1.Construction of lentiviral vectors for solute carrier family 1 member 5 overexpression and knockdown and stably transfected RAW264.7 cell line
Daxin GUO ; Susu FAN ; Zhendong ZHU ; Jianhong HOU ; Xuan ZHANG
Chinese Journal of Tissue Engineering Research 2025;29(7):1414-1421
BACKGROUND:Solute carrier family 1 member 5(SLC1A5)plays a potential role in a variety of diseases,but the exact mechanism of action is unclear.The construction of stable SLC1A5 overexpression and knockdown cell models can provide a powerful experimental tool for in-depth study of the exact role and mechanism of SLC1A5 in diseases and the discovery of potential therapeutic targets. OBJECTIVE:To construct lentiviral vectors for overexpression and knockdown of mouse SLC1A5 and establish stable transfected RAW264.7 cell lines,so as to provide an experimental foundation for further investigation of the role of SLC1A5 in inflammation. METHODS:Primers were designed and synthesized based on the SLC1A5 gene sequence,and the gene segment was amplified using polymerase chain reaction.Subsequently,the target gene segment was directionally inserted into the GV492 vector plasmid,which had been digested with AgeI/NheI enzymes,to construct recombinant lentiviral plasmids.Positive clones were further selected,and their sequences were confirmed.The pHelper1.0 plasmid vector and pHelper2.0 plasmid vector,along with the target plasmid vector,was co-cultured with 293T cells for transfection,resulting in the production and titration of lentiviral stocks.Furthermore,RAW264.7 cells were cultured in vitro,and the working concentration of puromycin was determined.Lentiviruses were separately co-cultured with RAW264.7 cells,and transfection efficiency was determined by measuring fluorescence intensity.Stable transfected cells were selected using puromycin,and real-time fluorescence quantitative PCR and western blot assay were employed to assess the gene and protein expression levels of SLC1A5 in stably transfected cell lines. RESULTS AND CONCLUSION:(1)Sequencing results indicated a perfect match between the sequencing and target sequences,confirming the successful construction of recombinant lentiviral vectors.(2)The titer for the overexpression SLC1A5 lentivirus was 1×109 TU/mL,while the titer for the knockdown SLC1A5 lentivirus was 3×109 TU/mL.(3)The working concentration of puromycin for RAW264.7 cells was determined to be 3 μg/mL.(4)The optimal conditions for transfecting RAW264.7 cells with overexpression/knockdown expression of SLC1A5 lentivirus involved the use of HiTransG P transfection enhancer with a multiplicity of infection value of 50.(5)A significant upregulation of the gene and protein expression levels of SLC1A5 was detected in cell lines stably overexpressing SLC1A5,while gene and protein expression levels of SLC1A5 were significantly decreased in the knockdown stable cell lines.These findings indicate that lentiviral vectors for mouse SLC1A5 overexpression and knockdown have been successfully constructed and a stably transfected RAW264.7 cell line has been obtained.
2."Integrated Internal and External Treatment with Harmonization of Body and Mind" for Tinnitus Caused by Constraint Syndrome
Wenli WU ; Ying ZHANG ; Daxin LIU ; Zhanfeng YAN ; Mengyu YIN
Journal of Traditional Chinese Medicine 2025;66(12):1281-1285
Based on the theories of "pathogenesis caused by constraint emotions" and "visceral orifices", it is believed that the pathogenesis of tinnitus caused by constraint syndrome follows the evolution pattern of "qi constraint (liver qi constraint) → fire disturbance (liver constraint transforming into fire) → deficiency impairment (liver constraint with spleen deficiency) → blood stasis (liver constraint leading to blood stasis)". The treatment is guided by the principle of integrated internal and external therapy with harmonization of both body and mind. Internally, the treatment focuses on rectifying zang-fu imbalances, primarily using the Xiaoyao Powder (逍遥散), with modifications based on syndrome differentiation. External therapies aim to unblock the meridians and orifices, commonly using auricular acupressure and Chinese herbal hot compresses. Additionally, traditional Chinese five-tone music therapy is applied to regulate emotional and mental disturbances. By integrating these three approaches, a comprehensive treatment strategy is formed that harmonizes both body and mind for managing tinnitus associated with constraint syndrome.
3.Assessment of perioperative pulmonary fluid volume using remote dielectric sensing (ReDSTM) non-invasive lung fluid measurement technology in transcatheter tricuspid valve-in-valve implantation: The first case report
Yuliang LONG ; Yuan ZHANG ; Xiaochun ZHANG ; Peng WANG ; Xiaotong CUI ; Wenzhi PAN ; Daxin ZHOU ; Junbo GE
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(04):571-574
One of its primary surgical treatments of tricuspid regurgitation is tricuspid valve biological valve replacement. Catheter tricuspid valve-in-valve implantation is a novel interventional alternative for biological valve failure. The non-invasive lung fluid measuring device remote dielectric sensing (ReDSTM) has been increasingly incorporated into clinical practice as a means of monitoring chronic heart failure in recent years. This report describes the process and outcomes of the first instance of perioperative lung fluid volume evaluation following transcatheter tricuspid valve implantation utilizing ReDSTM technology. The patient has a short-term, substantial increase in postoperative lung fluid volume as compared to baseline.
4.Annual report on transcatheter left atrial appendage closure in 2024
Yuan BAI ; Xiaochun ZHANG ; Jie ZENG ; Yongjian WU ; Daxin ZHOU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(06):746-751
After two decades of development, transcatheter left atrial appendage closure has emerged as a safe and effective intervention for stroke prevention in patients with atrial fibrillation. In 2024, significant advancements were made in the field of left atrial appendage closure in terms of evidence-based medicine, device research and development, and guideline consensus. The annual report on transcatheter left atrial appendage closure systematically reviews global academic progress in 2024, encompassing newly published clinical evidence, recently developed occlusion devices, and updated international guidelines/consensus statements. In the future, the development direction of transcatheter left atrial appendage closure mainly includes expanding surgical indications, optimizing imaging assistance technology, improving closure device design, and exploring individualized strategies for postoperative antithrombotic therapy.
5.2024 annual report of interventional treatment for congenital heart disease
Changdong ZHANG ; Yucheng ZHONG ; Geng LI ; Jun TIAN ; Gejun ZHANG ; Nianguo DONG ; Yuan FENG ; Daxin ZHOU ; Yongjian WU ; Lianglong CHEN ; Xiaoke SHANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(07):909-918
In recent years, with the continuous development and increasing maturity of interventional techniques, interventional treatment for congenital heart disease (CHD) has been progressively disseminated to county- and city-level hospitals in China. Concurrently, the standardized management of adult CHD (particularly patent foramen ovale) and the lifelong management of complex CHD are gaining increasing clinical attention, while the emergence of new techniques and products continuously advances the discipline. This article aims to review the new progress made in the field of interventional treatment for congenital heart disease in China during 2024. It specifically reviews and analyzes the following key aspects: (1) annual statistics on interventional closure procedures for CHD; (2) recent insights into patent foramen ovale closure; (3) advances in transcatheter pulmonary valve replacement; (4) interventional treatment and lifelong management strategies for complex CHD; (5) new interventional techniques for acquired heart disease; and (6) the application of artificial intelligence in CHD management. Through the synthesis and discussion of these topics, this article seeks to provide a detailed analysis of the current landscape of interventional treatment for CHD in China and project its future development trends.
6.Impact of the LAmbre device on left atrial appendage adjacent structures and left atrium
Zhengdan GE ; Dehong KONG ; Zhenyi GE ; Chunqiang HU ; Xiaochun ZHANG ; Haiyan CHEN ; Daxin ZHOU ; Xianhong SHU ; Cuizhen PAN
Chinese Journal of Ultrasonography 2024;33(2):142-150
Objective:To explore the effects of the LAmbre device and mitral annulus(MA), as well as left atrium(LA) in patients with non-valvular atrial fibrillation(NVAF) after left atrial appendage closure (LAAC) using real-time-three-dimensional transesophageal echocardiography (3D TEE).Methods:Fity-six consecutive patients who underwent LAAC with the LAmbre device in Zhongshan Hospital of Fudan University from June 2019 to March 2023 were retrospectively enrolled, with no or less than moderate mitral regurgitation (MR). All patients underwent pre-operative and follow-up two-and three-dimensional transesophageal echocardiography (2D TEE, 3D TEE) at 60 days after the operation. The quantitative parameters of MA and LA were obtained by offline analysis using QLab 13.0 (Philips Healthcare, Andover, MA). Importantly, mitral annular measurements were made at seven time points throughout the cardiac cycle: early diastole, mid-diastole, late diastole, mitral valve closure, early systole, mid-systole, and late systole, which facilitates constructing the dynamic model of MA to assess the annular morphology and dynamics.Results:The values of AP diameter (APD), AL-PM diameter (ALPMD), 3D annulus circumference (3DAC), 3D annulus area (3DAA) decreased significantly compared with pre-operative values at all time points of the cardiac cycle (all P<0.05), while non-planar angle (NPA) and AH/CD were not apparently changed (all P>0.05 ). Throughout the cardiac cycle, MA showed regular changes, gradually increased in systole with the saddle shape deepened, and gradually decreased in diastole with the saddle shape shallowed.During systole, there was an increase in the rate of change of AP in MA [pre-operative (3.01±2.64)%, post-operative (3.81±3.51)%, P=0.037] after LAAC, with no significant difference in the rate of change of ALPM, 3DAC, and 3DAA.Meanwhile, we observed an evident reduction in LA minimal volume (LAVmin) [pre-operative (78.36±25.16)ml, post-operative (70.73±22.78)ml, P=0.004] and an obvious increase in LA ejection function [pre-operative (22.88±10.09)%, post-operative (31.41±12.28)%, P<0.05] during follow-up. Conclusions:3D TEE can accurately assess the impact of LAAC on the MA and LA. The LAmbre device can affect the morphology of MA, as well as the structure and function of LA, while the change of the dynamics of MA is not so prominent.
7.Expert Consensus on Clinical Diseases Responding Specifically to Traditional Chinese Medicine:Aural Vertigo
Yingdi GONG ; Zhanfeng YAN ; Wei FENG ; Daxin LIU ; Jiaxi WANG ; Jianhua LIU ; Yu ZHANG ; Shusheng GONG ; Guopeng WANG ; Chunying XU ; Xin MA ; Bo LI ; Shuzhen GUO ; Mingxia ZHANG ; Jinfeng LIU ; Jihua GUO ; Zhengkui CAO ; Xiaoxiao ZHANG ; Zhonghai XIN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):215-222
Aural vertigo frequently encountered in the otolaryngology department of traditional Chinese medicine (TCM) mainly involves peripheral vestibular diseases of Western medicine, such as Meniere's disease, benign paroxysmal positional vertigo, vestibular neuritis, and vestibular migraine, being a hot research topic in both TCM and Western medicine. Western medical therapies alone have unsatisfactory effects on recurrent aural vertigo, aural vertigo affecting the quality of life, aural vertigo not relieved after surgery, aural vertigo with complex causes, and children's aural vertigo. The literature records and clinical practice have proven that TCM demonstrates unique advantages in the treatment of aural vertigo. The China Association of Chinese medicine sponsored the "17th youth salon on the diseases responding specifically to TCM: Aural vertigo" and invited vertigo experts of TCM and Western medicine to discuss the difficulties and advantages of TCM diagnosis and treatment of aural vertigo. The experts deeply discussed the achievements and contributions of TCM and Western medicine in the diagnosis and treatment of aural vertigo, the control and mitigation of the symptoms, and the solutions to disease recurrence. The discussion clarified the positioning and advantages of TCM treatment and provided guidance for clinical and basic research on aural vertigo.
8.Protocol for the Traditional Chinese Medicine Diagnosis and Treatment Guidelines for Children with Upper Airway cough Syndrome
Lingyun ZHANG ; Xiyou WANG ; Daxin LIU ; Qiang HE ; Xuefeng WANG ; Xun LI ; Yutong FEI ; Yi XIAO ; Xiaoxue LAN ; Yuanwen LIANG ; Xiaoxuan LIN ; Rong ZHOU ; Sirui GU ; Ying ZHANG ; Yue WANG ; Xingzhu YE ; Wenke LIU ; Hong CHEN ; Changhe YU
International Journal of Traditional Chinese Medicine 2024;46(8):961-966
In order to standardize the clinical diagnosis and treatment of upper airway cough syndrome (UACS) for children in China, Dongzhimen Hospital of Beijing University of Chinese Medicine and Affiliated Hospital of Liaoning University of Traditional Chinese Medicine initiated the development of this Traditional Chinese Medicine Diagnosis and Treatment Guidelines for Children with Upper Airway cough Syndrome based on evidence-based medical evidence. This guideline will process registration, write a plan, and develop relevant processes and writing norms, develop and publish official documents. This plan mainly introduces the scope of the guidelines, the purpose and significance, the composition of the guidelines working group, the management of conflicts of interest, the collection, selection and determination of clinical problems, the retrieval, screening and rating of evidence, and the consensus of recommendations. Registration information: This study has been registered in the international practice guidelines registry platform with the registration code of PREPARE-2023CN087.
9.Multicenter expert recommendations on interventional valve-in-valve technology for mitral bioprosthetic valve destruction in China
Haibo ZHANG ; Xiangbin PAN ; Yingqiang GUO ; Lai WEI ; Jian YANG ; Daxin ZHOU ; Yongjian WU ; Xu MENG ; Liming LIU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(08):1090-1095
Mitral valve replacement is one of the most common heart valve surgeries in China. In recent years, with the increase in degenerative valve diseases, older patients, and the progress of anti-calcification technology of biological valves, the proportion of mitral valve biological valve replacement has been increasing year by year. After the damage of traditional mitral valve biological valves, re-operation of valve replacement with thoracotomy is required. However, the adhesion between the heart and sternum, as well as the damage caused by cardiopulmonary bypass and cardiac arrest, can cause significant trauma to elderly patients and those with multiple organ dysfunction, leading to increased mortality and complication rates. In recent years, interventional valve surgery, especially transcatheter valve-in-valve surgery, has developed rapidly. This procedure can correct the damaged mitral valve function without stopping the heart, but there are still many differences between its technical process and conventional aortic valve replacement surgery. Therefore, organizing and writing multicenter expert recommendations on the technical process of transcatheter valve-in-valve surgery for damaged mitral valve biological valves is of great significance for the training and promotion of this technology.
10.Expression and clinical significance of interleukin-31 in plasma from patients with pulmonary arterial hypertension
Zhi ZHAN ; Xiaochun ZHANG ; Shiqiang HOU ; Lihua GUAN ; Dandan CHEN ; Wenzhi PAN ; Daxin ZHOU
Chinese Journal of Clinical Medicine 2024;31(3):394-401
Objective To investigate the plasma level of interleukin-31(IL-31)in patients with pulmonary arterial hypertension(PAH)and its clinical relevance.Methods The patients who were diagnosed as PAH in Zhongshan Hospital,Fudan University from January 1,2021 to December 30,2023(PAH group)and the healthy people in the same period(control group)were selected.The clinical data and follow-up records were collected.Plasma levels of IL-31,IL-1β,IL-6,IL-10,IL-12p70,monocyte chemoattractant protein-1(MCP-1),tumor necrosis factor-α(TNF-α)and transforming growth factor-β1(TGF-β1)were detected by enzyme-linked immunosorbent assay(ELISA).Pearson correlation test was used to evaluate the correlations between IL-31 and right cardiac catheterization parameters,echocardiography parameters and blood indices in patients with PAH.Cox proportional hazard model was used to analyze the prognostic factors of patients in PAH group.Results A total of 50 patients with PAH and 22 healthy controls were included.There was no significant difference in age,gender,body mass index and left ventricular ejection fraction between the two groups.Compared with the control group,the plasma level of IL-31 in the PAH group was significantly higher(168.82[149.14,177.26]pg/mL vs 152.76[145.58,159.41]pg/mL,P=0.001).Pearson correlation test showed that the plasma level of IL-31 in PAH patient was positively correlated with mean pulmonary artery pressure(r=0.652,P<0.001)and pulmonary vascular resistance(r=0.651,P<0.001),but was negatively correlated with tricuspid annular plane systolic excursion(r=-0.496,P<0.001).Cox proportional hazard model showed that higher plasma level of IL-31 was an independent predictor of readmission for heart failure/all-cause mortality in patients with PAH(HR=1.130,95%CI 1.052-1.214,P=0.001).Conclusions Plasma level of IL-31 may be significantly increased in patients with PAH and be positively correlated with the severity of PAH,and elevated level of IL-31 is predictive of poor prognosis in PAH patients.

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