ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.

ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.

Objective:To investigate the combined application of cytology, cell block histology and immunohistochemistry to improve the diagnostic accuracy of solid pancreatic lesions in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) samples.Methods:The pathological data of EUS-FNA in 311 cases of solid pancreatic lesions submitted to the Second Hospital of Hebei Medical University, Shijiazhuang, China from May 2019 to September 2023 were retrospectively analyzed. The cases included pancreatic ductal adenocarcinoma (PDAC, 172 cases), solid pseudopapillary neoplasm (SPN, 12 cases), neuroendocrine tumors (PNET, 14 cases) and chronic pancreatitis (113 cases). The cytological features of smears, the histology of cell block sections and the diagnostic markers in PDAC, SPN and PNET were analyzed. The diagnostic accuracies of cytology, cell block histology/immunohistochemistry and combination of the two methods for classifying these pancreatic solid lesions were evaluated.Results:Irregular arrangement of atypical (cancer) cells, anisonucleosis and nuclear atypia were the typical cytological features of PDAC, while presence of pseudopapillae with a myxoid/hyalinized fibrovascular core and low adhesion/salt-and-pepper chromatin were diagnostic features of SPN and NET, respectively. Immunohistochemical results showed that CK7 and CK19 were the most sensitive markers of pancreatic ductal epithelia, and the diffuse strong expression of S-100P (102/111, 91.9%) and aberrant expression of p53 (80/111, 72.1%) were important immunophenotypic markers of PDAC. Various degrees of CDX2 expression could be found in 66.4% PDAC. The expression of CD10, PR, vimentin, CD99 and cyclinD1 and the aberrant expression of β-catenin were the immunophenotypic features of SPN, while the expression of CgA, Syn and CD56 were indispensable immunemarkers for the diagnosis of PNET. Overall, cytology had higher sensitivity than cell block histology (93.9% versus 82.8%) and lower specificity (92.9% versus 99.1%), while the combination of the two methods significantly improved the sensitivity to 96.9% in solid pancreatic lesions. The combination of cytology and cell block histology could significantly improve the diagnostic efficacy of EUS-FNA in PDAC.Conclusions:Integrated diagnosis based on cytology (including rapid on-site evaluation), cell block histology and immunohistochemical findings could significantly improve the diagnostic yield of EUS-FNA in classifying solid pancreatic lesions.

Objective To investigate the role and potential mechanism of protein arginine methyltransferase 1(PRMT1)in regulating the proliferation,apoptosis and migration of human oral squamous cell carcinoma SCC-25 cells.Methods Normal oral mucosal cells S-G and oral squamous cell carcinoma cells SCC-25,OECM-1,and HSC-3 were collected and cultured,and the transcription level of PRMT1 was detected.SCC-25 cells were divided into the untransfected group(SCC-25 group),empty vector control group(SCC-25+NC group),and transfection group(SCC-25+si-PRMT1 group).Protein expression levels were detected by Western blot;cell proliferation and migration abilities were assessed by MTT assay and cell scratch wound healing assay,respectively;and the apoptosis rates were determined by flow cytometry.Exogenous TGF-β stimulation experiment was conducted on the basis of constructed SCC-25+si-PRMT1 cells to verify the role of TGF-β/Smad pathway in the PRMT1 silencing effect.Results Compared with normal oral mucosal cells S-G,PRMT1 was highly expressed in oral squamous cell carcinoma cells SCC-25,OECM-1,and HSC-3(P<0.05),with the highest expression observed in SCC-25 cells(P<0.05).After silencing the PRMT1 gene,the expressions of pro-apoptotic proteins Bax and C-Caspase-3 in SCC-25 cells significantly increased,the apoptosis rate significantly increased,the expression of anti-apoptotic protein Bcl-2 significantly decreased,and the cell proliferation and migration abilities were inhibited.Simultaneous silencing of the PRMT1 gene can significantly downregulated the expression levels of TGF-β and its downstream effector p-Smad.Compared with the SCC-25+si-PRMT1 group,the cell proliferation and migration abilities in the SCC-25+si-PRMT1+TGF-β group were enhanced,and the apoptosis rate decreased,which suggesting that TGF-β could partially reverse the tumor suppressor effect caused by PRMT1 silencing.Conclusion PRMT1 regulates the proliferation,apoptosis and migration of SCC-25 cells through TGF-β/Smad signaling pathway.

Objective:To investigate the combined application of cytology, cell block histology and immunohistochemistry to improve the diagnostic accuracy of solid pancreatic lesions in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) samples.Methods:The pathological data of EUS-FNA in 311 cases of solid pancreatic lesions submitted to the Second Hospital of Hebei Medical University, Shijiazhuang, China from May 2019 to September 2023 were retrospectively analyzed. The cases included pancreatic ductal adenocarcinoma (PDAC, 172 cases), solid pseudopapillary neoplasm (SPN, 12 cases), neuroendocrine tumors (PNET, 14 cases) and chronic pancreatitis (113 cases). The cytological features of smears, the histology of cell block sections and the diagnostic markers in PDAC, SPN and PNET were analyzed. The diagnostic accuracies of cytology, cell block histology/immunohistochemistry and combination of the two methods for classifying these pancreatic solid lesions were evaluated.Results:Irregular arrangement of atypical (cancer) cells, anisonucleosis and nuclear atypia were the typical cytological features of PDAC, while presence of pseudopapillae with a myxoid/hyalinized fibrovascular core and low adhesion/salt-and-pepper chromatin were diagnostic features of SPN and NET, respectively. Immunohistochemical results showed that CK7 and CK19 were the most sensitive markers of pancreatic ductal epithelia, and the diffuse strong expression of S-100P (102/111, 91.9%) and aberrant expression of p53 (80/111, 72.1%) were important immunophenotypic markers of PDAC. Various degrees of CDX2 expression could be found in 66.4% PDAC. The expression of CD10, PR, vimentin, CD99 and cyclinD1 and the aberrant expression of β-catenin were the immunophenotypic features of SPN, while the expression of CgA, Syn and CD56 were indispensable immunemarkers for the diagnosis of PNET. Overall, cytology had higher sensitivity than cell block histology (93.9% versus 82.8%) and lower specificity (92.9% versus 99.1%), while the combination of the two methods significantly improved the sensitivity to 96.9% in solid pancreatic lesions. The combination of cytology and cell block histology could significantly improve the diagnostic efficacy of EUS-FNA in PDAC.Conclusions:Integrated diagnosis based on cytology (including rapid on-site evaluation), cell block histology and immunohistochemical findings could significantly improve the diagnostic yield of EUS-FNA in classifying solid pancreatic lesions.

Objective To investigate the role and potential mechanism of protein arginine methyltransferase 1(PRMT1)in regulating the proliferation,apoptosis and migration of human oral squamous cell carcinoma SCC-25 cells.Methods Normal oral mucosal cells S-G and oral squamous cell carcinoma cells SCC-25,OECM-1,and HSC-3 were collected and cultured,and the transcription level of PRMT1 was detected.SCC-25 cells were divided into the untransfected group(SCC-25 group),empty vector control group(SCC-25+NC group),and transfection group(SCC-25+si-PRMT1 group).Protein expression levels were detected by Western blot;cell proliferation and migration abilities were assessed by MTT assay and cell scratch wound healing assay,respectively;and the apoptosis rates were determined by flow cytometry.Exogenous TGF-β stimulation experiment was conducted on the basis of constructed SCC-25+si-PRMT1 cells to verify the role of TGF-β/Smad pathway in the PRMT1 silencing effect.Results Compared with normal oral mucosal cells S-G,PRMT1 was highly expressed in oral squamous cell carcinoma cells SCC-25,OECM-1,and HSC-3(P<0.05),with the highest expression observed in SCC-25 cells(P<0.05).After silencing the PRMT1 gene,the expressions of pro-apoptotic proteins Bax and C-Caspase-3 in SCC-25 cells significantly increased,the apoptosis rate significantly increased,the expression of anti-apoptotic protein Bcl-2 significantly decreased,and the cell proliferation and migration abilities were inhibited.Simultaneous silencing of the PRMT1 gene can significantly downregulated the expression levels of TGF-β and its downstream effector p-Smad.Compared with the SCC-25+si-PRMT1 group,the cell proliferation and migration abilities in the SCC-25+si-PRMT1+TGF-β group were enhanced,and the apoptosis rate decreased,which suggesting that TGF-β could partially reverse the tumor suppressor effect caused by PRMT1 silencing.Conclusion PRMT1 regulates the proliferation,apoptosis and migration of SCC-25 cells through TGF-β/Smad signaling pathway.

Salvia miltiorrhiza is a perennial herb of the genus Salvia(Lamiaceae). As one of the earliest medicinal plants to undergo molecular biology research, it has gradually become a model plant for molecular biology of medicinal plants. With the gradual analysis of the genome of S. miltiorrhiza and the biosynthetic pathways of its main active components tanshinone and salvianolic acids, the transcriptional regulation mediated by transcription factors and related regulatory proteins has gradually become a new research focus. Due to the lack of scientific and unified naming of transcription factors and different research indexes in different literature, this paper systematically sorted out the transcription factors in different literature with the genomes of DSS3 from selfing for three generations and bh2-7 from selfing for six generations as reference. In total, 73 transcription factors and related regulatory proteins belonging to 13 gene families were identified. The effects of overexpression or gene silencing experiments on tanshinone and salvianolic acids were also analyzed. This study unified the identified transcription factors, which laid a foundation for further constructing the regulatory networks of secondary metabolites and insect or stress resistance and improving the quality of medicinal materials by using global transcriptional regulation engineering.
Salvia miltiorrhiza/chemistry* ; Plant Proteins/metabolism* ; Gene Expression Regulation, Plant ; Transcription Factors/metabolism* ; Abietanes/metabolism*

OBJECTIVE: To investigate the accuracy and safety of pedicle screw placement using 3D-printed auxiliary guides in scoliosis correction surgery for adolescents. METHODS: A retrospective analysis was conducted on the clinical data of 51 patients who underwent posterior scoliosis correction surgery from January 2020 to March 2023. Among them, there were 35 cases of adolescent idiopathic scoliosis and 16 cases of congenital scoliosis. The patients were divided into two groups based on the auxiliary tool used:the 3D-printed auxiliary guide screw placement group (3D printing group) and the free-hand screw placement group (free-hand group, without auxiliary tools). The 3D printing group included 32 patients (12 males and 20 females) with an average age of (12.59±2.60) years;the free-hand group included 19 patients (7 males and 12 females) with an average age of (14.58±3.53) years. The two groups were compared in terms of screw placement accuracy and safety, spinal correction rate, intraoperative blood loss, number of intraoperative fluoroscopies, operation time, hospital stay, and preoperative and last follow-up scores of the Scoliosis Research Society-22 (SRS-22) questionnaire. RESULTS: A total of 707 pedicle screws were placed in the two groups, with 441 screws in the 3D printing group and 266 screws in the free-hand group. All patients in both groups successfully completed the surgery. There was a statistically significant difference in operation time between the two groups (P<0.05). The screw placement accuracy rate of the 3D printing group was 95.46% (421/441), among which the Grade A placement rate was 89.34% (394/441);the screw placement accuracy rate of the free-hand group was 86.47% (230/266), with a Grade A placement rate of 73.31% (195/266). There were statistically significant differences in the accuracy of Grade A, B, and C screw placements between the two groups (P<0.05), while no statistically significant differences were observed in intraoperative blood loss, number of fluoroscopies, correction rate, or hospital stay (P>0.05). In the SRS-22 questionnaire scores, the scores of functional status and activity ability, self-image, mental status, and pain of patients in each group at the last follow-up were significantly improved compared with those before surgery (P<0.05), but there were no statistically significant differences in all scores between the two groups (P>0.05). CONCLUSION In scoliosis correction surgery, compared with traditional free-hand screw placement, the use of 3D-printed auxiliary guides for screw placement significantly improves the accuracy and safety of screw placement and shortens the operation time.
Humans ; Male ; Scoliosis/surgery* ; Female ; Adolescent ; Printing, Three-Dimensional ; Retrospective Studies ; Pedicle Screws ; Child

This study was aimed at understanding the etiological and genetic characteristics of Yersinia enterocolica isolated in Jiangsu Province between 2005 and 2019.All 110 identified strains of Y.enterocolica were from patients with foodborne diar-rhea in Jiangsu Province,or from pigs,dogs,cattle,sheep,poultry,flies,or food.Virulence genes,biological serotypes,drug resistance,multilocus sequence typing(MLST),and core genome multilocus sequence typing(cgMLST)based on whole-genome sequencing were performed.The strains included 27 pathogenic strains(24.5％)and 83 non-pathogenic strains(75.5％).Non-pathogenic strains accounted for a high proportion,particularly among strains from patients(15/16,93.8％).The biological serotypes of pathogenic strains were mainly type 3/O∶3(26/27,96.3％).Non-pathogenic strains included 1A/O∶8 type(23/83,27.7％),1 A/O∶5 type(14/83,16.9％),and the other four biological serotypes(excluding unknown se-rotypes).Pathogenic strains were dominated by type 3/O∶3(26/27,96.3％),and more than 80％of these strains were sensi-tive to 19 antibiotic types.Whole-genome sequencing indicated that the pathogenic strains were all ST135 type,whereas the non-pathogenic strains were more diverse and scattered.HierCC clustering analysis grouped all strains into three clus-ters:pathogenic strains were in one cluster,and strains from patients were found in all three clusters.In conclusion,the Y.enterocolica strains from patients were primarily non-patho-genic.Non-pathogenic strains showed richer epigenetic and ge-netic diversity than pathogenic strains.The monitoring of these strains should be strengthened to decrease the risk of human infection.

This study was aimed at understanding the etiological and genetic characteristics of Yersinia enterocolica isolated in Jiangsu Province between 2005 and 2019.All 110 identified strains of Y.enterocolica were from patients with foodborne diar-rhea in Jiangsu Province,or from pigs,dogs,cattle,sheep,poultry,flies,or food.Virulence genes,biological serotypes,drug resistance,multilocus sequence typing(MLST),and core genome multilocus sequence typing(cgMLST)based on whole-genome sequencing were performed.The strains included 27 pathogenic strains(24.5％)and 83 non-pathogenic strains(75.5％).Non-pathogenic strains accounted for a high proportion,particularly among strains from patients(15/16,93.8％).The biological serotypes of pathogenic strains were mainly type 3/O∶3(26/27,96.3％).Non-pathogenic strains included 1A/O∶8 type(23/83,27.7％),1 A/O∶5 type(14/83,16.9％),and the other four biological serotypes(excluding unknown se-rotypes).Pathogenic strains were dominated by type 3/O∶3(26/27,96.3％),and more than 80％of these strains were sensi-tive to 19 antibiotic types.Whole-genome sequencing indicated that the pathogenic strains were all ST135 type,whereas the non-pathogenic strains were more diverse and scattered.HierCC clustering analysis grouped all strains into three clus-ters:pathogenic strains were in one cluster,and strains from patients were found in all three clusters.In conclusion,the Y.enterocolica strains from patients were primarily non-patho-genic.Non-pathogenic strains showed richer epigenetic and ge-netic diversity than pathogenic strains.The monitoring of these strains should be strengthened to decrease the risk of human infection.