A cDNA sequence of Arnebia euchroma AP2/ERF named AeAP2/ERF was cloned by in silico cloning in this study, using ACX71873 sequence from Lithospermum erythrorhizon as the probe sequence. Some characters of the AP2/ERF gene and encoded protein sequences were predicted and analyzed by the bioinformatics methods, including general physical and chemical properties, hydrophobieity, signal peptide, secondary structure, localization sites in cells. Results showed that the 876 bp long gene included a 1 077 bp ORF and encoding 205 amino acid. The AeAP2/ERF protein had no signal peptide, it was a hydrophilic proteins located in nucleus. The function of the AP2/ERF protein was mainly involved with metabolism controlling and signal transduction.
Amino Acid Sequence ; Base Sequence ; Boraginaceae ; classification ; genetics ; Cloning, Molecular ; Computational Biology ; Computer Simulation ; Hydrophobic and Hydrophilic Interactions ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Protein Structure, Secondary ; Protein Structure, Tertiary ; Transcription Factors ; chemistry ; genetics

OBJECTIVETo establish an HPLC fingerprint to evaluate the quality of Polygalae Radix, root xylem, and those collected in different growth ages or harvest time.

METHODSeparation was performed at 30 °C on a Kromasil C18 column (4.6 mm x 250 mm, 5 μm); the mobile phases was acetonitrile and 0.05% H3PO4 water in the gradient elution; the flow rate was set at 1.0 mL · min(-1) and the detection wavelength at 314 nm; the quality discriminant analyses were accomplished by means of similarity analysis, cluster analysis, principal component analysis and neural network model.

RESULTIn 26 batches of Polygalae Radix, 24 batches fingerprint similarities were above 0.8. In 5 different growth or harvest time batches, 4 batches were above 0.8; in 8 batches root xylem samples, the similarities were all above 0.875. The similarity analysis was in accord with the quality discriminant analysis of cluster analysis, principal component analysis and neural network model.

CONCLUSIONFingerprint combined with chemical pattern recognition technique can effectively evaluate the quality of Polygalae Radix. The active substance species are all similar in cultivated, wild, different growth or harvest time Polygalae Radix and polygala root xylem, but the chromatography peak areas are different. The effective material contents are similar between wild and cultivated Polygalae Radix, but each chromatographic peak area of the root xylem is much smaller than that of Polygalae Radix. The chemical substance accumulation mainly depends on harvest month, but little growth time in Polygalae Radix.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Plant Roots ; chemistry ; classification ; Polygala ; chemistry ; classification ; Quality Control

The chemical investigation on Ganoderma philippii led to the isolation of sixteen compounds by silica gel and Sephadex LH-20 column chromatography. On the basis of spectroscopic data analyses, their structures were elucidated as 2, 5-dihydroxyacetophenone (1), methyl gentisate (2), (S) -dimethyl malate (3), muurola-4, 10 (14) -dien-11beta-ol (4), dihydroepicubenol (5), 5-hydroxymethylfuran carboxaldehyde (6), ergosta-7, 22E-dien-3beta-ol (7), ergosta-7, 22E-dien-3-one (8), ergosta-7, 22E-diene-2beta, 3alpha, 9alpha-triol (9), 6/beta-methoxyergo-sta-7, 22E-dien-3beta, 5alpha-diol (10), ergosta-4, 6, 8(14), 22E-tetraen-3-one (11), ergosta4, 6, 8-(14), 22E-etetraen-3beta-ol (12), 5alpha, 8alpha-epidioxy-ergosta-6, 22E-dien-3beta-ol (13), 7alpha-methoxy-5alpha, 6alpha-epoxyergosta-8-(14), 22E-dien-3beta-ol (14), ergosta-8, 22E-diene-3beta, 5alpha, 6beta, 7alpha-tetraol (15), and ergosta-5, 23-dien-3beta-ol, acetate (16). All the compounds were obtained from this fungus for the first time, and compounds 4 and 5 were isolated from the Ganoderma genus for the first time.
Ganoderma ; chemistry ; Medicine, Chinese Traditional ; Organic Chemicals ; analysis ; isolation & purification

Objective To observe the effects of comprehensive measures of changing grain,selenium supplementation,off-site education and resettlement on prevention of children's Kashin-Beck disease in Aba state.Methods Fifty eight villages in Aba Kashin-Beck disease areas were chosen as intervention points in Aba state Sichuan province from 2007 to 2011.Based on the implementation of prevention and control measures,the villages were divided into off-site education + changing grain + selenium supplementation group and resettlement + off-site education + changing grain + selenium supplementation group,Geletuo town of Seda county,Ganzi state was selected as a control point,and right-hand anteroposterior X-ray examination(including the wrist) was carried out on children aged 6-13 from 2007 to 2011 annually.Clinical and X-ray diagnosis of Kashin-Beck disease was made in accordance with the Diagnostic Criteria of Kaschin-Beck Disease(GB 16003-1995).The effects of prevention and control measures were evaluated by comparing the child X-ray detection rate before and after the implementation of the measures.Results The average X-ray positive detectable rate of children in the intervention points was 2.07％(66/3181),2.72％ (69/2540),1.16％ (35/3017),0.56％ (19/3397) and 0.56％ (24/4273),respectively from 2007 to 2011,with a downward trend (x2trend =66.74,P ＜ 0.01).There was a downward trend in the average X-ray positive detectable rate of children in off-site education + changing grain + selenium supplementation group [1.60％(29/1809),2.63％ (39/1484),1.29％ (25/1941),0.64％ (15/2332),0.42％ (10/2379)] and resettlement + off-site education + changing grain + selenium supplementation group [2.70％ (37/1372),2.84％ (30/1056),0.93％(10/1076),0.38％ (4/1065),0.74％(14/1894)] (x2trend=30.97,35.19,all P ＜ 0.01).The average X-ray positive detectable rate of children in the intervention group was 0 from 2007 to 2010,and was 1.61％ (1/62) in 2011.The difference of X-ray positive detectable rate was not statistically significant in the control group in the 5 years from 2007 to 2011.The difference of children's X-ray positive detectable rate was not statistically significant between control group and intervention group.Conclusions The effect of taking changing grain,selenium supplementation,off-site education and resettlement comprehensive measures to prevent children's Kashin-Beck disease is not significant in those places where the state of Kaschin-Beck disease is not active.

OBJECTIVETo identify the anatomical variability of the left spermatic vein (LSV) and determine its effect on the induction of experimental left varicocele (ELV) in adolescent rats.

METHODSWe equally randomized 30 adolescent male SD rats to groups A (LSV collaterals fully ligated and the left renal vein constricted), B (only the left renal vein constricted), and C (sham operation), observed the courses of the LSVs and measured their diameters. At 30 days after operation, we analyzed the changes in the left kidneys and the diameters of the LSVs.

RESULTSIrregular collaterals were observed in 90% of the LSVs and no abnormal changes were found in the left kidneys after surgery. The postoperative LSV diameter was remarkably increased in group A as compared with the baseline ([1.47 +/- 0.15 ] vs [0.16 +/- 0.08] mm, P < 0.01), but showed no significant difference in group B ([0.31 +/- 0.49] vs [0.15 +/- 0.07] mm, P > 0.05) and C ([0.17 +/- 0.07] vs [0.16 +/- 0.06] mm, P > 0.05), and it was significantly longer in A than in B (P < 0.01). The success rate of ELV induction was 100% in group A and 10% in group B, but no varicocele was observed in group C.

CONCLUSIONCorrect identification of the anatomical course of the LSV and ligation of its irregular collaterals are essential for the establishment of a stable and consistent ELV model.

Animals ; Disease Models, Animal ; Kidney ; pathology ; Ligation ; Male ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord ; blood supply ; Varicocele ; Veins ; abnormalities

OBJECTIVETo investigate the effect of rapid canine distalization through distraction of the periodontal ligament after reducing interseptal bone resistance.

METHODSTwenty canines in 11 patients who needed first premolar extractions were involved. A tooth-borne, custom-made distractor was bonded right after the first premolar extraction and the interseptal bone resistance reduction. Three days post-operatively, the distractor was activated 0.1 mm three times a day. Orthodontic models, panoramic radiographs, periapical radiographs, electrical vitality test were assessed pre- and post distraction procedure and 3 months after the completion of the procedure.

RESULTSThe distraction procedure was completed in 18 to 35 days [mean (25.6 +/- 4.7) days], with the distal displacement of the canines ranging from 3.53 to 8.29 mm [mean (5.56 +/- 1.32) mm]. The canines showed a mean of 12.20 degrees distal tipping and 18.53 degrees rotation. The anchorage teeth showed an average of (0.76 +/- 0.75) mm mesial movement. The mesial contact point of incisors showed a mean of (0.67 +/- 0.55) mm lingual movement. There was no significant root resorption or long-time change on pulp vitality after distraction.

CONCLUSIONSThe canine distalization through distraction of the periodontal ligament after reducing interseptal bone resistance was an effective approach to move canines rapidly.

Adolescent ; Cuspid ; surgery ; Female ; Humans ; Male ; Periodontal Ligament ; surgery ; Root Resorption ; surgery ; Tooth Movement Techniques ; methods

OBJECTIVETo investigate the degradation of artificial basement membrane (matrigel) co-cultured with oral carcinoma-associated fibroblasts (CAFs) and its possible mechanism.

METHODSCAFs and normal fibroblasts (NFs) were incubated on matrigel for 24, 48, 72 h. Equivalent amounts of conditioned medium were collected and assayed for total protein, hydroxyproline and matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) activity by gelatin zymography.

RESULTSOral CAFs were superior to oral NFs in total protein and hydroxyproline density, CAFs present more pro-MMP-2 and activated MMP-2.

CONCLUSIONCAFs were superior to NFs in degradation of matrigel. CAFs might play a key role in the reconstitution of extracellular matrix and the progression of tumor.

Basement Membrane ; Coculture Techniques ; Enzyme Precursors ; Fibroblasts ; Gelatinases ; Humans ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Membranes, Artificial ; Mouth Neoplasms

BACKGROUNDFew studies have examined the properties of human immunodeficiency virus type 1 (HIV-1) epitope-specific cytotoxic T lymphocyte (CTL) responses in children. To address this issue, we characterized epitope-specific CTL responses and analyzed the determinants that may affect CTL responses before and after highly active antiretroviral therapy (HAART) in children with HIV-1 infection.

METHODSA total of 22 HIV-1-infected children and 23 uninfected healthy children as control were enrolled in the study. Circulating CD4 T cells and HIV-1 RNA load in plasma were routinely measured. Peripheral HIV-1-specific CTL frequency and HIV-1 epitope-specific, interferon-gamma (IFN-gamma)-producing T lymphocytes were measured using tetramer staining and enzyme-linked immunospot (ELISPOT) assay, respectively. Circulating dendritic cell (DC) subsets were monitored with FACS analysis.

RESULTSMore than 80% of the children with HIV-1 infection exhibited a positive HIV-1-epitope-specific CTL response at baseline, but HIV-specific CTLs and IFN-gamma-producing lymphocytes decreased in patients who responded to HAART in comparison with non-responders and HAART-naive children. The duration of virus suppression resulted from HAART was inversely correlated with CTL frequency. While in HAART-naive children, HIV-1-specific CTL frequency was positively correlated with myeloid DC (mDC) frequency, although the cause and effect relationship between the DCs and CTLs remains unknown.

CONCLUSIONSHIV-1-epitope-specific CTL responses are dependent on antigenic stimulation. The impaired DC subsets in blood might result in a defect in DC-mediated T cell responses. These findings may provide insight into understanding the factors and related mechanisms that influence the outcome of HIV-1 carriers to HAART or future antiviral therapies.

Adolescent ; Antiretroviral Therapy, Highly Active ; CD8-Positive T-Lymphocytes ; immunology ; Child ; Epitopes ; immunology ; Female ; HIV Infections ; drug therapy ; immunology ; HIV-1 ; immunology ; Humans ; Male ; T-Lymphocytes, Cytotoxic ; immunology ; Viremia ; drug therapy

Objective To investigate the protective effect of modified Qianjin Weijing Decoction(MQJWJD)on TNF-α and NF-κB in rats with lung injury induced by particulate matter; To discuss relevant mechanism of action. Methods A tracheal drip 15 mg/kg fine particles of saline solution was used to establish modeling, every other day, three times. Thirty-two Wistar rats were randomly divided into normal group, model group and MQJWJD high-dose and low-dose groups, with eight rats in each group. Medication groups were given relevant medicine for gavage. The level of TNF-α in bronchoalveolar lavage fluid (BALF) was measured by ELISA. The expression of NF-κB protein in lung tissues was measured by immunohistochemistry. The histopathology of the lung injury was observed by light microscope. Results Compared with normal group, the level of TNF-α and the expression of NF-κB protein in the model group were higher than those in the normal group (P<0.01). Compared with model group, the level of TNF-α and the expression of NF-κB protein in MQJWJD low-dose and high-dose groups were lower than those in the model group (P<0.05, P<0.01). Pathological observation showed that, compared with normal group, model group showed intratracheal, alveolar and interstitial bacteria within a large number of fine particles calm, alveolar and pulmonary interstitial visible large amounts of phagocytic fine particles of macrophages and accompanied by more neutrophils and lymphocyte infiltration; Lung tissue pathological changes were significantly lighter in MQJWJD high-dose and low-dose groups than the model group. MQJWJD high-dose group showed mild inflammation, alveolar and pulmonary interstitial visible phagocytic fine particles of macrophages, a small amount of neutrophils and lymphocyte infiltration. Conclusion MQJWJD can reduce the pulmonary injury in rats induced by particulate matter and has protective effects on the rat model through decreasing the levels of TNF-α and the expressions of NF-κB protein in injured lung tissues.

OBJECTIVETo investigate the difference in the CD4+T lymphocytes activation between long term non progressors (LTNP) and typical progressors (TP) of HIV-1 infected patients.

METHODSTwenty-four HIV-1 infected patients and 15 heathy control adults were tested and flow cytometry was used to detect the activation marker CD38 and CD4 count in blood samples taken from the patients and control. bDNA method was used to test the virus load in the plasma of patients.

RESULTSThe activation of CD4+T cells was positively correlated with virus load and negatively correlated with CD4 counts. Compared with normal controls, the activation of CD4+T cells was obviously increased in TP patients but not obviously changed in LTNP patients.

CONCLUSIONCompared with healthy controls, the activation of CD4+T cells in LTNP did not obviously increase. This maybe partially accounts for LTNP patients keeping a good state for a long time.

ADP-ribosyl Cyclase 1 ; analysis ; Adult ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; Female ; Flow Cytometry ; HIV Infections ; blood ; immunology ; virology ; HIV-1 ; genetics ; physiology ; Host-Pathogen Interactions ; Humans ; Lymphocyte Activation ; immunology ; Male ; Middle Aged ; RNA, Viral ; blood ; Time Factors ; Viral Load ; Young Adult