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BACKGROUNDS: Kidney stone also known as urolithiasis or nephrolithiasis, is one of the oldest diseases known to medicine, however, the gene expression changes and related kidney injury remains unclear. METHODS: A calculi rat model was developed via ethylene glycol- and ammonium chloride-induction. Integrated proteomic and transcriptomic analysis was performed to characterize the distinct gene expression profiles in the kidney of calculi rat. Differential expressed genes (DEGs) were sub-clustered into distinct groups according to the consistency of transcriptome and proteome. Gene Ontology and KEGG pathway enrichment was performed to analyze the functions of each sub-group of DEGs. Immunohistochemistry was performed to validated the expression of identified proteins. RESULTS: Five thousand eight hundred ninety-seven genes were quantified at both transcriptome and proteome levels, and six distinct gene clusters were identified, of which 14 genes were consistently dysregulated. Functional enrichment analysis showed that the calculi rat kidney was increased expression of injured & apoptotic markers and immune-molecules, and decreased expression of solute carriers & transporters and many metabolic related factors. CONCLUSIONS: The present proteotranscriptomic study provided a data resource and new insights for better understanding of the pathogenesis of nephrolithiasis, will hopefully facilitate the future development of new strategies for the recurrence prevention and treatment in patients with kidney stone disease.
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Cálculos Renais , Transcriptoma , Ratos , Animais , Proteoma/genética , Proteômica , Cálculos Renais/genética , Rim/metabolismoRESUMO
OBJECTIVE：To optim ize ethanol extraction technology of Mongolian medicine Naru- 3. METHODS ：The L 9（34） orthogonal design was used to optimize ethanol extraction technology of Mongolian medicine Naru- 3 with solid-liquid ratio ，ethanol volume fraction and extraction time as factors ，using comprehensive scores for the contents of benzoylaconitine ，benzoylneoaconitine， benzoylhypoaconitine，aconitine，neoaconitine，hypoaconitine，piperine and gallic acid as indexes. RESULTS ：The optimal ethanol extraction technology was that solid-liquid ratio of 1∶10（g/mL），ethanol volume fraction of 75％，extracting for 1.5 h. After 3 times of validation tests ，average contents of above 8 components in ethanol extract from Naru- 3 were 1.69，1.48，14.69，0.28， 0.05，0.08，26.01，17.33 mg/g（RSDs were 0-4.96％，n＝3），respectively. Average comprehensive score was 19.03（RSD＝1.42％， n＝3）. CONCLUSIONS ：The optimal ethanol extraction technology of Mongolian medicine Naru- 3 is stable and feasible.
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Objective:To establish a method for rapid determination of iodine in whole blood by direct alkali dilution inductively coupled plasma mass spectrometry (ICP-MS).Methods:Totally 0.50 ml whole blood sample was collected, and 2% ammonia and 0.01% Triton X-100 solution were added to constitute a total volume of 10.0 ml. After shaking to uniformity, 1.0 μg/ml rhodium and 20% isopropanol were used as on-line internal standard solution. The flow ratio of internal standard solution to the solution to be measured was 1∶16. The sample was quantitatively determined by ICP-MS. The linear range, limit of detection (LOD), accuracy and precision of the method were evaluated.Results:Iodine in whole blood could be determined and had a good linear relationship within the range of 0-200 μg/L, with correlation coefficient ( r) > 0.999. The LOD of the method was 0.1 μg/L, the limit of quantitation (LOQ) was 0.3 μg/L, the recovery rate of iodine in whole blood was 88.5%-106.1%, and the relative standard deviation was 2.2%-4.7% ( n=7). Conclusions:A method for rapid determination of iodine in whole blood by direct alkali dilution ICP-MS is successfully established. This method is accurate, simple, rapid, and highly automatic, and it can be widely applied in determination of iodine in whole blood.
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OBJECTIVE： To establish a method for simultaneously determining the contents of 6 volatile components such as eugenol， methyl eugenol， isoeugenol， α-asarone， alkaloid and myristyl ether in Zhachong shisan. METHODS：Volatile oil was extracted by steam distillation and analyzed by gas chromatography. The chromatographic column was ov-1701， with hydrogen flame ionization detector. The carrier gas was nitrogen， the inlet temperature was 220 ℃， the detector temperature was 240 ℃， the flow rate was 1.0 mL/min， the injection volume was 1 μL， the split ratio was 10 ∶ 1， with programmed temperature. RESULTS： The linear ranges of eugenol， methyl eugenol， isoeugenol， α-asarone， costunolide and myristyl were 2.00-12.00， 1.00-6.00， 0.60-3.60， 0.05-0.30， 0.05-0.30， 0.60-3.60 mg/mL（r＝0.999 9）. The detection limits were 0.01， 0.01， 0.02， 0.01， 0.01， 0.01 mg/mL， respectively； the limits of quantification were 0.05， 0.03， 0.06， 0.02， 0.02， 0.03 mg/mL， respectively. RSDs of precision， stability and reproducible tests were all lower than 3％； average recovery rates were 99.33％-100.04％， RSDs were 0.02％-0.35％ （n＝6）； RSDs of durability test were all lower than 5％. CONCLUSIONS： The established method was simple and reproducible. It can be used for simultaneous determination of the contents of 6 volatile components in Zhachong shisan.
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OBJECTIVE:To establish HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry for rapid determination of aconitine,mesaconitine,hypaconitine,benzoylaconitine,benzoylmesaconine and benzoylhypacoitine in rat plasma. METHODS:Internal standard lappaconitine was added into plasma sample,and methanol precipitated protein was used for pretreatment. HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry was adopted. HPLC condition was as follows as Sinochrom ODS-BP C18column,mobile phase consisted of acetonitrile-1% formic acid solution(50:50,V/V),the flow rate of 0.6 mL/min,sample size of 10 μL,column temperature of 25 ℃,automatic sampler temperature of 4 ℃. Mass spectrum scanning mode was full ion monitoring model,positive ion acquisition,mass charge ratios(m/z)of ion were 646.32(aconitine), 632.30(mesaconitine),616.31(hypaconitine),604.31(benzoylaconitine),590.29(benzoylmesaconine),574.30(benzoylhypacoitine), 585.31(internal standard). Six male Wistar rats were collected and given single dose of total alkaloid extract of Aconitum carmichaeli(4 mg/kg)intragastrically. Blood samples were collected before medication(0 h)and 0.5,0.75,1.25,1.5,2,4,6, 8,10,24 h after medication. Plasma concentration was determined and pharmacokinetic parameters were calculated by using PK-Solver V2.0 software. RESULTS:The linear range of 6 kinds of aconitum alkaloids in plasma were 0.1-10 μ g/L(r>0.992). The limit of quantitation was 0.1 μ g/L. Average recovery was higher than 75%,RSDs of intra-day and inter-day,matrix effects,stability test were all lower than 15%. The tmaxof 6 kinds of aconite alkaloids were about 1.2 h;t1/2were about 10 h;cmaxof monoestertype aconite alkaloids were higher than those of diester-type aconite alkaloids. CONCLUSIONS:Established HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry is accurate,sensitive,simple and rapid, and can be used for plasma concentration monitoring of 6 kinds of aconitum alkaloids.
