{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002101","ANALYSIS_ID":"AN003434","VERSION":"1","CREATED_ON":"March 11, 2022, 5:11 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 lipidomics 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 Lipidomics)","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","GENDER":"Male"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"C-1",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"C-2",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"C-3",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"C-4",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"C-5",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"C-6",
"Factors":{"Treatment":"3 times per week with 4 consecutive respectively intraperitoneal injection (10 mL/kg of olive oil)"}
},
{
"Subject ID":"-",
"Sample ID":"AL-1",
"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":"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":"AL-3",
"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":"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":"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":"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":"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":"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":"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":"AH-4",
"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":"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":"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":"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":"CL-2",
"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":"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":"CL-4",
"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":"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":"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":"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":"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":"CM-3",
"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":"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":"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":"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":"CH-1",
"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":"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":"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":"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":"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":"CH-6",
"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":"200-220 mg liver tissues mixed with 1 mL 0.1×PBS were homogenized using the Mini-BeadBeater-16. 225 μL of methanol, 750 μL of MTBE and 188 μL of water were added into 20 μL of liver homogenate for lipid extraction. The supernatant was blown dry in nitrogen, reconstituted in 100 µL of organic solvent (IPA: ACN: water = 30: 65: 5, v/v/v) and transferred to auto-sampler vials."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"The resuspended samples were placed on the sample-tray at 4 ℃ with 1 μL-volume injection for targeted-lipidomics analysis, which was performed on an UHPLC Q-Trap/MS system (Qtrap 6500, SCIEX, USA). A Kinetex C18 column (100 mm × 3.0 mm, 2.6 μm; Phenomenex); mobile phase A was methanol: ACN: water = 1:1:1 (v/v/v) with 5 mM ammonium acetate, while mobile phase B was IPA with 5 mM ammonium acetate. The following gradient-elution program was 0-0.5 min, 80% A; 0.5-1.5 min, 80%-60% A; 1.5-3 min, 60%-40% A; 3-13 min, 40%-2% A; 13-13.1 min, 2%-80% A; 13.1-17 min, 80% A. The flow rate was at 0.3 mL/min and the column temperature was maintained at 50 ℃.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"SCIEX ExionLC AD","COLUMN_NAME":"Phenomenex Kinetex C18(100×3.0mm, 2.6μm)"},

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

"MS":{"INSTRUMENT_NAME":"ABI Sciex 6500 QTrap","INSTRUMENT_TYPE":"QTRAP","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Analyst® (version 1.7.1, AB Sciex, Monza, Italy) Multiquant™ (version 3.0.31721.0, AB Sciex, Monza, Italy)"},

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

"Data":[{"Metabolite":"24 OH 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