Objective To clone the full-length cDNA of MpCHS and analyze its expression pattern in different parts of Microcos paniculata and different growth periods of M. paniculata leaves. Methods Based on the transcriptome data of M. paniculata, we designed specific primers for MpCHS gene. The full-length cDNA of MpCHS was amplified by PCR and the positive clones were then sequenced, analyzed, and constructed prokaryotic expression vector. The bioinformatics analysis of MpCHS was also performed. Meanwhile, the mRNA expression of MpCHS was detected using real-time quantitative PCR. Results The relative molecular mass was 42 700, and its theoretical isoelectric point was 6.11, with three conserved functional active sites (165 C, 304 H, and 337 N) of the CHS family proteins and the tag sequence of RLMMYQQGCFAGGTVLR and GVLFGFGPGL. Phylogenetic tree analysis showed that MpCHS had close relationship with woody plants such as cocoa and upland cotton. We successfully cloned the full-length cDNA of MpCHS (GenBank: KY472608). It had an ORF of 1 176 bp which encoded a protein of 391 amino acid residues. RT-qPCR results showed that MpCHS was expressed in all parts of M. paniculata and its expression in leaves was gradually decreased along with its development. Conclusion MpCHS is cloned from M. paniculate for the first time, and the gene expression pattern of MpCHS in different parts of M. paniculata and different growth periods of M. paniculata leaves was analyzed. This study facilitates the further purification and functional validation of MpCHS protein and provides reference for further analysis of flavonoids biosynthesis pathway in M. paniculata.

Seven emotions generation relates to the interaction between individual and external objective matters,which is accompanied with interaction between internal desire and external matters.The activity of seven emotions bases on essence of zang-fu organs,being regulated by five zang viscera.Heart is the upstream controller,liver is the key organ to maintain normal emotionds,spleen and stomach is the hub of emotional activities,lung is the auxiliary organ, kidney is origin of emotional generation.Five zang viscera's cooperation and interaction generate the seven emotions.

Objective To explore the incidence,clinical status,risk factors and outcomes of invasive fungal infections(IFIs)after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) in pediatric patients.Methods Forty-one Patients who were underwent Allo-HSCT were selected from 2005 to 2006. Of 41 patients, 24 were boys and 17 were girls,aged 2-13 years old. Twenty-six cases with ?-thalassemia, 1 case with adrenoleukodystrophy,and the left 14 cases with other hematologic disorders.Twenty patients underwent bone marrow transplantation,19 patients underwent peripheral blood stem cell transplantation,2 patients underwent bone marrow transplantation and cord blood transplantation.Fourteen patients received Allo-HSCT from HLA-matched sibling donors or HLA mis-matched parents, 27 patients received Allo-HSCT from unrelated donors. Based on different types of transplant, patients were conditioned with busulfan, cyclophosphamide and Anti-thymocyte immune globulin. Fludalabine, total body irration, thiotepa or melphalan was used additionly in some cases. Cyclosporine A and mycophnolate mofetil were used as prophylaxis of graft versus host disease (GVHD).Results IFIs was observed in 5 cases(5/41 cases,12.2%),this comprised cases of proven,probable and possible IFIs at rates of 2.4%,4.9%,4.9%.The time of IFIs was 9-120 d after transplantation,the majority of IFIs in 3/5 cases(60%)children occurred within the first month.The difference of IFIs between patients who recived high-dose corticosteroid and those with no or conventional-dose corticosteroid was significant(?2=8.201 P=0.004);Regarding conditioning regimens,the IFIs of patients who with Thiotepa (TT) was significanthy higher than that of compared with those without TT(?2=9.549 P=0.002).The total effective rate was 40%.The effective rates of the patients with confimed diagnosis,cli-nical diagnosis,and with recommended diagnosis respectively were 100%,0 and 50% respectively.Conclusions IFIs is an important complication after Allo-HSCT,and the high-dose corticosteroid therapy and conditioning regimens with TT are the risky factors for IFIs.Aspergillus is the main pathogen bacteria.

Objective To explore effects of seasonal variation on blood lead in children.Methods Anodic stripping voltammetry(ASV) was used to detect blood lead level in children.The enrolled children were divided into 4 groups as follows:young children(ages 1 month to 3 years);preschool(ages 3 to 6 years);school(ages 6 to 12 years);teenagers(ages 12 to 18 years),and children′s blood lead level and lead poisoning rates were analyzed in the light of seasonality.Results Total 13 233 children were observed,aged from 1 month to 18 years old,8315 males,4918 females.The average blood lead levels were 60 ?g/L,the 5~(th) and 95~(th) percentage was 19,138 ?g/L.The rate of lead poisoning in children was 14.8%,with the majority of low-grade(?~2=116.3125 P

OBJECTIVETo supply basis for the establishment of quality standard of Cryptolepis buchanaii.

METHODCharacters of crude drugs, microscopic characteristic as well as UV spectrum of the herb were studied.

RESULTLaticifers were found in the cortex and pith of the stem; much papillary non-glandular hair was found covering the stomata in the sub-cuticle of the leaf.

CONCLUSIONThe results can be employed as the basis for identifying the herb.

Cryptolepis ; anatomy & histology ; cytology ; Drug Contamination ; Pharmacognosy ; Plant Leaves ; anatomy & histology ; cytology ; Plant Stems ; anatomy & histology ; cytology ; Plants, Medicinal ; anatomy & histology ; cytology ; Spectrophotometry, Ultraviolet

0.05).Conclusion Three Hybridoma cell lines which secrete the target antibodies with satisfied affinities and specificities have been successfully raised,which provides a basis to produce a domestic-made HCMVpp65 antigen diagnosis kit.

Objective To determine the expression of HPV16/18,31/33 DNA and p53,p21~(WAF1) and MDM2 proteins in invasive squamous cell carcinoma of cervix (ISCC)and to indicate the significance of them in the occurrence and development of ISCC.Methods Using tissue microarray,in situ hybridization (ISH) and immunohistochemical method,we detected the expression of HPV16/18,31/33 and investigate the expres- sion of p53,p21~(WAF1),MDM2 and proteins in ISCC,CIN and NCE.Results was analysed by SPSS vision 12.5. Results The positive expression rate of HPV16/18,p53,p21~(WAF1),MDM2 in ISCC was markedly higher than in CIN and NCE.We found the difference between HPV31/33 and lymph node transfer.Significant relation- ship was observed between p53 protein expression and histological grade and lymph node metastasis of the cancer.There was positive correlation between the expression of p21~(WAF1) protein and the depth of invasion(P

Objective To optimize the formulation of paeonol nanostructured lipid carrier (NLC) thermosensitive in situ gel through Box-Behnken response surface method; To investigate its release properties in vitro to provide references for the study of transdermal drug delivery system. Methods Taking mass fraction of poloxamer 407 and poloxamer 188 as the factors, the gelling temperature as the index, the mathematical relationship between the gelling temperature and two factors was established by binomial model and multivariate linear regression model. The Box-Behnken response surface method was used to optimize the formulation of paeonol NLC thermosensitive in situ gel, and the in vitro release characteristics of the preparation was investigated. Results There was a credible quantitative relationship between the gelling temperature and the 2 factors, and the binomial model was more reliable than the multivariate linear model. The best prescriptions of paeonol NLC thermosensitive in situ gel were 22.90% poloxamer 407 and 3.34% poloxamer 188; gelling temperature was (33.4±0.1)℃, and the cumulative release amount of paeonol in situ gel in 24 h was 51.19%. Conclusion This method is suitable for the formulation optimization of paeonol NLC thermosensitive in-situ gel, and the established mathematical model has good predictability. The optimized formulation can provide references for the development of paeonol transdermal preparation.

In-vitro assay methods were established to evaluate transactivation and binding activity of compounds on peroxisome proliferator-activated receptor y (PPARγ). Firstly, plasmids were constructed for transactivation assay of PPARγ response element (PPRE) triggered reporter gene expression, and for cell-based binding activity assay of the chimeric receptor, which was fused with PPARγ ligand binding domain (LBD) and yeast transcriptional activator Gal4. Secondly, by using PPARy competitive binding assay based on time resolved-fluorescence resonance energy transfer (TR-FRET), affinities of compounds and drugs to PPARγ were evaluated. In application of these above methods, the PPARγ activating potency and characteristics of different compounds were evaluated, and a novel benzeneselfonamide derivative, ZLJ01, was found to have comparable binding activity and affinity with the well-known PPARy agonist, but lack of PPRE mediated transactivation activity. In preliminary study on in-vitro hypoglycemic activity, ZLJ1 was found to promote insulin-stimulated glucose uptake by liver cells. Therefore, we believe that combining transactivation and binding activity as well as affinity evaluation, the system could be used to screen non-agonist PPARγ ligand as anovel PPARγ modulator
Genes, Reporter ; Hepatocytes ; Hypoglycemic Agents ; chemistry ; Ligands ; PPAR gamma ; agonists ; chemistry ; Plasmids ; Response Elements ; Sulfonamides ; chemistry ; Transcriptional Activation

Child ; Electroencephalography ; Epilepsy ; diagnosis ; Humans ; Syndrome ; Video Recording