Objective: Differential flora and differential metabolites shared by the intestinal and respiratory tracts of rats were screened to analyze the possible role of changes in intestinal flora and metabolites in the progression of pneumoconiosis in rats. Methods: In April 2020, 18 SD rats were randomly divided into three groups (control group, coal mine dust group and silica group, 6 in each group) , rats in the coal mine dust group and silica group were perfused with 1 ml of 50 mg/ml coal mine well dust suspension and silica suspension by nontracheal exposure, respectively. While rats in the control group were perfused with an equal dose of sterilized normal saline. Twenty four weeks after dust staining, rat feces, throat swabs, and lung lavages were collected. 16SrDNA gene sequencing and UHPLC-QTOF-MS untargeted metabolomics were used to analyze the flora and metabolites in feces, throat swabs and lung lavage fluid of rats in each group, to screen for shared differential flora and shared differential metabolites in intestinal and respiratory tract, and the correlation analysis between the differential flora and metabolites was performed using Spearman's statistics. Results: Compared with the control group, a total of 9 species shared differential flora between intestinal and respiratory tract were screened at phylum level, and a total of 9 species shared differential genus between intestinal and respiratory tract were screened at genus level in the coal mine dust group, mainly Firmicutes, Actinobacteria, Streptococcus, Lactobacillus, etc. Compared with the control group, a total of 9 shared differential flora were screened at the phylum level, and a total of 5 shared differential genus were screened at the genus level in the silica group, mainly Proteobacteria, Actinobacteria, Allobactera, Mucilaginibacter, etc. Compared with the control group, a total of 7 shared differential metabolites were screened for up-regulation of Stigmatellin, Linalool oxide and Isoleucine-leucine in both intestinal and respiratory tract in the coal mine dust group. Compared with the control group , a total of 19 shared differential metabolites werescreened in the silica group, of which Diethanolamine, 1-Aminocyclopropanecarboxylic acid, Isoleucine-leucine, Sphingosine, Palmitic acid, D-sphinganine, 1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine, and 1-Stearoyl-2-oleoyl-sn-glycerol 3-phosphocholine were up-regulated in both the intestinal and respiratory tract. Conclusion: There is a translocation of intestinal and respiratory flora in pneumoconiosis rats, and rats have an imbalance of lipid metabolism during the progression of pneumoconiosis.
Rats ; Animals ; Isoleucine ; Leucine ; Coal Mining ; Rats, Sprague-Dawley ; Pneumoconiosis ; Dust/analysis* ; Silicon Dioxide ; Coal

Ultra-high-performance liquid chromatography-Q exactive orbitrap tandem mass spectrometry(UHPLC-QEOrbitrap-MS/MS) was used to explore the inhibitory effect and mechanism of ginkgo flavone aglycone(GA) combined with doxorubicin(DOX) on H22 cells. The effects of different concentrations of GA and DOX on the viability of H22 cells were investigated, and combination index(CI) was used to evaluate the effects. In the experiments, control(CON) group, DOX group, GA group, and combined GA and DOX(GDOX) group were constructed. Then the metabolomics strategy was employed to explore the metabolic markers that were significantly changed after combination therapy on the basis of single medication treatment, and by analyzing their biological significance, the effect and mechanism of the anti-tumor effect of GA combined with DOX were explained. The results revealed that when 30 μg·mL~(-1) GA and 0.5 μmol·L~(-1) DOX was determined as the co-administration concentration, the CI value was 0.808, indicating that the combination of GA and DOX had a synergistic anti-tumor effect. Metabolomics analysis identified 23 metabolic markers, including L-arginine, L-tyrosine and L-valine, mostly amino acids. Compared with the CON group, 22 and 17 metabolic markers were significantly down-regulated after DOX treatment and GA treatment, respectively. Compared with the DOX and GA groups, the treatment of GA combined with DOX further down-regulated the levels of these metabolic markers in liver cancer, which might contribute to the synergistic effect of the two. Five key metabolic pathways were found in pathway enrichment analysis, including glutathione metabolism, phenylalanine metabolism, arginine and proline metabolism, β-alanine metabolism, and valine, leucine and isoleucine degradation. These findings demonstrated that the combination of GA and DOX remarkably inhibited the viability of H22 cells and exerted a synergistic anti-tumor effect. The mechanism might be related to the influence of the energy supply of tumor cells by interfering with the metabolism of various amino acids.
Arginine/therapeutic use* ; Doxorubicin/therapeutic use* ; Flavones/therapeutic use* ; Ginkgo biloba/chemistry* ; Glutathione ; Humans ; Isoleucine/therapeutic use* ; Leucine/therapeutic use* ; Liver Neoplasms/drug therapy* ; Metabolomics/methods* ; Phenylalanine/therapeutic use* ; Proline ; Tandem Mass Spectrometry/methods* ; Tyrosine/therapeutic use* ; Valine/therapeutic use* ; beta-Alanine/therapeutic use*

OBJECTIVE: To explore the serological characteristics and molecular biological mechanism of an a_el subtype specimen. METHODS: The ABO blood typing was identified by routine blood group serological and absorption/elution methods; PCR-SBT method for ABO genotyping: 7 exons of ABO gene were amplified by PCR, the amplified products were purified, and then sequencing primers were designed and the amplified products were sequenced directly for analysis; 3D molecular model was constructed and the difference of free energy (ΔΔG) was used to predict the GTA mutant stability. RESULTS: A antigen was not detected on erythrocytes through absorption and elution tests, which was not consistent with the serological characteristics of a_el, and the serological typing results were ambiguous. The ABO genotype was ABO*AEL.02/O.01.01, and there were two mutations in exon 7 of the gene, c.467C>T and c.646T>A, which could lead to the replacement of proline with leucine at position 156 (p.Pro156Leu) and phenylalanine with isoleucine at position 216 on the GTA, respectively. The 3D model predicts that the mutations do not introduce new hydrogen bonds to the GTA mutant and do not form a new secondary structure, but can lead to an increase in the ΔΔG value of the GTA mutant, suggesting a decrease in protein stability. CONCLUSION The serological characteristics alone is not reliable to determine the a_el subype; the a_el phenotype may be due to the GTA mutant that reduces enzyme stability.
ABO Blood-Group System/genetics* ; Alleles ; Genotype ; Isoleucine/genetics* ; Leucine/genetics* ; Phenotype ; Phenylalanine/genetics* ; Proline/genetics*

OBJECTIVE: To compare the analgesic effect of Jin Ling Zi Powder (JLZ) and its two single herbs. METHODS: The hot plate method was used to induce pain. Totally 36 mice were randomly divided into 6 groups by a complete random design, including control, model, aspirin (ASP, 0.14 g/kg body weight), JLZ (14 g/kg body weight), Corydalis yanhusuo (YHS, 14 g/kg body weight), and Toosendan Fructus (TF, 14 g/kg body weight) groups, 6 mice in each group. The mice in the control and model groups were given the same volume of saline, daily for 2 consecutive weeks. At 30, 60, 90, and 120 min after the last administration, the pain threshold of mice in each group was measured, and the improvement rate of pain threshold was calculated. Serum endogenous metabolites were analyzed by gas chromatography-mass spectrometry (GC-MS). RESULTS: There was no statistical difference in pain threshold among groups before administration (P>0.05). After 2 weeks of administration, compared with the model group, the pain threshold in JLZ, YHS, TF and ASP groups were increased to varying degrees (P<0.05). JLZ had the best analgesic effect and was superior to YHS and TF groups. A total of 14 potential biomarkers were screened in serum data analysis and potential biomarkers levels were all reversed to different degrees after the treatment with JLZ and its single herbs. These potential biomarkers were mainly related to glyoxylate and dicarboxylate metabolism, glycine, serine and threonine metabolism, valine, leucine and isoleucine biosynthesis, aminoacyl-tRNA biosynthesis and inositol phosphate metabolism. CONCLUSIONS The analgesic mechanism of JLZ and YHS was mainly due to the combination of glycine and its receptor, producing post-synaptic potential, reducing the excitability of neurons, and weakening the afferent effect of painful information.
Animals ; Mice ; Analgesics/therapeutic use* ; Aspirin/pharmacology* ; Biomarkers ; Body Weight ; Drugs, Chinese Herbal/therapeutic use* ; Glycine ; Glyoxylates ; Inositol Phosphates ; Isoleucine ; Leucine ; Metabolomics/methods* ; Powders ; RNA, Transfer ; Serine ; Threonine ; Valine

A 1-year-old female with maple syrup urine disease presenting with erythematous, partially eroded plaques on the trunk, anogenital area, and extremities experienced metabolic crisis. The skin lesions appeared at 11 months of age and was thought to result from amino acid imbalance secondary to erratic supplementation of specialized milk formula devoid of isoleucine, leucine, and valine. Serial urine monitoring showed persistent ketones and elevated serum leucine and valine. The patient was managed with emollients, intralipid 20%, and addition of isoleucine and valine supplements to counter the neurotoxic effect of leucine. After 8 days of proper feeding and continuous emollient application, the lesions improved and skin biopsy revealed superficial perivascular dermatitis. Although a decrease in erythema and desquamation was noted, the patient had persistent cerebral edema and continued to deteriorate.

Maple Syrup Urine Disease ; Isoleucine ; Leucine ; Valine ; Erythema

Two trials of external quality assessment (EQA) of conventional newborn screening tests for phenylketonuria, galactosaemia, congenital adrenal hyperplasia, maple syrup urine disease, homocystinuria, and congenital hypothyroidism, as well as of newborn screening tests using tandem mass spectrometry were performed in 2013. A total of 32 specimens in the form of dried blood spots were distributed to 16 laboratories and the response rate of these laboratories was 100%. Total T4, free T4, 17-hydroxyprogesterone, leucine, isoleucine, galactose, methionine, alanine, C8/C2, C8/C10, and C5-OH did not meet the accepted performance criteria. The mean, standard deviation, coefficient of variation, median, and cut-offs were evaluated for each analyte in the newborn screening tests. Two trials of EQA for the analyses of methylmalonic acid, vanillylmandelic acid, very long fatty acids, organic acids, and amino acids were also performed. A well-designed EQA program and continuous education would improve the performance of biochemical genetic tests.
17-alpha-Hydroxyprogesterone ; Adrenal Hyperplasia, Congenital ; Alanine ; Amino Acids ; Congenital Hypothyroidism ; Education ; Fatty Acids ; Galactose ; Homocystinuria ; Humans ; Infant, Newborn ; Isoleucine ; Korea ; Leucine ; Maple Syrup Urine Disease ; Mass Screening ; Methionine ; Methylmalonic Acid ; Molecular Biology* ; Phenylketonurias ; Tandem Mass Spectrometry ; Vanilmandelic Acid

Maple syrup urine disease (MSUD) is a disorder that involves the metabolism of branched chain amino acids, arising from a defect in branched-chain alpha-keto acid dehydrogenase complex. Mutations have been identified in the BCKDHA, BCKDHB, or DBT genes, which encode different subunits of the BCKDH complex. Although encephalopathy and progressive neurodegeneration are its major manifestations, the severity of the disease may range from the severe classic type to milder intermediate variants. We report two Korean siblings with the milder intermediate MSUD who were diagnosed with MSUD by a combination of newborn screening tests using tandem mass spectrometry and family genetic screening for MSUD. At diagnosis, the patients' plasma levels were elevated for leucine, isoleucine, valine, and alloisoleucine, and branched-chain alpha-keto acids and branched-chain alpha-hydroxy acids were detected in their urine. BCKDHA, BCKDHB, and DBT analysis was performed, and two novel mutations were identified in BCKDHB. Our patients were thought to have the milder intermediate variant of MSUD, rather than the classic form. Although MSUD is a typical metabolic disease with poor prognosis, better outcomes can be expected if early diagnosis and prompt management are provided, particularly for milder forms of the disease.
3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) ; Amino Acids ; Diagnosis ; Early Diagnosis ; Genetic Testing ; Humans ; Infant, Newborn ; Isoleucine ; Leucine ; Maple Syrup Urine Disease* ; Mass Screening ; Metabolic Diseases ; Metabolism ; Plasma ; Prognosis ; Siblings* ; Tandem Mass Spectrometry ; Valine

Resistance to thyroid hormone (RTH) is a rare inherited syndrome characterized by diminished response of the target tissue to thyroid hormone caused, in the majority of cases, by mutation of the thyroid hormone receptor beta (THRbeta) gene. Despite elevated serum levels of free thyroid hormones and thyroid stimulating hormone (TSH), the paucity of symptoms and signs of thyroid dysfunction suggest RTH. We report the case of a 9-year-old girl with goiter. Her thyroid function tests showed increased serum levels of free thyroxine, triiodothyronine, and TSH. The genetic analysis of THRbeta confirmed a novel mutation in exon 9; this was a heterozygous C-to-T change in the 327th codon, substituting threonine for isoleucine (T327I).
Child ; Codon ; Exons ; Female ; Goiter ; Humans ; Isoleucine ; Threonine ; Thyroid Function Tests ; Thyroid Gland* ; Thyroid Hormone Receptors beta* ; Thyroid Hormone Resistance Syndrome ; Thyroid Hormones ; Thyrotropin ; Thyroxine ; Triiodothyronine

OBJECTIVE: To determine the plasma amino acid metabolism of "same symptom for different diseases" in different cancer patients in Uyghur medicine. METHODS: Plasma amino acid concentration was tested by high-performance liquid chromatography (HPLC) in cancer patients with different symptom, and the spectral profiles were subjected to a t-test for statistical significance. RESULTS: Compared with the healthy group, lung cancer, cervical cancer, breast cancer and gastric cancer patients with abnormal Savda had lower concentration of plasma amino acids except some amino acids. Lung cancer patients with abnormal Savda had higher concentration of plasma phenylalanine, serine, cystine, valine, isoleucine, leucine and aspartic acid than Unsavda patients (P<0.05). Cervical cancer patients with abnormal Savda had low concentration of plasma arginine, but higher concentration of plasma cystine than Unsavda patients (P<0.05). Breast cancer patients with abnormal Savda had higher concentration of plasma leucine, serine, taurine, cystine, tyrosine, valine, isoleucine and asparagine than Unsavda patients (P<0.05). Gastric cancer patients with abnormal Savda had high concentration of plasma cystine but lower concentration of plasma phenylalanine, threonine and arginine than Unsavda patients (P<0.05). CONCLUSION Different tumor patients with abnormal Savda have common characteristics and significant differences.
Amino Acids ; blood ; Arginine ; Aspartic Acid ; Chromatography, High Pressure Liquid ; Cystine ; Humans ; Isoleucine ; Leucine ; Medicine, Chinese Traditional ; Neoplasms ; blood ; Serine ; Tyrosine ; Valine

OBJECTIVETo investigate the effect of 4-hydroxyisoleucine (4-HIL), an active component of Trigonella Foenum-graecum L. on high glucose induced insulin resistance (IR) in 3T3-L1 adipocytes, and to explore underlying molecular mechanisms.

METHODS3T3-L1 adipocytes were treated with 25 mmol/L glucose and 0.6 nmol/L insulin to induce IR. They were intervened by different concentrations of 4-HIL (at 5, 10, and 20 micromol/L). [3H]-Deoxy-D-glucose up-taking method was used to detect the glucose uptake. The mRNA expression of cellular tumor necrosis factor-alpha (TNF-alpha) was detected by polymerase chain reaction (PCR). The content of TNF-alpha in the culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). Palmitic acid (PA) acted as the control.

RESULTSAfter intervened by 25 mmol/L glucose and 0.6 nmol/L insulin for 18 h, the insulin-stimulated glucose transportation in 3T3-L1 adipocytes was inhibited by 63%. The mRNA expression of cellular TNF-alpha in adipocytes significantly increased, when compared with that in normal adipocytes (P < 0.05). The level of TNF-alpha secreted in the culture supernatant was increased by 70 pg/mL (P < 0.05). Similar changes occurred in the PA group. After exposure to 4-HIL (5, 10, or 20 micromol/L) for 24 h, the glucose transportation was increased by 35%, 50%, and 60%, respectively. PCR results showed that along with increasing 4-HIL concentrations, the mRNA expression of cellular TNF-alpha showed a decreasing trend, showing statistical difference when compared with the model group and the PA group (P < 0.05). Compared with the model group, the TNF-alpha level in the supernatant was respectively reduced by 10 pg/mL, 18 pg/mL, and 39 pg/mL after intervention (P < 0.05).

CONCLUSION4-HIL could remarkably improve high glucose-induced IR in 3T3-L1 adipocytes. Meanwhile, 4-HIL could inhibit the secretion of TNF-alpha.

3T3-L1 Cells ; Adipocytes ; drug effects ; metabolism ; Animals ; Glucose ; adverse effects ; metabolism ; Insulin ; metabolism ; Insulin Resistance ; Isoleucine ; analogs & derivatives ; pharmacology ; Male ; Mice ; Trigonella ; chemistry ; Tumor Necrosis Factor-alpha ; metabolism