Objective: To study the chemical constituents of Lonicerae Japonicae Caulis produced in Shandong Province. Methods: Compounds were isolated and purified by repeated column chromatographies including silica gel, MCI-gel resin, Sephadex LH-20, and so on. Their structures were elucidated by spectral data, including extensive 1D and 2D NMR techniques. Results: Four compounds were obtained and identified as 11,14,15-trihydroxy-12-octadecenoic acid methyl ester (1), ochnaflavone (2), β-daucosterol (3), and loganin (4). Conclusion: Compound 1 is a new organic acid named as loniceric acid ester, and compound 2 is isolated from Lonicerae Japonicae Caulis for the first time.

Objective To assess the effects of gonadotropin-releasing hormone analog on glucose and lipid metabolism in girls with idiopathic central precocious puberty(ICPP).Methods A total of 26 girls (aged 6-8 years,breast stage B2) with ICPP were followed up in Department of Endocrinology,Shenzhen Children's Hospital from Jan.2008 to Jun.2011.Those girls received triptorelin therapy for 12 months.Before and the end of the 6th month,the 12th month of the treatment,body mass index(BMI) was calculated,fasting plasma glucose(FPG),fasting plasma insulin(FPI),total cholesterol,triacylglycerols,high density lipoprotein cholesterol,low density lipoprotein cholesterol,apolipoprotein A,apolipoprotein B and estradiol(E2) were measured.Insulin sensitivity was estimated by homeostasis model assessment of insulin resistance(HOMA-IR).Twenty age-matched prepuberty girls were set as controls.Results 1.Before treatment,BMI,FPG,FPI and HOMA-IR in ICPP girls had no significant difference compared with the controls.2.After 6 months treatment of triptrolin,serum E2 concentration in ICPP girls declined from(30.5 ± 9.8) ng/L at the beginning of treatment to (11.2 ± 4.6) ng/L at the end of 6th month (P ＜ 0.01) ; The end of 12 th month of the treatment,FPG and FPI had no significant difference compared with that before of the treatment,but BMI increased from(16.46 ± 1.10) kg/m2 to(18.35 ± 1.30) kg/m2,the difference was significant(P ＜0.05),HOMA-IR increased from 1.24 ±0.30 to 2.08 ±0.40,the difference was significant(P ＜0.05).3.Lipid metabolism parameters remained unchangeable after 12 months of triptrolin treatment.Conclusion Triptorelin may lead to raise of BMI and HOMA-IR in girls with ICPP at 12 months after treatment.

Adult ; Aged ; Aged, 80 and over ; Carcinoma in Situ ; virology ; Carcinoma, Squamous Cell ; virology ; Female ; Human papillomavirus 16 ; isolation & purification ; Human papillomavirus 18 ; isolation & purification ; Humans ; Microarray Analysis ; methods ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; diagnosis ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Uterine Cervical Neoplasms ; virology

Objective To study the conventional MRI and dittusion weighted imaging(DWI) features of Japanese encephalitis(JE)in children.Methods Sixteen patients with JE were included. Conventional MRI and DWI sequences were performed in all patients.Seven patients received MRI within 10 days of onset and 9 after 10 days.The appearances on DWI and T_2 WI were recorded.The ADC values of lesions were calculated,and then were correlated with the corresponding time interval between onset of neurological symptoms and MRI scanning.Results The lesions of JE mainly showed long T_1 and long T_2 signal intensity on MRI.The thalami were the most frequently involved areas,and 15 out of 16 showed thalamic involvement and 6 patients only showed thalamic abnormalities without other lesions.Seven patients within 10 days of onset showed lesions with high signal intensity on both DWI and T_2WI,but whole or partial lesions showed clearer on DWI than on T_2WI,and 2 patients showed extra lesions that were invisible on T_2WI.As for the other 9 patients after 10 days of onset,the lesions showed clearer on T_2WI than on DWI. There was a direct correlations between thalamic ADC values and the disease duration (r=0.84,P

Objective To develop an early-warning indicator system of brucellosis outbreak. Methods The methods of literature review and expert discussion were used to formulate the initiatory framework and indicators, and then Delphi method was used to filter indicators, discuss the boundary of indicators and determine the weighting coefficient. Results The average length of service provided by experts who engaged in prevention and control of brucellosis was (25.10 ± 8.80) years and the positive coefficient of the consultant experts of the two-round results were 95% and 68%, respectively. Kendall coefficients were 0.35, 0.54 and X2R value were 81.31 and 285.27, respectively and P value was all less than 0.01. Five first-level indicators(the host animal, high-risk groups,social environment, index case and the level of previous disease) and 13 secondary indicators were selected to develop the early-warning indicator system of brucellosis outbreak. The weight coefficients of the five first-level indicators were 0.21, 0.22, 0.17, 0.21 and 0.19, respectively. Conclusions The early-waming indicator system of brucellosis outbreak is initially established. We propose to develop the early-warning programs according to local conditions and the indicator system.

OBJECTIVETo investigate the induction and culture of adventitious root of Panax notoginseng.

METHODThree ways, induction from the explants of three-year-old P. notoginseng. The explants of regenerated shoots and calluses, were used to induce adventitious roots. The effects of 2, 4-dichlorophenoxyacetic acid, indole-3-butyric acid and naphthylacetic acid on adventitious root induction were investigated respectively. The effects of four modes of separating adventitious roots from the parent tissues on culture in vitro were compared.

RESULTAdventitious roots were successfully induced by three methods, of which the young flower bud callus was the best material for the induction of adventitious root. Indole-3-butyric acid possessed the strongest potency for induction. The liquid culture system was established by continuous culture of adventitious roots together with their parent tissues before separated.

CONCLUSIONThe acquisition and culture in vitro in liquid culture system of adventitious roots of P. notoginseng lay a foundation for the next investigation.

2,4-Dichlorophenoxyacetic Acid ; pharmacology ; Indoles ; pharmacology ; Naphthaleneacetic Acids ; pharmacology ; Panax notoginseng ; drug effects ; growth & development ; Plant Growth Regulators ; pharmacology ; Plant Roots ; drug effects ; growth & development ; Plants, Medicinal ; drug effects ; growth & development ; Tissue Culture Techniques ; methods

HLA-A* 2402 is one of the most frequently encountered HLA-A alleles in East Asian populations. In order to study the CD8+ T cell responses in Chinese populations, we have described the generation and functional test of HLA-A* 2402 tetramer loaded with HCMV pp65(341-349) peptide (QYDPVAALF, QYD). The cDNA of HLA-A* 2402 heavy chain was cloned by RT-PCR from one of the donors. DNA fragment encoding the ectodomain of HLA-A* 2402 heavy chain fused at its carboxyl-terminal a BirA substrate peptide (BSP) was amplified by PCR with the cloned heavy chain cDNA as a template. The wild-type gene of HLA-A* 2402-BSP was not expressed in Escherichia coli (E. coli), while mutant HLA-A* 2402-BSP gene with optimized codons was overexpressed as inclusion bodies in E. coli. Furthermore, the soluble HLA-A* 2402-QYD monomers were generated by in vitro refolding of washed inclusion bodies in the presence of beta2-microglobulin and QYD peptide. The tetramer was subsequently formed by mixing HLA-A* 2402-QYD monomers with streptavidin-PE at a molar ratio of 4:1. Flow cytometry analysis indicated that this tetramer possessed binding activity with specific CTL from HLA-A24+ donors and the frequencies of tetramer-binding CTL were 0.09% - 0.37% within total CD8+ T cells. This tetrameric agent provides a powerful tool to explore the secrets of CTL responses against HCMV antigens in HLA-A* 2402 individuals.
Amino Acid Sequence ; CD8-Positive T-Lymphocytes ; cytology ; metabolism ; Carbon-Nitrogen Ligases ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Escherichia coli Proteins ; metabolism ; Flow Cytometry ; Gene Expression ; HLA-A Antigens ; chemistry ; genetics ; metabolism ; HLA-A24 Antigen ; Humans ; Oligopeptides ; genetics ; metabolism ; Phosphoproteins ; chemistry ; genetics ; metabolism ; Protein Multimerization ; Recombinant Fusion Proteins ; chemistry ; genetics ; metabolism ; Repressor Proteins ; metabolism ; Substrate Specificity ; T-Lymphocytes, Cytotoxic ; cytology ; metabolism ; Viral Matrix Proteins ; chemistry ; genetics ; metabolism

Background Leber hereditary optic neuropathy (LHON)is a common inherited eye disease,which generally affects young adults with bilateral loss of central vision.Mutation frequency of Leber hereditary has not been fully clarified. Objective This study was to investigate the mutation frequency of mitochondrial NDI gene associated with LHON in Chinese population. Methods The proposal of the study was approved by Ethic Committee of Wenzhou Medical College.Written informed consent was obtained from each subject initial of this trial.Eight hundred and ninety-four LHON patients and 134 normal subjects were collected.Genomic DNA was extracted from peripheral blood leukocytes of the all participants.Polymerase chain reaction (PCR) was used to amplify and sequence analysis of the mitochondrial ND1 gene was performed and aligned with revised Cambridge Reference Sequence(rCRS) of mitochondrial DNA.Then mutated gene frequency was screened and analyzed. Results Mutational analysis of mitochondrial ND1 gene in 894 LHON patients revealed the presence of G3316A,T3394C,G3460A,C3497T,G3635A,G3733A,and T4216C.11.19％ LHON patients (100/894 ) were found to be associated with the gene mutations mentioned above,and 3.24％ patients (29/894) showed the co-occurrence of three primary mutations.Mutation frequencies in LHON patients were 2.57％,2.23％,1.45％,3.80％,0.67％,0.11％,0.34％,respectively,and G3316A,T3394C,C3497T and T4216C also were detected in 134 normal controls with the mutation frequencies of 4.48％,2.99％,4.48％ and 1.49％,respectively.Mutation frequency analysis showed an insignificant difference in the mutations of G3316A,T3394C,C3497T and T4216C between LHON patients and normal controls (x2 =0.926,P=0.336;x2 =0.052,P=0.820; x2 =0.142,P=0.707;P=0.129).G3376A,G3496T,G3700A,A4136G,T4160C and C4171A were absent in Chinese LHON patients. Conclusions Mitoehondrial ND1 gene in LHON is a mutational hotspot in Chinese population,11.19％ (100/894)associated with LHON was caused by ND1 gene mutation.G3635A,G3733A may be rare pathological mutation in Chinese population.However,G3316A,T3394C,C3497T and T4216C are insufficient to produce the clinical phenotype,but they may play a synergic role for penetrance and phenotypic manifestation in LHON.

Objective:To investigate the immunomodulation of CCK8 on the Coxsackievirus B ( CVB )-attacked human peripheral blood plasmacytoid dendritic cells(pDC). Methods:Peripheral blood mononuclear cells of healthy volunteers were separated by Ficoll-Hypaque gradient density centrifugation. The pDC was separated and divided into five groups,which were the control group, CVB attacked group,the group of CCK8 treated after CVB attack,the group of PGE2 treated after CVB attack and the group of CCK8+PGE2 treated after CVB attack. 100-time TCID50 of CVB was applied for the attack on pDC. Real-time PCR and Immunofluorescence technique were employed to detect the expression of CCK1R/CCK2R mRNA and protein. Then,the expression levels of costimulatory molecules such as CD80,CD86,HLA-DR ligand,and the chemokine receptor CCR7 were evaluated by Flow Cytometry Analysis. The supernatants of pDCs were collected, and the content of IFN-α was determined by Enzyme-linked Immunosorbent Assay. Results:CCK1R and CCK2R were co-expressed in human peripheral blood pDC,and both were significantly upregulated after CVB attack in vitro. Expression of CD80,CD86,HLA-DR and IFN-α were decreased in the CVB+CCK8 group compared with the CVB group,which suggested that CCK8 may reduce the CVB activation of pDC. Whereas expression of CD80,CD86,HLA-DR,CCR7 and IFN-α were increased in the CVB+PGE2 group compared with the CVB group,which suggested that PGE2 may increase the CVB activation of pDC in vitro. Conclusion:CCK8 repressed the CVB-attacked pDC,while PGE2 activated the CVB-attacked pDC.

OBJECTIVETo report the diagnosis and treatment experience of pediatric gastric cancer.

METHODSClinicopathological data of a girl with gastric carcinoma in December 2011 was analyzed retrospectively. The literatures were reviewed. The disease onset, clinical manifestations, treatment, and prognosis of the patient with gastric carcinoma were studied.

RESULTSThe patient was a 12-year-old girl, and the tumor was located in greater curvature. The girl underwent laparoscopy-assisted radical total gastrectomy (D2). Pathological examination showed gastric ulcerative signet-ring cell cancer (pT4N0M0, stage II b). The outcome at the recent follow up was satisfactory. Literature review demonstrated 15 cases of pediatric gastric carcinoma, of whom 7 were boys and 8 girls. Their average age was 12.7 years (2.5-17.8 years). Tumor locations included cardia (n=5), gastric body (n=1), gastric antrum or stomach corner (n=4), gastric antrum and gastric body (n=1), and unknown (n=4). Histology showed signet-ring cell cancer in 4 patients, poorly differentiated in 6, moderate to poorly differentiated and moderate differentiated adenocarcinoma in 2. Operation was performed in 11 cases and perioperative chemotherapy in 8 cases. Nine patients had follow-up. Four patients died within 6 months and other 4 patients survived for 6 months to one year. The survival time of one patient who underwent radical surgery and perioperative chemotherapy was 102 months.

CONCLUSIONSPediatric gastric carcinoma is extremely rare. It is highly malignant and associated with poor prognosis. Radical surgery is the most important treatment. Perioperative chemotherapy may improve the prognosis.

Adolescent ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Male ; Retrospective Studies ; Stomach Neoplasms ; diagnosis ; therapy