Animals ; Glutamine ; blood ; metabolism ; Male ; Physical Conditioning, Animal ; physiology ; Physical Exertion ; physiology ; Rats ; Rats, Sprague-Dawley ; Sleep Deprivation ; metabolism ; physiopathology ; Thymus Gland ; metabolism ; Time Factors

To investigate me material basis of Mahuang Fuzi Xixin decoction (MFXD) for anti-inflammation and immune-suppression based on the combined method of serum chemical and serum pharmacological. The LC-MS/MS fingerprints of MFXD, drug-containing serum and blank serum were compared to define the components in plasma. Histamine, β-hexosaminidase released from RBL-2H3 cell infulenced by drug-containing serum at different time points were measured by ELISA. The effect of drug-containing serum on lipopolysaccharide-induced splenocyte proliferation at different time points were determined by MTT. A correlation analysis was made on components of MFXD and pharmacological indexes based the stepwise regression method. After the intragastrical administration with MFXD, 32 components were discovered in rat serum, including 27 prototype components (10 from Mahuang, 13 from Fuzi and four from Xixin) and five unknown components. Compared with blank serum, drug-containing serum could reduce the release of histamine from RBL-2H3 induced by antigen at different time points (P < 0.05); except the 4-hour drug-containing serum, all of the remaining drug-containing serums could inhibit the RBL-2H3 mastocyte degranulation induced by antigen at different time points (P < 0.05). Drug-containing serum could significantly lipopolysaccharide-induced mouse splenocyte proliferation at 15 and 30 min (P < 0.05). A regression analysis was made on the chemical data of components absorbed into blood and pharmacological indexes, i. e. release rate of histamine, release rate of β-hexosaminidase and inhibition rate of splenocyte. This suggested the close correlations among methyl pseudo-ephedrine, pseudoephedrine and histamine released from RBL-2H3 induced by antigen; pseudoephedrine, hypaconine, methyl pseudoephedrine and β-hexosaminidase released from RBL-2H3 induced by antigen; as well as benzoyl hypaconine, benzoylaconine, 14-benzoyl-10-OH-mesaconine, mesaconine and lipopolysaccharide-induced mouse splenocyte proliferation. Methylpseudoephedrine, pseudoephedrine, benzoyl hypaconine, benzoylaconine and mesaconine may be part of material basis of MFXD on anti-inflammation and immune suppression.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; Cell Degranulation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Histamine ; immunology ; Immunosuppressive Agents ; chemistry ; pharmacology ; Male ; Mass Spectrometry ; Mast Cells ; drug effects ; immunology ; Mice ; Rats ; Rats, Wistar ; Serum ; chemistry

Emergency medicine and critical care medicine are the new contents in standardized training of residency.The system of standardized training residency lacks for a traditional way to refer.According to the characteristic of the subjects and the practical situation,this paper discusses the methods of training in studying basic theory,idea education,improving technology and strain capability on improving the capability of standardized training residency in internal intensive medicine.

OBJECTIVETo investigate the enhancive effect of N,N'-dinitrosopiperazine (DNP) on induced carcinogenesis in nasal and/or nasopharyngeal epithelia among TgN(p53mt-LMP1)/HT transgenic mice to examine the underlying mechanism for the development of nasopharyngeal carcinoma (NPC).

METHODSTgN(p53mt-LMP1)/HT transgenic mice and the same strain of C(57)BL/6J wild-type mice both at the age of 5 months were randomly divided into 2 groups in parallel, respectively, i.e., TgN(p53mt-LMP1)/HT cancerous lesion-inducing group (TI), TgN(p53mt-LMP1)/HT control group (TC), C57BL/6J cancerous lesion-inducing group (CI), and C57BL/6J control group (CC). TI and CI mice were treated only with DNP for 16 weeks, twice each week, while TC and CC mice were given the same volume of saline as controls. At the end of treatment, animals were sacrificed to collect epithelial tissue samples from nasal cavity and nasopharynx for pathohistological evaluation by haematoxylin and eosin (HE) staining and for determination on the expression of TRAF2, c-Jun, and p16 by immunohistochemistry.

RESULTSAtypical hyperplasia was more significant in the samples of TI than in those of TC, CI, and CC, with the rates of lesions being 90%, 10%, 0, and 0 (P<0.01) respectively, though DNP was used alone in a much shortened inducing period at less dosage and without the use of carcinogenic promoter 12-O-tetradecanoylphorbol-13-acetate as usual. The expressions of tumor necrosis factor (TNF) receptor-associated factor 2 (TRAF2) and c-Jun in these samples were significantly up-regulated in TI (P<0.01), while the expression of p16 was significantly lower in TI than in the other groups (P<0.01).

CONCLUSIONTgN(p53mt-LMP1)/HT mice hold inherited constitutional defect in immune surveillance function, which can be aggravated by environmental carcinogens, such as DNP used even though in a much less strength. The enhanced carcinogenesis-inducing effect of DNP on TgN(p53mt-LMP1)/HT mice should be closely associated with abnormal signaling of activator protein-1 (AP-1) pathway, especially up-regulated expressions of TRAF2 and c-Jun, and down-regulated expression of p16.

Animals ; Epithelial Cells ; drug effects ; metabolism ; pathology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Mice ; Mice, Transgenic ; Mutation ; genetics ; Nasopharyngeal Neoplasms ; chemically induced ; genetics ; metabolism ; pathology ; Nitrosamines ; pharmacology ; Nose Neoplasms ; chemically induced ; genetics ; metabolism ; pathology ; Precancerous Conditions ; chemically induced ; genetics ; pathology ; TNF Receptor-Associated Factor 2 ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Viral Matrix Proteins ; genetics ; metabolism

AIMTo search for more bioactive compounds from the roots of Aconitum coreanum (Lèvl.) Rapaics.

METHODSHigh speed countercurrent chromatography was successfully applied to the separation of alkaloids from Aconitum coreanum. The structures were elucidated by their physicochemical properties and spectroscopic analysis.

RESULTSTwo-phase solvent system composed of CHCl3-CH3OH-0.2 mol x L(-1) HCl (10:3:3, volume ratio) was used in this experiment, eight alkaloids were obtained from the roots of Aconitum coreanum, which were identified as: 2alpha-propionyl-11alpha,13beta-diacetyl-14-hydroxyhetisine (I), Guanfu base P (II), Guanfu base G (III), Guanfu base F (IV), Guanfu base Z (V), Guanfu base O (VI), Guanfu base A (VII), Guanfu base B (VIII).

CONCLUSIONCompound I is a new alkaloid, named Guanfu base R.

Aconitum ; chemistry ; Countercurrent Distribution ; Heterocyclic Compounds, 4 or More Rings ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

Objective To study the proliferation of CRTH2 (CD4+ CD294+ Th2)cells promoted by pneumococcal vaccine through antigen presentation of dendritic cells (DCs),so as to provide a new approach for amplification and sorting of Th2 cells.Methods CDs induced from peripheral blood mononuclear cells were cocultured with T lym-phocytes after loading pneumococcal vaccine antigen,mixed lymphocyte reaction (MLR)was detected by cell count-ing kit-8(CCK8),DCs and CRTH2 cells were analyzed by flow cytometry.Results Pneumococcal vaccine could promote the maturation of DCs,together with TNF-a,it was adjuvant for maturation of DCs.Pneumococcal vaccine antigen-loaded DCs could increase the rate of subsets of CRTH2 cells on day 5([0.93±0.10]%)compared with day 1([0.70±0.02]%),and absolute number also increased (both P <0.05).Conclusion Amplification of CRTH2 cells can be greatly promoted by pneumococcal vaccine antigen-loaded DCs,which might be one of the effective way to induce amplification of Th2 cells.

OBJECTIVETo observe the distribution of toll-like receptor 4 (TLR4) in rats' respiratory tract. To study the influence of LPS and Eucalyptus globulus oil on the distribution of TMR4.

METHODThe Sprague-Dawley rats were intratracheally instilled with lipopolysaccharide (LPS,2 mg x kg(-1) per day) for two days to induce acute lung injury. The rats were sacrificed at 72 hours after LPS instillation. Lung morphology was studied. Leukocytes in Bronchoalveolar lavage fluid (BALF) were measured and TLR4 were detected by immunohistochemistry.

RESULTThe result of immunohistochemistry showed that TLR4 distributed widely in common rats' respiratory tract. In the group of acute lung injury, the number of leucocyte in BALF was increased apparently, the inflammation in bronchus and bronchioles was more apparently than that of the control group in morphology. And the expression of TLR4 was reinforced in main bronchus and bronchioles. In the group of E. globules oil (300 mg x kg(-1)), the leucocyte number was decreased apparently in BALF, the inflammation was lightened and the expression of TLR4 decreased as compared with the group of models.

CONCLUSIONThe expression of TLR4 distributes widely in rats' respiratory tract. The stimulation of LPS can reinforce the expression of TLR4. The E. globules oil can reduce the increase of TLR4 induced by LPS in bronchioles.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; isolation & purification ; pharmacology ; Bronchi ; metabolism ; Bronchoalveolar Lavage Fluid ; cytology ; Eucalyptus ; chemistry ; Leukocyte Count ; Lipopolysaccharides ; Lung ; pathology ; Male ; Oils, Volatile ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; chemically induced ; metabolism ; pathology ; Toll-Like Receptor 4 ; metabolism

This study was aimed to explore the role and mechanism of AML1a in abnormal hematopoiesis in mice. Plasmids pMSCV-FLAG-AML1a-IRES-YFP and pMSCV-IRES-YFP together with envelope-encoding plasmid pECO and packaging plasmid pGP were respectively transfected into 293T cells by using a method of calcium phosphate precipitation to produce retrovirus. Bone marrow mononuclear cells (BMMNC) from male C57BL/6J mice were transfected with the retroviral vector MSCV expressing FLAG-AML1a fusion protein and yellow fluorescent protein (YFP). The cells were cultured in M3434 semi-solid medium for colony formation assay and in M5300 fluid medium containing murine IL-3 (mIL-3), IL-6 (mIL-6) and SCF (mSCF) for long-term culture. The results showed that transfection of AML1a into BMMNC enhanced colony formation, colony size of the AML1a group was significantly larger than that of the control group, and the colonies were mainly composed of CFU-E and CFU-GEMM. In the long-term culture, AML1a-transfected BMMNC showed differentiation block, while the control cells were in a more mature stage. It is concluded that AML1a may block the normal hematopoiesis at the stage of primitive progenitors. At the same time, AML1a also enhances the proliferation activity of primitive progenitor cells.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Proliferation ; Colony-Forming Units Assay ; Core Binding Factor Alpha 2 Subunit ; genetics ; Genetic Vectors ; Hematopoietic Stem Cells ; cytology ; Male ; Mice ; Mice, Inbred C57BL ; Retroviridae ; genetics ; Transfection

OBJECTIVEThe 85th percentile of waist circumference (WC) is considered an appropriate cutoff for Chinese children in the prediction of cardiovascular risks in previous researches, but the risk for hypertension of moderate WC maybe underestimated. The purpose of this study was to understand the characteristics of the WC curve trajectory of children with hypertension and to determine the appropriate cutoffs for waist circumference for Chinese school-age children and adolescents for predicting hypertension risk factors.

METHODSA total of 8194 Han children aged 7 - 17 years were selected from schools in Suzhou, Hefei, and Chizhou cities in Anhui province by stratified cluster sampling. WC and blood pressure were accurately measured in all subjects. LMS (lambda-mu-sigma) method was used to construct WC centile curves. Optimal WC cutoffs were determined by comparing the performance of different percentile sets of WC cutoffs in predicting hypertension risk factors.

RESULTSThe fitted LMS curves of WC for the group of boys with hypertension deviate in parallel from that for the corresponding curve in the entire boy study population with higher WC. The WC curves for the group of girls with hypertension gradually upward deviated from the corresponding curve before 12 years of age and then gradually returned to the corresponding curve. WC of boys at the percentiles of 60 was already associated with an increased risk of hypertension and hazard ratio of WC at percentiles of 60, 70, 80, and 90 increased from 1.88(95%CI: 1.18 - 2.99) to 4.87 (95%CI: 3.31 - 7.16). WC of girls at the percentiles of 70 was already associated with an increased risk of hypertension and hazard ratio of WC at percentiles of 70, 80, and 90 increased from 1.71 (95%CI: 1.07 - 2.73) to 3.32 (95%CI: 2.16 - 5.09).

CONCLUSIONSThe WC growth trajectory of children with hypertension varies with gender. WC is an independent predictor of childhood hypertension, even when it is well within what is now defined as the normal range of WC. Thus, it is helpful to prevent the risk of cardiovascular disease of children using the parameter that WC cut-off level lower than 85th percentile.

Adolescent ; Age Distribution ; Body Mass Index ; Child ; China ; epidemiology ; Female ; Humans ; Hypertension ; epidemiology ; prevention & control ; Male ; Reference Values ; Risk Factors ; Waist Circumference

OBJECTIVE: To explore the molecular mechanism of Glanzmann thrombasthenia (GT). METHODS: All 45 exons of alphaIIb and beta3 subunit genes as well as their splicing sites were amplified by polymerase chain reaction(PCR) with 40 primer pairs, and then the PCR products were used to screen the gene mutation by single strand conformation polymorphism-polyacrylamide gel electrophoresis (SSCP-PAGE). The mutation was further confirmed by direct DNA sequencing. RESULTS: A DNA band alterated migration was detected after SSCP-PAGE. DNA sequencing showed that a base deletion within the band at the site of 540 in GPIIb gene(540A) was found. CONCLUSION The frame-shift mutation caused by the deletion of 540A in GPIIb gene is a novel mutation which is a genetic defect in patients with GT.
Base Sequence ; Child, Preschool ; Exons ; genetics ; Frameshift Mutation ; genetics ; Gene Deletion ; Humans ; Integrin beta3 ; genetics ; Male ; Molecular Sequence Data ; Platelet Membrane Glycoprotein IIb ; genetics ; Sequence Analysis, DNA ; Thrombasthenia ; genetics