Objective To investigate the expression characteristics of c-MET protein in colorectal cancer (CRC) and its associations with clinicopathological parameters, MET gene amplification status, and prognosis, and to evaluate the concordance between immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) detection strategies, thereby providing evidence for patient selection for MET-targeted therapy. Methods A total of 193 formalin-fixed paraffin-embedded (FFPE) tissue samples from CRC patients were collected. Immunohistochemistry (IHC) was used to detect c-MET protein expression, and fluorescence in situ hybridization (FISH) was performed to evaluate MET gene amplification status. The associations between c-MET protein expression and clinicopathological characteristics, including KRAS, NRAS, and BRAF mutation status, as well as progression-free survival (PFS), were analyzed. The concordance between IHC and FISH detection results was also evaluated. Results c-MET protein expression was significantly higher in tumor tissues than in adjacent normal tissues (P<0.05). Significant differences in c-MET expression were observed between primary tumors and liver or peritoneal/pelvic metastatic lesions (P<0.05). The proportion of high c-MET expression was significantly higher in KRAS-mutant patients than in KRAS wild-type patients (41.38% vs. 16.92%, P<0.05). The MET gene amplification rate was 8.29% (16/193), including clustered or diffuse amplification in 1.55% (3/193) and heterogeneous amplification in 6.74% (13/193). No MET exon 14 skipping mutation or amplification was detected by next-generation sequencing (NGS). Using an IHC H-Score≥150 as the threshold, the positive percent agreement (PPA) between IHC-detected c-MET protein expression and FISH-detected MET gene amplification was 100%. All clustered MET amplification regions exhibited diffuse strong c-MET expression (IHC 3+). No statistically significant association was observed between c-MET expression level and PFS (P>0.05). Conclusion High c-MET expression may represent a potential therapeutic target for KRAS-mutant CRC patients. An H-Score ≥150 is recommended as the initial screening threshold to rapidly identify patients suitable for MET-targeted therapy, with FISH testing when necessary. Diffuse strong c-MET expression (IHC 3+) shows high concordance with clustered MET gene amplification.

Objective:To investigate the effects of a matrix metalloproteinase(MMP) inhibitor (ilomastat, GM6001) on learning and memory function and hippocampal myelin-related proteins in epileptic model rats.Methods:Thirty 8-week-old male SPF Sprague-Dawley rats were randomly divided into control group, intervention control group, short-term model group, long-term model group and long-term intervention group, with 6 rats in each group.The lithium-pilocarpine method was used to induce status epilepticus (SE). SE was terminated respectively by intraperitoneal injection of diazepam(10 mg/kg) at 15 min in short-term model group and at 30 min both in the long-term model group and long-term intervention group. Rats in intervention control group and long-term intervention group were treated with GM6001 (20 μg/kg) via tail vein injection. Morris water maze test was used to assess the learning and memory function and the blood-brain barrier permeability was evaluated by Evans blue (EB) staining. The levels of serum MMP, the hippocampal myelin basic protein (MBP) and 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) were measured by ELISA and Western blot.Statistical analysis was performed by SPSS 21.0 software, with repeated measures ANOVA for Morris water maze data and one-way ANOVA with LSD post hoc test for other comparisons.Results:(1) In the water maze test, the interaction between the time and group on the latencies reached to the platform showed no significance among the 5 groups of rats ( F=0.430, P=0.970). The main effects of time and group on latencies were both significant among the 5 groups( F=144.521, 12.819, both P<0.001).From the second day to the fifth day, the latencies reached to the platform of rats in the long-term model group were significantly higher than those in the control group and in the long-term intervention group (both P<0.05). There were statistically significant differences in the latency of the first time reached the platform, the duration in the target quadrant, the frequency of crossing the platform and the distance to the platform among the 5 groups ( F=7.008, 7.306, 15.704, 11.559, all P<0.05). In the long-term intervention group, the latency of the first time reached the platform ((13.71±4.41) s) and the distance to the platform ((60.37±8.59) cm) were significantly lower than those in the long-term model group ((23.52±5.34) s, (73.06±12.01) cm) (both P<0.05), and the duration in the target quadrant ((10.93±2.99) s) and the frequency of crossing the platform (2.00(1.75, 2.75)times) were significantly higher than those of the long-term model group ((6.04±4.33) s, 1.00(0, 1.00)times) (both P<0.05).(2) The hippocampal EB fluorescence intensity and the serum levels of MMP-2 and MMP-9 were significantly different among the 5 groups ( F=15.249, 7.951, 33.299, all P<0.001). The EB fluorescence intensity of the long-term model group ((0.18±0.03) /100 μm 2) was significantly higher than that in control group ((0.09±0.02) /100 μm 2) and the long-term intervention group ((0.12±0.04) /100 μm 2) (both P<0.05). The serum MMP-2 and MMP-9 levels in the long-term model group ((10.91±1.36) ng/mL, (10.96±1.28) ng/mL) were significantly higher than those of the control group ((7.89±1.04) ng/mL, (4.20±1.48) ng/mL) and the long-term intervention group ((6.49±1.12) ng/mL, (8.06±0.91) ng/mL) (all P<0.05). (3) The expression levels of hippocampal MBP and CNPase were significantly different among the 5 groups ( F=8.757, 4.116, both P<0.05). The expression levels of MBP and CNPase in long-term model group ((1.00±0.19), (0.93±0.21)) were significantly lower than those in the control group ((1.57±0.27), (1.36±0.22)) and the long-term intervention group ((1.45±0.19), (1.23±0.26)) (all P<0.05). Conclusion:The MMP inhibitor GM6001 can ameliorate learning and memory deficits in epileptic model rats, potentially by reducing blood-brain barrier permeability and alleviating hippocampal myelin damage.

Objective::This study aimed to assess the impact of proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitor treatment immediately after percutaneous coronary intervention (PCI) on the myocardial salvage index (MSI) in patients with anterior ST-segment elevation myocardial infarction (STEMI) 5-10 d after the procedure.Methods::The early PCSK9 inhibitor thERapy Following pErcutaneous Coronary inTervention (PERFECT) trial is a prospective randomized controlled trial. From January 2021 to December 2023, 32 patients with anterior STEMI from Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, and Shanghai Tenth People’s Hospital were enrolled in the PERFECT trial. Patients were randomly assigned in a 1∶1 ratio to the PCSK9 inhibitor group ( n = 16) or the control group ( n = 16), and their baseline data were collected. Patients in the PCSK9 inhibitor group (ie, alirocumab group) received a subcutaneous injection of PCSK9 inhibitor (alirocumab, 75 mg) immediately after PCI based on conventional treatment. In the control group, patients received only conventional treatment. The primary endpoint was the MSI measured by cardiovascular magnetic resonance 5-10 d after PCI. The secondary endpoints included the left ventricular ejection fraction measured by cardiovascular magnetic resonance 5-10 d after PCI and the time to peak of creatine kinase isoenzyme-MB and high-sensitivity cardiac troponin T. Safety endpoints included any clinical adverse events that occurred during the 6-month follow-up period. Results::Baseline data during admission showed no intergroup significance. No significant difference in MSI (55.54% ± 14.80% vs. 44.72% ± 15.42%, P = 0.056) and left ventricular ejection fraction (51.24% ± 8.91% vs. 44.99% ± 8.84%, P = 0.060) was observed. Additional, there was no significant difference in the time to peak of creatine kinase isoenzyme-MB ((12.97 ± 5.67) h vs. (14.31 ± 7.04) h, P = 0.557) and high-sensitivity cardiac troponin T ((21.03 ± 12.46) h vs. (21.44 ± 9.99) h, P = 0.920) between the 2 groups. During the 6-month follow-up period, only 1 patient in the PCSK9 inhibitor group developed cerebral hemorrhage 6 months after PCI. Conclusions::Early treatment with alirocumab did not exhibit a significant increase in MSI at 5-10 d in patients with anterior STEMI. Larger trials are necessary to evaluate the impact of early administration of PCSK9 inhibitors after myocardial infarction.

Objective:To investigate the role and molecular mechanism of dual-specificity phosphatase 26(DUSP26)in the proliferation,migration and invasion of lung adenocarcinoma(LUAD)A549 cells.Methods:The expression profile of DUSP26 was retrieved from the GEPIA2 tumor database,and its differential expression in LUAD tissues and normal human lung tissues were analyzed.Twelve pairs of LUAD tissue and paracancerous tissue surgically resected at Pingxiang People's Hospital between October 2022 and October 2023 were collected.The difference in DUSP26 expression between LUAD tissues and paracancerous tissues was analyzed using immunohistochemical(IHC)staining and Western blotting(WB).Additionally,the expression of DUSP26 in four LUAD cells(A549,SK-LU-1,Calu-3,H1299)and two normal bronchial epithelial cells(BEAS-2B,HBEC)was detected using WB method.A549 cells stably overexpressing DUSP26(DUSP26-OE)or corresponding negative control(DUSP26-OENC)were constructed via lentiviral transfection.The effects of DUSP26 overexpression on cell proliferation,migration and invasion were detected using colony formation,scratch assay,and Transwell chamber assay,respectively.The expression levels of TGF-β1/SMAD2/3 pathway-and epithelial-mesenchymal transition(EMT)-related proteins were detected using WB method,and the expression levels of Ki-67 and cyclin D1 in cells were detected by immunofluorescence staining.Rescue experiments were conducted by adding 5 ng/mL recombinant TGF-β1.A nude mouse xenograft model was established using A549 cells to observe the effect of DUSP26 overexpression on the in vivo growth of transplanted tumors.The expression levels of TGF-β1/SMAD2/3 pathway-and EMT-related proteins in transplanted tumor tissues were assessed using WB method,and the expression levels of Ki-67 and cyclin D1 in transplanted tumor tissues were detected using immunofluorescence staining.Results:DUSP26 expression was downregulated in both LUAD tissues and cells(P<0.05,P<0.01,P<0.001 or P<0.000 1).Compared with the DUSP26-OENC group,the DUSP26-OE group showed significantly reduced proliferation,migration and invasion of A549 cells(P<0.01 or P<0.001).Furthermore,the protein levels of TGF-β1,p-SMAD2/3,vimentin,N-cadherin,snail,Ki-67,and cyclin D1 were significantly reduced(P<0.01,P<0.001 or P<0.000 1),while E-cadherin level was increased in the DUSP26-OE group(P<0.000 1).The addition of 5 ng/mL TGF-β1 recombinant protein partially reversed the effects of DUSP26 overexpression in vitro experiments.The nude mice A549 cell xenograft model was successfully constructed.The growth rate of transplanted tumors was significantly slower in the DUSP26-OE group,with reduced volume and mass(all P<0.001).The protein levels of TGF-β1,p-SMAD2/3,vimentin,N-cadherin,snail,Ki-67,and cyclin D1 in the transplanted tumor tissues were all reduced(P<0.01 or P<0.001),while E-cadherin level was increased(P<0.000 1).Conclusion:DUSP26 is downregulated in both LUAD tissues and cells.Upregulation of DUSP26 suppresses the proliferation,migration and invasion of A549 cells by inhibiting the TGF-β1/SMAD2/3 signaling pathway.

Objective:To investigate the effects of a matrix metalloproteinase(MMP) inhibitor (ilomastat, GM6001) on learning and memory function and hippocampal myelin-related proteins in epileptic model rats.Methods:Thirty 8-week-old male SPF Sprague-Dawley rats were randomly divided into control group, intervention control group, short-term model group, long-term model group and long-term intervention group, with 6 rats in each group.The lithium-pilocarpine method was used to induce status epilepticus (SE). SE was terminated respectively by intraperitoneal injection of diazepam(10 mg/kg) at 15 min in short-term model group and at 30 min both in the long-term model group and long-term intervention group. Rats in intervention control group and long-term intervention group were treated with GM6001 (20 μg/kg) via tail vein injection. Morris water maze test was used to assess the learning and memory function and the blood-brain barrier permeability was evaluated by Evans blue (EB) staining. The levels of serum MMP, the hippocampal myelin basic protein (MBP) and 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) were measured by ELISA and Western blot.Statistical analysis was performed by SPSS 21.0 software, with repeated measures ANOVA for Morris water maze data and one-way ANOVA with LSD post hoc test for other comparisons.Results:(1) In the water maze test, the interaction between the time and group on the latencies reached to the platform showed no significance among the 5 groups of rats ( F=0.430, P=0.970). The main effects of time and group on latencies were both significant among the 5 groups( F=144.521, 12.819, both P<0.001).From the second day to the fifth day, the latencies reached to the platform of rats in the long-term model group were significantly higher than those in the control group and in the long-term intervention group (both P<0.05). There were statistically significant differences in the latency of the first time reached the platform, the duration in the target quadrant, the frequency of crossing the platform and the distance to the platform among the 5 groups ( F=7.008, 7.306, 15.704, 11.559, all P<0.05). In the long-term intervention group, the latency of the first time reached the platform ((13.71±4.41) s) and the distance to the platform ((60.37±8.59) cm) were significantly lower than those in the long-term model group ((23.52±5.34) s, (73.06±12.01) cm) (both P<0.05), and the duration in the target quadrant ((10.93±2.99) s) and the frequency of crossing the platform (2.00(1.75, 2.75)times) were significantly higher than those of the long-term model group ((6.04±4.33) s, 1.00(0, 1.00)times) (both P<0.05).(2) The hippocampal EB fluorescence intensity and the serum levels of MMP-2 and MMP-9 were significantly different among the 5 groups ( F=15.249, 7.951, 33.299, all P<0.001). The EB fluorescence intensity of the long-term model group ((0.18±0.03) /100 μm 2) was significantly higher than that in control group ((0.09±0.02) /100 μm 2) and the long-term intervention group ((0.12±0.04) /100 μm 2) (both P<0.05). The serum MMP-2 and MMP-9 levels in the long-term model group ((10.91±1.36) ng/mL, (10.96±1.28) ng/mL) were significantly higher than those of the control group ((7.89±1.04) ng/mL, (4.20±1.48) ng/mL) and the long-term intervention group ((6.49±1.12) ng/mL, (8.06±0.91) ng/mL) (all P<0.05). (3) The expression levels of hippocampal MBP and CNPase were significantly different among the 5 groups ( F=8.757, 4.116, both P<0.05). The expression levels of MBP and CNPase in long-term model group ((1.00±0.19), (0.93±0.21)) were significantly lower than those in the control group ((1.57±0.27), (1.36±0.22)) and the long-term intervention group ((1.45±0.19), (1.23±0.26)) (all P<0.05). Conclusion:The MMP inhibitor GM6001 can ameliorate learning and memory deficits in epileptic model rats, potentially by reducing blood-brain barrier permeability and alleviating hippocampal myelin damage.

Bisphenol A(BPA)is an environmental endocrine disruptor commonly found in industri-al products such as plastics,resin coatings,and paper coatings,and it poses potential reproductive hazards.Despite extensive research,studies examining its effects on the F1 generation of rabbits are limited.This study established a BPA exposure model in pregnant female rabbits to investigate its impact on reproductive hormones,apoptosis,oxidative stress,inflammatory responses,and tissue integrity in weaned female offspring.The results indicate that BPA exposure induces oxidative stress and inflammation in F1 rabbits,disrupts hormonal balance,and affects antioxidant enzymes and inflammatory mediators through the modulation of the Nrf2 and NF-κB signaling pathways,ultimately leading to apoptosis and tissue damage.

Objective To establish orthotopic and ectopic pancreatic cancer models in C57BL/6N mice with normal immune function using in vivo imaging technology for visual characterization.Methods Orthotopic and ectopic pancreatic cancer models were established in Kunming mice by injecting a small volume of cell suspension containing firefly luciferase-expressing Panc02-luciferase pancreatic cancer cells into the head of the pancreas or the right axillary region.In vivo imaging technology was used to optimize the modeling method and timing in Kunming mice.Subsequently,the same method was applied to C57BL/6N mice using wild-type Panc02 pancreatic cancer cells to establish orthotopic and ectopic pancreatic cancer models with intact immune function.Key parameters,including body weight,inoculation positive rate,tumor growth time,tumor volume,and pathological characteristics across different organs,were compared be-tween the orthotopic and ectopic models in C57BL/6N mice to evaluate the applicability of these models.Results Both the small animal in vivo imaging experiments in Kunming mice and the tumor growth observation in C57BL/6N mice demonstrated that the construction periods for orthotopic and ectopic pancreatic cancer models were 20 days,with survival rates exceeding 90%.The inoculation positive rates in C57BL/6N mice were 92.3%for the orthotopic model and 78.6%for the ectopic model.On day 20 post-inoculation,the tumor volumes were(117.04±109.56)mm3 for the orthotopic model and(155.68±168.73)mm3 for the ectopic model,indicating high model success rates and consistent tumor growth.HE staining revealed pathological mitotic figures and poorly differentiated tumor tissues in both models of C57BL/6N mice,with no evidence of metastasis to other organs.Conclusions Orthotopic and ectopic pancreatic cancer models in immu-nocompetent mice were successfully developed in this study,mimicking early-stage pancreatic cancer characteristics.These models pro-vide a reliable platform for screening early diagnostic biomarkers and evaluating therapeutic interventions for pancreatic cancer.

Objective To establish orthotopic and ectopic pancreatic cancer models in C57BL/6N mice with normal immune function using in vivo imaging technology for visual characterization.Methods Orthotopic and ectopic pancreatic cancer models were established in Kunming mice by injecting a small volume of cell suspension containing firefly luciferase-expressing Panc02-luciferase pancreatic cancer cells into the head of the pancreas or the right axillary region.In vivo imaging technology was used to optimize the modeling method and timing in Kunming mice.Subsequently,the same method was applied to C57BL/6N mice using wild-type Panc02 pancreatic cancer cells to establish orthotopic and ectopic pancreatic cancer models with intact immune function.Key parameters,including body weight,inoculation positive rate,tumor growth time,tumor volume,and pathological characteristics across different organs,were compared be-tween the orthotopic and ectopic models in C57BL/6N mice to evaluate the applicability of these models.Results Both the small animal in vivo imaging experiments in Kunming mice and the tumor growth observation in C57BL/6N mice demonstrated that the construction periods for orthotopic and ectopic pancreatic cancer models were 20 days,with survival rates exceeding 90%.The inoculation positive rates in C57BL/6N mice were 92.3%for the orthotopic model and 78.6%for the ectopic model.On day 20 post-inoculation,the tumor volumes were(117.04±109.56)mm3 for the orthotopic model and(155.68±168.73)mm3 for the ectopic model,indicating high model success rates and consistent tumor growth.HE staining revealed pathological mitotic figures and poorly differentiated tumor tissues in both models of C57BL/6N mice,with no evidence of metastasis to other organs.Conclusions Orthotopic and ectopic pancreatic cancer models in immu-nocompetent mice were successfully developed in this study,mimicking early-stage pancreatic cancer characteristics.These models pro-vide a reliable platform for screening early diagnostic biomarkers and evaluating therapeutic interventions for pancreatic cancer.

BACKGROUND:The incidence of inflammatory bowel disease has been steadily rising,accompanied by a lack of definitive therapeutic strategies.Recent research endeavors have illuminated the promising potential of mesenchymal stem cells in mitigating the symptoms of inflammatory bowel disease,offering a glimmer of hope for afflicted patients.OBJECTIVE:To review the mechanisms of action of mesenchymal stem cells derived from various sources in the management of inflammatory bowel disease,aiming to provide insights for future research endeavors.METHODS:Utilizing the keywords"mesenchymal stem cells,MSCs,exosomes,extracellular vesicles,EVs,inflammatory bowel disease,IBD,ulcerative colitis,UC,Crohn's disease,CD"in English and their Chinese equivalents in CNKI and PubMed databases,a total of 89 eligible articles were selected for this review.RESULTS AND CONCLUSION:Currently,six types of mesenchymal stem cells are being explored for inflammatory bowel disease therapy:bone marrow-derived mesenchymal stem cells,adipose tissue-derived mesenchymal stem cells,perinatal tissue-derived mesenchymal stem cells,induced pluripotent stem cell-derived mesenchymal stem cells,embryonic stem cell-derived mesenchymal stem cells,and gingival mesenchymal stem cells.Five administration routes have been adopted,with intravenous and intraperitoneal injections being the most prevalent,followed by local,mesenteric,and intra rectal injections.Their therapeutic mechanisms encompass differentiation,regeneration,anti-inflammatory effects,immune modulation,neuroprotection,antioxidant stress response,homing,modulation of gut microbiota,autophagy,ferroptosis,and endoplasmic reticulum stress.While sharing functional similarities,mesenchymal stem cells from different sources exhibit unique characteristics that confer them with distinct advantages and therapeutic potentials.Nevertheless,research into the specific properties of these mesenchymal stem cells remains limited,necessitating deeper exploration of their nuanced differences to optimize their therapeutic efficacy in inflammatory bowel disease.Additionally,the clinical safety of mesenchymal stem cells-based therapies warrants further observation and evaluation.

In order to investigate the mitigating effect of Yin Chen aqueous extract on liver injury induced by LPS combined with D-GalN in mice,a mouse liver injury model was established by in-traperitoneal injection of LPS and D-GalN,and the mice were group-fed by instillation of saline,bi-phenyl dibenzyl ester,and Yin Chen aqueous extract with different concentrations of LPS and D-GalN.The liver index of mice was calculated,pathological tissue sections were observed,and the expression of ALT,AST,inflammatory cytokines,oxidative stress,hepatic enzymes,and IL-17/TNF pathway were detected in serum to investigate the effects and mechanisms of the aqueous ex-tracts of Yin Chen in alleviating the liver injury in mice.The results showed that the liver index of mice in the model group was significantly elevated,the serum levels of ALT and AST were signifi-cantly elevated,the levels of IL-iβ,IL-8 and TNF-α in liver tissue were significantly elevated,the levels of IL-4 were significantly reduced,the levels of GSH-Px,CAT and SOD were significantly reduced,the levels of ROS and MDA were significantly elevated,CYP2E1 and CYP1A2 protein content and mRNA expression were significantly up-regulated,and the expression levels of TNF,TNFR1,IL-17A,ACT1 and IL-6 mRNA were significantly up-regulated.The study showed that the aqueous extract of Yin Chen had a certain alleviating effect on the liver injury caused by LPS combined with D-GalN in mice.The mechanism of action includes decreasing the metabolic level of hepatic drug enzymes,alleviating oxidative stress,inhibiting the expression of IL-17/TNF pathway and down-regulating the level of inflammatory factors in mice.