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This article introduces the clinical approach and acupuncture characteristics of the traditional Chinese medicine practitioner Professor YU Hai-Bo in treating paediatric cerebral palsy using the"Jianpi Yishen Triple-Needle Grouping Acupoints".Guided by the theory of growth and development of"viscera-meridian-brain"growth and development,Professor YU believed that"insufficiency of spleen and kidney"is the core pathogenesis of paediatric cerebral palsy,and the treatment concept of"treating from the spleen and kidney"was proposed.He inherited and innovated the triple-needle grouping acupoints therapy and establishing the system of"Jianpi Yishen Triple-Needle Grouping Acupoints".Before regular acupuncture,the abdomen and dorsum are pricked to freely regulate the middle energizer,and the upper limbs are selected as"Hegu(LI4),Waiguan(SJ5),Quchi[(LI11),three acupoints on the hand]+ Neiguan(PC6)";the lower limbs are selected as"Zusanli(ST36),Sanyinjiao(SP6),Taichong[(LR3),three acupoints on the foot];"Shenmai(BL62),Zhaohai(KI6),Yongquan(KI1)",spleen and kidney are regulated simultaneously,and the head acupoints include Sishencong(EX-HN1),intelligence tri-needling,cerebral tri-needling,temporal tri-needling,mind-calming needling and bilateral Fengchi(GB20).In order to regulate the spirit and benefit the intellect,the matching acupoints are modified according to the disease and the syndromes.At the same time,it is supplemented with music therapy and auricular point seed-pressing.Emphasis is placed on the simultaneous regulation of"child-parent-doctor"and"treating the person"rather than the"treating the disease".
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Objective To identify inflammation-related genes in atrial fibrillation(AF)and explore the possible role and mechanism of these genes and infiltrating immune cells in the development of AF.Methods A series of bioinformatics analysis combined with machine learning algorithms to identify biomarkers of AF,the receiver operating characteristic(ROC)curves were used to verify the prediction and diagnostic value of key genes,and Spearman correlation analysis was used to clarify the correlation between key genes and infiltrating immune cells.Results 593 differential genes(| log2(fold change,FC)|>1,P<0.05),7 immune cell subtypes(P<0.05)were selected,190 immune-related differential genes were obtained,3 biomarkers(IGF1,PTGS 2 and PPARG),and the correlation analysis showed that 3 markers were significantly associated with infiltrating immune cells(P<0.05).Conclusion IGF1,PTGS2 and PPARG are inflammation-related genes of AF,which are speculated to be closely related to the process and pathway of immune cell infiltration.
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Objective@#To investigate fast food consumption behaviors among 4th and 5th grade primary school students in agricultural and pastoral areas in Qinghai Province, and to provide scientific basis for nutrition education and intervention measures for school age children in agricultural and pastoral areas.@*Methods@#Using multi stage stratified cluster random sampling method, a total of 969 fourth and fifth grade students were selected from 10 primary schools in 4 counties and districts in 3 cities of Haidong City, Hainan Prefecture and Haixi Prefecture from Qinghai Province. Self administered questionnaires were used to investigate the fast food consumption behaviors of participants.@*Results@#The proportions of senior primary school students in agricultural and pastoral areas of Qinghai Province who had consumed western fast food and traditioal fast food in the past week were 45.8% and 75.6%, respectively, with statistically significant differences ( χ 2=290.24, P <0.05). The times of traditional fast food consumption in the past week among boarding students were higher than that of non boarding students( Z =6.44,5.84, P <0.05). The main reasons for senior primary school students in agricultural and pastoral areas of Qinghai Province to choose to consume western fast food were that it was delicious (84.7%), nutritious (62.6%), clean and hygienic (57.4%), and a better environment (57.0%). The top 4 reasons for chousing Chinese fast food were yummy(83.8%),nutritious(82.8%),clean and healthy(67.4%),and good environment(53.5%). Among the surveyed primary school students, 64.7% believed that diet structure of Chinese fast food was reasonable, 43.0% believed that the nutritional value of Chinese fast food was high, and 39.4% believed that the energy content of western fast food was high.@*Conclusion@#Fast food consumption is a common dietary behavior of senior primary school students in agricultural and pastoral areas of Qinghai Province. Relevant departments should strengthen the nutrition education related to fast food, promote the dissemination of health knowledge, enable students to develop a good lifestyle and reduce fast food consumption.
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Objective:To observe the effects of Banxia Shumi Decoction on 5-HT 1AR, 5-HT 2AR, 5-HT, and 5-HIAA of chronic insomnia (CI) rats with internal obstruction of phlegm-damp (IOPD) type, to investigate the mechanisms of Banxia Shumi Decoction on resolving and draining dampness, guiding yang into yin and tranquilizing mind. Methods:A total of 48 Wistar rats were divided into control group, model group, Banxia Shumi Decoction low-dosage group, medium-dosage group, high-dosage group, and diazepam group according to random number table method, with 8 rats in each group. Except the control group, the CI with IOPD rats model were prepared by the method of "high-fat diet + single-platform water environment" in other groups. The rats in the Banxia Shumi Decoction low-, medium-, high-dosage group were treated with Banxia Shumi Decoction by gavage at the dose of 4.69, 9.38 and 18.75 g/kg respectively, the rats in the diazepam group were given 0.52 mg/kg diazepam aqueous solution by gavage, and the rats in the control group and model group were given the equal volume normal saline, once a day for consecutive 2 weeks. The mRNA expressions of 5-HT 1AR, 5-HT 2AR in rat brain stem were detected by qPCR, the protein expressions of 5-HT 1AR, 5-HT 2AR in rat brain raphe nucleus were detected by Western blot, and the contents of 5-HT and 5-HIAA in rat brain stem were determined by HPLC-MS. Results:Compared with model group, the expression of 5-HT 1AR mRNA significantly increased in the Banxia Shumi Decoction low-, medium-, high-dosage group, and diazepam group ( P<0.01); the expression of 5-HT 2AR mRNA significantly decreased in the Banxia Shumi Decoction high-dosage group and diazepam group ( P<0.05), and the expression of 5-HT 1AR and 5-HT 2AR significantly increased in the Banxia Shumi Decoction high-dosage group and diazepam group ( P<0.05 or P<0.01); 5-HT content significantly increased in the Banxia Shumi Decoction medium-, high-dosage group and diazepam group ( P<0.05 or P<0.01); 5-HIAA content significantly increased in the Banxia Shumi Decoction low-, medium-, high-dosage group, and diazepam group ( P<0.05 or P<0.01). Conclusion:Banxia Shumi Decoction may intervene CI with IOPD type and perform the actions of resolving and draining dampness, guiding yang into yin and tranquilizing mind by regulating the expressions of 5-HT 1AR, 5-HT 2AR, 5-HT and 5-HIAA.
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Abstract@#The growth and development of children is related to the future of the country and the nation. In recent years, there have been more and more cohort studies in the field of children s mental health. Bymainly introducing the advantages of cohort studies on children in distress and organizes domestic and foreign cohort studies in the field of mental health of children in distress, this article finds that it is mostly used in depression, anxiety, post traumatic stress disorder, suicidal ideation and attempts, etc, and mainly explores the risk and prevalence of mental health development in children in distress, and identifies long term negative damage. The research aims to promote the long term development and high quality development of such research by analyzing and summarizing the status quo and prospects of cohort research in the field of mental health of children in distress.
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Objective:To investigate the influencing factors for endoscopic retrograde cholangiopancreatography (ERCP)-related adverse events in novice trainees and establishment of its prediction model.Methods:The prospective study was conducted. The clinical data of 12 novice trainees of ERCP in the First Hospital of Lanzhou University from July 2016 to July 2019 were selected. The operation was performed by 12 novice trainees of ERCP under the guidance of the endoscopic experts. Observation indicators: (1) ERCP-related adverse events in novice trainees; (2) analysis of influencing factors for ERCP-related adverse events in novice trainees; (3) establishment of a prediction model for ERCP-related adverse events in novice trainees. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed by the t test. Measurement data with skewed distribution were represented as M(range), and com-parison between groups was analyzed by the Mann-Whitney U test. Count data were expressed as absolute numbers or percentages, and comparison between groups was analyzed by the chi-square test. The Logistic regression model was used for univariate and multivariate analyses. The regression coefficients were used to construct a prediction model. The receiver operating characteristic curve was drawn, and the area under curve was used to evaluate the predictive ability. Results:(1) ERCP-related adverse events in novice trainees. Of the 300 patients with ERCP operated by 12 novice trainees, 52 cases had ERCP-related adverse events and 248 cases had no ERCP-related adverse events. Cases in grade 1?2 or grade 3?4 of ERCP difficulty classification, score for intubation time, score for cannulation time, cases with or without completion of the cannulation, cases with or with-out basket stone removal, cases with or without stenosis expansion, score for contrast-enhanced interpretation, score for implementation of reasonable treatment and score for expected purpose reached were 22, 30, 8(range, 5?10), 20(rang, 9?20), 24, 28, 11, 41, 0, 52, 39±17, 39±19 and 44±23 for novice trainees with ERCP-related adverse events, versus 146, 102, 6(range, 4?9), 12(range, 8?20), 163, 85, 94, 154, 20, 228, 52±22, 80±20, 52±23 for novice trainees without ERCP-related adverse events, showing significant differences in the above indicators between them ( χ2=4.79, Z=?2.46, ?2.72, χ2=7.01, 5.30, 4.49, t=?4.00, ?2.97, ?2.29, P<0.05). (2) Analysis of influencing factors for ERCP-related adverse events in novice trainees. Results of univariate analysis showed that the diffi-culty classification of ERCP, intubation time, cannulation time, completion of cannulation, basket stone extraction, contrast-enhanced interpretation, implementation of reasonable treatment and expected purpose reached were related factors for ERCP-related adverse events in novice trainees ( odds ratio=1.95, 1.11, 1.08, 0.45, 0.44, 0.97, 0.98, 0.98, 95% confidence intervals as 1.07?3.58, 1.02?1.22, 1.02?1.14, 0.24?0.82, 0.22?0.90, 0.96?0.99, 0.96?0.99, 0.97?1.00, P<0.05). Results of multi-variate analysis showed that difficulty classification of ERCP and contrast-enhanced interpretation were independent influencing factors for ERCP-related adverse events in novice trainees ( odds ratio=2.08, 0.95, 95% confidence intervals as 1.10?3.96, 0.92?0.99, P<0.05). (3) Establishment of a predic-tion model for ERCP-related adverse events in novice trainees. According to the important outcome indicators of clinical training and results of multivariate analysis, 4 indicators including difficulty classification of ERCP, intubation time, cannulation time and contrast-enhanced interpretation were included to establish a prediction model for ERCP-related adverse events in novice trainees, which indicated that trainees with the predicted score >0.4 were in high risk of ERCP-related adverse events. The area under receiver operating characteristic curve of the prediction model was 0.72(95% confidence interval as 0.65?0.79, P<0.05), with the best cut-off value as 0.40, the sensitivity as 76.9% and the specificity as 63.3%. Conclusion:The difficulty classification of ERCP and contrast-enhanced interpretation are independent influencing factors for ERCP-related adverse events in novice trainees. Novice trainees with a predicted score >0.4 are high-risk groups of ERCP-related adverse events.
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Brain-computer interface (BCI) systems based on steady-state visual evoked potential (SSVEP) have become one of the major paradigms in BCI research due to their high signal-to-noise ratio and short training time required by users. Fast and accurate decoding of SSVEP features is a crucial step in SSVEP-BCI research. However, the current researches lack a systematic overview of SSVEP decoding algorithms and analyses of the connections and differences between them, so it is difficult for researchers to choose the optimum algorithm under different situations. To address this problem, this paper focuses on the progress of SSVEP decoding algorithms in recent years and divides them into two categories-trained and non-trained-based on whether training data are needed. This paper also explains the fundamental theories and application scopes of decoding algorithms such as canonical correlation analysis (CCA), task-related component analysis (TRCA) and the extended algorithms, concludes the commonly used strategies for processing decoding algorithms, and discusses the challenges and opportunities in this field in the end.
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Algorithms , Brain-Computer Interfaces , Electroencephalography , Evoked Potentials, Visual , Photic StimulationABSTRACT
OBJECTIVE@#To assess the association of single nucleotide polymorphisms (SNP) of aquaporin 7 ( AQP7) and aquaporin 9 ( AQP9) genes and type 2 diabetes mellitus (T2DM) among ethnic Han Chinese population.@*METHODS@#A case-control study involving 1194 subjects with T2DM and 1274 non-diabetic mellitus (NDM) subjects were enrolled. Genotypes of three SNPs (rs3758269 of AQP7 gene, rs16939881 and rs57139208 of AQP9 gene) were determined by using a MassArray method. The association of the three SNPs with T2DM was assess, and the correlation of glucose and lipid metabolism parameters with various SNP genotypes in the NDM group was analyzed.@*RESULTS@#The allelic and genotypic frequencies of the three SNPs did not differ significantly between the two groups (P>0.05). Nor was there significant difference between the two groups with different genetic models (P>0.05). No significant association of genotypes of AQP7 gene rs3758269, AQP9 gene rs16939881 and rs57139208 with glucose and lipid metabolism parameters were observed in the NDM group (P>0.05).@*CONCLUSION@#The rs3758269 in AQP7 gene and rs16939881 and rs57139208 in AQP9 gene are not associated with the genetic susceptibility of T2DM among ethnic Han Chinese population.
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Humans , Aquaporins/genetics , Case-Control Studies , China , Diabetes Mellitus, Type 2/genetics , Genetic Predisposition to Disease , Polymorphism, Single NucleotideABSTRACT
Objective:To introduce the improved "pull technique" and its preliminary application in Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine.Methods:Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine has started to implement the "pull technique" since March 2018.After one patient suffered from postoperative tunnel infection, we′ve improved the operation method: after successful extubation, small incision was made at the tunnel entrance, and the skin was properly trimmed and sutured to close the tunnel entrance.Results:Until May 2020, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine has implemented the modified tube removal for 15 patients.During the follow-up period (0-25 months), there was no secondary infection or peritoneal effusion.Conclusion:For patients who meet the indications of "pull technique" , the improved "pull technique" is a trial method, which can reduce the risk of secondary infection and peritoneal effusion.
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Objective:To evaluate the relationship between phosphorylation of Tau protein and apolipoprotein E (ApoE) containing 18 kDa fragments and investigate the mechanism of neuronal damage induced by sevoflurane.Methods:Primary neurons (ApoE3 and ApoE2 genotypes, 24 dishes for each genotype) of fetal mice cultured until the 5th day were divided into 4 groups ( n=12 each) using a random number table method: ApoE3 control group (A3C group), ApoE3 sevoflurane group (A3S group), ApoE2 control group (A2C group) and ApoE2 sevoflurane group (A2S group). Neurons were treated with 21% oxygen + 5% carbon dioxide + 4.1% sevoflurane for 4 h in A3S and A2S groups, while the neurons were only treated with 21% oxygen + 5% carbon dioxide in A3C and A2C groups.The cell proteins were then extracted to detect the expression of full-length ApoE and ApoE, AT8 and PHF1 containing 18 kDa fragments (by Western blot), expression of ApoE mRNA (by real-time polymerase chain reaction), and concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in the supernatant (by enzyme-linked immunosorbent assay). Results:Compared with A2C group, the expression of ApoE mRNA and full-length ApoE in neurons was up-regulated ( P<0.05), and no significant change was found in the expression of AT8 and PHF1 and concentrations of TNF-α and IL-6 in the supernatant in A2S group ( P>0.05). Compared with A3C group, the expression of ApoE mRNA, full-length ApoE, and ApoE, AT8 and PHF1 containing 18 kDa fragments was up-regulated, and the concentrations of TNF-α and IL-6 in the supernatant were increased in A3S group ( P<0.05). Conclusion:Sevoflurane may promote phosphorylation of Tau proteins and increase inflammatory responses through up-regulating the expression of ApoE containing 18 kDa fragments, thus leading to neuronal damage.
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Objective@#To understand the prevalence of HIV (human immunodeficiency virus) testing and associated factors among sexual active college students in Zhuhai City.@*Methods@#From November to December, 2019, an anonymous electronic questionnaire was administered among 12 235 students in six colleges and universities in Zhuhai City by multistage sampling. A total of 1 789 college students ever had sex were selected. Pearsons Chisquare test and nonconditional Logistic regression model were applied to analyze the factors associated with uptake of HIV testing.@*Results@#Among these students ever had sex, 7.55% (135/1 789) had been tested for HIV mainly through hospitals (71.85%). The main reasons for testing were regular testing (50.37%) and intending to know their infection status (23.70%). Multivariable Logistic regression showed that homosexual individuals (OR=4.62, 95%CI=1.07-19.95) and those who had heterosexual commercial sex in the past year (OR=3.77, 95%CI=1.96-7.26) were more likely to test for HIV, while female (OR=0.41, 95%CI=0.24-0.69) were less likely to test for HIV.@*Conclusion@#The proportion of HIV testing was low among sexual active college students in Zhuhai City. Interventions should be tailored including strengthening the HIV testing propaganda education and enhancing students awareness of HIV testing, and the influencing factors such as gender, sexual orientation and commercial sexual behavior should be taken into consideration, so as to improve the HIV testing coverage of this population.
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@#AIM: To isolate the retinal ganglion cells(RGCs)of neonatal Sprague-Dawley(SD)rats <i>in vitro</i>, and to establish the RGCs primary culture method and high glucose model of neonatal SD rats.<p>METHODS: The retinal tissues of SD rats from 1-3d as the materials were taken, from which the RGCs were isolated and purified for primary culture. Toluidine blue and immunofluorescence staining methods were adopted to identify the cultured cells. After 48-72h of continuous culture, RGCs were randomly divided into 6 groups and cultured in different glucose concentrations of 5.5mmol/L(normal control group), 20mmol/L, 25mmol/L, 30mmol/L, 35mmol/L and 40mmol/L for 24, 48 and 72h, respectively. Finally, the CCK8 method and TUNEL method were adopted to determine the cell survival rate and apoptosis rate. <p>RESULTS: The primary RGCs purified and cultured <i>in vitro</i> represented typical cell morphology and grew well, the cells were confluent and aggregated in small lamellar manner, while the axons crisscrossed into a network, in addition, cell halo could be seen around the cell body. Nissl bodies with clear structure were found in the cytoplasm of toluidine blue stained cells, the percentage of neurons was more than 95%. RGCs specific antibodies Thy-1 and Brn-3a were employed to identify the purified cells <i>in vitro</i>, and the positive rate reached more than 90%. The CCK8 results showed that the survival rate of cells decreased(OD value decreased)with the increase of culture time and glucose concentration. When the cells were treated with different glucose concentrations for 24h, the OD values of each group were lower than those of the normal control group, but there was no significant difference between the OD values of each group and the normal control group(all <i>P</i>>0.05). With the extension of culture time, the OD values of 35mmol/L and 40mmol/L glucose concentration intervention RGCs 48h, 30mmol/L, 35mmol/L, 40mmol/L intervention RGCs 72h were significantly lower than those of the normal control group, the difference was statistically significant compared with the normal control group(all <i>P</i><0.05). TUNEL results revealed that the apoptosis rate of RGCs increased with the increase of glucose concentration and time, among them, the apoptosis rate of RGCs cultured in glucose concentration of 30mmol/L, 35mmol/L and 40mmol/L for 48h and 72h was significantly statistical different from that of normal control group(all <i>P</i><0.05). <p>CONCLUSION: The RGCs primary culture method established in this study is capable of separating typical RGCs with high purity. With the increase of glucose concentration in the medium, the survival rate of RGCs have been decreased while the apoptosis rate increased. Notably, the 35mmol/L glucose intervention for 48h can be employed as the optimal intervention concentration and time to effectively induce RGCs to establish the high glucose model.
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This paper describes how to apply UpToDate ? system can be applied into the evidence-based teaching of difficult and critical clinical problems of nephrology, combined with the practical case of standardized training for specialists. The treatment difficulties can be put forward by teachers or students, and appropriate terms are selected to search in UpToDate ?. The students are required to learn the content of the searched items, and then give their treatment choices and clarify reasons according to the condition of patients. After that, the instructing doctor will comment on the statements of each training specialist, and give treatment plans. Promotion the application of UpToDate ? system is helpful to improve the teaching quality of the standardized training for specialists.
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Objective:To evaluate the role of hippocampal ApoE in age factors-induced effect on long-term cognitive function in mice undergoing multiple exposure to sevoflurane anesthesia using the gene knockout method.Methods:Seventy-two juvenile (6 days old, weighing 3-5 g) and adult (60 days old, weighing 20-25 g) wild-type (WT) C57BL/6J mice and 56 ApoE gene knockout (ApoE-KO) C57BL/6J mice were studied.WT mice were divided into 4 groups ( n=18 each) using a random number table method: juvenile control group (P6+ C group), juvenile sevoflurane group (P6+ S group), adult control group (P60+ C group) and adult sevoflurane group (P60+ S group). ApoE-KO mice were randomly divided into 4 groups ( n=14 each) according to a random number table method: juvenile control group (P6+ C group), juvenile sevoflurane group (P6+ S group), adult control group (P60+ C group) and adult sevoflurane group (P60+ S group). Mice inhaled 3% sevoflurane in 60% O 2 for 2 h every day for 3 consecutive days in sevoflurane group, while mice were placed in the same environment and inhaled 60% O 2 for 2 h every day for 3 consecutive days in control group.Four mice were selected at the end of sevoflurane anesthesia (8 or 62 days after birth) for determination of IL-1β, IL-6 and TNF-α contents in hippocampus by enzyme-linked immunosorbent assay.Four mice in each group of WT mice were randomly selected to detect the expression of full-length ApoE and ApoE fragments in hippocampus by Western blot.The remaining 10 mice in each group of WT mice received standardized feeding, and contextual fear conditioning test was performed at 22 days after sevoflurane anesthesia (30 or 84 days after birth). Results:WT mice Compared with P6+ C group, IL-1β, IL-6 and TNF-α contents were significantly increased, the expression of full-length ApoE and ApoE fragments was up-regulated, and the standstill time was shortened in P6+ S group ( P <0.05), and no significant change was found in the parameters mentioned above in P60+ C and P60+ S groups ( P>0.05). ApoE-KO mice Compared with P6+ C and P6+ S groups, no significant change was found in IL-1β, IL-6 and TNF-α contents or standstill time in P60+ C group and P60+ S group ( P>0.05). Conclusion:The mechanism of age influencing the long-term cognitive dysfunction is related to up-regulating ApoE expression in hippocampus and inducing neuroinflammatory response in mice undergoing multiple exposure to sevoflurane anesthesia.
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@#Lysyl oxidase(LOX), an amino oxidase with copper binding site outside the cell, has been confirmed that may be involved in the pathogenesis of many diseases by a large number of studies. At present, although the potential mechanism of common variants of LOX in the development of disease is still unknown, but the potential role may be to affect the biological activity of LOX through tissue-specific alternative splicing or expression regulation.However, no matter under normal or pathological conditions, there is still little information about LOX in eye tissue yet.In recent years, it has been found that LOX may be involved in the occurrence and development of many eye diseases in related studies. Based on this, the review will summarize the research progress of eye diseases in which LOX is involved of recent years.
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Objective Alzheimer's disease (AD) is the most common cause of dementia. The pathophysiology of the disease mostly remains unearthed, thereby challenging drug development for AD. This study aims to screen high throughput gene expression data using weighted co-expression network analysis (WGCNA) to explore the potential therapeutic targets.Methods The dataset of GSE36980 was obtained from the Gene Expression Omnibus (GEO) database. Normalization, quality control, filtration, and soft-threshold calculation were carried out before clustering the co-expressed genes into different modules. Furthermore, the correlation coefficients between the modules and clinical traits were computed to identify the key modules. Gene ontology and pathway enrichment analyses were performed on the key module genes. The STRING database was used to construct the protein-protein interaction (PPI) networks, which were further analyzed by Cytoscape app (MCODE). Finally, validation of hub genes was conducted by external GEO datasets of GSE 1297 and GSE 28146.Results Co-expressed genes were clustered into 27 modules, among which 6 modules were identified as the key module relating to AD occurrence. These key modules are primarily involved in chemical synaptic transmission (GO:0007268), the tricarboxylic acid (TCA) cycle and respiratory electron transport (R-HSA-1428517).
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Objective To investigate the key role of nuclear factor E2-related factor 2 (Nrf 2) in the treatment of lung injury in sepsis mice by regulating Nrf 2/heme oxygenase-1 (HO-1)/high mobility group protein B1 (HMGB1) pathway. Methods 120 male wild type (WT) and 120 Nrf 2 knockout (Nrf 2-KO) ICR mice were randomly divided into Sham group, H2 control group (Sham+H2 group), cecal ligation and puncture (CLP) induced sepsis model group (CLP group) and H2 intervention group (CLP+H2 group), with 30 mice in each group. The sepsis model was reproduced by CLP. The same operation was done in Sham group and Sham+H2 group except CLP. The mice in Sham+H2 group and CLP+H2 group were challenged by 2% H2 for 1 hour at 1 hour and 6 hours after operation respectively, while the mice in Sham group and CLP group only inhaled air. Twenty mice in each group were collected to observe the 7-day survival. The other mice were sacrificed at 24 hours after the reproduction of model, and the lung tissues were harvested. The activities of superoxide dismutase (SOD) and catalase (CAT) and malondialdehyde (MDA) contents were determined by enzyme-linked immunosorbent assay (ELISA). The expressions of HO-1 and HMGB1 were determined by Western Blot, and the positive expression of HO-1 was also detected by immunofluorescence. Results Compared with Sham groups, the 7-day survival rates of WT and Nrf 2-KO mice in CLP groups were significantly lowered [WT: 0% (0/20) vs. 100% (20/20), Nrf 2-KO: 0% (0/20) vs. 100% (0/20), both P < 0.05]; the 7-day survival rates of CLP+H2 group in WT mice were significantly higher than those of CLP group [40% (8/20) vs. 0% (0/20), P < 0.05], but there was no significant difference between CLP+H2 group and CLP group in Nrf 2-KO mice [0% (0/20) vs. 0% (0/200), P > 0.05]. In WT mice, compared with Sham group, the activities of SOD and CAT in lung tissue of CLP group were decreased significantly [SOD (kU/g): 131.30±28.21 vs. 251.00±22.84, CAT (kU/g): 13.43±1.52 vs. 20.76±1.63, both P < 0.01], the MDA content, the expressions of HO-1 and HMGB1 were increased significantly [MDA (μmol/g): 6.26±1.18 vs. 4.16±0.58, HO-1/β-actin: 0.160±0.045 vs. 0.023±0.005, HMGB1/β-actin: 0.656±0.055 vs. 0.005±0.001, all P < 0.05]. Compared with CLP group, the activities of SOD, CAT and HO-1 expression in lung tissue of CLP+H2 group were significantly increased [SOD (kU/g): 220.32±35.06 vs. 131.30±28.21, CAT (kU/g): 18.95±2.49 vs. 13.43±1.52, HO-1/β-actin: 0.376±0.025 vs. 0.160±0.045, all P < 0.01], while the MDA contents and HMGB1 expressions were significantly decreased [MDA (μmol/g): 4.26±0.75 vs. 6.26±1.18, HMGB1/β-actin: 0.343±0.040 vs. 0.656±0.055, both P < 0.05]. In Nrf 2-KO mice, compared with Sham group, the activity of CAT in CLP group was significantly lowered (kU/g: 12.28±1.49 vs. 19.11±1.53, P < 0.01), MDA contents and the expressions of HO-1 and HMGB1 were significantly increased [MDA (μmol/g): 6.85±0.54 vs. 4.59±0.50, HO-1/β-actin: 0.063±0.005 vs. 0.021±0.003, HMGB1/β-actin: 0.713±0.035 vs. 0.005±0.001, all P < 0.01], while there was no significant difference in SOD activity (kU/g: 114.19±9.94 vs. 135.75±28.10, P > 0.05). There was no significant difference in above parameters between CLP+H2 group and CLP group. Conclusions H2 inhibits lung injury in septic mice through Nrf 2/HO-1/HMGB1 pathway. Nrf 2 plays a major role in the treatment of septic lung injury by H2.
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Objective To evaluate the effect of hydrogen on the expression of matrix metalloproteinase-9 (MMP-9) in brain tissues of mice with sepsis-associated encephalopathy (SAE).Methods A total of 212 clean-grade healthy male C57BL/6J mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups (n =53 each) using a random number table method:sham operation group (group Sham),sham operation plus hydrogen group (group Sham+H2),group SAE and SAE plus hydrogen group (group SAE+ H2).The sepsis model was established by intraperitoneally injecting human fecal suspension.Sham+H2 and SAE+H2 groups inhaled 2% hydrogen for 1 h at 1 and 6 h after establishing the model,respectively.Postoperative 14-day survival rate was recorded.Evans blue (EB) was injected into the tail vein at 24 h after establishing the model,and the content of EB in brain tissues was calculated.Brain tissues were taken for determination of brain water content.The expression of tumor necrosis factor-alpha (TNF-α),interleukin-6 (IL-6) and high-mobility group box 1 protein (HMGB1) in hippocampal tissues was detected by enzymelinked immunosorbent assay.The expression of MMP-9 and ZO-1 protein and mRNA in hippocampal tissues was determined by Western blot and real-time polymerase chain reaction,respectively,at 6,12 and 24 h after establishing the model.Results Compared with Sham group,the 14-day survival rate was significantly decreased,the EB content in brain tissues,brain water content,and contents of TNF-α,IL-6 and HMGB1 were significantly increased,the expression of MMP-9 protein and mRNA was up-regulated,and the expression of ZO-1 protein and mRNA was down-regulated in SAE and SAE+H2 groups (P<0.05),and no significant change was found in the parameters mentioned above in group Sham+H2 (P>0.05).Compared with group SAE,the 14-day survival rate was significantly increased,the EB content in brain tissues,brain water content,and contents of TNF-α,IL-6 and HMGB1 were significantly decreased,the expression of MMP-9 protein and mRNA was down-regulated,and the expression of ZO-1 protein and mRNA was up-regulated in group SAE+H2 (P<0.05).Conclusion The mechanism by which hydrogen attenuates the blood-brain barrier damage may be related to inhibiting MMP-9 expression and to reducing inflammatory responses in brain tissues of SAE mice.
ABSTRACT
Objective@#To evaluate the effect of hydrogen on mitochondrial biosynthesis in the hippocampus of mice with sepsis-associated encephalopathy (SAE).@*Method@#Two hundred and twenty-four healthy clean-grade male C57BL/6J mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups (n=56 each) using a random number table method: sham operation group (group SH), sham operation plus hydrogen group (group SH+ H2), group SAE, and SAE plus hydrogen group (group SAE+ H2). The model of SAE was established by cecal ligation and puncture in anesthetized mice.Group SH+ H2 and group SAE+ H2 inhaled 2% hydrogen for 1 h starting from 1 and 6 h after operation, respectively.Twenty mice were selectde to record the postoperative 7-day survival rate.The remaining animals were sacrificed at 24 h after operation, and brain tissues were taken for examination of the pathological changes in hippocampal CA1 region (with a light microscope) and for determination of neuronal apoptosis (by TUNEL), mitochondrial membrane potential (MMP) (by fluorescence spectrophotometry) and ATP content (by a bioluminescence assay). The apoptosis rate was calculated.The expression of peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) in hippocampus was determined by Western blot at 6, 12 and 24 h after operation.@*Results@#Compared with group SH, the postoperative 7-day survival rate was significantly decreased, the apoptosis rate of hippocampal neurons was increased, the contents of MMP and ATP were decreased, and the expression of PGC-1α was up-regulated in SAE and SAE+ H2groups (P<0.05), and no significant change was found in the parameters mentioned above in group SH+ H2(P>0.05). Compared with group SAE, the postoperative 7-day survival rate was significantly increased, and the apoptosis rate of hippocampal neurons was decreased, the contents of MMP and ATP were increased, and the expression of PGC-1α was up-regulated (P<0.05), and the pathological changes of hippocampal tissues were significantly attenuated in group SAE+ H2.@*Conclusion@#The mechanism by which hydrogen mitigates SAE may be related to promoting mitochondrial biosynthesis in mice.
ABSTRACT
Objective To evaluate the effect of hydrogen on mitochondrial biosynthesis in the hippocampus of mice with sepsis-associated encephalopathy (SAE).Method Two hundred and twenty-four healthy clean-grade male C57BL/6J mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups (n=56 each) using a random number table method:sham operation group (group SH),sham operation plus hydrogen group (group SH+H2),group SAE,and SAE plus hydrogen group (group SAE+H2).The model of SAE was established by cecal ligation and puncture in anesthetized mice.Group SH+H2 and group SAE+H2 inhaled 2% hydrogen for 1 h starting from 1 and 6 h after operation,respectively.Twenty mice were selectde to record the postoperative 7-day survival rate.The remaining animals were sacrificed at 24 h after operation,and brain tissues were taken for examination of the pathological changes in hippocampal CA1 region (with a light microscope) and for determination of neuronal apoptosis (by TUNEL),mitochondrial membrane potential (MMP) (by fluorescence spectrophotometry) and ATP content (by a bioluminescence assay).The apoptosis rate was calculated.The expression of peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) in hippocampus was determined by Western blot at 6,12 and 24 h after operation.Results Compared with group SH,the postoperative 7-day survival rate was significantly decreased,the apoptosis rate of hippocampal neurons was increased,the contents of MMP and ATP were decreased,and the expression of PGC-1α was up-regulated in SAE and SAE+H2groups (P<0.05),and no significant change was found in the parameters mentioned above in group SH+H2 (P>0.05).Compared with group SAE,the postoperative 7-day survival rate was significantly increased,and the apoptosis rate of hippocampal neurons was decreased,the contents of MMP and ATP were increased,and the expression of PGC-1α was up-regulated (P<0.05),and the pathological changes of hippocampal tissues were significantly attenuated in group SAE+H2.Conclusion The mechanism by which hydrogen mitigates SAE may be related to promoting mitochondrial biosynthesis in mice.