{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002097","ANALYSIS_ID":"AN003426","VERSION":"1","CREATED_ON":"March 10, 2022, 7:01 am"},

"PROJECT":{"PROJECT_TITLE":"Functional metabolomics-based molecular profiling of acute and chronic hepatitis","PROJECT_TYPE":"Targeted MS quantitative analysis","PROJECT_SUMMARY":"Characteristics of liver metabolomics in acute and chronic hepatitis","INSTITUTE":"Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University","DEPARTMENT":"Shanghai Center for Systems Biomedicine","LABORATORY":"Lu Group","LAST_NAME":"Lu","FIRST_NAME":"Haitao","ADDRESS":"800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China","EMAIL":"haitao_lu@sjtu.edu.cn","PHONE":"15221478139"},

"STUDY":{"STUDY_TITLE":"Functional metabolomics-based molecular profiling of acute and chronic hepatitis (Liver Metabolomics)","STUDY_SUMMARY":"Non-alcoholic steatohepatitis (NASH) is a metabolic dysregulation triggered by an overload disrupting the hepatic tolerance to external molecules. With the complexity and diversity of hepatitis triggers, no effective clinical classification and treatment are available, and even using the same strategies or approaches for acute and chronic hepatitis. For us, it is really difficult to precisely diagnose and treat hepatitis accordingly. To overcome this challenge, we integrated metabolomic, lipidomics, transcriptomics and other life science frontier technologies for functional metabolomics studies, and pioneered the redefinition of hepatitis at the molecular level. Our findings suggested that acute hepatitis mainly interferes with purine metabolism and amino acids metabolism, while chronic hepatitis mainly causes disruption of hepatic bile acids and lipidome, especially glycerolipids. Based on the liver-gut axis, we also found that the metabolic regulation of the gut microbiota is another key factor for chronic hepatitis development. In conclusion, functional metabolomics enables the cognition of disease occurrence, development and regression from small molecule metabolic modifications and modulations, realizing the ultimate goal of treating diseases and improving population health through regulation of dysregulated metabolism","INSTITUTE":"Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University","DEPARTMENT":"Shanghai Center for Systems Biomedicine","LABORATORY":"Lu Group","LAST_NAME":"Lu","FIRST_NAME":"Haitao","ADDRESS":"800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China","EMAIL":"haitao_lu@sjtu.edu.cn","PHONE":"15221478139"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"20210522-C-3",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-C-4",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-C-5",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-C-6",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-C-7",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-C-8",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AL-1-0524",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 1% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AL-2",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 1% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AL-4",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 1% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AL-5",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 1% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AL-6",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 1% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AL-7",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 1% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AH-1",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 5% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AH-2",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 5% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AH-3",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 5% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AH-5",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 5% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AH-6",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 5% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-AH-8",
"Factors":{"Treatment":"Only intraperitoneally (i.p) injected with 5% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil."}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CL-1",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 1% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CL-3",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 1% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CL-5",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 1% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CL-6",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 1% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CL-7",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 1% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CL-8",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 1% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CM-1",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 5% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CM-2",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 5% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CM-3-2",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 5% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CM-4",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 5% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CM-5",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 5% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CM-6",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 5% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CH-2",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 10% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CH-3",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 10% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CH-4",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 10% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CH-5",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 10% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CH-6",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 10% CCl4 (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"20210522-CH-8",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection with 10% CCl4 (10 mL/kg of olive oil)"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Animals were killed at 24h post the last CCl4 treatment and the liver tissue samples were harvested for further assays.","SAMPLE_TYPE":"Liver"},

"TREATMENT":{"TREATMENT_SUMMARY":"Prior to experimental, animals were acclimatized breeding to laboratory conditions for 1 week. The mice were randomly allocated into 6 groups (n=8 each), as follows: a control group (C); two acute hepatitis groups, low and high concentrations (AL/ AH); three chronic hepatitis groups, low, middle and high concentration (CL/ CM/ CH). Mice in chronic groups were treated 3 times per week with 4 consecutive respectively intraperitoneal (i.p) injection with 1% / 5% / 10% CCl4 (10 mL/kg, dissolved in olive oil). However, in the acute groups, mice were only intraperitoneally (i.p) injected with 1% / 5% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil. Animals were killed at 24h post the last CCl4 treatment and serum, stool and liver tissue samples were harvested for further assays."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"100-130 mg liver tissue samples were weighted, 1.2 mL of 80% ice-cold aqueous methanol, which contains 0.001 mg/mL internal standard, and approximately 1 g of glass beads (1.0 mm i.d.) was added into a 2 mL screw-cap plastic microvial. The Mini-BeadBeater-16 (Biospec Products, USA) was used for tissue disruption. Samples were homogenized 3 times for 2 min each, with cooling procedure in between, centrifuged at 20,000 g for 10 min at 4 °C, and the supernatants were mixed with 800 μL of ice-cold acetonitrile and centrifuged again for protein precipitation. The supernatant was filtered through a 0.22 µm organic phase membrane, blown dry in nitrogen, reconstituted in 100 µL of water and transferred to vials for metabolomics assay."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"In this study, a newly developed precision-targeted metabolomics method with a UPLC-TQ/MS system (Agilent 1290 Infnity, Agilent Technologies, USA; Agilent 6495 QQQ, Agilent Technologies, USA) in a DMRM scan-mode was applied to analyze the metabolome of interest from trial samples (serum, liver tissues and stool). Briefly, the method was performed with an ACQUITY UPLC HSS T3 column (2.1 mm i.d. × 100 mm, 1.8 μm; Waters); mobile phase A and B were water and acetonitrile with 0.1% formic acid (v/v) respectively. The flow rate was at 0.3 mL/min and the column temperature was maintained at 40 ℃. The samples were placed in an auto-sampler maintained at 4 °C with a 5 μL injection volume. The optimized gradient-elution program, as follows: 0-2 min, 98% A; 2-10 min, 98-65% A; 10-12 min, 65-20% A; 12-14 min, 20-2% A; 14-30 min, 2% A.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 1290 Infinity","COLUMN_NAME":"ACQUITY UPLC HSS T3 column (2.1 mm i.d. × 100 mm, 1.8 μm; Waters)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Agilent 6495 QQQ","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Agilent MassHunter Workstation Data Acquisition Agilent MassHunter"},

"MS_METABOLITE_DATA":{
"Units":"counts",

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acid","Polarity":"Positive","m/z":"104.13","RT":"0.96"},{"Metabolite":"L-Proline","Polarity":"Positive","m/z":"116.14","RT":"1"},{"Metabolite":"L-Methionine","Polarity":"Positive","m/z":"150.22","RT":"1.44"},{"Metabolite":"L-Leucine","Polarity":"Positive","m/z":"132.18","RT":"2.21"},{"Metabolite":"L-Threonine","Polarity":"Positive","m/z":"120.13","RT":"1.07"},{"Metabolite":"L-Phenylalanine","Polarity":"Positive","m/z":"166.2","RT":"4.31"},{"Metabolite":"L-Glutamate","Polarity":"Positive","m/z":"148.06","RT":"1.08"},{"Metabolite":"his","Polarity":"Positive","m/z":"134.11","RT":"1.11"},{"Metabolite":"L-Valine","Polarity":"Positive","m/z":"118.09","RT":"1.25"},{"Metabolite":"Hippuric acid","Polarity":"Positive","m/z":"180.16","RT":"6.84"},{"Metabolite":"6-Aminocaproic acid","Polarity":"Positive","m/z":"132.11","RT":"1.01"},{"Metabolite":"Beta-Sitosterol","Polarity":"Positive","m/z":"415.72","RT":"13"},{"Metabolite":"Trans-4-Hydroxy-L-proline","Polarity":"Positive","m/z":"132.14","RT":"2.25"},{"Metabolite":"L-Ascorbic acid","Polarity":"Positive","m/z":"177.04","RT":"1.19"},{"Metabolite":"Sarcosine","Polarity":"Positive","m/z":"90.06","RT":"0.99"},{"Metabolite":"2-Deoxyguanosine","Polarity":"Positive","m/z":"268.27","RT":"3.53"},{"Metabolite":"Deoxycytidine","Polarity":"Positive","m/z":"228.23","RT":"1.38"},{"Metabolite":"Dopamine","Polarity":"Positive","m/z":"154.15","RT":"2.76"},{"Metabolite":"Isonicotinic acid","Polarity":"Positive","m/z":"124.12","RT":"1.53"},{"Metabolite":"Reduced glutathione","Polarity":"Positive","m/z":"308.09","RT":"1.46"},{"Metabolite":"2-Deoxyadenosine","Polarity":"Positive","m/z":"252.1","RT":"4.57"},{"Metabolite":"Cytidine monophosphate","Polarity":"Positive","m/z":"324.21","RT":"1.04"},{"Metabolite":"Biotin","Polarity":"Positive","m/z":"245.32","RT":"7.32"},{"Metabolite":"Adenosine","Polarity":"Positive","m/z":"268.25","RT":"2.68"},{"Metabolite":"L-Homocysteine","Polarity":"Positive","m/z":"136.05","RT":"1.06"},{"Metabolite":"Inosine-5'-monophosphate","Polarity":"Positive","m/z":"349.06","RT":"1.33"},{"Metabolite":"Cytidine","Polarity":"Positive","m/z":"244.23","RT":"1.27"},{"Metabolite":"N-Acetylglutamine","Polarity":"Positive","m/z":"189.19","RT":"1.24"},{"Metabolite":"Betaine","Polarity":"Positive","m/z":"118.16","RT":"0.95"},{"Metabolite":"Indole-3-acetic acid","Polarity":"Positive","m/z":"176.1","RT":"11.07"},{"Metabolite":"Histamine","Polarity":"Positive","m/z":"112.09","RT":"1.26"},{"Metabolite":"L-Arginine","Polarity":"Positive","m/z":"175.21","RT":"1.05"},{"Metabolite":"L-Lysine","Polarity":"Positive","m/z":"147.2","RT":"1.07"},{"Metabolite":"Thymine","Polarity":"Positive","m/z":"127.12","RT":"3.84"},{"Metabolite":"Putrescine","Polarity":"Positive","m/z":"89.08","RT":"1.66"},{"Metabolite":"Serotonin","Polarity":"Positive","m/z":"177.19","RT":"3.65"},{"Metabolite":"L-Carnitine hydrochloride","Polarity":"Positive","m/z":"162.1","RT":"0.95"},{"Metabolite":"5-Aminolevulinic acid","Polarity":"Positive","m/z":"132.07","RT":"1"},{"Metabolite":"5-aminosalicylic acid","Polarity":"Positive","m/z":"154.15","RT":"2.69"},{"Metabolite":"Adenylosuccinic acid","Polarity":"Positive","m/z":"464.08","RT":"3.83"},{"Metabolite":"Hexanoylcarnitine","Polarity":"Positive","m/z":"260.1","RT":"8.53"},{"Metabolite":"Palmitoyl-l-carnitine","Polarity":"Positive","m/z":"400.34","RT":"13.25"},{"Metabolite":"3-Guanidinopropionic acid?","Polarity":"Positive","m/z":"132.14","RT":"1.04"},{"Metabolite":"Acetylcarnitine","Polarity":"Positive","m/z":"204.21","RT":"1.39"},{"Metabolite":"Adenosine 5'-monophosphate","Polarity":"Positive","m/z":"348.07","RT":"1.33"},{"Metabolite":"Sphinganine","Polarity":"Positive","m/z":"302.52","RT":"12.97"},{"Metabolite":"Taurine","Polarity":"Positive","m/z":"126.16","RT":"1.06"},{"Metabolite":"Oxidized glutathione","Polarity":"Positive","m/z":"613.2","RT":"1.72"},{"Metabolite":"2'-Deoxycytidine 5'-monophosphate","Polarity":"Positive","m/z":"308.07","RT":"1.18"},{"Metabolite":"Uridine 5'-monophosphate","Polarity":"Positive","m/z":"325.05","RT":"1.13"},{"Metabolite":"S-(5'-Adenosyl)-L-homocysteine","Polarity":"Positive","m/z":"385.1","RT":"1.69"},{"Metabolite":"Dephospho-CoA","Polarity":"Positive","m/z":"688.2","RT":"4.95"},{"Metabolite":"Choline","Polarity":"Positive","m/z":"105.1","RT":"0.93"},{"Metabolite":"Creatine","Polarity":"Positive","m/z":"132.08","RT":"0.99"},{"Metabolite":"Spermidine","Polarity":"Positive","m/z":"146.17","RT":"0.71"},{"Metabolite":"Thiamine","Polarity":"Positive","m/z":"266.12","RT":"0.96"},{"Metabolite":"Inosine","Polarity":"Positive","m/z":"269.09","RT":"3.08"},{"Metabolite":"Cytosine","Polarity":"Positive","m/z":"112.05","RT":"1.26"},{"Metabolite":"Guanosine 5'-monophosphate","Polarity":"Positive","m/z":"364.07","RT":"1.37"},{"Metabolite":"Alanine","Polarity":"Positive","m/z":"90.1","RT":"0.99"},{"Metabolite":"Uracil","Polarity":"Positive","m/z":"113.1","RT":"1.76"},{"Metabolite":"Tryptophan","Polarity":"Positive","m/z":"205.1","RT":"5.73"},{"Metabolite":"Adenine","Polarity":"Positive","m/z":"136.1","RT":"2.68"},{"Metabolite":"Phosphoenolpyruvic acid","Polarity":"Positive","m/z":"168.99","RT":"0.91"},{"Metabolite":"Taurolithocholic acid","Polarity":"Positive","m/z":"484.31","RT":"12.24"},{"Metabolite":"Taurohyodeoxycholic acid","Polarity":"Positive","m/z":"500.31","RT":"12.27"},{"Metabolite":"Aminoadipic acid","Polarity":"Positive","m/z":"162.08","RT":"1.02"},{"Metabolite":"Norvaline","Polarity":"Positive","m/z":"118.09","RT":"1.22"},{"Metabolite":"Pyroglutamic acid","Polarity":"Positive","m/z":"130.05","RT":"1.25"},{"Metabolite":"Pipecolic acid","Polarity":"Positive","m/z":"130.09","RT":"1.27"},{"Metabolite":"L-Glutamine","Polarity":"Positive","m/z":"147.08","RT":"1.07"},{"Metabolite":"L-Ornithine","Polarity":"Positive","m/z":"133.1","RT":"2.02"}]
}

}