To compare the chemical change of Paeoniae Radix Alba (PRA) after vinegar-baking processing, as well as the effect of vinegar types exerted on the processing, 1H NMR-based metabolomic approach combined with multivariate statistical analysis was used to investigate the different metabolites between the raw and two vinegar-baked PRA. More than thirty metabolites were identified in the 1H NMR spectrum of PRA, and the multivariate statistical analysis showed that raw and two vinegar-baked PRA could be separated obviously. After vinegar-baking, the contents of isoleucine, lactate, alanine, arginine, albiflorin, and 5-hydroxymethyl furfural (5-HMF) elevated, while those of sucrose, paeoniflorin and its analogues (calculated by benzoate) decreased. The chemical compositions of two vinegar-baked PRA were also different. Shanxi vinegar- baked PRA showed higher levels of leucine, isoleucine, valine, and albiflorin, while rice vinegar-baked PRA contained more sucrose and paeoniflorin's analogues (calculated by benzoate). And the chemical changes in Shanxi vinegar-baked PRA were greater than those of rice vinegar-baked PRA. The results revealed the chemical differences between raw and vinegar-baked PRA, as well as the influence of vinegar type on processing, in a holistic manner, the results obtained suggested that the correlations between the chemical change and the drug action after processing, as well as the vinegar type used in processing, should be further studied.
Acetic Acid ; Benzoates ; Bridged-Ring Compounds ; Cooking ; Drugs, Chinese Herbal ; Furaldehyde ; analogs & derivatives ; Glucosides ; Metabolome ; Metabolomics ; Monoterpenes ; Paeonia ; chemistry ; Plant Roots ; chemistry

Objective Bevacizumab ( BM ) is an angiogenesis inhibitor widely used in cancer therapy, but its off-target effect of proteinuria may lead to discontinuation of treatment.This study was to explore the mechanisms of BM inducing proteinuria in mice. Methods Twenty-four healthy mice were randomly divided into four groups, saline control, low-dose BM, medium-dose BM, and high-dose BM, treated by injection of normal saline and BM at 10, 35, and 60 mg per kg of the body weight, respectively, though the tail vein.At 4 weeks after injection, 24-hour urine was collected to determine the total urine protein and blood obtained from the eyeballs for biochemical analysis.Then all the mice were sacrificed and the kidneys harvested for observation of pathologic changes in the glomeruli as well as for immunohistochemistry, Western blotting, and real-time PCR analysis. Results Compared with normal saline,BM obviously elevated the level of 24-hour urine protein, with statistically significant differences between the control and the medium-and high-dose BM groups (0.23 ±0.02 vs 1.14 ±0.13 and 1.43 ±0.10, P<0.01), but not between the control and the low-dose BM (0.23 ±0.02 vs 0.29 ±0.07, P>0.05).No significant differences were observed among the four groups in the levels of Cr, BUN, AST and ALT (P>0.05).Under the optical microscope, the kidneys showed normal structures in the control group, no signifi-cant pathologic changes in the low-dose BM, and vacuolus-like alteration with atrophic glomerular endothelial cells in the medium-and high-dose BM groups.Immunohistochemical analysis demonstrated the expressions of VEGF and podocin were moderately or strongly positive in the control and low-dose BM groups, by weakly positive or negative in the medium-and high-dose BM groups.Compared with the control group, the expression of the VEGF protein in the renal tissue was significantly decreased in the high-dose BM group (0.76 ±0.09 vs 0.39 ±0.05, P<0.01) but had no remarkable difference from that in the low-dose (0.81 ±0.10) or medium-dose BM (0.64 ±0.08) group (P>0.05), and the expression of the podocin protein was significantly reduced in the medium-dose BM (0.67 ±0.07 vs 0.43 ±0.10, P<0.05) and high-dose BM (0.67 ±0.07 vs 0.19 ±0.04, P<0.01), but not in the low-dose BM group (0.67 ±0.03) (P>0.05).The mRNA expressions of VEGF and podocin were not significantly changed in the low-dose BM group as compared with the control (1.07 ±0.61 and 1.12 ±0.09 vs 1.23 ±0.25 and 1.17 ±0.19, P>0.05) but remarkably de-creased in the medium-dose (0.82 ±0.38 and 0.71 ±0.18) and high-dose BM groups and (0.47 ±0.64 and 0.42 ±0.09) groups (P<0.01). Conclusion Bevacizumab damages glomerular filtration membrane and induce proteinuria partially by down-regulating the protein and mRNA expressions of VEGF and podocin.

Objective To explore the mediating effect and moderating effect of positive mental characters in the relationship between childhood abuse and depression among recruits.Methods 1925 recruits aged from 16 ～24 were investigated using the Childhood Trauma Questionnaire-28 item Short Form (CTQ-SF),Self-Rating Depression Scale (SDS) and Positive Mental Characters Scale for Recruits (PMCS-R) through random cluster sampling.Results The scores of emotional abuse,physical abuse,sex abuse,emotional neglect,physical neglect,depression,positive mental characters were 6.11±1.69,5.49±1.32,5.61±1.45,9.15±3.66,9.57±2.93,0.45±0.11,3.80±0.64,respectively.There were correlations among childhood abuse,depression and positive mental characters (P＜0.05).Emotional neglect,physical neglect and emotional abuse could explain 28.6％ of the total variance of depression (F=256.72,P＜0.05).The positive mental characters partially mediated the relationship between emotional neglect,physical neglect,emotional abuse and depression (mediating effect were 0.19,0.15 and 0.09,respectively),and it could only moderate the relationship between emotional neglect and depression (F=24.73,P＜0.05).Conclusion Childhood abuse not only directly but also indirectly affects depression through the positive mental characters;meanwhile the positive mental characters can change the relationship between childhood abuse and depression.

Objective To investigate the prevalence of mutation in the locus 306 of embB gene in multi-drug resistant (MDR) Mycobacterium tuberculosis (TB) and evaluate the prospects for using it as a molecular marker to detect MDR-TB.Methods The 291 strains enrolled in this study were from the reference laboratory of Shanghai municipal centers for disease control and prevention, all of which had been tested for drug susceptibility.Mutation in embB 306 was screened both by amplification refractory mutation system (ARMS) and DNA sequencing.The mutation frequencies of embB 306 in the sample groups varied in drug resistance were statistically analyzed.Results 38(51.4% ) of the 74 MDR-TB were embB 306-mutant (X2 =93.8,P＜0.01).Of the 24 TB resistant to at least two drugs but not MDR, 9(37.5% ) were embB 306 mutant (X2 =60.1 ,P＜0.01 ).But only two(4.9% ) embB 306-mutant strains were found in 41 strains resistant to only one drug (X2 =6.8,P=0.0093).None embB 306-mutant strains were found in 152 pansensitive strains.The specificity of using embB 306 as a molecular marker for detecting multi-drug resistant TB was 94.9% (206/217).Conclusions As a molecular marker for screening drug resistant TB,especially MDR-TB, the gene locus embB 306 shows a relatively high sensitivity and specificity, promising a sound future for its application in clinics to realize fast screening of patients infected with MDR-TB and to provide evidence for appropriate medication.

OBJECTIVETo research serum level of interleukin-17 (IL-17) associated with the progression of hepatic injury in the chronic patients with hepatitis B.

METHODSThe serum level of IL-17 was measured by ELISA and the serum levels of IL-6, IL-8 were measured by RIA in patient groups and healthy group, the patient groups including 42 mild patients, 37 moderate patients and 38 severe patients.

RESULTSIL-17,IL-6 and IL-8 levels in patient patients were significantly higher than those in healthy people (P < 0.01). There is no significant difference among mild patients and moderate patients. Compared with mild patients and moderate patients,the cytokines lever were significantly higher in severe patients (P < 0.01).

CONCLUSIONIL-17 as a new cytokine probably play a multiple role as immune factor and inflammation element in the progression of the chronic hepatitis B disease. Maybe, it can provide a new approach to the therapy of the chronic hepatitis B.

Adult ; Aged ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; blood ; pathology ; Humans ; Interleukin-17 ; blood ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Male ; Middle Aged

1H NMR metabonomics approach was used to reveal the chemical difference of urine between patients with Xiao-Chaihu Tang syndrome (XCHTS) and healthy participants (HP). The partial least square method was used to establish a model to distinguish the patients with Xiao-Chaihu-Tang syndrome from the healthy controls. Thirty-four endogenous metabolites were identified in the 1H NMR spectrum, and orthogonal partial least squares discriminant analysis showed the urine of patients with Xiao-Chaihu Tang syndrome and healthy participants could be separated clearly. It is indicated that the metabolic profiling of patients with Xiao-Chaihu Tang syndrome was changed obviously. Fifteen metabolites were found by S-pot of OPLS-DA and VIP value. The contents of leucine, formic acid, glycine, hippuric acid and uracil increased in the urine of patients, while threonine, 2-hydroxyisobutyrate, acetamide, 2-oxoglutarate, citric acid, dimethylamine, malonic acid, betaine, trimethylamine oxide, phenylacetyl glycine, and uridine decreased. These metabolites involve the intestinal microbial balance, energy metabolism and amino acid metabolism pathways, which is related with the major symptom of Xiao-Chaihu Tang syndrome. The patients with Xiao-Chaihu Tang syndrome could be identified and predicted correctly using the established partial least squares model. This study could be served as the basis for the accurate diagnostic and reasonable administration of Xiao-Chaihu-Tang syndrome.
Humans ; Least-Squares Analysis ; Medicine, Chinese Traditional ; Metabolome ; Metabolomics ; Proton Magnetic Resonance Spectroscopy ; Syndrome ; Urinalysis

Axons ; chemistry ; Myelin Sheath ; chemistry ; Staining and Labeling ; methods

BACKGROUND:As a noticeable tissue engineering material of polyhy droxyalka noates family, poly (3-hydroxybutyrate-co-4-hydroxybutyrate)(P3HB4HB) exhibitsgood biocompatibility, adhesion and mechanicalproperties, presenting aextensive application future in tissue-engineered research.
OBJECTIVE:To investigate the biocompat ibilityin vitroand ectopic osteogenic differentiationin vivoof P3HB4HB and human bone marrow mesenchymal stem cels.
METHODS:Passage 5human bone marrow mesenchymal stem cels transplanted ontothe three-dimensional P3HB4HB scaffoldwereincubated with osteogenic induction medium (test group)or with no osteogenic induction(control group), respectively. After 5-day incubation, thecelgrowth was assessed by acridine orange staining and scanning electron microscopy; after14-day incubation, both kinds of cel-scaffold composites were subcutaneously implanted into the nude mice. At 16 weeks after implantation, the cel-scaffold composites were removed to observeectopic osteogenic differentiationin vivousing hematoxylin-eosin staining, von Kossa staining and colagen type I immunohistochemical staining.
RESULTS AND CONCLUSION:Acridine orange staining showed that cels adhered wel on the surface of the scaffold;under thescanning electron microscope, induced celsgrew wel on the P3HB4HB scaffold and produced abundant extracelular matrixes. In addition, at 16 weeks after implantation, there were osteoidtissues in the test group, positive for von Kossa staining as wel as colagen type I immunohistochemical staining;furthermore, hematoxylin-eosin staining showednumerous osteoblasts and bone lacunas. In contrast, no bone tissues appeared in the control group. To conclude, P3HB4HB is a suitable material for bone tissue engineering.

Objective To investigate the histological changes of the right ventricular remodeling in Xinjiang Mar-mota baibacina which are adapted to high-altitude hypoxia .Methods Hematoxylin and eosin and Masson staining was used to examine the pathological changes and degree of fibrosis , and immunohistochemical technique was used to detect connexin 43 ( CX43) expression in the right ventricular myocardium .Results In the right ventricular myocardium , cardio-myocyte hypertrophy and increased amount of collagen fibers were observed .The right ventricular hypertrophy index and weight index of Marmota baibacina were significantly higher , and expression of CX43 was significantly reduced than that of normal rats ( P<0.05 for both ) .Conclusions High-altitude hypoxia induces right ventricular remodeling in Xinjiang Marmota baibacina.It can be used as an ideal animal model for studies on the mechanisms of hypoxia adaptation on high -altitude plateau .

Objective To investigate the effects of p300/CBP on histone acetylation of Foxp3 gene and its roles in the immunological pathogenesis of Kawasaki disease (KD).Methods Forty-six children with KD and twenty-eight age-matched health children were consented to participate in this study.Co-immunoprecipitation and real-time PCR were performed to detect Foxp3-associated acetylation levels of histone H4 and binding abilities of p300, CBP, pSmad3 (phosphorylated mothers against decapentaplegic homolog 3) and NF-AT (nuclear factor of activated T cells) with Foxp3 gene in CD4+ T cells.The percentages of CD4+CD25high Foxp3+ cells (Treg) and the expression of Foxp3, CTLA4 (cytotoxic T-lymphocyte-associated protein 4), p300, CBP, TGF-βRⅡ (transforming growth factor β receptor Ⅱ) and pLAT1 at protein level were analyzed by flow cytometry.Quantitative real-time PCR was used to measure the expression of Foxp3, IL-10, TGF-β, TGF-βRⅠ, Egr-1 (early growth response protein 1), RARα (retinoic acid receptor α) and PLCγ1 (phospholipase C-γ1) in Treg cells at mRNA level.Plasma concentrations of TGF-β and retinol acid (RA) were measured by enzyme-linked immunosorbent assay.Results (1) The percentages of Treg cells, levels of Foxp3 and molecules associated with suppressive function of Treg cells (TGF-β, IL-10 and CTLA4), acetylation levels of histone H4 associated with promoter, conserved non-coding DNA sequence 1 (CNS1) and CNS2 of Foxp3 gene decreased remarkably during acute KD (P<0.05), but were restored after IVIG therapy (P<0.05).Meanwhile, all of the aforementioned items in KD patients with coronary artery lesions (KD-CAL+) were lower than those without coronary artery lesions (KD-CAL-) (P<0.05).No significant differences in histone H4 acetylation associated with CNS3 were found among different groups (P>0.05).(2) The levels of p300 and CBP in Treg cells and their binding abilities with Foxp3 gene were down-regulated significantly during acute KD (P<0.05), but were restored to some extent after IVIG treatment (P<0.05).The Foxp3-associated histone acetylation was positively correlated with the expression of p300 and CBP at mRNA level during acute KD (r=0.65, 0.42, P<0.05).Furthermore, the expression of p300 and CBP and their binding abilities with Foxp3 gene in KD-CAL+ group were lower than those in KD-CAL-group (P<0.05).(3) Compared with healthy subjects, plasma concentrations of TGF-β and RA and the expression of TGF-βRⅠ/Ⅱ/Egr-1, RARα and pLAT1/PLCγ1 were down-regulated during acute KD (P<0.05);the binding abilities of pSmad3 and NFAT with Foxp3 gene were reduced remarkably in patients with acute KD (P<0.05).All the items mentioned above were restored after IVIG treatment (P<0.05).Moreover, the ten items aforementioned in KD-CAL+ group were lower than those in KD-CAL-group (P<0.05).(4) Higher acetylation levels of histone H4 associated with promoter, CNS1 and CNS2, and enhanced binding abilities of p300 and CBP with Foxp3 gene were found in CD4+ T cells isolated from patients with acute KD after co-stimulation with TGF-β, RA and anti-CD3/CD28 antibodies as compared with those in CD4+ T cells without stimulation (P<0.05).However, no statistical difference in the acetylation level of histone H4 associated with CNS3 was found between the two groups (P>0.05).Conclusion Hypoacetylation of histone H4 associated with Foxp3 gene caused by insufficient expression of p300/CBP and their impaired binding abilities might be involved with immune dysfunction in KD.IVIG therapy regulates the expression of p300/CBP and their binding abilities with Foxp3 gene through up-regulating TGF-β signal.