1.Glutathione Detection Method Based on Electron Paramagnetic Resonance Spectroscopy
Zhi-Wen WANG ; Jian KUANG ; Ao-Kun LIU ; Ruo-Tong WEI ; Lu YU ; Chang-Lin TIAN
Progress in Biochemistry and Biophysics 2024;51(11):3034-3045
ObjectiveGlutathione (γ-glutamyl-L-cysteinylglycine, GSH) is the most abundant non-protein compound containing sulfhydryl (―SH) groups in cells. It serves as a source of reducing equivalents, effectively neutralizing harmful reactive substances, and playing a crucial role in maintaining cellular redox balance. Therefore, sensitive detection and accurate measurement of GSH levels in tissues are of great importance. In this work, we presents a novel method for GSH detection utilizing electron paramagnetic resonance (EPR) spectroscopy. MethodsInitially, ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate acid)) solution was mixed with K2S2O8 solution and reacted in the dark for 12 to 16 h to prepare ABTS·+ solution, which was then quantified using UV-Vis spectroscopy. Subsequently, the concentration of glutathione (GSH) was determined based on the changes in the EPR signal of ABTS·+. On this basis, the optimal reaction time and temperature were explored to establish a standard equation correlating the EPR signal intensity of ABTS·+ with GSH concentration. Finally, the derived standard curve was employed to quantitatively analyze the GSH concentration in whole blood from C57BL/6J mice, and the results were compared with those reported in the literature to verify the accuracy of the method. ResultsThe experimental results demonstrate that this method has a linear detection range from50 nmol/L to 15 μmol/L for GSH, spanning two orders of magnitude, with a limit of detection (LOD) at0.50 nmol/L. The measured GSH content in mouse whole blood is (10 660±706) nmol/g Hb, which agrees with the value of (11 200±237) nmol/g Hb as previously reported. Furthermore, a similar method was developed for detection of glutathione disulfide (GSSG) at higher reaction temperature. ConclusionThis article presents a novel assay for the rapid detection of GSH using the intensity of EPR signal from ABTS·+ as indicator. This method demonstrates enhanced detection sensitivity and a broader linear range compared to conventional colorimetric methods. Furthermore, we have extended the application of this method to detect GSH content in blood samples efficiently and accurately, offering valuable information for assessing tissue redox balance, thus holding significant potentials.
2.Study on drug properties of Arisaematis Rhizoma and Arisaema Cum Bile based on substance and energy metabolism in normal and cold/heat syndrome model rats.
Fa-Zhi SU ; Chen-Xi BAI ; Wen-Sen ZHANG ; Jing YANG ; Jian-Ping HU ; Yan-Ping SUN ; Bing-You YANG ; Hai-Xue KUANG ; Qiu-Hong WANG
China Journal of Chinese Materia Medica 2022;47(17):4682-4690
This paper clarified the scientific connotation of the changes in cold and heat properties of Arisaematis Rhizoma and Arisaema Cum Bile through investigating the changes of substance and energy metabolism after drug intervention in the rats with normal and cold/heat syndrome, so as to improve the method of evaluating the drug properties of Chinese medicine. After one week of adaptive feeding, healthy male SD rats were randomly divided into three parts: normal rats, heat syndrome rat models, and cold syndrome rat models. Through ice water bath and oral euthyrox(120 μg·kg~(-1)), the models of cold syndrome and heat syndrome were induced, respectively. The models were made at 9:00 am. and administrated by gavage at 3:00 pm. every day. All administration groups were administrated with Arisaematis Rhizoma and Arisaema Cum Bile decoction, respectively, and the blank group was given the same dose of normal saline. After continuous administration for 15 d, the rats were anesthetized by chloral hydrate, blood was taken from abdominal aorta, and the hearts and livers were removed and stored at-80 ℃. The changes in the body weight and anal temperature of rats during administration were detected, and the liver coefficient of rats was detected after removing the liver. Enzyme-linked immunosorbent assay(ELISA) was adopted to detect the expression level of the indexes related to substance and energy metabolism in liver and heart of rat, and Western blot was used to detect the expression of key proteins in AMPK/mTOR signaling pathway for further verification. The results showed that Arisaematis Rhizoma enhanced the expression level of enzymes related to substance and energy metabolism in the normal and cold and heat syndrome rat models, and increased anal temperature, which exhibited warm(hot) drug property. Arisaema Cum Bile inhibited the level of substance and energy metabolism in rats, and reduced anal temperature, which showed cold(cool) drug property. Chinese Pharmacopoeia has recorded "Arisaematis Rhizoma has warm property and Arisaema Cum Bile has cool property", which is consistent with the phenomenon in this study. Therefore, it is feasible to evaluate the drug properties of Chinese medicine based on the substance and energy metabolism of normal and cold/heat syndrome model rats, which completes the method of evaluating drug properties of Chinese medicine.
AMP-Activated Protein Kinases
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Animals
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Arisaema/chemistry*
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Bile
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Chloral Hydrate
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Cold-Shock Response/drug effects*
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Drugs, Chinese Herbal/therapeutic use*
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Energy Metabolism
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Heat Stroke/therapy*
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Hot Temperature
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Male
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Rats
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Rats, Sprague-Dawley
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Saline Solution
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Syndrome
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TOR Serine-Threonine Kinases
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Thyroxine
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Water
3.Best evidence summary of percutaneous pericardial drainage tube nursing
Xuemei LIU ; Yuqin GU ; Hongjing YU ; Wanting LI ; Xiaocui CAO ; Liying YUAN ; Jian SONG ; Yongyan KUANG ; Caiyin ZHOU ; Lin YANG ; Wen LI ; Donglan LING ; Wei GUO
Chinese Journal of Modern Nursing 2021;27(32):4453-4458
Objective:To select the relevant evidence of percutaneous pericardial drainage tube nursing and summarize the best evidence.Methods:Evidence-based questions were established based on PIPOST model. BMJ Best Clinical Practice, China Biology Medicine disc (CBM) , UpTodate, Cochrane Library, Joanna Briggs Institute Evidence-based Health Care Database, China Guide Network, British Guide Network, National Guide Line Clearing House (NGC) , PubMed, EMbase, Evidence-based Medicine (EBM) , Registered Nurses' Association of Ontario (RNAO) , The European Society of Cardiology (ESC) , American Heart Association (AHA) , Chinese Journal Full-text Database and Wanfang Database were conducted computer retrieval. The search time was from the establishment of the database to December 31, 2020. Two researchers respectively evaluated the quality of the included literature and extracted data and summarized and summarized the evidence that met the standards.Results:Finally, 12 articles were included, including 1 evidence summary, 2 systematic reviews, 1 systematic assesment, 2 guidelines, 1 expert consensus and 5 case series studies. Finally, 11 pieces of evidence were formed, including 6 themes such as drainage tube selection, puncture wound nursing, drainage flow control, flushing and sealing of the tube, observation and recording points, extubation indications and care.Conclusions:This study summarizes the best evidence for percutaneous pericardial drainage tube nursing, which provides evidence-based basis for improving the quality of percutaneous pericardial drainage tube care.
4.Application of Intermittent Lung Inflation Combined with Rigid Ureteroscopy in Treating Upper Ureteral Stones That Were Not Fully Visible.
Feng Jin WANG ; Hua Jian GUO ; Guo Wei ZHENG ; Hua Qian KUANG ; Wei Wei QU ; Bin Hong ZHU ; Zhao Zhen HUANG ; Yun Wen SHU ; Pei HUANG ; Hua Jian WU
Acta Academiae Medicinae Sinicae 2019;41(6):793-798
To analyze the effectiveness and safety of intermittent lung inflation combined with rigid ureteroscopy in the treatment of upper ureteral stones that were not fully visible. The clinical and imaging data of 56 patients with upper ureteral stone undergoing rigid ureteroscopic lithotripsy combined with intermittent lung inflation in Zhejiang Quhua Hospital from March 2016 to October 2017 were retrospectively analyzed.Intermittentt lung inflation was used to change and stabilize the position of ureteral calculi during the operation,so as to ensure the visual field of ureteroscopy.Holmium laser lithotripsy was performed to remove the stones.Urinary tract abdominal plain X-ray or CT urography was performed 1 and 3 months after the operation to evaluate the residual stones and the clinical efficacy. Stones were successfully removed after a single attempt in 48 patients.In 5 patients,stones escaped into the kidney during ureteroscopic lithotripsy,and thus flexible ureteroscopy were performed.In 3 patients,a second session of auxiliary procedure was required,among whom 2 patients received extracorporeal shock wave lithotripsy and 1 patient underwent extracorporeal shock wave lithotripsy+ureteroscopic lithotripsy.The stone-free rates 1 and 3 months after surgery were 94.6%(53/56)and 100%(56/56),respectively.No severe complication such as ureter perforation,gross hematuria,septic shock,or pneumothorax occurred during and after surgery. Intermittent lung inflation in tracheal intubation under general anesthesia in patients with proximal ureteral stones that can not be fully visible during rigid ureteroscopic lithotripsy was feasible and reliable.It can effectively change the location of stones and thus enable safe and effective lithotripy.It expands the indications of rigid ureteroscopy for treating upper ureteral stones.
Humans
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Lithotripsy, Laser
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Retrospective Studies
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Ureteral Calculi
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diagnostic imaging
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Ureteroscopes
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Ureteroscopy
5.Downregulation of iASPP Expression Suppresses Proliferation, Invasion and Increases Chemosensitivity to Paclitaxel of Head and Neck Squamous Cell Carcinoma .
Zheng-Zheng LIU ; Wei-Lu KUANG ; Wen-Jing ZENG ; Jian-Yun XIAO ; Yong-Quan TIAN
Chinese Medical Sciences Journal 2019;34(3):184-193
Objective Our previous study has revealed that iASPP is elevated in human head and neck squamous cell carcinoma (HNSCC) and iASPP overexpression signifcantly correlates with tumor malignant progression and poor survival of HNSCC. This study investigated the function of iASPP playing in proliferation and invasion of HNSCC . Methods HNSCC cell line Tu686 transfected with Lentiviral vector-mediated iASPP-specific shRNA and control shRNA were named the shRNA-iASPP group and shRNA-NC group, respectively. The non-infected Tu686 cells were named the CON group. CCK-8 assay, flow cytometry, transwell invasion assay were performed to detect the effects of iASPP inhibition . Results Our results demonstrated that the proliferation of shRNA-iASPP cells at the time of 72 h (=32.459, =0.000), 96 h (=51.407, =0.000), 120 h (=35.125, =0.000) post-transfection, was significantly lower than that of shRNA-NC cells and CON cells. The apoptosis ratio of shRNA-iASPP cells was 9.42% ± 0.39% (=299.490, =0.000), which was significantly higher than that of CON cells (2.80% ± 0.42%) and shRNA-NC cells (3.18% ± 0.28%). The percentage of shRNA-iASPP cells in G0/G1 phase was 74.65% ± 1.09% (=388.901, =0.000), which was strikingly increased, compared with that of CON cells (55.19% ± 1.02%) and shRNA-NC cells (54.62% ± 0.88%). The number of invading cells was 56 ± 4 in the shRNA-iASPP group (=84.965, =0.000), which decreased significantly, compared with the CON group (111 ± 3) and the shRNA-NC group (105 ± 8). The survival rate of shRNA-iASPP cells administrated with paclitaxel was highly decreased, compared with CON cells and shRNA-NC cells (=634.841, =0.000). Conclusion These results suggest iASPP may play an important role in progression and aggressive behavior of HNSCC and may be an efficient chemotherapeutic target for the treatment of HNSCC.
6. Analysis of mortality trend and related factors for aged in-patients with diabetes mellitus
Li-na ZHANG ; Yu-qiong WEN ; Xiao-ying FU ; Jin-xin LIN ; Shui-qing LAI ; Hong-mei CHEN ; Si-min LIU ; Jian KUANG
Chinese Journal of Practical Internal Medicine 2019;39(09):809-812
OBJECTIVE: To find out the mortality trend and related factors in aged hospitalized patients with diabetes mellitus(DM). METHODS: The case information diabetic in patients who died during the period from 2005 to 2014 were collected and the mortality and causes of death were analyzed. RESULTS: From 2005 to 2014, 1297 diabetic patients died, and the mortality of elderly DM inpatients was 4.44%(1162 cases), significantly higher than that of the non-elderly of 0.94%(P<0.001). The death rate of elderly diabetic patients was significantly higher in males than in females(5.22% vs. 3.47%, P<0.001). The mortality of the aged diabetic patients decreased within 10 years(P<0.001), decreasing from 4.75% in 2005 to 3.01% in 2009(P<0.001) in the year of 2005-2009, while there were no differences in the year of 2010-2014. The main death causes of the aged diabetic in-patients were as follows: infections(27.71%), cardiovascular diseases(25.22%), tumor(21.34%), cerebral vascular diseases(10.41%) and diabetic complications(5.51%). The first death cause in the 60-79 yrs group was cardiovascular diseases, while in the ≥80 yrs group, it was infections. The constituent ratio of infection as death cause in the aged during 2010-2014 significantly increased(22.60% vs. 32.50%, P<0.001), increasing by 43.81%, and it became the first cause of death in 2010. CONCLUSION: The death rate of the elderly DM in-patients has decreased significantly within 10 years, from 2005 to 2014, while the rate has kept steady from 2010. Infections and cardiovascular diseases are the main cause of death. So it's important to prevent the elderly hospitalized DM patients from infection, in addition to cardiovascular diseases, and to control in time.
7.Assessment of hypoglycemic status among hospitalized elderly patients with type 2 diabetes.
Xiang-Hua WENG ; Yu-Qiong WEN ; Shu-Ting ZHANG ; Xiao-Ying FU ; Hong-Mei CHEN ; Liang CHEN ; Jian-Hao PEI ; Si-Min LIU ; Jian KUANG
Journal of Southern Medical University 2018;38(5):591-595
OBJECTIVETo investigate the hypoglycemic characteristics of hospitalized elderly patients with type 2 diabetes mellitus (T2DM).
METHODSFrom January, 2014 to December, 2015, the data of 58 565 blood measurements using a standard blood glucose monitoring system (BGMS) were collected from 1187 cases of patients with type 2 diabetes during hospitalization in the Department of Endocrinology, Guangdong General Hospital (Guangzhou, China). Stratified analyses were conducted by dividing the patients into 3 age groups, namely <45 years group (128 cases), 45-64 years group (594 cases), and ≥65 years group (465 cases). The incidence and time distribution of hypoglycemia in these patients were compared among the 3 age groups.
RESULTSThe risk of hypoglycemia increased with age. Compared with those below 45 years of age, the patients beyond or equal to 65 years had a significantly increased hypoglycemic density (0.95% vs 0.40%, P<0.001), a higher proportion of patients with hypoglycemia (28.17% vs 10.94%, P<0.001), and greater patient-days with hypoglycemia (4.48% vs 1.76%, P<0.001). In the elderly patients, hypoglycemia occurred most frequently before dawn, at which time the hypoglycemic density was 2.66% in patients ≥65 years of age, significantly higher than that in patients below 45 years (1.09%, P<0.05) and between 45 and 64 years (1.90%, P<0.05); the proportion of patients with hypoglycemia was also significantly higher in the elderly patients (14.57%) than in those below 45 years (3.77%, P<0.02) and between 45 and 64 years (9.42%, P<0.02). The proportion of patients with recurrent hypoglycemia (≥2 times) was significantly higher in patients ≥65 years (13.33%) than in younger patients (2.34% in <45 years group and 9.43% in 45-64 years group, P<0.05).
CONCLUSIONThe hypoglycemic risk in hospitalized elderly patients with T2DM is significantly higher than that in younger patients, especially before dawn and in terms of recurrent hypoglycemia. Clinicians should develop differential blood glucose monitoring and management strategies for these elderly patients to improve the clinical safety.
8.Contrast Researches of Liver Biopsy and Ultrasound in the Diagnosis of Fatty Liver
Lin ZHUANG ; Ming-Fen HU ; Xing-Mei PANG ; Bao-Li BAI ; Lin WANG ; Hong-Jie YANG ; Yun-Li LI ; Zhi-Jian DONG ; Wen-Qing HUAN ; Qing-Qing WANG ; Wei-Qin YU ; Xiao-Lin KUANG
Journal of Kunming Medical University 2018;39(1):85-91
Objective To evaluate the significance of liver biopsy and B ultrasonograpgy in the diagnosis of fatty liver. Methods The results of 62 patients with liver steatosis diagnosed by liver puncture biopsy but not by B-ultrasonograpgy were contrastively analyzed and combined with liver function, blood lipids, blood glucose, and body mass index. Results The 62 cases which were not diagnosed as fatty liver by B-ultrasonograpgy were proved to be 5%-33%liver steatosis after liver puncture biopsy. Among the 62 cases, 23 cases were indicated by the B-ultrasonograpgy that the liver parenchyma echo did not see abnormalities, 18 cases showed the liver parenchyma echo slightly was enlarged, 17 cases showed the liver parenchyma echo density was a bit enhanced and 4 cases were diffuse liver damage,which respectively were 37.01%、29.03%、27.42%and 6.45%. Pathologically it indicated that 45 cases were 5%≤liver steatosis≤19%. Among the 45 cases, 18 cases were indicated by the B-ultrasonograpgy that the liver parenchyma echo was not seen abnormalities, 8 cases showed the liver parenchyma echo slightly was enlarged, 17 cases showed the liver parenchyma echo density was a bit enhanced, and 2 cases were diffuse liver damage, and the change of ultrasound was mainly showed by the liver parenchyma echo not seen abnormalities and the enhanced liver parenchyma echo density. Pathologically it indicated that 17 cases were 20%≤liver steatosis≤33%, 6 cases were indicated by the B-ultrasonograpgy that the liver parenchyma echo did not see abnormalities, 5 cases showed the liver parenchyma echo slightly was enlarged, 5 cases showed the liver parenchyma echo density was a bit enhanced, and 1 case was diffuse liver damage, and the change of ultrasound was mainly showed by the liver parenchyma echo not seen abnormalities, the slightly enlarged liver parenchyma echo and the enhanced liver parenchyma echo density. By analyzing the influence to the ultrasound changes by patients' liver function, body mass index, blood fat and blood sugar, and with logistic regression analysis through a disorderly classification, it was found that the larger value of the glutamine transferase, alkaline phosphatase, body mass index, triglyceride and low density lipoprotein cholesterol, the higher possibility of diffuse liver damage, and the higher level of fatty degeneration, the higher possibility of diffuse liver damage. Conclusion In the diagnosis of fatty liver, when the fatty degeneration is below 1/3, B-ultrasonic examination can't show characteristic changes of fatty liver. It should be closely observed or take liver puncture biopsy to make a definite diagnosis of fatty liver.
9.Gene Analysis for the Sudden Death of Hypertrophic Cardiomyopathy by Whole Exome Sequencing
chao Chuan XU ; zhi Yun BAI ; shu Xin XU ; li Guo L(U) ; ping Xiao LAI ; Rui CHEN ; guang Han LIN ; jian Wen KUANG
Journal of Forensic Medicine 2017;33(4):339-343
Objective To analyze the related pathogenicity gene mutations in a sudden death of hypertrophic cardiomyopathy (HCM) on whole exome level.Methods Whole exome sequencing (WES) was been performed on a sudden death case sample with pathological features of HCM by Illumina(R) Hiseq 2500 platform.Using hgl9 as the reference sequences,the sequencing data were analyzed.Suspicious single nucleotide variants (SNV) were screened,and the conservatism and function were analyzed by the software such as PhyloP,PolyPhen-2,SIFT,etc.Results After screening,a heterozygous mutation C719R was finally identified in the gene MYBPC3 of this case.Conclusion The molecular anatomy on whole exome level by second generation sequencing technology can help to define the molecular mechanism of HCM and provide a new mothed and thought for analysis of death cause.
10.Application of SEMG to study the effects of imagery training on back-style high jump.
Wen-Feng LIU ; Yong-Ling CHANG ; Chang-Fa TANG ; Zhen-Zhen HONG ; Li-Qin YIN ; Jin CHEN ; Wen-Ning REN ; Long JIANG ; Jian KUANG
Chinese Journal of Applied Physiology 2013;29(3):260-270
Adolescent
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Adult
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Athletic Performance
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psychology
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Back
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physiology
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Electromyography
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Exercise
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physiology
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Humans
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Imagery (Psychotherapy)
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Male
;
Young Adult

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