1.Preliminary serum pharmacological exploration of method to preparate Ganlong capsule-containing serum
Keke ZHANG ; Hao BAI ; Jinyu LÜ ; Yue JIN ; Rui LUO
Chinese Journal of Pathophysiology 2024;40(5):956-960
AIM:To investigate the optimal time of blood collection and the best concentration of Ganlong capsule(GLC)-containing serum for steatohepatitis.METHODS:Sixty C57BL/6J mice were gavaged with GLC at a dose of 90 mg/kg body weight,twice a day for 7 days.Control mice were administered the same volume of 0.9%saline.After 0.5,1,2 and 4 h from the last gavage,blood was collected to prepare GLC-containing serum.The LC-MS/MS was used to detect the inosine content of blank and drug-containing serum at different blood collection times to determine the optimal time.Subsequently,NCTC1469 cells stimulated with palmitic acid(PA)were treated with different concentrations of the GLC-containing serum.Nile Red staining was used to detect lipid droplet formation,and RT-qPCR was employed to as-sess the expression of intracellular fatty acid synthesis(Fabp1,Scd1)and fatty acid β-oxidation gene(Pparα).RE-SULTS:(1)LC-MS/MS detection revealed that the inosine concentration peaked 2 h after the last administration.(2)Nile red staining demonstrated that the GLC-containing serum exerted a dose-dependent decreasing trend of lipid droplets in hepatocytes induced by PA.(3)GLC-containing serum significantly downregulated the mRNA expression levels of lipo-genesis-genes(Fabp1 and Scd1)with a dose-dependent decreasing trend in the PA-induced NCTC1469 cells.The mRNA expression of genes associated with fatty acid β-oxidation(Pparα)were upregulated with a dose-dependent increasing trend.CONCLUSION:The 10%GLC-containing serum collected 2 hours after the last administration in mice has the best therapeutic effect for steatohepatitis.
2.Non-targeted Metabolomics Analysis of Fuling Yunhua Granules in Treatment of Type 2 Diabetes Mellitus Rats
Mengyao TIAN ; Keke LUO ; Mengxiao WANG ; Tianbao HU ; Hongmei LI ; Zongyuan HE ; Lixin YANG ; Liyu HAO ; Nan SI ; Yuyang LIU ; Baolin BIAN ; Hongjie WANG ; Yanyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):195-204
ObjectiveBased on non-targeted metabolomics, to analyze the regulation of endogenous differential metabolites in serum of type 2 diabetes mellitus(T2DM) rats by Fuling Yunhua granules, and to clarify the metabolic pathways through which this granules exerted its effect on improving T2DM. MethodSeventy SD rats, half male and half female, were randomly divided into the control group, model group, and high, medium, low dose groups of Fuling Yunhua granules(20.70, 10.35, 5.18 g·kg-1 in raw drug amount) and the positive drug group(pioglitazone hydrochloride tablets, 8.1 mg·kg-1). Except for the control group, other groups were fed with high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin(STZ) to establish a T2DM rat model. After successful modeling, the treatment groups were administered the corresponding drugs by gavage, and the control group and model group were treated with an equal volume of saline by gavage, once/d, for 28 d. Fasting blood glucose(FBG) and glycosylated hemoglobin A1c(GHbA1c) levels were measured in all groups of rats during the administration period, and hematoxylin-eosin(HE) staining was used to observe the pathomorphological changes in the pancreatic tissues of rats at the end of the administration period. The endogenous metabolite levels in rat serum were detected by ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-LTQ-Orbitrap MS), and the data were processed using principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Differential metabolites were identified by the Human Metabolome Database(HMDB) and the Kyoto Encyclopedia of Genes and Genomes(KEGG), and screened for differential metabolites with variable importance in the projection(VIP) value>1, P<0.05, and fold change(FC)<0.6 or FC>1. And the metabolic pathway enrichment analysis of the screened differential metabolites was performed by MetaboAnalyst 5.0, then the screened differential metabolites were diagnosed and evaluated by the receiver operating characteristic(ROC) curves. ResultCompared with the control group, the FBG level of rats in the model group increased significantly(P<0.01), the GHbA1c content tended to increase, but the difference was not statistically significant, and the pancreatic tissue of rats was obviously damaged, the number of pancreatic islets decreased, and the pancreatic β-cells were obviously reduced, atrophied and enlarged. Compared with the model group, the FBG levels of rats in the high dose group of Fuling Yunhua granules and the positive drug group were significantly reduced after 2 weeks of administration(P<0.05, P<0.01), the GHbA1c content of rats in the high dose group of Fuling Yunhua granules was significantly reduced(P<0.05), and the pancreatic tissue lesions of rats in the different dose groups of Fuling Yunhua granules were reduced. The results of non-targeted metabolomics showed that 46 differential metabolites were significantly changed in the model group compared with the blank group. Pathway enrichment analysis found that T2DM mainly affected biological processes including biosynthesis of primary bile acid, D-amino acid metabolism, steroid hormone biosynthesis, and glycerophospholipid metabolism in rats. Compared with the model group, the levels of 8 differential metabolites in the high dose group of Fuling Yunhua granules were significantly adjusted, and the pathway enrichment analysis found that D-amino acid metabolism, retinol metabolism, glycine, serine and threonine metabolism, tryptophan metabolism and other metabolic pathways were mainly involved. ROC curves further analysis revealed that the four characteristic differential markers of 11-cis-retinol, D-piperidinic acid, D-serine, and p-cresol sulfate had high diagnostic value for the treatment of T2DM with Fuling Yunhua granules. ConclusionFuling Yunhua granules can improve the symptoms of T2DM rats by regulating the amino acid metabolic and retinol metabolic pathways through the modulation of endogenous differential metabolites.
3.Research status and prospects of research on intraspecific differentiation of Cannabis sativa L.
Keke FU ; Delan WANG ; Jinyuan HU ; Hao NIE ; Ying ZHANG ; Ning AN ; Fangru LIU ; Xingchun ZHAO ; Shan GAO ; Baishi WANG
Chinese Journal of Forensic Medicine 2024;39(5):600-605
The DNA characterization of Cannabis sativa L.has been one of the key directions of anti-drug research at home and abroad.Previous research mainly focused on the identification of cannabis-species and gender differentiation,and have constructed a number of corresponding composite amplification systems.With the rapid development of high-throughput sequencing technology,the whole genome of C.sativa and the sequences of key enzyme genes for its major physicochemical components have been sequenced successively,and intra-species differentiation studies of C.sativa based on specific molecular markers have gradually emerged.However,due to the high variability of cannabis subspecies-and variety-specific molecular markers,relevant foreign studies failed to provide ideal molecular marker support for the identification of intra-specific distinctions of Cannabis sativa in China.Based on this,this paper comprehensively analyzes the current situation and shortcomings of domestic and international research on intra-specific differentiation of C.sativa,and combines the previous research results of this group to elaborate on how to use high-throughput sequencing technology to solve the problem of the lack of intra-specific molecular markers of C.sativa in China.
4.Microbial monitoring of urban drinking water in Jiangxi Province from 2016 to 2020
Keke LU ; Jiafen HE ; Junjie FU ; Hao WU ; Wenxin HE ; Min XU ; Feibao LU ; Wenbin JIANG
Shanghai Journal of Preventive Medicine 2023;35(3):253-257
ObjectiveTo monitor the microbes in urban drinking water in Jiangxi from 2016 to 2020, to analyze the change in microbial qualification rate, and to provide a scientific basis for government decision-making. MethodsAccording to the Standard Examination Method for Drinking Water (GB/T 5750‒2006) and the Standards for Drinking Water Quality (GB 5749‒2006), the water samples were collected, tested and evaluated for hygienic safety. The chi-square test was used to compare the qualification rates among different water periods, water source types, water supply modes, water samples, treatment processes, and disinfection methods. ResultsA total of 10 584 water samples were collected and examined from 2016 to 2020,with a qualification rate of 97.72%. The qualified rate of the microbiological index increased gradually over the years. There was no statistically significant difference in the microbiological qualification rate of water samples monitored in different water periods (χ2=0.718,P=0.398), and the qualification rates were 97.85% and 97.60% in dry and abundant water periods respectively. There was a statistically significant difference in the qualification rates of water samples monitored in different water source types (χ2=79.560,P=0.398), with groundwater having a higher qualification rate of 98.83% than surface water (97.70%). The microbiological pass rate of water samples differed among different water supply methods (χ2=201.836,P<0.001). The microbiological pass rates of municipal centralized water supply, self-built centralized water supply and secondary water supply were 97.51%, 88.94% and 96.56% respectively. The microbiological pass rates of water samples differed among different treatment processes. There were differences in the qualification rates of the water samples between different treatment processes(χ2=855.034,P<0.001), with 98.22%, 67.19%, 95.60% and 53.13% of the conventionally treated, precipitated filtered, disinfected only and untreated water samples, respectively. The differences in the microbiological qualification rates of the monitored water samples between different disinfection methods were statistically significant (χ2=904.993,P<0.001). Chlorination disinfection, chlorine dioxide disinfection, other disinfection methods and non-disinfected water samples had the pass rates of 98.35%, 98.19%, 84.38% and 60.16%, respectively. ConclusionThe qualified rate of the microbiological index of drinking water in Jiangxi increases year by year, but there are still some drinking water without appropriate disinfectants and water quality treatment processes. There is still a need to strengthen the purification, disinfection and monitoring of drinking water.
5.Advances in the study of the roles of mitochondrial dysfunction and neuroinflammation in the pathogenesis of schizophrenia
Keke HAO ; Huiling WANG ; Gaohua WANG
Sichuan Mental Health 2023;36(1):74-79
This paper aims to present a review of the research progress on the roles of mitochondrial dysfunction and microglia-mediated inflammation in the pathophysiology of schizophrenia. Schizophrenia is a serious psychiatric disorder in which the immuno-inflammatory pathway is considered to be one of the main pathogenic mechanisms of disease, and manifested by microglia activation and elevated levels of inflammatory cytokines in the brain. However, the biological mechanism underlying immune disorder in schizophrenia has not been fully elucidated. In recent years, accumulating evidence has indicated that mitochondrial dysfunction may play a key role in the pathogenesis of schizophrenia. Therefore, this article reviews the research progress on the roles of mitochondrial days function and microglia-mediated inflammation in the pathophysiology of schizophrenia, so as to inform the study of the pathogenesis of schizophrenia.
6.Feasibility and safety of bipolar-plasmakinetic transurethral enucleation and resection of the prostate in day surgery mode.
Zhihui ZOU ; Ligang ZHANG ; Keke CAI ; Yongtao HU ; Shuchen LIU ; Jia CHEN ; Qintao GE ; Xiaohu ZHAO ; Zongyao HAO ; Chaozhao LIANG
Journal of Zhejiang University. Medical sciences 2023;52(2):148-155
OBJECTIVES:
To evaluate the feasibility and safety of bipolar-plasmakinetic transurethral enucleation and resection of the prostate (B-TUERP) in day surgery.
METHODS:
From January 2021 to August 2022, 34 patients with benign prostatic hyperplasia (BPH) underwent B-TUERP in day surgery in the First Affiliated Hospital of Anhui Medical University. Patients completed the screening and anesthesia evaluation before admission and received the standard surgery which implements "anatomical enucleation of the prostate" and "absolute bleeding control" on the same day of admission, and by the same doctor. Bladder irrigation was stopped, catheter was removed and the discharge evaluation was performed on the first day after operation. The baseline data, perioperative conditions, time of recovery, treatment outcomes, hospitalization costs, and postoperative complications were analyzed.
RESULTS:
All operations were successfully conducted. The average age of the patients was (62.2±7.8) years, average prostate volume was (50.2±29.3) mL. The average operation time was (36.5±19.1) min, the average hemoglobin and blood sodium were decreased by (16.2±7.1) g/L and (2.2±2.0) mmol/L, respectively. The average postoperative length of hospital stay, and total length of hospital stay were (17.7±2.2) and (20.8±2.1) h, respectively, and the average hospitalization cost was (13 558±2320) CNY. All patients were discharged on the day after surgery except for one patient who was transferred to a general ward. Three patients received indwelling catheterization after catheter removal. The 3-month follow-up results showed a substantial improvement in the International Prostate Symptom Score, quality of life score and maximum urinary flow rate (all P<0.01). Three patients experienced temporary urinary incontinence, 1 patient experienced urinary tract infection, 4 patients were diagnosed with urethral stricture and 2 patients experienced bladder neck contracture. No complications above Clavien grade Ⅱ occurred.
CONCLUSIONS
The preliminary results showed that B-TUERP ambulatory surgery is a safe, feasible, economical and effective treatment for appropriately selected patients with BPH.
Male
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Humans
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Middle Aged
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Aged
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Prostate/surgery*
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Prostatic Hyperplasia/surgery*
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Ambulatory Surgical Procedures
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Quality of Life
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Feasibility Studies
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Retrospective Studies
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Treatment Outcome
7.Association between residual cholesterol and the risk of subclinical renal damage
Xi ZHANG ; Zejiaxin NIU ; Guilin HU ; Mingfei DU ; Ting ZOU ; Xiaoyu ZHANG ; Lan WANG ; Chao CHU ; Yueyuan LIAO ; Qiong MA ; Dan WANG ; Keke WANG ; Hao JIA ; Chen CHEN ; Yu YAN ; Yue SUN ; Tongshuai GUO ; Jie ZHANG ; Weihua GAO ; Ziyue MAN ; Ke GAO ; Wenjing LUO ; Jianjun MU ; Yang WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2023;44(1):6-13
【Objective】 Dyslipidemia has shown to be associated with cardiovascular, metabolic and renal diseases. This study aimed to investigate the association between residual cholesterol and the risk of subclinical renal damage (SRD). 【Methods】 A total of 2 342 participants were recruited from the previously established Hanzhong Adolescent Hypertension Study cohort. According to estimated glomerular filtration rate(eGFR) and urinary albumin-to-creatine ratio(uACR), the subjects were divided into SRD group and non-SRD group. The associations of residual cholesterol with eGFR, uACR, and the risk of SRD were analyzed by multiple linear and Logistic regression analyses. 【Results】 Residual cholesterol was positively correlated with uACR(r=0.081, P<0.001) but negatively correlated with eGFR (r=-0.091, P<0.001). Multiple linear regression analysis revealed that residual cholesterol was an influencing factor of uACR (β=0.075, P<0.001) and eGFR (β=-0.027, P<0.001) after adjustment for gender, age, smoke, alcohol, exercise, BMI, hypertension, diabetes and serum uric acid. In addition, Logistic regression analysis revealed that residual cholesterol was significantly associated with the risk of SRD independently of potential confounders [OR(95% CI)=1.387 (1.113-1.728), P<0.001]. Further subgroup analysis showed that residual cholesterol was significantly associated with the risk of SRD in women but not in men. 【Conclusion】 Residual cholesterol is a contributing factor in the risk of subclinical renal damage with gender-specific association.
8.Associations of genetic polymorphisms in Corin with blood pressure responses to salt and potassium intake
Lan WANG ; Zejiaxin NIU ; Yanjie GUO ; Nairong LIU ; Yanni YAO ; Beibei YANG ; Jiaxin WANG ; Chuang LI ; Panpan LIU ; Chang’e YANG ; Mingfei DU ; Guilin HU ; Xi ZHANG ; Dan WANG ; Xiaoyu ZHANG ; Chao CHU ; Yueyuan LIAO ; Qiong MA ; Keke WANG ; Hao JIA ; Yue SUN ; Tongshuai GUO ; Weihua GAO ; Jianjun MU ; Yang WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2023;44(1):22-29
【Objective】 Corin, a transmembrane serine protease that can cleave atrial natriuretic peptide precursor (pro-ANP) into atrial natriuretic peptide with smaller bioactive molecules, participates in the pathophysiological process of hypertension and cardiac hypertrophy. The purpose of this study was to explore the relationship of Corin gene variation with blood pressure responses to sodium and potassium dietary interventions. 【Methods】 In 2004, we recruited 514 participants from 124 families in 7 villages of Baoji, Shaanxi Province, China. All the subjects received a 3-day normal diet, a 7-day low-salt diet, a 7-day high-salt diet, and finally a 7-day high-salt and potassium supplementation. Fifteen single nucleotide polymorphisms (SNPs) of Corin gene were selected for final analysis. 【Results】 SNPs rs12509275 were significantly associated with diastolic blood pressure (DBP) response to low-salt diet, while rs3749584 was associated with pulse pressure (PP) response to low-salt diet.SNP rs3749584 and rs10517195 were significantly associated with PP response to high-salt diet. In addition,rs17654278 were significantly associated with systolic blood pressure (SBP) response to high-salt and potassium supplementation, rs2271037 was significantly correlated with DBP responses to high-salt and potassium supplementation, and rs4695253, rs12509275, rs2351783, rs36090894 were significantly associated with PP response to high-salt and potassium supplementation. 【Conclusion】 Corin gene polymorphisms were associated with blood pressure response to sodium and potassium, suggesting that Corin gene may be involved in pathophysiological process of salt sensitivity and potassium sensitivity.
9.Association of genetic variants in NEDD4L with blood pressure responses to dietary salt and potassium intake
Zejiaxin NIU ; Mingfei DU ; Guilin HU ; Xi ZHANG ; Dan WANG ; Lan WANG ; Wenjing LUO ; Mingke CHANG ; Ting ZOU ; Xiaoyu ZHANG ; Yu YAN ; Chao CHU ; Yueyuan LIAO ; Qiong MA ; Keke WANG ; Hao JIA ; Chen CHEN ; Yue SUN ; Tongshuai GUO ; Jie ZHANG ; Weihua GAO ; Ziyue MAN ; Ke GAO ; Jianjun MU ; Yang WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2023;44(1):30-37
【Objective】 4-like protein with down-regulated expression and development in neural precursor cells (NEDD4L) plays an important role in blood pressure (BP) regulation and sodium homeostasis by regulating epithelial sodium channel protein. In this study, we aimed to explore the relationship of NEDD4L gene polymorphisms with BP responses to sodium and potassium intake. 【Methods】 In 2004, 514 subjects from 124 families in Meixian County, Shaanxi Province, were recruited to establish a salt-sensitive hypertension study cohort. All the subjects received a 3-day baseline survey, a 7-day low-salt diet, a 7-day high-salt diet, and finally a 7-day high-salt and potassium supplementation. Their BP was measured and peripheral blood samples were collected at different intervention periods. The 14 gene polymorphisms of NEDD4L gene were genotyped and analyzed by MassARRAY platform. 【Results】 BP decreased on a low-salt diet, and significantly increased on a high-salt diet, and decreased again after potassium supplementation. NEDD4L SNPs rs74408486 were significantly associated with systolic BP, diastolic BP and mean arterial pressure responses to the low-salt diet. SNPs rs292449 and rs2288775 were significantly associated with pulse pressure response to the high-salt diet. In addition, SNPs rs563283 and rs292449 were significantly associated with diastolic BP, mean arterial pressure, and pulse pressure responses to high-salt and potassium supplementation diet. 【Conclusion】 NEDD4L gene polymorphisms were significantly associated with BP responses to sodium and potassium intake, suggesting that NEDD4L gene may be involved in the development of salt sensitivity and potassium sensitivity.
10.Association of genetic variants in uromodulin with blood pressure responses to dietary salt and potassium intake
Yan WU ; Mingfei DU ; Xi ZHANG ; Lan WANG ; Guilin HU ; Zejiaxin NIU ; Ting ZOU ; Xiaoyu ZHANG ; Chao CHU ; Yueyuan LIAO ; Qiong MA ; Dan WANG ; Keke WANG ; Hao JIA ; Chen CHEN ; Yu YAN ; Yue SUN ; Tongshuai GUO ; Jie ZHANG ; Weihua GAO ; Ziyue MAN ; Ke GAO ; Wenjing LUO ; Jianjun MU ; Yang WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2023;44(1):38-45
【Objective】 Based on our previously established salt-sensitive hypertension cohort, we aimed to examine the association of genetic variants in uromodulin with blood pressure(BP) responses to dietary interventions of sodium and potassium intake. 【Methods】 In 2004, 514 subjects from 124 families in Mei County, Shaanxi Province, were recruited to establish the salt-sensitive hypertension study cohort. Among them, 333 non-parent subjects were selected and sequentially maintained on a normal-diet for 3 days, low-salt diet for 7 days, then a high-salt diet for 7 days and a high-salt diet with potassium supplementation for another 7 days. Thirteen single nucleotide polymorphisms(SNPs) in the uromodulin gene were genotyped on the MassARRAY platform. 【Results】 BP levels decreased from the baseline to low-salt diet, increased from low-salt to high-salt diet, and decreased again from the high-salt diet to the high-salt plus potassium supplementation intervention. SNPs rs77875418 and rs4997081 of the uromodulin gene were significantly associated with diastolic BP(DBP) and mean arterial pressure(MAP) responses to high-salt diet. In addition, SNPs rs77875418, rs79245268, rs4293393, rs6497476, rs4997081, rs13333226, and rs12917707 were significantly associated with systolic BP(SBP), DBP, and MAP responses to high-salt diet with potassium supplementation. 【Conclusion】 Genetic variants in uromodulin gene are significantly associated with BP responses to sodium and potassium supplementation, suggesting that uromodulin may be mechanistically involved in BP sodium-sensitivity and potassium-sensitivity.

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