1.Effect of azathioprine on ferroptosis in spermatocytes of mice induced by RSL3
Yanjue YE ; Ziyi TANG ; Yupei TAN ; Shiying YANG ; Yong LIU ; Li YIN
Journal of Jilin University(Medicine Edition) 2024;50(5):1217-1226
		                        		
		                        			
		                        			Objective:To discuss the effect of azathioprine(AZA)on ferroptosis in spermatocytes of the mice induced by reduced glutathione(GSH)peroxidase 4 inhibitor RSL3,and to clarify the potential mechanism.Methods:The spermatogonia GC-2 cells of the mice were randomly divided into control group(no treatment),RSL3 group(treated with 10 nmol·L-1 RSL3 for 24 h),RSL3+ferroptosis inhibitor(Ferrostatin-1,Fer-1)group(treated with 10 nmol·L-1 RSL3 for 24 h+2 μmol·L-1 Fer-1 for 12 h),RSL3+low dose of AZA group(treated with 10 nmol·L-1 RSL3 for 24 h+5 μmol·L-1 AZA for 12 h),RSL3+medium dose of AZA group(treated with 10 nmol·L-1 RSL3 for 24 h+10 μmol·L-1 AZA for 12 h),and RSL3+high dose of AZA group(treated with 10 nmol L-1 RSL3 for 24 h+20 μmol·L-1 AZA for 12 h).The MTT method was used to detect the activities of the GC-2 cells in various groups after treated with different concentrations of AZA and RSL3;the GSH and GSSG levels in the GC-2 cells were detected by GSH and oxidized glutathione(GSSG)detection kits;the malondialdehyde(MDA)levels in the GC-2 cells in various groups were detected by MDA detection kit;Western blotting method was used to detect the expression levels of long-chain acyl-CoA synthetase 4(ACSL4),heme oxygenase-1(HO-1),and glutathione peroxidase 4(GPX4)proteins in the cells in various groups;immunofluorescence staining was used to detect the expressions of ACSL4 protein in the cells in various groups.Results:Compared with control group,the differences in activities of the GC-2 cells in 5,10,and 20 μmol·L-1 AZA groups had no significant differences(P>0.05),while the activities of the GC-2 cells in 30 and 40 μmol·L-1 AZA groups were significantly decreased(P<0.01);therefore,the AZA concentration was selected to be within 20 μmol·L-1.Compared with control group,the differences of the activities of the GC-2 cells in 1,5,and 10 nmol·L-1 RSL3 groups had no significant differences(P>0.05),while the activities of the GC-2 cells in 50,100,500,and 1 000 nmol·L-1 RSL3 groups were significantly decreased(P<0.05 or P<0.01);therefore,the RSL3 concentration was set to be within 10 nmol·L-1.The GSH and MDA detection kits results showed that compared with control group,the levels of GSSG and MDA in the GC-2 cells in RSL3 group were significantly increased(P<0.05),while the GSH levels were significantly decreased(P<0.05);compared with RSL3 group,the levels of GSSG and MDA in the GC-2 cells in RSL3+Fer-1 group and RSL3+AZA group were significantly decreased(P<0.01),while the GSH levels were significantly increased(P<0.01).The Western blotting results showed that compared with control group,the expression levels of ACSL4 and HO-1 proteins in the GC-2 cells in RSL3 group were significantly increased(P<0.05),while the expression level of GPX4 protein was significantly decreased(P<0.01);compared with RSL3 group,the expression levels of GPX4 protein in the GC-2 cells in RSL3+Fer-1 group and RSL3+AZA group were significantly increased(P<0.05),while the expression levels of ACSL4 and HO-1 proteins were significantly decreased(P<0.01).The immunofluorescence staining results showed that compared with control group,the expression amount of ACSL4 protein in the GC-2 cells in RSL3 group was significantly increased,and compared with RSL3 group,the expression amounts of ACSL4 protein in the cells in RSL3+Fer-1 group and RSL3+AZA group were significantly decreased.Conclusion:AZA can alleviate the ferroptosis-induced by RSL3 in spermatocytes of the mice.
		                        		
		                        		
		                        		
		                        	
2.Feasibility and efficacy of pre-management of superior laryngeal artery in endoscopic surgery for hypopharyngeal cancer
Shisheng LI ; Shiying ZENG ; Qinglai TANG ; Gangcai ZHU ; Danhui YIN ; Xia PENG ; Qian YANG ; Xinming YANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2024;59(2):127-132
		                        		
		                        			
		                        			Objective:To explore the feasibility and efficacy for the dissection and ligation of the superior laryngeal artery in endoscopic surgery for hypopharyngeal cancer.Methods:Eight cadaveric heads were selected, and the laryngopharynxes were harvested. The positions of the superior laryngeal arteries entering the larynxes were dissected and observed under endoscopic vision, and their anatomical characteristics were summarized. Twenty-nine patients (all were male, aged 39-74 years old) with hypopharyngeal cancer who underwent transoral endoscopic surgery at the Department of Otorhinolaryngology Head and Neck Surgery of the Second Xiangya Hospital, Central South University from January 2018 to December 2019 were selected, and the patients were randomly divided into two groups by drawing lots, namely, the superior laryngeal artery was actively dissected and occluded during surgery in observation group ( n=15) or not in control group ( n=14). The differences in surgical time, bleeding volume, postoperative complications, and postoperative disease-free survival rate were compared between the two groups. Statistical analysis was conducted using SPSS 25.0 software. Results:The entry point of the superior laryngeal artery into the larynx was approximately at the level of the superior edge of the thyroid cartilage, and entered the larynx at the posterior one-third of the lateral wall of the pyriform fossa. The superior laryngeal artery might be determined through endoscopic exploration in all patients of observation group. The endoscopic surgery time [(40.00±7.56) minutes] and intraoperative bleeding volume [(24.00±8.28) ml] in the observation group were respectively less than those [(48.57±14.06) minutes and (42.86±15.41) ml] in the control group, and the differences were statistically significant ( t=-2.064, P=0.049; t=-4.064, P=0.001). There was no case with postoperative bleeding in the observation group, but with one case of postoperative bleeding in the control group. Total disease free survival rate was 86.2% and there was no significant difference in disease free survival rates between the two groups during a follow-up period of at least 36 months ( P=0.986). Conclusion:Dissection of the superior laryngeal artery during endoscopic surgery for hypopharyngeal cancer is feasible, and pre-management and occlusion of the superior laryngeal artery can effectively reduce intraoperative bleeding.
		                        		
		                        		
		                        		
		                        	
3.Reconstitution of double-negative T cells after cord blood transplantation and its predictive value for acute graft-versus-host disease
Tianzhong PAN ; Peng DING ; Aijie HUANG ; Baolin TANG ; Kaidi SONG ; Guangyu SUN ; Yue WU ; Shiying YANG ; Xingchi CHEN ; Dongyao WANG ; Xiaoyu ZHU
Chinese Medical Journal 2024;137(10):1207-1217
		                        		
		                        			
		                        			Background::With an increasing number of patients with hematological malignancies being treated with umbilical cord blood transplantation (UCBT), the correlation between immune reconstitution (IR) after UCBT and graft-versus-host disease (GVHD) has been reported successively, but reports on double-negative T (DNT) cell reconstitution and its association with acute GVHD (aGVHD) after UCBT are lacking.Methods::A population-based observational study was conducted among 131 patients with hematological malignancies who underwent single-unit UCBT as their first transplant at the Department of Hematology, the First Affiliated Hospital of USTC, between August 2018 and June 2021. IR differences were compared between the patients with and without aGVHD.Results::The absolute number of DNT cells in the healthy Chinese population was 109 (70-157)/μL, accounting for 5.82 (3.98-8.19)% of lymphocytes. DNT cells showed delayed recovery and could not reach their normal levels even one year after transplantation. Importantly, the absolute number and percentage of DNT cells were significantly higher in UCBT patients without aGVHD than in those with aGVHD within one year ( F = 4.684, P = 0.039 and F = 5.583, P = 0.026, respectively). In addition, the number of DNT cells in the first month after transplantation decreased significantly with the degree of aGVHD increased, and faster DNT cell reconstitution in the first month after UCBT was an independent protective factor for aGVHD (HR = 0.46, 95% confidence interval [CI]: 0.23-0.93; P = 0.031). Conclusions::Compared to the number of DNT cells in Chinese healthy people, the reconstitution of DNT cells in adults with hematological malignancies after UCBT was slow. In addition, the faster reconstitution of DNT cells in the early stage after transplantation was associated with a lower incidence of aGVHD.
		                        		
		                        		
		                        		
		                        	
4.Expert consensus on implementation strategy of awake prone positioning for non-intubated patients in China (2023).
Yuanyuan MI ; Zheyi CAI ; Jing LIU ; Fei TIAN ; Liping YANG ; Lei BAO ; Shanbing HOU ; Su GU ; Li LI ; Xueli ZHOU ; Yun XU ; Shumei ZHANG ; Xiaoxia FU ; Xiaodi LI ; Chuansheng LI ; Liang SUN ; Xiaohong ZHANG ; Hong QI ; Shiying YUAN ; Liqun ZHU ; Haiyan HUANG ; You SHANG
Chinese Critical Care Medicine 2023;35(4):337-351
		                        		
		                        			
		                        			The awake prone position plays an important role in the treatment of hypoxemia and the improvement of respiratory distress symptoms in non-intubated patients. It is widely used in clinical practice because of its simple operation, safety, and economy. To enable clinical medical staff to scientifically and normatively implement prone position for awake patients without intubation, the committees of consensus formulation, guided by evidence-based methodology and Delphi method, conducted literature search, literature quality evaluation and evidence synthesis around seven topics, including indications and contraindications, evaluation, implementation, monitoring and safety management, termination time, complication prevention and health education of awake prone position. After two rounds of expert letter consultation, Expert consensus on implementation strategy of awake prone positioning for non-intubated patients in China (2023) was formulated, and provide guidance for clinical medical staff.
		                        		
		                        		
		                        		
		                        			Humans
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		                        			Consensus
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		                        			Prone Position
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		                        			Wakefulness
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		                        			China
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		                        			Dyspnea
		                        			
		                        		
		                        	
5.Investigation on medical practice of overseas acupuncturists based on the top 5 general hospitals in the United States of America and the inspiration
Shuaishuai LIU ; Shiying HUANG ; Lu GAN ; Yang LUO ; Leyao XIAN ; Luwen ZHANG
International Journal of Traditional Chinese Medicine 2023;45(6):667-672
		                        		
		                        			
		                        			This study selected the top five general hospitals in the United States of America, collected the practice information of acupuncturists through the official website, and analyzed the professional department, expertise, professional title, education background, residency, fellowship, board certification, etc. of these acupuncturists to understand the practice situation of acupuncturists in the hospitals. The results of the study showed the practice of acupuncturists in the United States of America is improving. With further localization, locally trained acupuncturists have gradually become the main body of acupuncturists; acupuncture treatment is still mainly for analgesia, but the scope of treatment continues to expand, and departments that accept acupuncturists are gradually increasing. However, the group of licensed acupuncturists is still a minority in the group of acupuncturists in the United States of America. Native American physicians still use acupuncture as an alternative option and neglect to obtain a license. The entry barrier for acupuncturists still needs to be raised.
		                        		
		                        		
		                        		
		                        	
6.Expression of cold-inducible RNA-binding protein in radiation-induced lung injury model
Shiying NIU ; Changsheng CONG ; Meili SUN ; Yifan JIANG ; Tong YANG ; Zhaopeng WANG ; Yueying ZHANG
Chinese Journal of Radiological Health 2022;31(1):33-38
		                        		
		                        			
		                        			Objective To investigate the changes in the expression of cold-inducible RNA-binding protein (CIRBP) in a radiation-induced lung injury model. Methods Thirty male C57BL/6 mice were randomly divided by body weight into control group (no intervention) and model group (single chest X-ray irradiation with a dose of 20 Gy to build a radiation-induced lung injury model). The mice were dissected five weeks after irradiation. Hematoxylin-eosin staining and Masson staining were used to observe the pathological changes of the lung tissue and the deposition of collagen fibers. Immunohistochemistry was used to measure the expression of the inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the lung tissue. qRT-PCR was used to measure the expression of CIRBP mRNA in the lung tissue. The expression of CIRBP protein in the lung tissue was determined by the immunofluorescence assay and Western blot. Results Compared with the control group, the model group showed significant pulmonary vascular congestion, significant inflammatory cell infiltration, significant thickening of some alveolar septa, significantly increased IL-6 expression [(129.41 ± 5.58) vs (187.22 ± 34.77), t = 3.179, P < 0.05], significantly increased TNF-α expression [(137.52 ± 23.53) vs (187.02 ± 19.16), t = 5.069, P < 0.05], significantly increased CIRBP mRNA expression [(1 ± 0.08) vs (1.97 ± 0.39), t = 3.45, P < 0.05], and significantly increased CIRBP protein expression [(9.32 ± 1.26) vs (14.76 ± 1.61), t = 3.751, P < 0.05], by the immunofluorescence assay; [(1.13 ± 0.17) vs (1.49 ± 0.14), t = 2.819, P < 0.05], by Western blot). Conclusion The expression of CIRBP is significantly increased in the radiation-induced lung injury model, which may be an important pro-inflammatory factor in radiation-induced lung injury.
		                        		
		                        		
		                        		
		                        	
8.Enlightenment of the cooperative mode between social and commercial of Medicare in the United States on development of " City-customized Medical Insurance" in China
Yangyang ZHENG ; Meng YANG ; Minglong FU ; Shiying JIANG ; Kai LIAN
Chinese Journal of Hospital Administration 2022;38(12):896-900
		                        		
		                        			
		                        			As a powerful attempt by government to promote the construction of the multi-level healthcare security system and social and commercial integration in China, " City-customized Medical Insurance" still has many problems to be solved at the beginning of its development, such as unclear boundary between government and enterprises, limited coverage and strength of security. On the basis of clarifying the current situation of " City-customized Medical Insurance", and combing the management experience of social and commercial integration in Medicare Part C plan of the United States, the authors put forward that China should make full use of the advantages of the combination of promising government and efficient market, guide differentiated product design, and establish market access and evaluation mechanism, so as to promote the effective connection between China′s commercial health insurance and basic healthcare insurance, and further reduce the people′s medical burden.
		                        		
		                        		
		                        		
		                        	
9.Feasibility and efficacy of preserving internal branch of superior laryngeal nerve in endoscopic surgery for hypopharyngeal squamous cancer: an observational study
Qian YANG ; Shisheng LI ; Qinglai TANG ; Xinming YANG ; Zi′an XIAO ; Xia PENG ; Gangcai ZHU ; Danhui YIN ; Peiying HUANG ; Shiying ZENG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2022;57(12):1463-1469
		                        		
		                        			
		                        			Objective:This study was performed to investigate the feasibility of preservation of internal branch of superior laryngeal nerve(ibSLN) during transoral endoscopic surgery for hypopharyngeal squamous cancer(HSCC) and the influence on patient′s swallowing function after operation.Methods:From May 2020 to June 2021, the data of 29 HSCC patients who required for transoral endoscopic surgery in the Department of Otorhinolaryngology Head and Neck Surgery, the Second Xiangya Hospital of Central South University were prospectively included, and the included patients were divided into two groups randomly by lottery. According to whether ibSLN was actively dissected during operation, they were divided into ibSLN preservation group ( n=15) and control group ( n=14, without ibSLN preservation). Operation time, intraoperative hemorrhage, intraoperative neck dissection, postoperative radiotherapy, postoperative recurrence within 1 year, retention and swallowing function, the recovery of oral soft diet and the quality of life were compared between two groups. SPSS 25.0 software was used for statistical analysis. Results:The study included 29 eligible patients, including 25 males and 4 females.The age ranged from 42 to 67 (56.07±5.93) years. There were no significant differences( P>0.05) between 2 groups in the following data,including age( t=-0.56), gender( χ2=0.01), TNM stage(T stage χ2=0.29, N stage χ2=0.02), pathological diagnosis( χ2=0.03), preoperative swallowing function( χ2=0.00) and M. D. Anderson Dysphagia Inventory(MDADI) score(global t=0.55, emotional t=0.16, functional t=0.60, physical t=0.64), operation time( t=1.62) and intraoperative hemorrhage( t=-1.46), intraoperative neck dissection( χ2=0.01), postoperative radiotherapy( χ2=0.32), postoperative recurrence within 1 year( P>0.050). The swallowing function was evaluated by water swallowing test after operation. The swallowing function of ibSLN preservation group was better than control group, and the difference between two groups was statistically significant on the 1st ( χ2=4.44, P=0.035), 5th ( χ2=4.24, P=0.039) and 7th ( χ2=4.55, P=0.033) day after operation. On the 14th day after operation, the MDADI scores of patients in the ibSLN preservation group were higher than those in the control group in global ( t=2.45, P=0.021), functional ( t=2.54, P=0.017) and physical ( t=2.24, P=0.034) dimensions, except for emotional dimension ( t=1.89, P=0.070). The median time of oral soft diet( U=23.00, P<0.001), normal oral diet( U=21.00, P<0.001) and the nasogastric tube removal time ( U=18.50, P<0.001) in ibSLN preservation group was 2 days, 5 days and 6 days respectively, earlier than that in control group, which had statistically significant difference. Conclusion:Our results show that it is feasible to preserve the ibSLN during HSCC transoral endoscopic surgery, which can achieve rapid recovery of postoperative swallowing function.
		                        		
		                        		
		                        		
		                        	
10.Mechanism of Talibin-1 regulating vascular remodeling of aortic dissection in mice
Yudong SUN ; Shiying WANG ; Jiang ZHU ; Tonglei HAN ; Dihao WEN ; Jing YANG ; Zaiping JING ; Jian ZHOU ; Zhiqing ZHAO ; Xiaolong WEI
Chinese Journal of General Surgery 2021;36(1):25-29
		                        		
		                        			
		                        			Objective:To explore the role and mechanism of Talin-1 in mouse aortic dissection.Methods:Sixty male FVB mice were evenly divided into groups of blank, model, Talin-1 up-regulation, Talin-1 up-regulation control, Talin-1 down-regulation, and Talin-1 down-regulation control. Except mice in the blank group, mice were treated with β-aminopropionitrile (BAPN) combined with angiotensin to construct a mouse aortic dissection model. Hematoxylin-eosin and vascular elastic fiber staining (EVG) were used to observe the aorta and elastic fiber morphology and structure. Western blot was used to detect the phosphorylation levels of FAK and ERK1 / 2 in mouse aortic tissue.Results:The success rate of aortic dissection in model mice was 70%, and there was no aortic dissection appeared in the blank group.No mice died during the experiment. The incidence of aortic dissection in the Talin-1 down-regulated group was 100%, which was significantly higher than that in the Talin-1 down-regulated control group( P<0.05). The incidence of aortic dissection in the Talin-1 up-regulated group was 20%, significantly lower than that in the Talin-1 up-regulated control group. The wall thickness of the aorta of mice in the Talin-1 down-regulated group was accompanied by hematoma or pseudocavity formation. The median elastic fiber content was higher than that in the Talin-1 downregulation control group( P<0.05). The content of elastic fibers in the blood vessel wall of mice in the Talin-1 up-regulation group was significantly higher than that in the Talin-1 up-regulation control group.The down-regulation of Talin-1 significantly inhibited FAK phosphorylation, and instead promoted ERK1/2 phosphorylation( P<0.05). Conclusions:Down-regulation of Talin-1 may reduce the elastic fiber content in the aorta of mice by activating the ERK1/2 signaling pathway, leading to vascular remodeling of the aortic wall and promoting the occurrence of aortic dissection.
		                        		
		                        		
		                        		
		                        	
            
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