg of pertussis toxin.The anterior segment and fundus of mice in each group were observed and photographed under a slit-lamp microscope.The clinical and histopatho-logical scoring of these mice was conducted according to the Caspi grading scale based on the severity of inflammation.The serum levels of IFN-γ and IL-17 were measured using the enzyme-linked immunosorbent assay(ELISA),while the mRNA expression of Tim-3 in the spleen and ocular tissues was detected using the real-time quantitative polymerase chain reaction(RT-qPCR).Western blot was employed to detect the protein expression of Tim-3,and immunohistochemistry was used to examine the protein expression of Tim-3 in the spleen tissue.Statistical analysis was performed using GraphPad Prism 9.0.Results The clinical scores of the anterior segment,fundus,and histopathology of the mice increased over time after modeling,with statistically significant differences among these groups(P＜0.05).The serum levels of IFN-γ and IL-17 in the mice also increased over time after modeling,with statistically significant differences among these groups(P＜0.05).The relative mRNA expression of Tim-3 in the spleen and ocular tissues of the mice decreased over time after modeling,with statistically significant differences among these groups(P＜0.05).The protein expression of Tim-3 in the ocular and spleen tissues showed the same pattern as its mRNA expression.Conclusion The expression of Tim-3 decreases with the exacerbation of inflammation in the progression of EAU,suggesting that Tim-3 may play a negative immunoregulatory role in the development of uveitis.

Objective To study the basic features,judicial identification and trial characteristics of medical malpractice cases related to gastrointestinal endoscopy,enhance the awareness of risks of gastrointestinal endoscopy among medical staffand patients.,emphasize the importance of preoperative education and provide judicial identification and trial personnel with ideas for handling such cases,ensuring standardization and consistency in handling similar cases.Methods A total of 100 medical dispute cases involving gastrointestinal endoscopy from May 2010 to May 2024 were included from the China Judgments Online database.The analysis focused on document types,years,regions,court levels,details of judicial expertise,the proportion of court decision responsibility.Results The number of cases has increased significantly since 2017,with the highest incidence in 2020 and 2021(collectively accounting for 39%);Regions with the most disputes included Fujian,Guangdong,Shandong,Shanghai,Liaoning,Beijing,Jiangsu,collectively accounting for 58%of cases.The primary method of forensic expertise was medical damage identification,accounting for 82.0%of cases;Among these,4 cases underwent reappraisal,all of which resulted in modifications to the original expertise.Male patients outnumbered female patients,with"Digestive tract discomfort"being the main reason for consultation(63%).The most common medical injury was digestive tract perforation(37%),among which colon perforation was the most common;Of the 100 cases,60 cases were found to have errors in gastrointestinal endoscopy,and the medical errors were concentrated in"Improper operation during Operation"(42 cases),The contributory roles of various negligence types in the damage consequence ranged from"no causation"to"complete causation"in varing proportions.Notably,negligence categories such as"improper intraoperative manipulation"and"delayed treatment due to untimely examination"consistently demonstrated contributory roles of"secondary causation or higher."Regarding court rulings,the majority of cases(23 cases)assigned liability proportions of 31%-50%,followed by 20 cases with liability proportions of 71%-100%.

Objective To investigate the expression and mechanistic role of the M2-type pyruvate kinase(PKM2)/signal transducer and activator of transcription 3(STAT3)signaling pathway in the development of experimental autoim-mune uveitis(EAU).Methods Eighteen 4-to 5-week-old C57BL/6J mice were selected and randomly divided into a control group(normal breeding,set as the 0-day state post-modeling),Experimental Group A,and Experimental Group B(both established as stable EAU mouse models using the interphotoreceptor retinoid-binding protein peptide 651-670,com-plete Freund's adjuvant,and pertussis toxin,designated as the 14-day and 21-day states post-modeling,respectively),with 6 mice in each group.The anterior segment of the mice was observed using a slit-lamp microscope,fundus findings were collected using a fundus camera,and clinical and histopathological scores were evaluated after hematoxylin-eosin stai-ning.The protein expression level of serum interleukin(IL)-17A was detected by ELISA.The expression level of PKM2 protein in retinal tissue was detected by immunohistochemistry.The protein expression levels of PKM2,phosphorylated STAT3(p-STAT3),and STAT3 in mouse retinal tissue were detected by Western blot.Results The clinical scores of the anterior segment and fundus,as well as the retinal histopathological scores in Experimental Group A and Experimental Group B,were all significantly higher than those in the control group(all P＜0.05).The serum IL-17A protein expression levels in the control group,Experimental Group A,and Experimental Group B were(69.05±0.45)ng·L-1,(75.06±0.46)ng·L-1,and(72.04±0.82)ng·L-1,respectively.The optical density values of PKM2 protein expression in retinal tissue were(18.51±2.59)％,(37.35±4.67)％,and(29.75±2.17)％,respectively.The expression levels of serum IL-17A protein,retinal PKM2 protein,and retinal STAT3 and p-STAT3 proteins in Experimental Group A and Experimental Group B were all significantly higher than those in the control group(all P＜0.05).Conclusion The expression levels of key factors in the PKM2/STAT3 signaling pathway are positively correlated with the severity of EAU,indicating that this sig-naling pathway,as a positive regulator of the immune response,is involved in the pathological process of EAU.

Objective To differentiate disguised visual acuity,(VA)by analyzing discrepancies across diversified visual acuity tests.Methods Volunteers were recruited,and VA(V,and V2)was measured before and after the experiment.Volunteers independently selected an experimental VA(Vt)and were assigned to either the disguised group or the control group.A specially designed heterogeneous optotype chart and a single-optotype chart with variable test distances were used to measure VA(t1,t2,t3),while a standard logarithmic VA chart was used to measure VA(t4).Based on the maximum VA discrepancy(△t),t4 was classified as"real"(≤ 1 line),"disguised"(≥ 2 lines),or"suspicious"(＞1 and＜2 lines with anomaly).Results A total of 126 valid cases were collected,including 30 in the control group and 96 in the disguised group,with VA ranging from 0.2～1.5.All 88 opinions classified as"disguised"were from the disguised group,and all 29 classified as"real"were from the control group.Of the 7 cases deemed"suspicious,"6 were from the disguised group.The sensitivity,specificity,and overall diagnostic accuracy were 91.7％,96.7％(P＜0.0001),and 92.9％(P＜0.0001),respectively.In the disguised group,38 cases exhibited unexplained identifying anomalies.Conclusion A VA discrepancy of ≥ 2 lines is a reliable indicator for detecting disguised visual acuity.Identifying anomalies may serve as a novel and sensitive marker for recognizing visual disguise.

Objective:To study the risk factors and the molecular epidemiology characteristics for Carbapenem-resistant Enterobacteriaceae(CRE) colonization in neonatal inpatients in Shenzhen region, China, which provide reference for the prevention and control of clinical CRE infection.Methods:This study is a prospective case-control study.Anal samples from inpatients between January 2023 and December 2023 at Longgang Maternity and Child Institute of Shantou University Medical College and Shenzhen Children's Hospital were collected for screening CRE strain. Drug susceptibility test, modified Carbapenem Inactivation Method (mCIM) test, drug resistance-related gene sequencing and multilocus sequence typing (MLST) were performed for isolated CRE strains.Meanwhile, the clinical data were collected for analyzing the risk factors of CRE intestinal colonization by multivariate regression analysis.Results:A total of 1 517 patients were screened, 26 CRE(1.7%, 26/1 517) were identified which including 14 Escherichia coli(53.8%, 14/26), 11 Klebsiella pneumoniae(42.3%, 11/26), 1 Enterobacter cloacae(3.9%, 1/26). The predominant carbapenemase gene was New Delhi Metallo(NDM) (92.4%, 24/26), followed by Imipenem (IMP) (3.8%, 1/26) and Guiana extended spectrum gene (GES) (3.8%, 1/26).Among the carried NDM resistance genes, New Delhi Metallo 5 (NDM5) was the main one, accounting for 84.6% (22/26).The MLST typing of Escherichia coli was mainly Sequence Type 48 (ST48) (6/14), while that of Klebsiella pneumoniae was mainly Sequence Type 35 (ST35) (10/11). All CRE isolates were resistant to penicillin, penicillinase inhibitors, cephalosporins, ertapenem and imipenem.The resistance rates of Escherichia coli to amikacin, levofloxacin was 1/14, 4/14, respectively. All isolates of Klebsiella pneumoniae were sensitive to amikacin, and the resistance rate to levofloxacin is 1/11. Risk factors for CRE colonization include the older age, length of hospital stay, tracheal intubation, invasive respiration, lumbar puncture, Apgar <7 score, and exposure to antibiotics.Conclusions:NDM5 is the predominant resistant gene in CRE isolated from neonatal patients feces in Shenzhen region.It is necessary to strengthen the screening of CRE colonization in neonate for prevention and control of CRE infection.