To investigate the role and mechanism of ceramide (Cer) regulation in alcohol-induced neuronal proliferation and the newborn neurons formation, we used sphingomyelin synthase 2 (predominant enzyme of Cer metabolism) knockout (SMS2(-/-)) and wild type (WT) female mice to establish the model of prenatal alcohol exposure. In 24 h after being given birth (postnatal day 0, P0), the offspring of model mice received blood sphingomyelin (SM) measurement with enzymatic method. On P0, P7, P14 and P30, the proliferation of granule cells in the dentate gyrus and newborn neurons were investigated with immunofluorescent labeling. The expression of protein kinase Cα (PKCα) in the hippocampus was tested with Western blot analysis. The results showed that the SM level of blood in SMS2(-/-) pups was significantly lower than that in WT pups. No matter in SMS2(-/-) or WT mice, the prenatal alcohol exposure down-regulated the SM levels in pups with dose-dependency. In both SMS2(-/-) and WT pups, the number of proliferative neurons and newborn neurons in the dentate gyrus gradually decreased with the growing age. Compared with the WT pups, SMS2(-/-) pups showed significantly more proliferative neurons and newborn neurons in the dentate gyrus. Notably, prenatal alcohol exposure dose-dependently increased proliferative neurons and newborn neurons in the dentate gyrus in both WT and SMS2(-/-) pups. The hippocampal expression of PKCα protein in SMS2(-/-) mice was lower than that in WT mice, and prenatal alcohol exposure could up-regulate the PKCα protein expression in both WT and SMS2(-/-) mice with dose dependency. These results suggest that alcohol exposure during pregnancy can induce the compensatory neural cell proliferation and the production of newborn neurons in offspring, and the Cer-ceramide-1-phosphate (C1P) pathway is involved in alcohol-induced neural cell proliferation. The activation of PKCα may be a key step to start the Cer-C1P pathway and up-regulate the alcohol-induced neural cell proliferation and the newborn neurons formation.
Animals ; Animals, Newborn ; Cell Proliferation ; drug effects ; Cells, Cultured ; Ceramides ; metabolism ; Dentate Gyrus ; cytology ; Ethanol ; toxicity ; Female ; Mice ; Mice, Knockout ; Neurons ; cytology ; Pregnancy ; Prenatal Exposure Delayed Effects ; physiopathology ; Protein Kinase C-alpha ; metabolism ; Signal Transduction ; Transferases (Other Substituted Phosphate Groups) ; genetics

Objective To analyze clinical diagnosis and management of 5 patients with actinomycete keratitis.Design Retro- spective case series.Participants 5 patients (5 eyes) with actinomycete keratitis.Methods The clinical features and microbiologic da- ta of 5 culture-proven cases of actinomycete keratitis recorded between October 2004 to March 2006 were analyzed.Main Outcome Measures clinical characteristics,isolations identification,drug susceptibility test and treatments.Results All patients were males and farmers.Of the 5 cases presented in this study,4 cases were followed by minor trauma as a predominant risk factor,and were pre- sented by a chronic progressive corneal ulcer with a wreath pattern of infiltrate.The diagnosis of all cases was based on laboratory in- vestigations,by which 4 cases of nocardia and one case of streptomyce were identified.A variable drug sensitivities were presented in nocardia isolates,which including TMP-SMZ,amicasin,gentamicin and fluorine-quinolones.Conclusions Nocardia keratitis is mainly followed by a minor trauma.It is identified predominantly by laboratory investigations.Tropical and systemically sensitive biotic are the initial choice,while debridement and amnionic transplantation could be an effective alternative.

OBJECTIVETo investigate the relation of heparanase gene and angiogenesis with the progress of gastric carcinoma (GC).

METHODSExpression of heparanase mRNA in 52 GC tissue was detected by in situ hybridization assay. Microvessel density (MVD) was examined by immunohistochemical method. MVD and heparanase mRNA expression were analysed with their relation to histological grade, invasion depth,lymph node metastasis and organ metastasis.

RESULTSMVD was 73.2 +/- 22.8 in 25 (48.1%) tissue with positive heparanase mRNA. It was 44.8 +/- 11.9 in 27 (51.9%) tissues with negative heparanase mRNA, between which the difference was significant (P < 0.001). MVD and heparanase mRNA expression were related with lymph node metastasis and depth of serosal invasion in GC (P < 0.005).

CONCLUSIONHeparanase, being related to angiogenesis in gastric cancer, promotes growth, invasion and metastasis. Heparanase mRNA expression is an important predictor of the biological behavior of human gastric carcinoma.

Adenocarcinoma ; blood supply ; enzymology ; pathology ; Adult ; Aged ; Female ; Gastric Mucosa ; enzymology ; Gene Expression Regulation, Neoplastic ; Glucuronidase ; biosynthesis ; genetics ; Humans ; Lymphatic Metastasis ; Male ; Microcirculation ; pathology ; Middle Aged ; Neoplasm Invasiveness ; Neovascularization, Pathologic ; enzymology ; RNA, Messenger ; biosynthesis ; genetics ; Stomach Neoplasms ; blood supply ; enzymology ; pathology

OBJECTIVETo find new serum tumor markers for ovarian epithelial cancers by 2-DE DIGE and MALDI-TOF/TOF proteomic methods, in order to improve the diagnostic sensitivity and specificity.

METHODSSerum samples from 103 cases of ovarian epithelial cancers, 60 cases of healthy women, 63 cases of benign ovarian tumors and 63 cases of benign pelvic diseases were collected. Sera of 20 cases of ovarian epithelial cancers (A), 20 cases of ovarian benign tumors (B), 20 cases of pelvic benign diseases (C) and 20 cases of health control (D) were matched by age and pooled, respectively. After depletion of high abundance serum albumin and IgG, the samples were assayed by 2-DE DIGE. The test was repeated three times. Analysis with DeCyder software revealed significant differential protein spots which were identified by MAIDI-TOF/TOF. Western blot and ELISA were used to validate the candidate serum markers.

RESULTS1) There were 41 proteins having significant differences between the groups. MAIDI-TOF/TOF successfully identified 28 proteins. Haptoglobin (Hp) was the most significantly up-regulated protein, and transferrin (Tf) was the most significantly down-regulated protein. 2) Western blot and ELISA proved that there were significant differences in Hp and Tf between ovarian epithelial cancers and normal controls (P = 0.000), between ovarian epithelial cancers and ovarian benign tumors (P = 0.000), between ovarian epithelial cancers and benign pelvic disease sera (P = 0.000). 3) CA125 + Hp + Tf combined detection of ovarian cancer had higher sensitivity and specificity than CA125, Hp or Tf detection alone.

CONCLUSIONHp and Tf are differently expressed in the sera of patients with ovarian epitheliual cancers. They can be used as serum biomarkers for ovarian epithelial cancers. CA125 + Hp + Tf combined detection may improve the sensitivity and specificity of diagnosis of ovarian epithelial cancers.

Adenocarcinoma, Clear Cell ; blood ; Biomarkers, Tumor ; blood ; Cystadenocarcinoma, Mucinous ; blood ; Cystadenocarcinoma, Serous ; blood ; Electrophoresis, Gel, Two-Dimensional ; Endometriosis ; blood ; Female ; Haptoglobins ; analysis ; Humans ; Ovarian Neoplasms ; blood ; Pelvic Inflammatory Disease ; blood ; Proteins ; analysis ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Teratoma ; blood ; Transferrin ; analysis

Human placental decidua basalis-mesenchymal stem cells (PDB-MSCs) are multipotent cells from the human term placenta, which are ethically conducive, easily accessible and high-yielding source. PDB-MSCs can differentiate into adipogenic, osteogenic and neurogenic cells under appropriate conditions, which may be an attractive and alternative source of seed cells for tissue engineering. To investigate the effect of hypoxia (1% O2) on human PDB-MSCs and the expression of cytokine, PDB-MSCs were isolated from human placenta by density gradient centrifugation and cultured in the Dulbecco's modified Eagle's medium-high glucose (DMEM-HG) containing 10% fetal bovine serum (FBS), and the fifth passage of PDB-MSCs were taken. PDB-MSCs were divided into 4 groups according to the concentrations of O2 and FBS: 20% O2, 10% FBS; 20% O2, 0% FBS; 1% O2, 10% FBS; 1% O2, 0% FBS. The proliferation and apoptosis of PDB-MSCs were detected by MTT and flow cytometric analysis at the time points of 6 h, 12 h, 24 h, 48 h, 72 h, and 96 h, respectively. Vascular endothelial growth factor (VEGF) released from PDB-MSCs was detected by enzyme-linked immunosorbent assay (ELISA) at the same time points. The results showed that hypoxia enhanced the proliferation of PDB-MSCs at 12 h under the condition of 10% FBS, while at 24 h under the condition of 0% FBS (P<0.01, n=3). In normoxia, the cells cultured in 10% FBS displayed a significant proliferation compared to those cultured in 0% FBS. However, in hypoxia, the number of cells cultured in 0% FBS (serum deprivation) increased significantly compared to that cultured in 10% FBS at 24 h and 96 h respectively (P<0.05, P<0.01, n=3). With the flow cytometric analysis of cell apoptosis under the condition of hypoxia and serum deprivation, we found that hypoxia and serum deprivation did not induce PDB-MSCs apoptosis (P>0.05, n=3). This conclusion may relate to the expression of VEGF which needs further research. In conclusion, the results obtained indicate that PDB-MSCs are able to bear hypoxia and serum deprivation, suggesting that PDB-MSCs can be used as seed cells for ischemia related tissue engineering.
Apoptosis ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Decidua ; cytology ; Female ; Humans ; Mesenchymal Stromal Cells ; cytology ; Placenta ; cytology ; Pregnancy ; Tissue Engineering ; Vascular Endothelial Growth Factor A ; metabolism

Objective To investigate the frequency of CD4~+CD25~+FoxP3~+regulatory T cells (Treg)and the expression of the functional protein,FoxP3,in patients with active tuberculosis and the relationship between Treg and the pathogenesis of tuberculosis.Methods Forty-five patients with active tuberculosis(including 25 cases of pulmonary tuberculosis and 20 tuberculous lymphadenitis), 20 healthy controls,20 recovered tuberculosis patients and 6 patients with reactive hyperplasia in cer- vical lymph node were enrolled.The frequency of CD4~+ CD25~+ FoxP3~+ Treg in the peripheral blood was measured by flow cytometry.FoxP3 mRNA expression was determined by real-time reverse transcriptase-polymerase chain reaction(RT-PCR)and the expression of FoxP3 protein in lymphoid tissues was measured by immunohistochemistry.Results The frequency of natural Treg in the peripheral blood from the patients with active tuberculosis was 2.91%?0.23%,which was signifi- cantly higher than that of healthy control group(1.22%?0.18%)and recovered tuberculosis patients(1.50%?0.17%,P

AIMTo separate and purify the anti-myocardial ischemic polysaccharide fraction with a homogenous molecular weight from Ophiopogon japonicus, then study the chemical structure of the parts.

METHODSCrude polysaccharides were prepared by extracting the tube root fraction of Ophiopogon japonicus with water, then precipitation with ethanol. From the crude polysaccharides, the polysaccharide of MDG-1 was separated and purified using ultrafiltration, DEAE Sepharose FF and Sephadex G-25 column chromatography. Its structure was studied by complete hydrolysis, periodate oxidation, Smith degradation, methylation analysis, 1H NMR and 13C NMR analysis etc.

RESULTS AND CONCLUSIONMDG-1 was a water-soluble beta-D-fructosan, containing a backbone composed of Fruf (2 --> 1), and a branch of Fruf (2 --> 6) Fruf (2 --> per average 2. 8 of main chain residues. Mn, Mw and Mp of MDG-1 were 3 400, 4 800 and 5 000, respectively. MDG-1 contains trace of Glc, which maybe connect to its reducing terminal. Molar ratio of Fru and Glc is approximately 35: 1.

Methylation ; Molecular Structure ; Molecular Weight ; Ophiopogon ; chemistry ; Plant Tubers ; chemistry ; Plants, Medicinal ; chemistry ; Polysaccharides ; chemistry ; isolation & purification

OBJECTIVE: To investigate the relationship between serum level of homocysteine and the development of collaterals in patients with severe coronary artery stenosis (SCAS). METHODS: Eighty patients with at least one vessel stenosis over 90% among the 3 main vessels of coronary artery were consecutively enrolled into the study according to angiographic estimation. The development of collaterals was classified by Rentrop's method. RESULTS: The serum levels of homocysteine among the single-vessel, bi-vessel and tri-vessel coronary artery disease groups had no significant difference; there was no linear correlation between the serum level of homocysteine and Gensini's score. The level of homocysteine in the poorly developed collaterals was significantly higher than that in the well-developed collaterals in the SCAS patients (P<0.001). Multiple stepwise logistic analysis revealed that homocysteine negatively correlated with the development of collaterals (P<0.001, odds ratio=0.353; 95% confidence interval=0.201 - 0.620), whereas it positively correlated with the number of stenosis vessels. CONCLUSION The serum level of homocysteine is independently and negatively associated with the development of collateral circulation in severe SCAs patients.
Adult ; Aged ; Aged, 80 and over ; Collateral Circulation ; Coronary Angiography ; Coronary Circulation ; Coronary Stenosis ; blood ; physiopathology ; Female ; Homocystine ; blood ; Humans ; Logistic Models ; Male ; Middle Aged

OBJECTIVETo evaluate the quality difference between Coptis chinensis planted with ecological techniques and shelf planted Coptis chinensis.

METHODSUltraviolet-visible spectrophotometry, alcohol extract mensuration, moisture mensuration, and ash mensuration were used to determine the contents of total alkaloids, alcohol extract, water, and total ash of Coptis chinensis, which were planted in shelf, Rhus chinensis wood, M mulbery wood, corn wood, Magnolia officinalis wood, fruiter wood, shading net, and firry wood, respectively.

RESULTSThe contents of total alkaloids and alcohol extract of Coptis chinensis Table planted with ecological techniques were higher than those of Coptis chinensis planted in shelf. The contents of water and total ash were less than 12% and 5%, respectively, which met the provisions of the pharmacopoeia.

CONCLUSIONThe quality of Coptis chinensis planted with ecological techniques is similar to that of Coptis chinensis planted in shelf. These ecological techniques for Coptis chinensis have become mature and practical.

Agriculture ; methods ; Alkaloids ; analysis ; Conservation of Natural Resources ; Coptis ; chemistry ; growth & development ; Ecosystem ; Forestry ; methods ; Plant Extracts ; analysis ; Plants, Medicinal ; chemistry ; growth & development ; Quality Control ; Water ; analysis

OBJECTIVE: To evaluate the short and mid-term changes of the cardiac morphology after percutaneous transcatheter closure of ventricular septal defects (VSD) with transthoracic echocardiography (TTE). METHODS: The left ventricular end-diastolic diameter (LVEDD), left ventricular end-diastolic volume (LVEDV), left atrial diameter (LAd), and right ventricular diameter (RVd) in 30 VSD patients were measured before the VSD closure,and on the 3rd day, 3rd month, and 6th month after the VSD closure by TTE. RESULTS: LVEDD and LVEDV significantly decreased on the 3rd day after the VSD closure compared with pre-VSD closure. LVEDD and LVEDV continuously decreased on the 3rd month and 6th month after the VSD closure. LAd was smaller on the 3rd month and 6th month after the VSD closure, but there was not significant difference between the 3rd and 6th month. RVd increased on the 3rd day after the VSD closure, while no significant difference was found among the 3rd month and 6th month before and after VSD closure. CONCLUSION Percutaneous transcatheter VSD closure may effectively improve the cardiac remodeling in VSD patients in the short and mid-term follow-up.
Adolescent ; Adult ; Cardiac Catheterization ; methods ; Child ; Child, Preschool ; Echocardiography ; Female ; Follow-Up Studies ; Heart Septal Defects, Ventricular ; diagnostic imaging ; therapy ; Humans ; Male ; Prosthesis Implantation ; methods ; Time Factors ; Ventricular Remodeling