BACKGROUND:Patch clamp technique has been developed for more than 40 years as the"gold standard"for the study of ion channels.However,the research content of scientific research institutions is relatively independent,and the existing research results are not systematically summarized,which leads to the phenomenon of high repeatability and weak innovation in the existing research.Therefore,it is urgent to make a comprehensive review of patch clamp technology to clarify the current research status,hot spots,and future development direction.OBJECTIVE:To summarize the research status and development trend of patch clamp technique in recent 10 years.METHODS:Publications on patch clamp technology from 2013 to 2023 were collected using the Web of Science core collection database.CiteSpace and VOSviewer software were used to quantify the number of publications and analyze the network of literature entries,including countries,institutions,journals,authors,keywords,highly cited literature,and co-cited references.RESULTS AND CONCLUSION:(1)In recent 10 years,the research in the field of patch clamp technology has gradually entered a stage of stable development.(2)China and the United States are the leading countries in this regard.The Chinese Academy of Sciences is an institution with core influence.Journal of Neuroscience is the main publication.Park,Won Sun team(Jeonbuk National University)and Chu,Li team(Hebei Key Laboratory of Chinese Medicine Research on Cardio-Cerebrovascular Disease)have made outstanding contributions in this field,but there is less collaboration and communication between the teams and no network cooperation model has been formed.(3)Patch clamp technology is mainly used in the electrophysiological characteristics of the nervous system and the pathological mechanism of the disease,which is the focus of researchers'continuous attention.(4)In the study of electrophysiological characteristics of cardiovascular system and its pathological mechanism,the electrophysiological characteristics of primary cardiomyocytes and induced pluripotent stem cell-derived cardiomyocytes and the pathological mechanism of atrial fibrillation,cardiotoxicity,sudden cardiac death,hypertension and other cardiovascular diseases have been the focus of research in recent years.(5)In the application of patch clamp technology combined with other biotechnology,it will be an important research direction to focus on the cross fusion with optogenetics,two-photon calcium imaging and other technologies.(6)In the research of drug screening and identification of therapeutic targets,especially the research of patch clamp technology and traditional Chinese medicine compound,it will become a great help in the future research of component traditional Chinese medicine.

BACKGROUND:Patch clamp technique has been developed for more than 40 years as the"gold standard"for the study of ion channels.However,the research content of scientific research institutions is relatively independent,and the existing research results are not systematically summarized,which leads to the phenomenon of high repeatability and weak innovation in the existing research.Therefore,it is urgent to make a comprehensive review of patch clamp technology to clarify the current research status,hot spots,and future development direction.OBJECTIVE:To summarize the research status and development trend of patch clamp technique in recent 10 years.METHODS:Publications on patch clamp technology from 2013 to 2023 were collected using the Web of Science core collection database.CiteSpace and VOSviewer software were used to quantify the number of publications and analyze the network of literature entries,including countries,institutions,journals,authors,keywords,highly cited literature,and co-cited references.RESULTS AND CONCLUSION:(1)In recent 10 years,the research in the field of patch clamp technology has gradually entered a stage of stable development.(2)China and the United States are the leading countries in this regard.The Chinese Academy of Sciences is an institution with core influence.Journal of Neuroscience is the main publication.Park,Won Sun team(Jeonbuk National University)and Chu,Li team(Hebei Key Laboratory of Chinese Medicine Research on Cardio-Cerebrovascular Disease)have made outstanding contributions in this field,but there is less collaboration and communication between the teams and no network cooperation model has been formed.(3)Patch clamp technology is mainly used in the electrophysiological characteristics of the nervous system and the pathological mechanism of the disease,which is the focus of researchers'continuous attention.(4)In the study of electrophysiological characteristics of cardiovascular system and its pathological mechanism,the electrophysiological characteristics of primary cardiomyocytes and induced pluripotent stem cell-derived cardiomyocytes and the pathological mechanism of atrial fibrillation,cardiotoxicity,sudden cardiac death,hypertension and other cardiovascular diseases have been the focus of research in recent years.(5)In the application of patch clamp technology combined with other biotechnology,it will be an important research direction to focus on the cross fusion with optogenetics,two-photon calcium imaging and other technologies.(6)In the research of drug screening and identification of therapeutic targets,especially the research of patch clamp technology and traditional Chinese medicine compound,it will become a great help in the future research of component traditional Chinese medicine.

OBJECTIVE To investigate the in vitro inhibitory mechanism of berberine on the proliferation of tumor stem cells and evaluate its in vivo safety. METHODS Flow cytometry was used to select tumor stem cells from mouse skin melanoma B16F10 cells； CD44， CD133， Nanog homologous box protein （NANOG） and octamer-binding transcription factor 4 （OCT4） were used as indicators to characterize tumor stem cells. Tumor stem cells were divided into control group， all-trans retinoic acid （ATRA） group， and berberine group， and the CCK-8 method was used to detect the effects of berberine on the viability of tumor stem cells； flow cytometry was adopted to detect cell apoptotic rate， the proportion of CD44+/CD133+ and the positive cell rate of sex determining region Y box protein 2 （SOX2）； the morphological changes of tumor balls were recorded after treatment with berberine； the morphology of cell pyroptosis in each group was recorded， and the release rate of lactate dehydrogenase （LDH） was detected； Western blot assay was adopted to detect the expressions of pyroptosis-related protein gasdermin E （GSDME）， GSDME- N， caspase-3 and cleaved caspase-3. Preliminary evaluation of in vivo safety of berberine was conducted by using zebrafish embryo toxicity experiments. RESULTS Compared with B16F10 cells， the proportion of CD44+/CD133+ cells in tumor stem cells and the fluorescence intensity of NANOG and OCT4 were significantly increased （P＜0.000 1）. The half-inhibitory concentration of berberine to tumor stem cells was 50.98 μmol/L. Compared with the control group， the apoptotic rate of cells in the berberine group was significantly increased， while the proportion of CD44+/CD133+ cells and the rate of SOX2 positive cells were reduced significantly （P＜0.000 1）； tumor stem cell spheroids were atrophied， with partial cell death. After treatment with berberine， tumor stem cells exhibited swelling in their outermost layer， the release rate of LDH of cells was significantly increased and the release rate of LDH increased with increasing dose； the protein expressions of GSDME-N and cleaved-caspase-3 of cells in berberine 20， 40 μmol/L groups were significantly increased， and the protein expressions of GSDME and caspase-3 were significantly reduced （except for berberine 20 μmol/L group， P＜0.05）. The embryonic development of zebrafish treated with berberine was almost unaffected， and the survival rate of embryo reached 100%， with no obvious abnormalities observed. CONCLUSIONS Berberine has good activity against the proliferation of tumor stem cells， and its mechanism of action may be related to activating GSDME and promoting cell pyroptosis； berberine has good in vivo safety.

ObjectiveTo employ the effective components and antiarrhythmic mechanism of Wenxin Granules （WXKL） by cell membrane chromatography （CMC） and ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF/MS）， combined with network pharmacology. MethodIn this study， the CMC/UPLC-Q-TOF/MS technique was employed to identify the components in WXKL that could specifically bind to myocardial cell membranes. By utilizing databases such as SwissTarget Prediction and GeneCards， the targets of WXKL's effective components and arrhythmia-related targets were mined. Cytoscape software was used to construct a "component-target-disease" network. Gene ontology（GO） function and Kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment analyses were carried out， and molecular docking of key components and targets was performed. Finally， further verification was conducted through in vivo experiment of rats. ResultA total of 39 effective components were identified in WXKL. These included 13 components derived from Panax notoginseng， 15 components from Codonopsis pilosula， seven components from Glycyrrhizae Radix et Rhizoma， one component from Succinum， one component from Polygonatum odoratum， one component shared by both P. odoratum and C. pilosula， and one component shared by both Panax notoginseng and C. pilosula. Network pharmacology predicted that WXKL had 16 core antiarrhythmic targets and 79 related pathways， mainly involving adrenergic signaling in cardiomyocytes， cyclic guanosine monophosphate （cGMP）/protein kinase G （PKG）， calcium signal， cyclic adenosine monophosphate （cAMP）， interleukin （IL）-17， mitogen-activated protein kinase （MAPK）， and tumor necrosis factor （TNF） signaling pathways. The results of in vivo experiment of rats showed that WXKL significantly improved the expression of β₁-adrenergic receptor （β₁-AR）， cAMP， TNF-α， and calcium voltage-gated channel subunit alpha 1C （CACNA1C）. ConclusionWXKL can exert its antiarrhythmic effects through multiple components， multiple targets， and multiple pathways. This study provides a scientific basis for explaining the potential pharmacodynamic substance foundation and mechanism of action of traditional Chinese medicine in treating arrhythmia.

Objective To evaluate the maturity and metabolic status of heterotopic ossification(HO)by single-photon emission computed tomography(SPECT)/CT fusion bone imaging.Methods The clinical and SPECT/CT fusion bone imaging data of 57 patients with HO confirmed by pathology or follow-up were analyzed retrospectively.HO was graded by CT,and the characteristics of radioactive concentration of HO were analyzed.Results Of 57 cases,single lesion in 52 cases,and multiple lesions in 5 cases,with a total of 63 lesions,mostly located in the hip joint(55.6%,35/63)and thigh(19.0%,12/63).There were 41 lesions in the middle stage and 22 lesions in the late stage.In the visual evaluation of SPECT/CT fusion bone imaging,the middle stage lesions were mostly clumps or flakes,with moderate or high radioactive concentration(75.6%,31/41),furthermore,the concentration range was larger than or equal to the total or limited range of CT ossification(21/41,50.1%),with high concentration mainly located in the mixed areas of ossification density.The concentration of the late stage lesions was mostly non-radioactive(72.7%,16/22).Conclusion SPECT/CT fusion bone imaging can show the range,degree and maturity of HO radioactive concentration,and can accurately locate the area with osteoblastic activity,which provides scientific basis for the selection of surgical timing.

RNAs are involved in the crucial processes of disease progression and have emerged as powerful therapeutic targets and diagnostic biomarkers. However, efficient delivery of therapeutic RNA to the targeted location and precise detection of RNA markers remains challenging. Recently, more and more attention has been paid to applying nucleic acid nanoassemblies in diagnosing and treating. Due to the flexibility and deformability of nucleic acids, the nanoassemblies could be fabricated with different shapes and structures. With hybridization, nucleic acid nanoassemblies, including DNA and RNA nanostructures, can be applied to enhance RNA therapeutics and diagnosis. This review briefly introduces the construction and properties of different nucleic acid nanoassemblies and their applications for RNA therapy and diagnosis and makes further prospects for their development.

【Objective】 We combined the concept of traditional medicine with magnetic induction technology, originally brought up the research concept of magnetic hyperthermia to cure KOA, explored the mechanism and constructed a new treatment of KOA with modern medical features. 【Methods】 Through establishing a primary KOA model in rats and constructing ferrimagnetic vortex domain iron oxide nanorings (FVIOs) as a platform for highly efficient magnetic hyperthermia agent, the lesions of KOA were heated accurately under the low-intensity magnetic field. We confirmed the curative effect through the results of pain perception, histopathology, knee joint morphology and microscopic bone structure and the content of serum inflammatory factor, to study the therapeutic mechanism of magnetic hyperthermia for KOA. 【Results】 Compared with the model group, the recovery of mechanical pain threshold after magnetic hyperthermia improved by approximately 48.9%; the degree of hyperemia and edema of joint capsule and synovial tissue and the wear degree of joint cartilage surface, were significantly reduced; the Mankin and OARSI scores decreased by about 33% and 20%, respectively; the MicroCT results indicated that the degree of hardening of the subchondral bone also improved; the expression of inflammatory factors in the serum was reduced. 【Conclusion】 In this study, we utilized the FVIOs as a high-efficiency magnetic hyperthermia platform for the treatment of KOA. The efficacy of magnetic hyperthermia on KOA is clarified, and the mechanism is related to the inhibition of inflammatory factors.

Recent studies have suggested that long-term application of anti-angiogenic drugs may impair oral mucosal wound healing. This study investigated the effect of sunitinib on oral mucosal healing impairment in mice and the therapeutic potential of Bifidobacterium breve (B. breve). A mouse hard palate mucosal defect model was used to investigate the influence of sunitinib and/or zoledronate on wound healing. The volume and density of the bone under the mucosal defect were assessed by micro-computed tomography (micro-CT). Inflammatory factors were detected by protein microarray analysis and enzyme-linked immunosorbent assay (ELISA). The senescence and biological functions were tested in oral mucosal stem cells (OMSCs) treated with sunitinib. Ligated loop experiments were used to investigate the effect of oral B. breve. Neutralizing antibody for interleukin-10 (IL-10) was used to prove the critical role of IL-10 in the pro-healing process derived from B. breve. Results showed that sunitinib caused oral mucosal wound healing impairment in mice. In vitro, sunitinib induced cellular senescence in OMSCs and affected biological functions such as proliferation, migration, and differentiation. Oral administration of B. breve reduced oral mucosal inflammation and promoted wound healing via intestinal dendritic cells (DCs)-derived IL-10. IL-10 reversed cellular senescence caused by sunitinib in OMSCs, and IL-10 neutralizing antibody blocked the ameliorative effect of B. breve on oral mucosal wound healing under sunitinib treatment conditions. In conclusion, sunitinib induces cellular senescence in OMSCs and causes oral mucosal wound healing impairment and oral administration of B. breve could improve wound healing impairment via intestinal DCs-derived IL-10.
Animals ; Mice ; Interleukin-10 ; Bifidobacterium breve ; Up-Regulation ; Angiogenesis Inhibitors ; Sunitinib ; X-Ray Microtomography ; Administration, Oral ; Wound Healing ; Antibodies, Neutralizing

Objective:To qualitatively analyze the cardiac rehabilitation information need based on KANO model in patients after percutaneous coronary intervention (PCI) and to obtain the satisfaction and importance of every need so as to provide a basis for optimizing clinical rehabilitation health guidance.Methods:From May 2019, this study selected 144 patients with selective PCI as subjects by convenience sampling. The KANO two-factor questionnaire was designed based on 10 aspects of needs including the basic knowledge, nutrition, exercises, medications, works and professions, pressure and psychology, social concern, crisis and safety, diagnosis and therapy, risk factor management of cardiac rehabilitation information need, to gain the need class and draw the need satisfaction-importance magic quadrant.Results:Among 51 items in the cardiac rehabilitation information need questionnaire, there were 5 of them (9.8%) with essential attribute, 14 of them (27.5%) with expected attribute, 8 of them (15.7%) with glamour attribute and 24 of them (47.1%) with no difference attribute. The satisfaction-importance matrix diagram showed that 4 aspects of need were in the area for improvement and 20 of them in the area for the secondary improvement among cardiac rehabilitation information needs after PCI.Conclusions:The need importance matrix directly reflects the patient satisfaction and indirectly reflects patients' need importance.

Objective To investigate the MRI characteristics of primary myxofibrosarcoma (MFS) in the extremity soft-tissues.Methods The MRI of 14 cases of MFS confirmed by operation and pathology were analyzed retrospectively.Results In the 14 cases,9 were in the thigh,3 in the lower leg,and 2 in the upper arm.The volumes of the tumors were relatively big and all located in the subcutaneous fat or superficial inter-muscular space.The tumor showed expanding growth and had well-defined boundary.On T1 WI,one case showed isosignal intensity,one showed equal signal intensity mostly,and 12 cases showed patch slightly low signal intensity;On fat suppression T2 WI and STIR,tumors showed high-low mixed signal intensity,which mainly showed high signal intensity;On MRI enhanced study,inhomogeneous enhancement was observed in most of the tumors,and "tail sign" was showed in 11 cases.Conclusion MFS is of characteristics on MRI,the position of tumor is shallow,the tumor is relatively big and has clear boundary.On T1 WI,most tumors show iso-low mixed signal intensity.On fat suppression T2WI and STIR,tumors show high-low mixed signal intensity;On enhanced MRI,inhomogeneous enhancement and "tail sign" are its characteristic signs.