ObjectiveTo explore the mechanism by which Tangbikang granules improve diabetic peripheral neuropathy based on ferroptosis mediated by the adenosine monophosphate-activated protein kinase/nuclear factor erythroid 2-related factor 2 （AMPK/Nrf2） signaling pathway. MethodsA diabetes model was established using spontaneous male Zucker diabetic fatty （ZDF） rats. After successful modeling， the rats were divided into a normal group， a model group， high-， medium-， and low-dose Tangbikang granules groups， and a metformin hydrochloride group. The high-， medium-， and low-dose Tangbikang granules groups were administered by gavage at doses of 2.5， 1.25， 0.625 g·kg^-1， respectively. The metformin hydrochloride group received 0.135 g·kg^-1 by gavage， while the remaining groups received an equal volume of deionized water. Administration continued for 12 weeks. Blood glucose levels were measured after administration， and at 4， 8， 12 weeks. Following the 12-week intervention， the thermal pain threshold and the sciatic nerve conduction velocity （SNCV） were measured. The levels of malondialdehyde （MDA）， superoxide dismutase （SOD）， and adenosine triphosphate （ATP） in the sciatic nerve were measured using enzyme-linked immunosorbent assay （ELISA）. Morphological changes in the sciatic nerve were observed using hematoxylin and eosin （HE） staining， and the ultrastructural changes were examined using transmission electron microscopy. The levels of glutathione peroxidase 4 （GPx4） were detected using immunofluorescence （IF） assay. The protein expression levels of p-AMPK， Nrf2， GPx4， and acyl-CoA synthetase long-chain family member 4 （ACSL4） were detected using Western blot. ResultsCompared with the normal group， the model group had significantly higher blood glucose levels after administration and at weeks 4， 8 and 12 （P<0.01）. The thermal pain threshold was significantly prolonged （P<0.01）， and the SNCV was significantly slowed down （P<0.01）. The SOD and ATP levels significantly decreased （P<0.01）， while the MDA levels significantly increased （P<0.01）. Pathologically， the sciatic nerve fibers in the model group showed a dispersed structure， disordered and sparse arrangement， axonal atrophy， irregular myelin sheath halo， increased and swollen Schwann cell nuclei， obvious endoneurial fibrosis， and collagen hyperplasia. Immunofluorescence assay revealed fragmented red fluorescence and significantly reduced expression of GPx4 （P<0.01）. Western blot analysis showed significantly decreased protein expression levels of p-AMPK， Nrf2， and GPx4 （P<0.01）， and significantly increased expression of ACSL4 （P<0.01） in the model group. Compared with the model group， fasting blood glucose level decreased significantly in the high-dose Tangbikang granules group at weeks 4 and 12 （P<0.05）. The thermal pain threshold was significantly shortened in the high- and medium-dose Tangbikang granules groups （P<0.01）. The SNCV was significantly accelerated in the high- and medium-dose Tangbikang granules groups （P<0.01）. The SOD levels were significantly elevated in the high-dose Tangbikang granules group （P<0.01）. The MDA levels significantly decreased in all Tangbikang granules groups （P<0.01）. Both the metformin hydrochloride group and the high-dose Tangbikang granules group exhibited relatively orderly and densely arranged sciatic nerve fibers with more regular myelin sheath halos. The GPx4 expression significantly increased in both the metformin hydrochloride group and all Tangbikang granules groups （P<0.01）. The protein expression levels of p-AMPK， Nrf2， and GPx4 were significantly increased （P<0.01）， while ACSL4 protein expression significantly decreased （P<0.01）. ConclusionTangbikang granules may improve peripheral neuropathy by suppressing ferroptosis through the regulation of the AMPK/Nrf2 signaling pathway.

Objective:To explore the doctors′ perceived value driving of internet medical services, and provide references for promoting the sustainable development of internet medical services.Methods:From August to December 2023, purposive sampling and typical sampling methods were used to select 23 doctors from 8 tertiary hospitals in a certain province as interviewees for semi-structured interviews. Based on the grounded theory, the interview data were encoded in three levels, and the value motivation model of internet doctors was constructed.Results:Through three-level coding, 68 formal concepts, 17 categories, and 8 main categories. The value motivation model of internet doctors showed that functional value, emotional value and performance value constitute service driven value, and efficiency value and resource value constitute connection driven value, which were the internal driving factors of the doctors′ perceived value; Personal structure factors and platform construction were the external driving factors of the doctors′ perceived value.Conclusions:The doctors′ perceived value driving in internet medical services was the " service-connection" dual drive type. The internal driving factors determined the degree of doctors′ perceived value, while the external driving factors affectted the strength and direction of doctors′ perceived value of internet medical services. It was recommended that relevant departments should promote the release of service driven value through online and offline medical complementarity and the increase of service net utility. Multiple technology embeddings and long tail demand response should be used to create connection driven value. The intensity of value creation should be increased through the two-way adjustment of individual structural factors and the competitive drive of platform construction.

Objective:To explore the doctors′ perceived value driving of internet medical services, and provide references for promoting the sustainable development of internet medical services.Methods:From August to December 2023, purposive sampling and typical sampling methods were used to select 23 doctors from 8 tertiary hospitals in a certain province as interviewees for semi-structured interviews. Based on the grounded theory, the interview data were encoded in three levels, and the value motivation model of internet doctors was constructed.Results:Through three-level coding, 68 formal concepts, 17 categories, and 8 main categories. The value motivation model of internet doctors showed that functional value, emotional value and performance value constitute service driven value, and efficiency value and resource value constitute connection driven value, which were the internal driving factors of the doctors′ perceived value; Personal structure factors and platform construction were the external driving factors of the doctors′ perceived value.Conclusions:The doctors′ perceived value driving in internet medical services was the " service-connection" dual drive type. The internal driving factors determined the degree of doctors′ perceived value, while the external driving factors affectted the strength and direction of doctors′ perceived value of internet medical services. It was recommended that relevant departments should promote the release of service driven value through online and offline medical complementarity and the increase of service net utility. Multiple technology embeddings and long tail demand response should be used to create connection driven value. The intensity of value creation should be increased through the two-way adjustment of individual structural factors and the competitive drive of platform construction.

Background and purpose:Colorectal cancer(CRC)is one of the major malignant tumors threatening human health worldwide,with long-term high incidence and mortality rate.Potassium channel modulatory factor 1(KCMF1)is a member of the E3 ubiquitin ligase family.It binds to target proteins through the RING domain and participates in the regulation of a variety of biological processes in vivo.However,the function of KCMF1 in CRC remains unclear.This study aimed to investigate the expression level of E3 ubiquitin ligase KCMF1 in colorectal tumor,and to explore the effects of KCMF1 on the proliferation of CRC cells and its underlying molecular mechanism.Methods:The The Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)databases were used to analyze the expression level of KCMF1 in CRC tissues and adjacent tissues and the association between the KCMF1 expression and the prognosis of CRC patients.Furthermore,immunohistochemical staining was performed to detect the protein level of KCMF1 in 90 paired human CRC tissues and adjacent non-tumor tissues.Lentiviral shRNA delivery system was employed to specifically target the KCMF1 gene(shKCMF1)in HCT116 and HCT15 CRC cell lines.The effects of KCMF1 knockdown on cell proliferation,apoptosis and cell cycle distribution were assessed by methyl thiazoyl terazolium(MTT)assay,colony formation assay,Western blot and flow cytometry.Changes in the transcriptional profile in HCT116 cells upon KCMF1 knockdown were identified by RNA sequencing(RNA-Seq),and the affected signaling pathways were evaluated by bioinformatics analysis.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR),Western blot,luciferase reporter assay and cell immunofluorescence assay were utilized to validate the alteration of the affected signaling pathway.Results:The TCGA and GTEx databases and IHC results showed that the mRNA and protein expression levels of KCMF1 in CRC tissues were significantly upregulated compared with adjacent tissues(P＜0.01).KCMF1 expression level was negatively correlated with the survival time of patients with CRC(P＜0.01),and was positively associated with CRC clinical stage(P＜0.05).Compared with control cells,KCMF1 knockdown significantly inhibited the proliferation of HCT116 and HCT15 cells(P＜0.001),induced cell apoptosis(P＜0.001),and led to cell cycle arrest in G1 phase(P＜0.01).RNA-Seq analysis showed that KCMF1 was involved in the regulation of several signaling pathways,including nuclear factor-κB(NF-κB)signaling pathway.KCMF1 knockdown reduced the transcription levels of the target genes of NF-κB signaling pathway,including BCL-XL,XIAP and CIAP(P＜0.05),and suppressed the expression of phosphorylated p65 and nuclear translocation of p65(P＜0.01).Meanwhile,the activity of NF-κB reporter was reduced in tumor cells upon KCMF1 knockdown(P＜0.01).Conclusion:The expression of KCMF1 is significantly upregulated in human CRC tissues and positively associated with advanced clinical stage and poor prognosis.KCMF1 may promote the proliferation of CRC cells by activating the NF-κB signaling pathway.KCMF1 may be a potential new therapeutic target for CRC.

Objective To observe the changes of liver function,blood cell,and lung function of healthy young and middle-aged men before and after fast-advancing short-term high altitude exposure(FSHAE);and to explore the effects and possible mechanisms of FSHAE on the function of liver,blood cells,and lung tissues.Methods This study included 48 healthy young and middle-aged male volunteers,who collected physiological indicators,tested liver function indicators,blood cell indicators,and lung function-related indicators 1 day before entering the plateau(100m above sea level),and 15 days after FSHAE(3000m above sea level).Differences in the relevant parameters of each system were compared before and after FSHAE.Results Compared with those before entering the plateau,the physiological parameters of young and middle-aged men after 15 days of FSHAE heart rate increased significantly,respiratory rate increased,systolic blood pressure increased,mean arterial blood pressure increased,oxygen saturation decreased(P＜0.01),and diastolic blood pressure increased(P＜0.05),all of which were statistically significant;and the indicators of liver function:glutamic oxaloacetic aminotransferase,glutamic alanine aminotransferase increased(P＜0.01),glutamylamine aminotransferase,glutamate aminotransferase,glutaminase,and pulmonary function were increased(P＜0.01),glutamyl transpeptidase,alkaline phosphatase,and total bile acids were elevated,and total protein decreased(P＜0.05),and the differences were statistically significant.Hemocyte-related indexes:erythrocyte count,erythrocyte pressure volume,mean erythrocyte volume,mean hemoglobin volume,mean hemoglobin concentration,and hemoglobin were elevated,and platelet count decreased(P＜0.01),and the differences were statistically significant.although there was an elevation of leukocyte count(P＞0.05);Lung function-related indexes:decreased exertion lung volume(P＜0.05).There were decreased exertion expiratory volume in the first second,increased one-second rate(P＞0.05).Conclusion FSHAE can lead to oxidative stress in the organism,and acute hypoxic multisystemic injury will occur,with the simultaneous emergence of hypoxic adaptive regulation of various systems,self-compensatory repair of various organs of the organism,and there may be the possibility of interactions between various systems.

Objective:To investigate the value of three-dimensional (3D) T 1WI structural images using different acceleration methods including parallel acquisition technique, joint compressed sensing (uCS) technique, and artificial intelligence-assisted compressed sensing (ACS) technique for brain region segmentation. Methods:In this cross-sectional study, fifty patients (female: n=25, age range: 13 to 87 years old) at Corning Hospital of Ningbo University from July to September 2023 were prospectively and consecutively collected. All the subjects underwent brain MRI. Six groups of 3D T 1WI structural images were obtained using different acceleration technique and parameters, including 3D T 1WI without acceleration factor (3D-T 1WI group), 3D T 1WI with parallel acquisition technique with acceleration factor 3 (3D-T 1WI-PI-3 group), 3D T 1WI with uCS technique with acceleration factor 4.5 and 6.9 (3D-T 1WI-uCS-4.5 group, 3D-T 1WI-uCS-6.9 group), 3D T 1WI by ACS technique with acceleration factors of 3 and 5 (3D-T 1WI-ACS-3 group, 3D-T 1WI-ACS-5 group). T 2WI fluid-attenuated inversion recovery (FLAIR) images were also acquired. Subjective scores (cerebral grey matter and white matter clarity scores, clarity scores of cerebral white matter degeneration lesions in relation to the surrounding white matter, and Gibbs artifact scores) and objective metrics [signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), cerebrospinal fluid signal homogeneity, peak signal-to-noise ratio (PSNR), structural similarity index (SSIM), and natural image quality evaluator (NIQE)] were used to evaluate image quality in different groups. Totally 109 brain regions were segmented and volumes were measured using the uAI Research Portal image analysis tool. Kappa or intraclass correlation coefficient ( ICC) was used to evaluate the agreement of subjective and objective evaluation indexes between the 3D-T 1WI-PI-3 group, 3D-T 1WI-uCS-4.5 group, 3D-T 1WI-uCS-6.9 group, 3D-T 1WI-ACS-3 group, 3D-T 1WI-ACS-5 group, and 3D-T 1WI group. Kappa or ICC value>0.70 was considered as good agreement. Results:The acquisition time for the 3D-T 1WI group, 3D-T 1WI-PI-3 group, 3D-T 1WI-uCS-4.5 group, 3D-T 1WI-uCS-6.9 group, 3D-T 1WI-ACS-3 group, and 3D-T 1WI-ACS-5 group were 527, 204, 169, 95, 133, 90 s, respectively. Subjective evaluation showed that the 3D-T 1WI-uCS-4.5, 3D-T 1WI-ACS-3, and 3D-T 1WI-ACS-5 groups had excellent agreement with the 3D-T 1WI group in terms of the distribution of cases of cerebral grey matter and white matter clarity scores, respectively (all Kappa value=1.000); The distribution of cases of clarity score of cerebral white matter lesions and surrounding white matter in the 3D-T 1WI-PI-3 group, 3D-T 1WI-uCS-4.5 group, and 3D-T 1WI-ACS-3 group were in good agreement with that of the 3D-T 1WI group ( Kappa values of 0.775, 0.701, and 0.777, respectively); the distribution of the number of cases of the Gibbs artifact score of the 3D-T 1WI-uCS-4.5, 3D-T 1WI-ACS-3, and 3D-T 1WI-ACS-5 groups was in good agreement with the 3D-T 1WI group (all Kappa value=1.000). Objective evaluation showed the CNR of the images in the 3D-T 1WI-PI-3, 3D-T 1WI-uCS-4.5, and 3D-T 1WI-uCS-6.9 groups were in good agreement with those of the 3D-T 1WI group ( ICC of 0.720, 0.759, and 0.752, respectively); PSNR and SSIM were in good agreement among the 3D-T 1WI-PI-3 group, 3D-T 1WI-uCS-4.5 group, 3D-T 1WI-uCS-6.9 group, 3D-T 1WI-ACS-3 group, and 3D-T 1WI-ACS-5 group (PSNR: ICC=0.854; SSIM: ICC=0.851). NIQE of 3D-T 1WI-PI-3 group, 3D-T 1WI-uCS-4.5 group, and 3D-T 1WI-ACS-3 group images were in good agreement with the 3D-T 1WI group ( ICC value of 0.866, 0.727, 0.753, respectively). The ICC values of the volume of each segmented brain region among the 3D-T 1WI-PI-3, 3D-T 1WI-uCS-4.5, 3D-T 1WI-uCS-6.9, 3D-T 1WI-ACS-3, 3D-T 1WI-ACS-5 group and the 3D-T 1WI group images showed decreased in order (all ICC≥0.62). Conclusions:The uCS and ACS techniques used in 3D-T 1WI show high agreement with 3D-T 1WI in terms of brain segmentation. The application of these accelerating techniques can significantly shorten the acquisition time with obtaining images with good image quality, displaying great value.

Background::Long non-coding RNAs (lncRNAs) plays an important role in the progression of gastric cancer (GC). Their involvement ranges from genetic regulation to cancer progression. However, the mechanistic roles of RP11-789C1.1 in GC are not fully understood.Methods::We identified the expression of lncRNA RP11-789C1.1 in GC tissues and cell lines by real-time fluorescent quantitative polymerase chain reaction. A series of functional experiments revealed the effect of RP11-789C1.1 on the proliferation of GC cells. In vivo experiments verified the effect of RP11-789C1.1 on the biological behavior of a GC cell line. RNA pull-down unveiled RP11-789C1.1 interacting proteins. Western blot analysis indicated the downstream pathway changes of RP11-789C1.1, and an oxaliplatin dosing experiment disclosed the influence of RP11-789C1.1 on the drug sensitivity of oxaliplatin. Results::Our results demonstrated that RP11-789C1.1 inhibited the proliferation of GC cells and promoted the apoptosis of GC cells. Mechanistically, RP11-789C1.1 inhibited checkpoint kinase 1 (CHK1) phosphorylation by binding ataxiatelangiectasia mutated and Rad3 related (ATR), a serine/threonine-specific protein kinase, promoted GC apoptosis, and mediated oxaliplatin sensitivity.Conclusion::In general, we discovered a tumor suppressor molecule RP11-789C1.1 and confirmed its mechanism of action, providing a theoretical basis for targeted GC therapy.

Objective:To explore the standardization of totally implantable venous power port of nursing process in CT enhancement and application effect of contrast medium injection, so as to provide a safer and more efficient way for contrast medium injection in CT enhanced examination for patients with malignant tumors.Methods:A non-randomized prospective study was conducted, 358 patients with malignant tumors were selected in Sun Yat-sen Memorial Hospital, Sun Yat-sen University who underwent CT enhanced examination from August 1, 2022 to July 31, 2023, 179 patients who had been implanted totally implantable venous power port were selected as the experimental group, and the standardized nursing procedure was given. The other 179 patients were the control group, using radiology routine high-pressure intravenous indwelling needle as the contrast medium access, with routine peripheral venous nursing process. The incidence of contrast medium extravasation during CT enhanced examination was observed and compared between the two groups.Results:All the patients were included. There were 85 males and 94 females, aged (55.50±11.72) years old in the control group. There were 83 males and 96 females, aged (54.50±12.24) years old in the experimental group. The incidence of contrast medium extravasation was 0 in the experimental group and 3.35%(6/179) in the control group. The difference between the two groups was statistically significant (Fisher exact probability, P<0.05). Conclusions:The application of standardized nursing procedure of totally implantable venous power port to the injection of contrast medium in CT enhanced examination of malignant tumor patients, can significantly reduce the incidence of contrast medium extravasation.

Serine/arginine-rich splicing factor 7 (SRSF7), a known splicing factor, has been revealed to play oncogenic roles in multiple cancers. However, the mechanisms underlying its oncogenic roles have not been well addressed. Here, based on N⁶-methyladenosine (m⁶A) co-methylation network analysis across diverse cell lines, we find that the gene expression of SRSF7 is positively correlated with glioblastoma (GBM) cell-specific m⁶A methylation. We then indicate that SRSF7 is a novel m⁶A regulator, which specifically facilitates the m⁶A methylation near its binding sites on the mRNAs involved in cell proliferation and migration, through recruiting the methyltransferase complex. Moreover, SRSF7 promotes the proliferation and migration of GBM cells largely dependent on the presence of the m⁶A methyltransferase. The two m⁶A sites on the mRNA for PDZ-binding kinase (PBK) are regulated by SRSF7 and partially mediate the effects of SRSF7 in GBM cells through recognition by insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2). Together, our discovery reveals a novel role of SRSF7 in regulating m⁶A and validates the presence and functional importance of temporal- and spatial-specific regulation of m⁶A mediated by RNA-binding proteins (RBPs).
Humans ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Glioblastoma/genetics* ; Methyltransferases/metabolism* ; RNA Splicing Factors/metabolism* ; RNA, Messenger/genetics* ; RNA-Binding Proteins/metabolism* ; Serine-Arginine Splicing Factors/metabolism* ; RNA Methylation/genetics*

Objective:To investigate the effects of fasudil hydrochloride(FH) on Rho-associated kinase 2(ROCK2) protein and ferroptosis in hippocampal area during early brain injury in rats with subarachnoid hemorrhage(SAH).Methods:Total 36 SPF grade Sprague-Dawley rats were divided into 3 groups by random number table method: Sham group, SAH group and SAH+ FH (a ROCK2 protein inhibitor) group (FH goup) with 12 rats in each group.SAH animal model was established by internal carotid artery perforation.The rats in FH group were injected intraperitoneally with FH(15 mg/kg) 30 minutes after successful modeling, and rats in Sham group and SAH group were injected intraperitoneally with the same volume of 0.9% sodium chloride solution.Twenty-four hours after the intervention, shuttle box test was used to observe the learning and memory ability of rats.The Fe 2+ content in rat hippocampus tissue was detected by colorimetry, and the protein levels of ROCK2 and ferroptosis-related long-chain acyl-CoA synthetase 4(ACSL4) and glutathione peroxidase 4(GPX4) in hippocampus were detected by immunohistochemistry and Western blot.Statistical analysis was performed by SPSS 20.0 software.One-way ANOVA was used for multigroup comparison, and LSD test was used for further pairwise comparison. Results:(1)In the shuttle box test, there were statistically significant differences in the number of avoidance reactions and avoidance reaction time of rats among the three groups( F=20.348, 22.316, both P<0.05). The number of avoidance reaction in SAH group was less than that in Sham group ((17.92±2.94) times, (27.13±3.48) times, P<0.05), the time of avoidance reaction in SAH group was longer than that in Sham group ((9.15±2.87) s, (3.68±1.09) s, P<0.05), while the number of avoidance reaction in FH group ((21.63±4.11) times) was more than that in SAH group, and the time of avoidance reaction ((6.08±1.76) s) was shorter than that in SAH group (both P<0.05). (2) The colorimetry results showed that there was a statistically significant difference in the content of Fe 2+ in hippocampus of rats among the three groups( F=7.965, P<0.05). The Fe 2+ content in SAH group was significantly higher than that of Sham group((0.091±0.032) nmol/mg, (0.038±0.024) nmol/mg, P<0.05), and the Fe 2+ content in the FH group ((0.065±0.021) nmol/mg) was lower than that of SAH group ( P<0.05). (3) There were significant differences in the number of ROCK2, ACSL4 and GPX4 positive cells in hippocampus of rats among the three groups in immunohistochemistry ( F=7.602, 14.171, 36.077, all P<0.05). The positive cells of ROCK2 and ACSL4 in SAH group ((21.63±4.72), (55.13±19.41)) were significantly higher than those of Sham group ((11.63±3.62), (23.38±3.74)) (both P<0.05), and the positive cells of ROCK2 and ACSL4 in FH group ((15.88±6.64), (44.75±8.29)) were both lower than those of SAH group(both P<0.05), while the number of GPX4 positive cells in SAH group (25.38±6.30) was significantly lower than that of Sham group (60.25±10.36) ( P<0.05), and the number of GPX4 positive cells in FH group (45.13±7.51) was higher than that of SAH group( P<0.05). (4)The results of Western blot showed that there were significant differences in the expression levels of ROCK2, ACSL4 and GPX4 proteins in the hippocampus of rats among the three groups( F=4.812, 12.573, 10.849, all P<0.05). The protein expression levels of ROCK2 and ACSL4 in SAH group were significantly higher than those in Sham group(both P<0.05), and the protein expression levels of ROCK2 and ACSL4 in FH group were lower than those in SAH group (both P<0.05), while the expression level of GPX4 protein in SAH group (0.27±0.09) was significantly lower than that in Sham group( P<0.05), and the expression level of GPX4 protein in FH group was higher than that of SAH group ( P<0.05). Conclusion:FH can inhibit ferroptosis in the hippocampus and improve the learning and memory ability of rats, and the mechanism may be related with down-regulation of ROCK2 protein.