LC-Q-TOF-MS and LC-IT-MS in positive and negative ion mode were applied to simultaneously characterize the constituents in Suanzaoren tang. Analysis was performed on an Agilent Zorbax SB-C18, Rapid Resolution HT column(4.6 mmx 50 mm, 1. 8 micro m) with gradient elution of acetonitrile(A) -aqueous solution containing 0. 05% formic acid(B) at a flow rate of 0. 6 mL min(-1) and the column temperature was 30 degreesC. By comparing MS fragmentation, accurate molecular weight, literature date and standard compounds information, a total of48 compounds were successfully identified or speculated. The origins of these compounds were assigned to the corresponding Chinese medicine. Thirty-one compounds were reported in Suanzaoren tang for the first time. LC-Q-TOF-MS combined with LC-IT-MS is a simple and rapid tool for the identification of constituents of Suanzaoren tang, and the results could provide evidence for the research on quality combined and effective constituents of Suanzaoren tang.
Chromatography, Liquid ; Drugs, Chinese Herbal ; chemistry ; Mass Spectrometry ; Organic Chemicals ; analysis

Objective To analyze the gene expression profiles in relation to the migration ability of adult human bone marrow-derived neural stem cells (Md-NSCs), and identify the genetic basis of the high migration potential of Md-NSCs in the central nervous system (CNS). Methods Adult human bone marrow stromal celIs(BMSCs) obtained from adult healthy volunteers were induced to differentiate into Md-NSCs in vitro, and the expressions of the genes related to cell invasiveness and metastasis were investigated by microarray analysis. Quantitative real-time PCR (RT-PCR) was used to verify the microarray results. Results The results of microarray analysis revealed significant overexpressions of the genes MMP1, MMP2, MMP17, ITGA3, RhoB, RhoC and RhoD in the Md-NSCs as compared with the expressions in fresh normal human adult bone marrow cells depleted of red blood cells. Quantitative RT-PCR verified the overexpression ofMMP2 gene by 2.84×100 folds, ITGA3 gene by 2.22×102folds, and RhoC gene by 4.92×100 folds. Conclusion The high migration potential of the Md-NSCs in the CNS is probably associated with the overexpression of the genes that promote cell invasiveness and metastasis. These overexpressed genes are also important oncogenes, and therefore the tumorigenicity of the Md-NSCs warrants further investigation.

Background Our previous study determined that expressions of matrix metalloproteinases (MMPs) and tissue matrix metalloproteinase inhibitors (TIMPs)change in the conjuntival fibroblasts of conjunctivochalasis in vitro.To seek a suitable drug is very important in the prevention and treatment of conjunctivochalasis.Objective This study was to explore the effect of qijingmingmu soup on the expressions of MMPs and TIMPs in human conjunctival fibroblasts of conjunctivochalasis.Methods Twenty-four SD rats were randomized into two groups.Qijingmingmu soup was administration gastrically for consecutive 3 days,and normal saline solution was given in the same way in the control group.The blood was collected from aortaventralis and drug serum was prepared.Human conjunctival samples were obtained during the surgery of conjunctivochalasis relaxation and cultured in the DMEM containing 10％ fetal bovine serum,20％,15％,10％,5％ of drug serum and 8 ml/L epidermal growth factor(EGF) was added into the medium respectively.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expressions of MMP1,MMP3,TIMP1 and TIMP3in conjunctival fibroblasts.Results Cultured cells grew well with the fusiform shape and showed the positive response for vimentin.The expression value of MMP1 (A value)in the cells was declined after administration of qijingmingmu soup.A significant difference was found in the expression of MMP1 among the control group,20％,15％,10％,5％ drug serum groups and EGF group(F=466.664,P＜0.05),and that in the 10％ ([9.92±0.14] mg/L) and 20％ ([11.87 ±0.11] mg/L) drug serum groups was significantly lowed in comparison with the control group([16.31±0.10] mg/L)(t=99.974,87.394,P＜0.05).The expression value of the MMP3in the cells in the various drug serum groups,EGF group and the control group was significantly different(F=158.168,P＜0.05),with a lower value in the 20％ drug serum group compared with the control group ([3.50±0.03] mg/L vs.[4.44 ± 0.11] mg/L) (t =21.991,P ＜ 0.05).Also,the significantly different expressing value of TIMP1 was seen among all the groups (F=183.508,P＜0.05),and expressing value of TIMP1 in the 15％ drug serum group was(1.88±0.06)mg/L,which was lower than(3.20±0.32) mg/L of the control group(t=10.353,P＜0.05).Furthermore,the expressing value of the TIMP3 in the cells was significantly different among the various groups(F=54.503,P＜0.05),and that of the 20％ drug serum group was (1.743±0.065)mg/L and it was significantly higher than (1.54 ± 0.05) mg/L of the control group (t =5.046,P =0.004).However,the expressing value of TIMP3of the 15％,10％ and 5％ drug serum groups was lower than that of the control group,respectively all at(P＜0.05).Conclusions Qijingmingmu soup drug serum at the concentration of 20％ can down-regulate the expressions of MMP1,MMP3,TIMP1 and up-regulate the expression of TIMP3 in human conjunctivochalasis bulbar conjunctival fibroblastsin vitro,which probably plays preventive and therapeutic effects on conjunctivochalasis.

Objective:To investigate gender differences in the attentional network in 5-year-old children.Methods:A totally of 70 healthy children [31 girls,(62.5 ± 2.6 months old;39 boys,(63.3 ± 2.8) months old] from the top class of kindergarten were enrolled in the study.A modified version of the Attention Network Test (ANT) was used to assess the attentional networks (alerting,orienting,and executive control).Results:The mean rate of accuracy was higher than 90％.A significant gender difference was found in the alerting network,boys scored higher than girls [(139 ± 54) ms vs.(109 ± 49) ms,P ＜ 0.05].No gender difference was found in the orienting or executive control networks (P ＞ 0.05).Conclusion:It suggests that 5-year-old children are fully capable of completing the attentional networks test,and the 5-year-old boy's alert function may be stronger than that of girls.

OBJECTIVETo evaluate the expression of N-cadherin and β-catenin protein and their relationship with clinicopathological characteristics of osteosarcoma.

METHODSThe expressions of N-cadherin and β-catenin at protein level were detected by immunohistochemical staining in 54 cases of osteosarcoma, 11 cases of osteoid osteoma, 7 cases of osteoblastoma and 8 cases of newly formed bone in nonmalignant bone diseases. The relationship between the two indexes and clinicopathological characteristics of osteosarcoma was analyzed.

RESULTSIn newly formed bone, osteoblastoma, osteoid osteoma and osteosarcoma, the positive expression rate of N-cadherin protein was 75.0%, 71.4%, 63.6% and 35.2%, respectively. The positive expression rate of N-cadherin protein in osteosarcoma was significantly lower than that in osteoid osteoma, osteoblastoma and newly formed bone in nonmalignant bone diseases (P = 0.035). The positive expression rate of N-cadherin protein in osteosarcoma cases with pulmonary metastasis was lower than that in cases without (21.7% vs. 56.3%, P = 0.027). The positive expression rate of N-cadherin protein in osteosarcoma cases died in two years was lower than that in cases lived for more than two years (18.2% vs. 50.0%, P = 0.024). In newly formed bone, osteoblastoma, osteoid osteoma and osteosarcoma, the aberrant expression rate of β-catenin protein was 12.5%, 28.6%, 27.3% and 66.7%, respectively. The aberrant expression rate of β-catenin protein in osteosarcoma was significantly higher than that in osteoid osteoma, osteoblastoma and newly formed bone (P = 0.002). Aberrant expression rate of β-catenin in osteosarcoma cases with pulmonary metastasis was higher than that without (82.6% vs. 43.8%, P = 0.011). An inverse correlation was found between the aberrant expression of β-catenin and N-cadherin expression in osteosarcoma(r = -0.302, P = 0.027).

CONCLUSIONThe positive expression rate of N-cadherin is decreased in osteosarcoma while aberrant expression rate of β-catenin increased. The expression of N-cadherin protein is closely correlated with the metastasis and prognosis of osteosarcoma, but the expression of β-catenin protein is merely correlated with the metastasis of osteosarcoma.

Adolescent ; Adult ; Bone Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Cadherins ; metabolism ; Child ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; metabolism ; secondary ; Male ; Middle Aged ; Osteoblastoma ; metabolism ; Osteoma, Osteoid ; metabolism ; Osteosarcoma ; drug therapy ; metabolism ; secondary ; surgery ; Survival Rate ; Young Adult ; beta Catenin ; metabolism

To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.
Administration, Oral ; Animals ; Arylamine N-Acetyltransferase ; genetics ; metabolism ; Caffeine ; metabolism ; urine ; Cytochrome P-450 CYP1A2 ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Male ; Medicine, Tibetan Traditional ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Theophylline ; urine ; Uracil ; analogs & derivatives ; urine ; Uric Acid ; analogs & derivatives ; urine ; Xanthines ; urine

Objective To determine the changes of cerebrospinal fluid(CSF) ?-endorphin(?-EP) and C-reactive protein(CRP) in children with central nervous system(CNS) infection.Methods Sixty-five children suffered from CNS infection were determined the plasma and CSF ?-EP and CRP concentration during the acute and recovering stage with radioimmunoassay, which included 48 viral encephalitis, 12 purulent meningitis and 5 tuberculou meningitis,and 24 non-CNS disease children were as control group.Results The concentrations of plasma and CSF ?-EP of every experimental group were obviously higher than those of control group during the early stage of CNS infection and these were obviously lower during the recovering stage. The serum concentration of CRP during acute stage was significantly higher than that during recovering stage. No change of serum and CSF CRP concentration was determined during either the acute or recovering stage in the other two experimental groups.Conclusions Determining the plasma and CSF ?-EP is mea-(ningful) in early diagnosis of CNS infection,and determining the serum CRP at the same time may be helpful in differentiating septic and inseptic infection.

OBJECTIVETo study the expression of E-cadherin and beta-catenin in neuroblastomas of various degrees of differentiation, and to investigate their molecular mechanisms in correlation with clinicopathologic parameters.

METHODSImmunohistochemistry EnVision method was used to detect E-cadherin and beta-catenin expression in 90 paraffin-embedded tissue samples of neuroblastomas. The methylation status of CpG islands of E-cadherin promoter was investigated by MSP in 7 fresh tissue and 24 paraffin-embedded tissue samples. The mutation status of exon 3 of beta-catenin gene was studied by PCR in 7 fresh tissue samples. Statistical analysis of the data was performed by SPSS software.

RESULTSE-cadherin and beta-catenin were abnormally expressed in neuroblastomas in general. The expression of beta-catenin in well-differentiated neuroblastoms was markedly higher (47/70, 67.1%) than that of the poorly differentiated tumors (8/20, 40.0%). There was a markedly decreased expression of both genes in tumors with lymph node metastasis than those without. Demethylation was seen in some regions of the promoter of E-cadherin in 31 cases of nuroblatomas. PCR of the exon 3 of beta-catenin followed by DNA sequencing demonstrated rearrangements and mutations in 7 cases, including 2 cases harboring identical point mutation at gene position 27184, leading to a T-->A alteration.

CONCLUSIONSThe abnormal over-expression of E-cadherin in neuroblastomas is independent of the methylation status of their promoter sequences. The abnormal expression of beta-catenin may be related to mutational changes at exon 3 of the gene.

Cadherins ; genetics ; metabolism ; Child ; Child, Preschool ; CpG Islands ; genetics ; DNA Methylation ; DNA, Neoplasm ; genetics ; Exons ; Female ; Ganglioneuroblastoma ; genetics ; metabolism ; pathology ; Gene Rearrangement ; Humans ; Infant ; Lymphatic Metastasis ; Male ; Mediastinal Neoplasms ; genetics ; metabolism ; pathology ; Neuroblastoma ; genetics ; metabolism ; pathology ; Point Mutation ; Promoter Regions, Genetic ; genetics ; Retroperitoneal Neoplasms ; genetics ; metabolism ; pathology ; Sequence Analysis, DNA ; beta Catenin ; genetics ; metabolism

OBJECTIVETo investigate the clinical utility of multiplex ligation-dependent probe amplification (MLPA) for detecting 22q11 deletion and duplication in congenital heart disease (CHD) cases and to study the incidence of 22q11 deletion and duplicaton in different kinds of CHD.

METHODSForty eight probes of which 25 located in 22q11 low copy number region (LCR 22s A-H), 7 in 22q11 surrounding region (CES, 22q13) and 16 in chromosomes 4, 8, 10 and 17 were selected to detect 22q11 deletion and duplication in 181 preoperative children with CHD and 14 fetuses with serious CHD or CHD with multiple malformations. In these cases, karyotype analysis was also performed.

RESULTSMLPA demonstrated that 7 cases had 22q11 deletion [6 cases from CLTCL1 to LZTR1(LCR A-D) and 1 case from CLTCL1 to PCQAP (LCR A-C)] and that 1 case had 22q11 duplication,spanning from ZNF74 to LZTR1(LCR B-D). The phenotypes of heart defect included ventricular septal defect, atrioventricular septal defect, pulmonary stenosis and tetralogy of Fallot. Karyotype analysis showed that 1 case had 21q deletion [46, XY, 21q], 1 case had mosaic trisomy 8 [47,XY, +8/46, XY(1:2)] and 4 cases had trisomy 21. One of the 4 cases with trisomy 21 had concurrent 22q11 duplication.

CONCLUSIONSMLPA is a rapid, sensitive, site specific and relatively inexpensive method for diagnosis of 22q11 deletion and duplication in CHD. 22q11 deletion and duplication may cause various kinds of CHD, suggesting that genetic detection should be performed routinely in CHD patients.

Adolescent ; Chromosome Deletion ; Chromosomes, Human, Pair 22 ; Female ; Gene Duplication ; Heart Defects, Congenital ; genetics ; Humans ; Infant, Newborn ; Male ; Nucleic Acid Amplification Techniques ; methods

OBJECTIVETo evaluate the ability of a polycaprolactone/polylactic acid (PCL/PLA) membrane to inhibit epidural scar adhesion after laminectomy, and observe the responsive changes of the pain media in the spinal cord.

METHODSL(1), L(3) laminectomies were performed on 96 Wistar rats. The rats were divided into 3 groups: None-implant Control Group (NC), Autologous free fat graft group (AFFG) and PCL/PLA membrane group (PCL/PLAm). The rats were killed at 1, 3, 6, and 12 weeks postoperatively. Epidural scar formation and adhesion were observed grossly and histologically. Reverse transcription polymerase chain reaction (RT-PCR) were used to analyses the expression of Transforming growth factor beta (TGF-beta) in the epidural scar. Immunohistochemistry stain and RT-PCR were performed to evaluate the expression of the substance P and the c-fos gene in the relevant spinal cord, and the results were analyzed statistically.

RESULTSGross evaluation and histological evaluation showed that in the NC lamina defect site had much scar tissue and had wide and tight adhesions to the dura; in the AFFG, with the fat degrading gradually, the adhesions were increased; whereas in the PCL/PLAm group, there were slightly adhesions to the dura. RT-PCR showed that the expression of the TGF-beta was much less in the PCL/PLAm group than in the NC group. The insertion of the PCL/PLA membrane and the fat patch reduced the expression of the substance P and the c-fos gene in the spinal cord.

CONCLUSIONThe insertion of the PCL/PLA membrane reduces scar formation and separates fibrosis tissue from the dura, the results indicate that PCL/PLA membrane is an effective way of reducing peridural scar formation and preventing the failed back surgery syndrome.

Animals ; Biocompatible Materials ; Cicatrix ; prevention & control ; Female ; Lactic Acid ; Laminectomy ; adverse effects ; Membranes, Artificial ; Polyesters ; Polymers ; Postoperative Complications ; prevention & control ; Prosthesis Implantation ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Rats ; Rats, Wistar ; Spinal Cord ; metabolism ; Spinal Diseases ; prevention & control ; Substance P ; biosynthesis ; Tissue Adhesions ; prevention & control