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.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. 
		                        		
		                        		
		                        		
		                        	
3.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.
		                        		
		                        		
		                        		
		                        	
4.Relationship between serum HMGB1,GP73,IL-37 and HBV-DNA load in patients with HBV-ACLF and their prognostic value
Xiuhong LIU ; Qingling SONG ; Miao GUO ; Cong WANG ; Daxin LEI
International Journal of Laboratory Medicine 2024;45(18):2207-2212
		                        		
		                        			
		                        			Objective To investigate the relationship between serum high mobility group protein B1(HMGB1),Golgi protein 73(GP73),interleukin-37(IL-37)and HBV-DNA load in patients with hepatitis B virus-associated chronic plus acute liver failure(HBV-ACLF)and the value of predicting prognosis.Methods A total of 112 HBV-ACLF patients from the 980th Hospital of PLA Joint Logistic Support Force from July 2018 to July 2023 were retrospectively analyzed,and were divided into death group(n=42)and sur-vival group(n=70)according to the clinical outcomes of HBV-CALF patients within 90 days after admis-sion.The clinical data and serum levels of HMGB1,GP73 and IL-37 of the two groups were compared,the prognotic factors of the HBV-CALF patient were analyzed,and the relationship between serum HMGB1,GP73 and IL-37 and HBV-DNA load was analyzed.The prognostic value of serum HMGB1,GP73 and IL-37 in HBV-CALF patients were evaluated.Results There were significant differences in disease stage,peritonitis,hepatic encephalopathy,pulmonary infection,HBV-DNA load and MELD score between the death group and the survival group(P<0.05).Serum levels of HMGB1,GP73 and IL-37 in death group were higher than those in survival group(P<0.05).Disease stage,peritonitis,hepatic encephalopathy,pulmonary infection,HBV-DNA load,MELD score and serum HMGB1,GP73 and IL-37 levels were all independent prognoses of HBV-ACLF patients(P<0.05).The levels of serum HMGB1,GP73 and IL-37 in HBV-ACLF patients were negatively correlated with HBV-DNA load(P<0.05).The area under the curve(AUC)values of serum HMGB1,GP73 and IL-37 in predicting the prognosis of HBV-ACLF patients were 0.781,0.790 and 0.782,re-spectively.The AUC value of the combined prediction was the largest(0.944),and the sensitivity and speci-ficity were 86.33%and 91.43%,respectively.Conclusion Serum HMGB1,GP73 and IL-37 levels are closely related to HBV-DNA load and prognosis of HBV-CALF patients,and the combined detection of these three indexes is helpful to improve prognosis prediction efficiency.
		                        		
		                        		
		                        		
		                        	
5.Diagnosis and treatment of hypertonic dehydration in the elderly
Chinese Journal of Geriatrics 2023;42(5):598-602
		                        		
		                        			
		                        			Hypertonic dehydration is prevalent in the elderly, increases the risk of adverse events, reduces quality of life, and increases hospitalization and mortality.Current methods of diagnosing dehydration, such as physical signs, urine tests, urea nitrogen/creatinine, bioelectrical impedance and saliva, are not suitable for the diagnosis of dehydration in the elderly.There is no gold standard for the diagnosis of hypertonic dehydration in the elderly.The European Society for Clinical Nutrition and Metabolism(ESPEN)recommends the direct measurement of plasma/serum osmotic pressure or using an osmotic pressure formula[osmolarity(mmol/L)=1.86 ×(Na + + K + )+ 1.15×glucose+ urea+ 14](all measured in mmol/L)for the diagnosis of hypertonic dehydration.
		                        		
		                        		
		                        		
		                        	
6.Standard for monitoring and evaluation of two-dimensional- and three-dimensional-transesophageal echocardiography during transcatheter tricuspid valve replacement
Cuizhen PAN ; Wei LI ; Daxin ZHOU ; Yuan ZHANG ; Wenzhi PAN ; Shasha CHEN ; Jing SHI ; Haiyan CHEN ; Dehong KONG ; Yu LIU ; Zhenyi GE ; Chunqiang HU ; Kefang GUO ; Xianhong SHU ; Junbo GE
Chinese Journal of Ultrasonography 2023;32(5):449-454
		                        		
		                        			
		                        			Transcatheter tricuspid valve intervention is the new frontier of interventional cardiology. The LuX-Valve is a radial force-independent orthotopic tricuspid valve replacement device developed in China. The LuX-Valve Plus transcatheter tricuspid valve replacement (TTVR) system is changed from the trans-atrial to the transjugular approach, which further reduces trauma and pulmonary complications compared with the first generation LuX-Valve. The first-in-human study has been completed at Zhongshan Hospital, Fudan University and an exploratory multicentre clinical study is underway. Echocardiography plays an important role in pre-TTVR screening, intraoperative guidance and postoperative evaluation and follow-up, especially two-dimensional transoesophageal echocardiography (2D-TEE) and three-dimensional transoesophageal echocardiography (3D-TEE). However, there is a lack of appropriate intraoperative guidance and assessment protocols. In this study, we briefly described the protocols and imaging considerations for intraoperative 2D-TEE and 3D-TEE to ensure the successful implantation of TTVR.
		                        		
		                        		
		                        		
		                        	
7.Advantages of Traditional Chinese Medicine in Treating Dominant Disease: Allergic Rhinitis
Lili LIU ; Daxin LIU ; Jinfeng LIU ; Shuzhen GUO ; Zhonghai XIN ; Renzhong WANG ; Li TIAN ; Kuiji WANG ; Mingxia ZHANG ; Shirui YANG ; Shufan GUO ; Yonggang LIU ; Wei ZHANG ; Lingyan JIANG ; Hui CHEN ; Xing LIAO ; Geng LI ; Chenyu CHI ; Xiaoxiao ZHANG ; Zhanfeng YAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(2):203-211
		                        		
		                        			
		                        			In response to the Opinions of the CPC Central Committee and the State Council on Promoting the Inheritance, Innovation, and Development of traditional Chinese medicine(TCM) and the spirit of the National Conference on TCM, Chinese Association of Chinese Medicine organized experts in Otorhinolaryngology Head and Neck Surgery of traditional Chinese and western medicine to discuss the clinical advantages of TCM and integrated traditional Chinese and western medicine in the treatment of allergic rhinitis (AR) and they reached a basic consensus. In recent years, the prevalence of AR has been on the rise, threatening the quality of life of patients and giving rise to a heavy burden to both the patients and the society. AR is resulted from immune imbalance rather than reduced immunity or hyperimmunity, and the imbalance is similar to the Yin-yang disharmony in TCM. In the treatment of this disease, western medicine features rapid onset. However, it is cost-intensive and causes severe surgical trauma, and the recurrence is common. TCM boasts diverse methods for AR, which can be used in all stages of this disease. It has advantages in controlling symptoms such as nasal congestion, runny nose, or dysosmia in the attack stage, preventing recurrence in the remission stage, and treating refractory AR or steroid-resistant AR. In particular, acupuncture enjoys a reputation in treatment of AR, which has been supported by evidence-based medicine and recommended by guidelines. While treating local symptoms of AR, TCM regulates the psychosomatic conditions, which facilitates chronic disease management and long-term follow-up. We should integrate the advantages of TCM and western medicine, give full play to the unique nonnegligible and irreplaceable advantages of TCM, formulate a comprehensive diagnosis and treatment scheme for learning and promotion, and summarize the research outcomes to promote the theoretical innovation of TCM on AR from the perspective of integrated traditional Chinese and western medicine. 
		                        		
		                        		
		                        		
		                        	
8.Real-time monitoring and step-by-step guidance for transcatheter tricuspid annuloplasty using transesophageal echocardiography
Cuizhen PAN ; Daxin ZHOU ; Xiaochun ZHANG ; Wei LI ; Shasha CHEN ; Yuan ZHANG ; Jing SHI ; Haiyan CHEN ; Dehong KONG ; Yu LIU ; Zhenyi GE ; Kefang GUO ; Xianhong SHU ; Junbo GE
Chinese Journal of Ultrasonography 2022;31(7):626-630
		                        		
		                        			
		                        			Tricuspid regurgitation (TR) interventions are under rapid development. The K-Clip? system is the first domestic transcatheter tricuspid annuloplasty system with unique clamping procedure to achieve annular reduction.Intraoperative echocardiographic monitoring procedures for transcatheter tricuspid annuloplasty have not been reported yet in China. Thus, this review aimed to propose the standard two-dimensional and three-dimensional transesophageal echocardiographic workplanes and procedures to guide and monitor the implantation of K-Clip system based on our experience in Zhongshan Hospital, Fudan University to provide a reference point for the intraoperative echocardiographic monitoring of future transcatheter tricuspid annuloplasty devices in China.
		                        		
		                        		
		                        		
		                        	
9.Relationship between SUVmax in 18F-FDG PET/CT and PD-L1 expression in invasive lung adenocarcinoma
GUO Daxin ; HUANG Wenxia ; HUANG Xiaoli ; HE Fuqian ; SU Minggang ; JIANG Yong
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2020;27(03):290-296
		                        		
		                        			
		                        			Objective    To investigate the relationship between the expression of programmed cell death ligand-1 (PD-L1) and the maximal standardized uptake value (SUVmax) in 18F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) and the correlation of clinical factors between SUVmax values and PD-L1. Methods    The clinical data of 84 patients with invasive lung adenocarcinoma diagnosed pathologically in West China Hospital, Sichuan University from August 2016 to November 2018 were analyzed retrospectively, including 38 males and 46 females, aged 60 (32-85) years. The tumor was acinar-predominant in 37 patients, papillary in 20, lepidic in 19, solid in 5 and micropapillary in 3. Multivariate analysis of the relationship between SUVmax value and other clinicopathological features was performed by linear regression. Logistic regression analysis was used to analyze the relationship between PD-L1 protein expression and other pathological features. Results    The SUVmax of the PD-L1 expression group was significantly higher than that of the non-PD-L1 expression group in the whole invasive lung adenocarcinoma group (P=0.002) and intermediate-grade histologic subtype (P=0.016). The SUVmax cut-off value of PD-L1 expression in the whole invasive lung adenocarcinoma group and intermediate-grade histologic subtype was 5.34 (AUC: 0.732, P=0.002) and 5.34 (AUC: 0.720, P=0.017), respectively. Multivariate analysis showed that pleura involvement, vascular tumor thrombus and the increase of tumor diameter could cause the increase of the SUVmax value, while the SUVmax value decreased in the moderately differentiated tumor compared with the poorly differentiated tumor. The SUVmax cut-off value between low-grade histologic subtype and intermediate-grade histologic subtype, intermediate-grade histologic subtype and high-grade histologic subtypes was 1.54 (AUC: 0.854, P<0.001) and 5.79 (AUC: 0.889, P<0.001), respectively. Multivariate analysis of PD-L1 expression showed pleura involvement (P=0.021, OR=0.022, 95%CI 0.001 to 0.558) and moderate differentiation (opposite to poor differentiation) (P=0.004, OR=0.053, 95%CI 0.007 to 0.042) decreased the expression of PD-L1. Conclusion    The SUVmax of the PD-L1 expression group is significantly higher than that of the non-PD-L1 expression group in the whole invasive lung adenocarcinoma group and intermediate-grade histologic subtype. The level of SUVmax and the expression of PD-L1 in invasive lung adenocarcinoma are related to many clinical factors.
		                        		
		                        		
		                        		
		                        	
10.Clinical role of transthoracic and transesophageal echocardiography in transapical mitral valve repair
Zhenyi GE ; Cuizhen PAN ; Wei LI ; Haiyan CHEN ; Dehong KONG ; Daxin ZHOU ; Wenzhi PAN ; Lai WEI ; Kefang GUO ; Junbo GE ; Xianhong SHU
Chinese Journal of Ultrasonography 2019;28(5):382-386
		                        		
		                        			
		                        			Objective To evaluate the role of transthoracic echocardiography ( T T E ) and transesophageal echocardiography( T EE) in the process of transapical mitral valve repair using a novel edge‐to‐edge device( ValveClamp) and this device′s efficacy and safety in a preliminary clinical trial . Methods Six patients with moderate to severe or severe degenerative mitral regurgitation ( DM R) confirmed by T T E and T EE were enrolled . T T E was performed pre and post procedure as well as 30 days post procedures . Related cardiac structure and hemodynamic parameters ,including mitral regurgitation area ( MRA‐max ) , vena contracta width ( VCW ) ,mitral valve effective orifice area ( M VEOA ) ,left ventricular end diastolic diameter ( LVEDD ) , left ventricular end systolic diameter ( LVESD ) , left ventricular ejection fraction ( LVEF) ,max and mean mitral valve pressure gradient ( M VPG‐max and M VPG‐mean) were recorded and evaluated in a central core laboratory . Results All the procedures were successfully performed .M RA‐max , VCW and M VEOA decreased significantly post procedures ( all P < 0 .000 ) , and they remained no significant changes within 30 days post procedures ( all P > 0 .05 ) . M eanwhile ,M VPG‐max and M VPG‐mean slightly increased ( all P <0 .01 ) and left atrial anterior‐posterior dimension attenuated 30 days post procedures( P <0 .05) ,but all M VPG‐mean were lower than 5 mm Hg ( 1 mm Hg=0 .133 kPa) . T here were no significant changes in other hemodynamic parameters ( all P > 0 .05) . Conclusions T ransapical mitral valve repair using ValveClamp can be performed safely and a significant reduction in mitral regurgitation can be achieved in patients with DM R . T EE and T T E facilitate the patient selection for ValveClamp procedures as well as perioperative navigation and assessment .
		                        		
		                        		
		                        		
		                        	
            
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