Objective To investigate association between human leukocyte antigen DQB1 (HLADQB1 ) gene polymorphisms and bronchial asthma among Mongolian and Han nationalities. Methods Sequence-specific primer polymerase chain reaction (PCR-SSP) was used to detect frequencies of HLA DQB1 genotypes and alleles in 50 cases of Han and 68 Mongolian asthmatic patients, and 50 Han and 54 Mongolian healthy controls, respectively. Difference in gene frequencies between the two nationalities was estimated by odds ratio (OR) and chi-square test. Results Frequency of the HLA-DQB1 0602 allele was significantly higher in Han patients with bronchial asthma than that in healthy Han nationality (OR = 6.163,P ＜0.01 ). Frequency of the HLA-DQB1 0603/0608 allele decreased in Mongolian asthmatic patients, as compared to that in healthy Mongolians ( OR = 0.199, P ＜ 0.05 ). Frequency of the HLA-DQB1 0301/4 allele was significantly higher in Mongolian asthmatic patients as compared to that in healthy Mongolians ( OR =2.074,P ＜0.05). Frequency of the HLA-DQB1 0301/4 allele was significantly higher in Mongolian than that in Han asthmatic patients ( OR = 2.482 ,P =0.05). Frequency of the HLA- DQB1 0602 allele was significantly higher in healthy Mongolians than that in healthy Han nationality ( OR = 3.341, P ＜ 0.05 ), in contrast, frequency of the HLA-DQB1 0402 allele was significantly lower in healthy Mongolians than that in healthy Han nationality ( OR = 0.209, P ＜ 0.05 ). Conclusions The HLA-DQB1 0603/0608 allele is possibly a protective gene and the HLA-DQB1 0301/4 allele a susceptible gene for bronchial asthma in Mongolians, and the HLA-DQB1 0602 allele is possibly a susceptible gene for bronchial asthma in Han nationlity.

Objective To construct sigF deletion mutant of Bacillus anthracis and the complementary strain of sigF deletion mutant in order to analyze the effect of losing sigF on formation of spores.Methods The spectinomycinadenyltransferase gene(spc) was inserted to replace sigF of B.anthracis by homologous recombination.A plasmid which contained sigF and sigF promotor was constructed and then transferred to the mutant to get a complementary strain of sigF deletion mutant.The characters of the mutant were analyzed by measuring growth curves, the ability of carbohydrate metabolism was compared, and spore formation was observed under a microscope.Results The sigF deletion strain A16D2△sigF was constructed from A16D2,which had a similar growth rate to the wild type A16D2 in logarithmic phase, but was not significantly different from the initial strain in the ability to use carbohydrates,although unable to form spores.The strain was found to maintain the state of asymmetric division by microfluidics experiment.Conclusion It is showed by this study that sigF is the essential gene of B.anthracis for spore formation, but not essential for vegetative growth.

"Omics" and bioinformatics have brought new ideas to the study of traditional Chinese medicine. This study used metabonomics and network pharmacology to investigate the pharmacodynamic basis and regulation of Qishen Yiqi dropping pill (QDP) improving cardiac energy metabolism in rats with heart failure (HF). ¹H NMR metabonomics analysis showed that eight metabolites, including carnitine, glutamine, creatine, proline, homocitrulline, lactic acid, taurine and alanine appeared significant callback after QDP treatment for HF. The results indicate that QDP regulates the metabolism of carbohydrate, lipid, ATP and protein. The animal experiment was conducted in accordance with the regulations of the Ethics Committee for Experimental Animal Management and Animal Welfare of Institute of Materia Medica, Chinese Academy of Medical Sciences. A "drug-component-target-disease" network was established using network pharmacology, and the "component-target" sub-network related to the above energy metabolism processes was extracted by combining metabonomics results. Results revealed 79 chemical compounds and 47 potential targets of QDP involved in the regulation of energy metabolism, and identified key chemical components including ursolic acid, notoginsenoside G, ginsenoside-Rh1, and core targets such as INS, PPARG, and AKT1. The results also demonstrated the complex multi-target and multi-component relationship between QDP and HF from the perspective of energy metabolism. The molecular docking technique verified a strong interaction between some targets and chemical compounds, with affinities less than -5 kcal·mol^-1. The results of this study provide useful information for the clinical application, development, and utilization of QDP.

OBJECTIVETo analysis the genetic mode of Rh DEL phenotype and RHD 1227A allele in Zhejiang Han population through family investigations.

METHODSRh DEL phenotypes were identified by a serologic adsorption-elution method. Two polymerase chain reaction-sequence specific prime (PCR-SSP) methods which detectED RHD 1227A allele and Rhesus hybrid box, respectively, and a nucleotide sequencing method focused on the exon 9 of RHD 1227A allele were employed to determine the zygosity of RHD allele.

RESULTSAll five probands with Rh DEL phenotype harbored a RHD 1227A allele and had a RHD allele deletion, and they were RHD 1227A/RHd heterozygote. One of the parent members was found to contain a RHD 1227A allele and a normal RHD allele in pedigree 1, 2 and 3, respectively. Thus, they were RHD 1227A/RHD heterozygotes and presented normal D positive phenotype. The son of proband No 1. inherited the RHD 1227A allele and presented a normal D positive phenotype due to a RHD 1227A/RHD heterozygote; The offsprings of proband No. 2, No. 4, and No. 5 did not inherit RHD 1227A allele and presented a normal D positive phenotype.

CONCLUSIONRHD 1227A allele is an important genetic marker of Rh DEL phenotype; RHD 1227A is recessive to normal RHD allele and dominant to RHd allele; RHD 1227A allele is an ancestral, but not a spontaneously mutated allele.

Alleles ; China ; Female ; Genotype ; Humans ; Male ; Pedigree ; Phenotype ; Polymerase Chain Reaction ; methods ; Rh-Hr Blood-Group System ; genetics

OBJECTIVETo assess the skeletodental changes after extracting mandibular third molars in skeletal Class III patients in both post-puberty and adult.

METHODSNineteen skeletal Class III patients in post-puberty or adult (16 female, 3 male) successfully treated with extracting mandibular third molars were included in the study. Cephalograms were analyzed to assess the skeletal and dental changes before and after treatment. Paired t-test was performed for statistical analysis of the data.

RESULTSThe results revealed that the main changes were upright [(12.37 ± 6.81)°] and distal movement [(3.16 ± 1.23) mm] of the mandibular molars, and retraction [(1.98 ± 1.36) mm] of the mandibular incisors. There was a significant improvement in overjet [(2.75 ± 1.65) mm].

CONCLUSIONSThe lower molars and incisors could be distalized significantly with extraction of the lower third molars.

Adolescent ; Adult ; Cephalometry ; Female ; Humans ; Incisor ; pathology ; Male ; Malocclusion, Angle Class III ; therapy ; Mandible ; Molar ; pathology ; Molar, Third ; surgery ; Orthodontics, Corrective ; methods ; Puberty ; Tooth Extraction ; Tooth Movement Techniques ; Young Adult

OBJECTIVETo explore a method for objectively and accurately evaluating cervical vertebral bone age among Chinese growing subjects.

METHODSUsing cephalometric and hand-wrist radiographs of 90 boys and 90 girls who were during pubertal period, we measured the third and fourth cervical vertebral bodies and calculated hand bone age using Liaokawa's method. A formula for calculating cervical vertebral bone age was determined by stepwise multiple regression analysis.

RESULTSA method for objectively evaluating cervical vertebral bone age among Chinese growing subjects during pubertal period was established. Y (Y: cervical vertebral bone age) = -20.189 + 24.666X(1) (X(1): AH(4)/AP) +46.468X(2) (X(2): AH(3)/AP(3)) +39.854X(3) (X(3): AH(4)/H) (r = 0.901). The R square of the equation was 0.81. The mean and standard deviation were as follows: constant = -20.19 +/- 4.89 (t = -4.13, P < 0.01), AH(4)/AP(4) = 24.67 +/- 8.32 (t = 2.97, P < 0.01), AH(3)/AP(3) = 46.47 +/- 6.60 (t = 7.04, P < 0.01), AH(4)/H(4) = 39.85 +/- 7.04 (t = 5.66, P < 0.01).

CONCLUSIONSThis method could be used to evaluate Chinese growth stages objectively.

Adolescent ; Age Determination by Skeleton ; methods ; Cervical Vertebrae ; diagnostic imaging ; growth & development ; Female ; Humans ; Male ; Puberty ; Regression Analysis ; Wrist ; diagnostic imaging ; growth & development

OBJECTIVETo study the effects of TGF-beta/Smad signaling on the growth and apoptosis of human rhabdomyosarcoma cell line RD.

METHODSBiosynthesized short hairpin RNA (shRNA) was transfected into RD cells by cation liposome vector to suppress Smad4 expression. The mRNA and protein expression of Smad4 in RD after shRNA-transfection were examined by RT-PCR and Western blot respectively. Immunofluorescent staining was used to detect the location of Smad2/3 in RD by laser scanning confocal microscopy. The viability of RD cells was examined by MTT method and (3)H-thymidine incorporation assay. The apoptosis of RD was examined by flow cytometry analysis and fluorescent staining.

RESULTSThe expression of mRNA and protein of Smad4 in RD were effectively suppressed by shRNA interference. Such suppression effectively interrupted the endogenous TGF-beta/Smad signaling and consequently blocked the translocation of Smad2/3. The interruption of endogenous TGF-beta/Smad signaling not only inhibited the growth of RD but also induced apoptosis of RD. Exogenous TGF-beta1 inhibited the growth of RD but did not influence the apoptosis of RD.

CONCLUSIONshRNA interference can effectively interrupt the TGF-beta/Smad signaling by suppressing the expression of Smad4. TGF-beta/Smad signaling subtly regulates the growth and apoptosis of RD.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Cell Proliferation ; drug effects ; Cytoplasm ; metabolism ; Humans ; Protein Transport ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; pharmacology ; Rhabdomyosarcoma ; genetics ; metabolism ; pathology ; Signal Transduction ; Smad2 Protein ; metabolism ; Smad3 Protein ; metabolism ; Smad4 Protein ; biosynthesis ; genetics ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

BACKGROUNDTransforming growth factor-beta (TGF-beta) can inhibit the growth of most epithelial and endothelial cells. The growth regulative role of TGF-beta on soft tissue sarcoma was seldom reported. Here we examined TGF-beta1 effects on the growth of human rhabdomyosarcoma cell RD and searched the relative molecular mechanism.

METHODSThe viability of RD was examined by [(3)H]-thymidine incorporation and [3-(4,5-dimethylthiazol-z-yl)-2,5-diphenyl tetrazolium bromide] (MTT) assay. RD cell cycle was analysed by flow cytometry. The protein and mRNA of cell cycle regulative factors in RD were detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. The kinase activity of cdk2 or cdk4 was examined by immunoprecipitation and kinase assay. Immunofluorescent staining was used to detect the location of cell cycle regulative factors in RD by laser scanning confocal microscope.

RESULTSTGF-beta1 inhibits RD proliferation by G1-arrest in cell cycle progression. TGF-beta1 can prominently up-regulate P27 of RD, then augment P27 to bind cyclinE-cdk2 complexes, which effectively suppress cdk2 kinase activity. P21 increased and c-myc decreased in RD due to TGF-beta1. Both P15 and cdk4 have not been involved in the growth inhibitory event. TGF-beta1 treatment induced P27 to congregate around nucleus. P21 pervaded from nucleus to both nucleus and cytoplasm by TGF-beta1 treatment.

CONCLUSIONTGF-beta1 inhibits the proliferation of human rhabdomyosarcoma cell line RD and induces RD G1-arrest. This course is accomplished by TGF-beta1 up-regulating P27 to suppress cdk2 kinase activity. The induction of P21 and down-regulation of C-myc might participate in the growth-arrest event.

Cell Cycle Proteins ; analysis ; genetics ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; G1 Phase ; drug effects ; Genes, myc ; Humans ; RNA, Messenger ; analysis ; Rhabdomyosarcoma ; pathology ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1 ; Tumor Suppressor Proteins ; analysis ; genetics

OBJECTIVETo study the regulatory effect of TGF-beta1 on growth of rhabdomyosarcoma RD cell line.

METHODSAfter various durations of TGF-beta1 treatment, the viability of RD cell line was examined by growth rate measurement, MTT assay and (3)H-thymidine incorporation. The cell cycle was analyzed by flow cytometry. Immunofluorescent staining was used to localize p15, p21 and p27 in RD cell line under laser scanning confocal microscope. The protein and mRNA of p15, p21 and p27 in RD cell line were detected by western blot and reverse transcriptase-polymerase chain reaction respectively.

RESULTSThe viability of RD cell line treated with TGF-beta1 was obviously decreased. RD cell line was arrested in G(1) phase by TGF-beta1. There was increased expression of p21 and p27 in RD cell line with TGF-beta1 treatment at protein and mRNA levels. The expression of p21 in RD cell line was seen in both nucleus and cytoplasm after 24 hours of TGF-beta1 treatment. The expression of p15 showed no obvious changes upon TGF-beta1 treatment.

CONCLUSIONSTGF-beta1 inhibits growth of RD cell line and induces G(1)-arrest. It up-regulates protein and mRNA of p21 and p27 and shows no obvious influence on p15 expression. The growth arrest of RD cell line may result from the up-regulation of p21 and p27 by TGF-beta1.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p15 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p27 ; biosynthesis ; genetics ; G1 Phase ; drug effects ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Rhabdomyosarcoma ; pathology ; Transforming Growth Factor beta1 ; pharmacology

BACKGROUNDTransforming growth factor beta (TGF-beta) plays an essential role in the regulation of normal physiologic processes of cells. TGF-beta has been shown to regulate several mitogen-activated protein kinases (MAPK) pathways in several epithelial cells. However, the effects of TGF-beta on soft tissue sarcoma are seldom reported. Our previous studies suggested that there should be some other signal transduction pathways besides Smads, which are important to regulate the growth of human embryonal rhabdomyosarcoma (RMS) cells. In the present study, we examined the expression and functional relations of extracellular signal-regulated kinase 2 (ERK2) and Smad4 in human RMS tissue and a RMS cell line, RD.

METHODSRD cells and normal human primary skeletal myoblasts (Mb) were treated with TGF-beta1 to establish the expression profile of ERK2 at the mRNA and protein levels detected by RT-PCR and immunofluorescence. Immunohistochemistry was used to detect the expression of ERK2 and Smad4 in 50 tissue specimens of human RMS and 23 specimens of normal skeletal muscles. Follow-up of specimens was performed 6 months to 70 months later.

RESULTSRD cells and human RMS tissues showed the higher expression of ERK2 and Smad4 than the normal control, either the protein level or the mRNA level. And, exogenous TGF-beta1 stimulation can lead to higher expression of ERK2 and its nuclear translocation, so TGF-beta1 can also activated MAPK (ERK2) pathway, resulting in a sustained activation of ERK2 for at least 2 hours. Immunohistochemistry analysis, however, showed that there was no correlation between ERK2 and Smad4 protein. The overexpression of ERK2 and Smad4 had no indicative effects on histological subtypes, histological grading, gender, age, and prognosis.

CONCLUSIONSIn RMS, signaling of TGF-beta1 from cell surface to nucleus can also be directed through the MAPK (ERK2) pathway besides the TGF-beta1/Smads pathway. The activation of ERK2 by TGF-beta1 may be Smad4 independent. Moreover, there may be some other tanglesome relationships between the TGF-beta1/Smads pathway and the MAPK pathway which takes part in the development, invasion and metastasis of tumor cells.

Cells, Cultured ; Humans ; Mitogen-Activated Protein Kinase 1 ; physiology ; Muscle, Skeletal ; metabolism ; RNA, Messenger ; analysis ; Rhabdomyosarcoma ; metabolism ; Signal Transduction ; Smad4 Protein ; physiology ; Transforming Growth Factor beta1 ; pharmacology