Objective To study the current research and future focus of transcutaneous electrical nerve stimulation(TENS)for pain treatment.Methods We retrieved the literatures on TENS for pain treatment from Jan.1,2013 to Dec.31,2022 from CNKI and Web of Science,and Bibliometrix package in R language and the VOSviewer software in Java language were used to analyze the literatures,identifying the current research and future focus on TENS for pain treatment.Results A total of 143 articles in Chinese and 769 articles in English were included in this study,and both showed an increasing trend from 2013 to 2022;the highest citation for Chinese article was 130 and the highest for English article was 235.Total frequency analysis of keywords of English literatures showed that effectiveness and mechanism research were the research focus.The frequency analysis of keywords over years showed that brain-stem,prevention,and shoulder pain were the most frequent keywords in 2021.The strategic coordinate map results showed 2 major future development directions:(1)postoperative pain,analgesia,and opioid receptors,and(2)neuropathic pain,spinal cord,and activation.Total frequency analysis of keywords of Chinese literatures showed that transcutaneous nerve electrical stimulation,pain,analgesia,labor analgesia,and transcutaneous acupoint electrical stimulation were the research focuses.Conclusion TENS therapy for pain is gradually gaining popularity,with a focus on the clinical effectiveness and mechanisms of various types of pain.Future research may focus on further need to anisms study of opioid receptors and spinal cord.

Cysteine protease inhibitor SN(Cystatin SN , CST1), a member of the type 2 cystatins superfamily, regulates tumor biological activities by modulating cysteine protease activity. CST1 is abnormally expressed in digestive system neoplasms, such as esophageal cancer, gastric cancer, hepatocellular carcinoma, colorectal cancer, and pancreatic cancer. CST1 influences tumor cell proliferation, migration, and epithelial-mesenchymal transition (EMT) through signaling pathways including Wnt, STAT3, and PI3K-AKT. Its expression levels significantly correlated with tumor stage, metastasis, and patient prognosis, suggesting its potiential as a novel biomarker for early diagnosis, therapeutic evaluation, and prognosis in digestive system neoplasms. This article systematically reviews the molecular characteristics of CST1, its mechanisms of action in digestive system neoplasms, clinical significance, and explores its potential as a therapeutic target.

Objective To explore the value of multiparameter MRI for follow-up observation on changes of spinal cord microstructures in patients with hereditary spastic paraplegias type 5.Methods Eleven patients with SPG5 who underwent cervico-thoracic spinal cord MR examination and spastic paraplegia rating scale(SPRS)were prospectively enrolled.The second MR examination and SPRS were completed after 1 year follow-up,and the changes of SPRS score,the overall structures and microstructures of spinal cord were compared.Results No significant difference of SPRS scores was found(P＞0.05).Compared to the first cervico-thoracic spinal cord MR examination,atrophy of spinal cord aggravated in the second time MRI.Significant difference of axial diffusivity of C4 right corticospinal tract(CST)was found(t=3.987,P＜0.01),but not of the other parameters of C4(all P＞0.05)between the first and the second time MRI.No significant difference of fractional anisotropy(FA),mean diffusivity(MD),AD,radial diffusivity(RD)nor T1 value of the other centrums'white matter,posterior funiculus or bilateral CST in spinal cord was found between the first and the second time MRI(all P＞0.05).Meanwhile,no significant difference of CSA,left and right diameter nor anteroposterior diameter of C1-T9 was found between the first and second time MRI(all P＞0.05).FA value of white matter,posterior funiculus and bilateral corticospinal tract in cervical spinal cord were all lower,whereas RD value at the above position were all higher in the second time MRI than those in the first time MRI(all P＞0.05).Conclusion Multiparameter MRI could be used for follow-up observation on changes of microstructure spinal cord in patients with hereditary spastic paraplegias type 5.

Objective:To explore the effects of coenzyme Q10(CoQ10) on high-fat diet-induced obesity, lipid disorders, and bile acid metabolism in mice.Methods:Eight-week-old C57BL/6J mice were randomly divided into control group(regular chow), high-fat diet group(45% high-fat chow), and CoQ10 intervention group(45% high-fat chow+ 100 mg·kg -1·d -1CoQ10) based on their body weights according to the randomized block design. The body weight and food intake of mice in each group were collected. The levels of serum total cholesterol, triglyceride, low density lipoprotein-cholesterol, high density lipoprotein-cholesterol, alanine aminotransferase, and aspartate aminotransferase were detected. The contents of 17 bile acids in serum, liver, and colon contents of mice were detected by ultra-performance liquid chromatography-tandem mass(UPLC-MS/MS). The protein expressions of cholesterol 12α-hydroxylase(CYP8B1) and oxysterol 7α-hydroxylase(CYP7B1) in liver were detected by Western blotting. Results:CoQ10 significantly reduced body weight and ameliorated lipid metabolism disorders in mice fed a high-fat diet. Compared with the control group, serum total bile acid levels were reduced in the high-fat diet group( P<0.05); CoQ10 intervention elevated serum and colonic total bile acid levels( P=0.021, P=0.014) and increased liver, colon, and serum deoxycholic acid and ursodeoxycholic acid levels( P<0.05) in the mice compared with the high-fat diet group. Both colonic and serum deoxycholic acid levels in the CoQ10 intervention group were negatively correlated with body weights( P=0.024, P=0.019), and colonic deoxycholic acid and total cholesterol levels were also negatively correlated( P=0.006). CoQ10 increased the expression of CYP8B1 and CYP7B1 proteins in the liver of mice. Conclusion:CoQ10 can modulate bile acid metabolism in high-fat diet-fed mice and alleviate their obesity and lipid metabolism disorders.

Objective:To document any effect of a pulsed electromagnetic field (PEMF) on the regulation of neuropeptide Y (NPY) in nucleus pulposus (NP) tissue, NP cell apoptosis and matrix degradation using rats with intervertebral disc degeneration (IDD).Methods:Eighteen Sprague-Dawley rats were randomly divided into a control group, an IDD model group (the model group), and a PEMF group. IDD was induced in both the model and PEMF groups. Right after the modeling, the PEMF group received 14 days of PEMF treatment, while the control group and model group were given no special treatment. Meanwhile, the primary rat nucleus pulposus cells (NPCs) were cultured using Dulbecco′s Modified Eagle Medium at 37℃ and 5% CO 2. When the fusion rate reached 90% after passage, the NPCs were divided into a control group, a TNF-α model group (referred to as model group) and TNF-α + PEMF group (referred to as PEMF group) and treated accordingly. Eight weeks after the modeling, safranin-o/fast green staining was used to assess any pathological morphology changes. The expression of NPY, neuropeptide Y receptor Y2 (NPY2R), bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), collagen type II (Col-II) and matrix metalloproteinase-3 (MMP3) in the intervertebral disc and the cultivated nucleus pulposus cells of the 3 groups were determined. Results:The intervertebral disc cells in the model group were ruptured and folded, with significantly increased polysaccharide and protein components, and significantly increased bone fibers. In the PEMF group the cell boundaries were clearer, with less fibrin fracture and increased cartilage tissue. NPY was expressed in the fibrous annulus and the nucleus pulposus of the intervertebral disc in the model group. The average expression levels of NPY and NPY2R were significantly higher than in the control group and the model group. Compared with the control group, there was a significant increase in the level of Bax and a significant decrease in the expression of Bcl-2 in the model group, and there was a significant decrease in the level of Bax in the PEMF group. Compared with the control group, there was a significant decrease in the Col-II level but a significant increase in the MMP3 protein expression in the model group. The average Col-II mRNA expression was significantly higher in the PEMF group compared with the model group, but the average MMP3 protein expression was significantly less. Those results are consistent with observations in vivo.Conclusion:PEMF may reverse the imbalance of ECM metabolism and delay IDD degeneration by up-regulating the expression of NPY and Bcl-2, as well as blocking the Bax/Bcl-2 signaling pathway to inhibit apoptosis of nucleus pulposus cells.

Objective:To study the effects of different single cell phenotypes on the prognosis of patients with intrahepatic cholangiocarcinoma by using spatial analysis, providing clues for obtaining potential immunotherapeutic targets.Methods:This study was a retrospective cohort study. A total of 41 intrahepatic cholangiocarcinoma (ICC) patients who underwent surgery in Beijing Youan Hospital Affiliated to Capital Medical University from February 2013 to June 2019 were enrolled. According to the 5-year survival situation, the patients were divided into survival group ( n=10) and death group ( n=31). A metal label-based tissue imaging mass panel containing 36 related markers was designed and constructed for staining different components in tumor samples. Through the analysis of the type and quantity of different metacluster and spatial location information and combined with the clinical outcomes of patients with information, certain metaclusters were found related to the prognosis of patients. Measurement data with normal distribution were expressed as mean±standard deviation ( ± s), paired t-test was used for comparison between groups. Measurement data with skewed distribution were expressed as median, and rank sum test was used for comparison between groups. The chi-square test was used to compare the counting data between groups. Results:36 biomarkers of 41 ICC patients were located and quantified to generate 1 476 single-cell resolution histological images. The expression information of various markers was analyzed by t-distributed Stochastic Neighbor Embedding (tSNE), and subgroups annotations (1-29) were added. It revealed that the density of metacluster 7(CD8 + T cells) was lower in survival group. The density of metacluster 16(Bcl-2 + CK7 + cancer cells) within tumors, as well as the density of metacluster 3(Vista + GB + CD11b + neutrophils) within stroma were higher in death group. Conclusion:The density of metacluster 7(Activated CD8 + T cells), metacluster 16(Bcl-2 + CK7 + tumor cells) and a novel neutrophil metacluster 3(Vista + GB + CD11b + neutrophils) correlated with ICC patients prognosis.

Objective The aim of this study was to draw single-cell transcriptome profiles of high-grade serous ovarian cancer(HGSOC),borderline ovarian cancer(OC),and normal ovaries in order to identify biomarkers that can diagnose and predict the prognosis of OC.Methods The differentially expressed genes between HGSOC,borderline OC,and normal ovarian tissues were ana-lyzed using single-cell data sequenced(SRA database:PRJNA756768).The cell subsets associated with tumor progression were screened by functional enrichment,cell communication between different subsets was analyzed by Cellchat,and cell differentiation traj-ectories were explored by pseudotime analysis to finally determine the subsets most relevant to tumor progression.Combined with OC transcriptome data of OC from the Cancer Genome Atlas(TCGA)with patient prognosis,biomarkers for diagnosing and predicting sur-vival of OC patients were ultimately screened.Results After using t-distribution stochastic neighbor embedding(t-SNE)for di-mensionality reduction,nine cell subpopulations were obtained:endothelial cells,myeloid cells,fibroblasts,T cells,stromal cells,B cells,and 3 epithelial cell subpopulations(C1,C4,and C7).Further analysis revealed that copy number variation(CNV)in the C4 group had the highest score in HGSOC,higher than those of borderline OC and normal ovaries,and was negatively correlated with prognosis.DMKN was a key marker gene in this group.Transcriptome analysis of OC in the TCGA database showed a close correlation between DMKN and poor prognosis(P=0.026),and the diagnostic efficacy of DMKN for OC was significant(A UC=0.906).Con-clusion This study is based on single-cell sequencing data to screen for DMKN,which can effectively diagnose and predict the prognosis of OC.This study provides new ideas for the diagnosis and prognosis prediction of OC.

Objective:To explore any effect of pulsed electromagnetic field (PEMF) stimulation on intervertebral disc degeneration (IDD).Methods:Forty Sprague-Dawley rats were randomly divided into a control group, an IDD model group, a PEMF group and an observation group. An IDD model was induced in all except those in the control group. Both the PEMF and observation groups were given PEMF stimulation, while the latter was additionally injected with the A2AR agonist CGS-21680. Eight weeks after the modelling any pathological changes in the morphology of the rats′ intervertebral disc tissues were evaluated using saffron solid green staining. The expression of A2AR, cyclic adenosine phosphate (cAMP), protein kinase A (PKA), cysteine aspartate proteolytic enzyme-3 (Caspase-3), type II collagen (COL-II) and matrix metalloproteinase-3 (MMP3) in the intervertebral discs were evaluated.Results:The nucleus pulposus had shrunk, while fibrous tissues and chondrocytes had increased in the IDD model group. In the observation group the nucleus pulposus was intact and of basically normal shape. A2AR mRNA and protein levels were higher in the intervertebral disc tissue of the model group than among the control group, on average, while the levels in the observation group were significantly higher than in the other groups. In the PEMF and observation groups cAMP and PKA mRNA were significantly higher than in the IDD model group. The p38 MAPK and P-P38 MAPK levels of the IDD model group and its average P-P38 MAPK/p38 MAPK ratio were significantly higher than in the control group. In the PEMF and observation groups those indices had decreased to varying degrees, with those of the observation group significantly lower than among the model and PEMF groups on average, except for the p38 MAPK values. Caspase-3 and its mRNA were significantly higher in the model group than in the control group, on average, and those values were significantly lower in the PEMF and observation groups than in the IDD model group. The average MMP3 contents of the IDD model group had increased significantly compared with the control group, while the Col-Ⅱ level had decreased significantly. Compared with the IDD model group, the MMP3 level had decreased but Col-Ⅱ expression had increased in both the PEMF and observation groups, with significant differences between the IDD model and observation groups.Conclusions:The activation of the p38 MAPK signaling pathway by inflammatory factors to induce apoptosis is one of the important reasons for the aggravation of IDD lesions. PEMF combined with A2AR agonists can activate the A2AR/cAMP/PKA signaling pathway, inhibit p38 MAPK phosphorylation, reduce apoptosis of nucleus pulposus cells, and relieve IDD damage.

Objective:To observe any regulatory effect of a pulsed electromagnetic field (PEMF) on A2A adenosine receptors (A2ARs) in the nucleus pulposus of rats with intervertebral disc degeneration (IDD), and to explore any combination with the A2ARs′ agonist-mediating ROS/PI3K/Akt signaling pathway.Methods:Fifty Sprague-Dawley rats were randomly divided into a control group, an intervertebral disc degeneration group (the model group), an A2AR agonist CGS-21680 treatment group (the agonist group), a PEMF group and a PEMF combined with CGS-21680 treatment group (the observation group). IDD was modeled in all except the rats in the control group. 100μL of CGS-21680 (100μg/kg) was injected into the L 5-6 intervertebral discs of the agonist group, while the PEMF group was given 30 minutes of PEMF intervention daily for 14 days at 1.5mT and 75Hz with a pulse width of 150μs. The observation group was injected with CGS-21680 and then given the same PEMF intervention. Primary nucleus pulposus cells from each group (50ng/mL) were cultured and the expressions of 8-OHDG, SOD, MDA and ROS were detected by immunohistochemistry, immunofluorescence or with an ELISA kit. The A2AR, PI3K, AKT and p-AKT protein levels were detected using western blotting. Results:The nucleus pulposus cells and the annulus fibrosus were obviously wrinkled, necrotic and broken in the model group but the annulus fibrosus was intact and the nucleus pulposus was almost normal in the observation group. Compared with the model group, the levels of SOD and A2AR, PI3K, p-AKT and AKT protein were higher in the agonist, PEMF and observation groups, while the expressions of MDA, ROS and 8-OHDG were weaker. The ROS level in the observation group was significantly lower than that in the agonist and PEMF groups, and the phosphorylation level of p-AKT in the observation group was significantly higher than in the agonist and PEMF groups. The average levels of SOD, A2AR, PI3K, p-AKT and AKT protein in the nucleus pulposus cells of the agonist, PEMF and observation groups were significantly higher than the IL-1β group′s average, while the average levels of MDA, ROS and 8-OHDG were significantly lower. The ROS levels in the observation group were significantly lower than in the agonist and PEMF groups, while the A2AR protein content and p-AKT phosphorylation in the observation group were significantly greater. The average Bax levels in the nucleus pulposus cells of the agonist, PEMF and observation groups were significantly lower than that in the IL-1β group, and the expression of Bcl-2 was significantly increased. There was significantly less apoptosis observed in the observation group than in the agonist and PEMF groups, while the expression of Bcl-2 was significantly higher.Conclusions:PEMF plays an anti-oxidative stress role by up-regulating A2AR activity and reducing ROS generation. Treatment with PEMF and A2AR agonist could further activate the phosphorylation of PI3K/Akt, down-regulate Bax and up-regulate Bcl-2, thus inhibiting the apoptosis of nucleus pulposus cells and alleviating the malignant progression of IDD.

ObjectiveTo investigate the biological function and molecular mechanism of long non-coding RNA Linc‑pint in colorectal cancer. MethodsQuantitative real‑time quantitative （qRT‑PCR） was performed to detect the expression level of Linc‑pint in 31 pairs of colorectal cancer tumor and adjacent normal tissues； correlation between the expression level of Linc‑pint and the clinicopathological characteristics was analyzed by the chi‑square test. Kaplan-Meier survival analysis was used to assess the relationship between Linc‑pint expression level and the prognosis of patients. Cox regression model was used to analyze the relationship between clinicopathological characteristics and the prognosis of patients. Expression level of Linc‑pint were detected by qRT‑PCR in 5 common colorectal cancer cell lines. Effect of Linc‑pint on cell proliferation， invasion and migration was measured by cell counting kit‑8 assay， Transwell assay and harvested xenografts from nude mice. qRT‑PCR was performed to detect the expression level of Linc‑pint's target gene micro RNA（miR）‑21 in 31 pairs of colorectal cancer tumor tissues and adjacent normal tissues. Pearson correlation coefficient was used to assess the correlation between Linc‑pint and miR‑21. qRT‑PCR was used to detect the expression of overexpression of Linc‑pint on miR‑21 in colorectal cancer cells. ResultsExpression level of Linc‑pint in normal tissues （3.95±1.16） was significantly higher than that in colorectal cancer tissues （2.74±0.95） （t=6.17， P<0.05）. Overall survival rate of patients with high expression of Linc‑pint was 62.5%， which was significantly higher than that of patients with low expression of Linc‑pint （34.3%， P<0.05）. The proliferation， invasion and migration of CRC cells were inhibited after overexpression of Linc‑pint. In colorectal cancer tumor and adjacent normal tissues， Linc‑pint and miR‑21 showed opposite expression in tumor tissues and were negatively correlated （r=-0.288 and -0.908， both P<0.05）. ConclusionLinc‑pint acts as a tumor suppressor by down‑regulating the expression level of miR‑21 to inhibit the proliferation， invasion and migration of colorectal cancer.