Three butylphthalide derivatives were isolated from the Rhizome of Ligusticum chuanxiong using a series of isolation and purification approaches including macroporous resin, ODS-A column, Sephadex LH-20 and preparative HPLC. These structures were elucidated based on extensive spectroscopic data (UV, IR, HR-ESI-MS and NMR) and identified as (3Z,3aE)-(6R,7R,2'S)-6-hydroxy-7-(2-carboxyl-2-hydroxyethylthio)-3-(2-hydroxybutylidene)-4,5,6,7-tetrahydro-phthalide (1), (3Z,3aZ)-3-butylidene-6,7-dihydroxy-4,5,6,7-tetrahydro-phthalide 7-O-α-D-glucopyranosyl-(1→2)-β-D-fructo-furanoside (2) and 3-(3-β-D-glucopyranosyloxy-butylidene)-7-hydroxy-phthalide (3).

Objective To investigate the presence of occult brain tissue damage in patients with relapsing neuromyelitis optica(RNMO)and its possible mechanism by using diffusion tensor imaging (DTI).Methods DTI scans were performed in 16 patients with RNMO and 16 sex-and age-matched healthy controls.Histogram analysis of mean diffusivity (MD)and fractional anisotropy (FA)was performed in brain tissue (BT),white matter (WM)and gray matter (GM)to detect the presence of occult brain tissue damage in RNMO patients.Region of interest(ROI )analysis of MD and FA was also performed in 6 dedicated regions with or without direct connection with spinal cord or optic nerve to determine the relationship between occult brain tissue damage and the damage of spinal cord and optic nerve.Results Patients with RNMO had a significantly higher average MD of the BT[RNMO(0.95?0.02)? 10~(-3)mm~2/s,controls (0.91?0.03)?10~(-3)mm~2/s,t = 3.940,P

OBJECTIVETo investigate the pathogenesis of globozoospermia, fertilization ability of round-headed sperm, and the application value of assisted oocyte activation in intracytoplasmic sperm injection (ICSI) for the wives of glohozoospermia men.

METHODSWe collected oocytes from the wives of 2 globozoospermia patients and randomly divided them into two groups after ICSI to receive calcium ionophore A23187-activation and conventional treatment, respectively. We reviewed the relevant literature published at home and abroad, and discussed the etiology of globozoospermia, fertilization ability of round-headed sperm, and treatment options for this disease.

RESULTSQuality embryos were obtained in the A23187-activation group while no fertilized oocytes, oocyte cleavage, quality embryos, or blastular formation were found in the conventional treatment group. Both women achieved pregnancy and gave birth to healthy neonates after transfer of the quality embryos from the A23187-activation group.

CONCLUSIONCalcium ionophore A23187 can be applied to ICSI for the wives of globozoospermia men and bring about desirable clinical outcomes. Meanwhile, attention should be paid to its safety.

Calcimycin ; therapeutic use ; Calcium Ionophores ; therapeutic use ; Female ; Humans ; Infertility, Male ; drug therapy ; Male ; Oocytes ; Pregnancy ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; abnormalities

A method for the simultaneous determination of oxygen and nitrogen in synthetic diamonds by inert gas high temperature extraction-impulse heating method was proposed. The sample weight, the selection of analysis power and the calibration curves of oxygen and nitrogen were discussed. Oxygen and nitrogen in analytical samples are determined. Values of RSDs (n=7) for oxygen and nitrogen were less than 4. 5% and 4. 0% respectively. The analytical results of oxygen and nitrogen obtained by the proposed method were in good agreement with those by inert gas fusion-impulse heating method.

A method for the determination of Fe, Co, Mn and Ni in synthetic diamonds by inductively coupled plasma atomic emission spectrometry ( ICP-AES) was proposed. The synthetic diamond sample was decomposed completely, while the sample was burned in air at 1000 ℃ for 10 h, and then a mixed acid of H2 SO4 , aqua regia and HClO4 was used for the dissolving the residue of the sample. In this method, the limits of detection of Fe, Co, Mn and Ni were 0. 0147, 0. 0018, 0. 0006 and 0. 0027 mg/L, respectively. Under the optimum condition, Fe, Co, Mn and Ni in synthetic diamond sample were determined. The values of RSDs (n=7) were less than 0. 5%. The recoveries of added standard were 94. 0%-105. 0%.

OBJECTIVETo study the toxic effects of aqueous extract of Crotalariae Assamicae Semen (CAS), one of the pyrrolizidine alkaloid-containing Chinese herbal medicines, in rats and the possible mechanism in association with liver damage.

METHODThe aqueous extract of CAS (CASE) was prepared by the conventional water extracting-alcohol precipitating method. The LD50 value of CASE in rats was determined by Kärber method. Rats were randomly divided into four groups in which three groups were orally administered with different doses of the CASE and one group with distilled water as control. Toxic effects were assessed by morphological, biochemical and histopathological changes. Moreover, in vitro metabolism using rat liver microsomes was also conducted and applied for the exploration of the underlying mechanism of liver damage.

RESULTThe LD50 value of CASE in Wistar rats was (2.36 +/- 0.26) g x kg(-1). The toxic effects were found in all groups of rats dosed with CASE, in which serum levels of ALT and AST were significantly elevated, and the obvious and dose-dependent damages in liver and lung were observed by histopathological examination. Moreover, the liver tissue-bound pyrroles were detected and generated in a dose-dependent manner, and the pyrrole metabolites observed in the in vitro microsomal metabolism. All the evidences suggested a strong correlation between metabolism and toxicity of CASE in rats.

CONCLUSIONCASE could induce the acute toxicity in rats, of which liver and lung were the major targets. Toxic effects were strongly correlated with pyrrolizidine alkaloids in CAS. The possible mechanism for its liver toxicity may be related to the formation of pyrrole metabolites as well as the corresponding tissue-binding products.

Alanine Transaminase ; metabolism ; Animals ; Crotalaria ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; toxicity ; Lethal Dose 50 ; Liver ; drug effects ; enzymology ; injuries ; Male ; Microsomes, Liver ; drug effects ; enzymology ; Pyrrolizidine Alkaloids ; administration & dosage ; toxicity ; Rats ; Rats, Wistar

OBJECTIVETo investigate the association of -141C insert/delete polymorphism with schizophrenia in Wuhan of Hubei province.

METHODSA case-control study was conducted to analyze the polymorphism in the D(2) receptor gene promoter region with schizophrenia. A total of 120 cases of schizophrenia diagnosed according to CCMD-II R criteria and 100 normal controls were recruited in the study.

RESULTSIn this sample, the allele and genotype showed statistically significant differences between patients and normal controls (P<0.05).Especially, the frequency of -141C del was 11% in patients and 18% in control(OR 0.55, 95% CI 0.30-0.96; P<0.05). This allele was less common in schizophrenia than in normal controls (P<0.05).

CONCLUSIONThe -141C del polymorphism is associated with schizophrenia.The polymorphism may modify the association with other factors. Possibly -141C del in the DRD(2) promoter region is a strong candidate for a protective factor for this trait.

Adult ; Alleles ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Promoter Regions, Genetic ; genetics ; Receptors, Dopamine D2 ; genetics ; Schizophrenia ; diagnosis ; genetics

BACKGROUNDInteractions of tumor cells with the microenvironment were deemed to promote the tumor invasion and metastasis. CXC chemokine receptor 4 (CXCR4) and extracellular matrix metalloproteinase inducer (EMMPRIN) had reported to participate in this process. However the roles of bone marrow microenvironment in leukemic infiltration were not well investigated.

METHODSA co-culture system between SHI-1 cells and bone marrow stromal cells (BMSCs) is used to simulate the interactions of leukemic cells with their microenvironment. The trans-matrigel invasion was used to detect the capability of SHI-1 cells invasion. The BMSCs and SHI-1 cells were mixed in a ratio of 1:10 and added to the millicell chamber coated with matrigel. Either the co-culture supernatant or the functional blocking peptide of CXCR4 and EMMPRIN were added to the trans-matrigel invasion system. The expressions of EMMPRIN in SHI-1 cells and BMSCs were detected by RT-PCR. The changes of the expression of matrix metalloproteinase-2, 9 (MMP-2, MMP-9), tissue inhibitor of metalloproteinase 2 (TIMP-2), and CXCR4 mRNA in SHI-1 cells were determined by real-time PCR. The concentration of stromal cell derived factor 1 (SDF-1) in serum free supernatant was measured by ELISA.

RESULTSBoth SHI-1 cells and BMSCs express EMMPRIN. SHI-1 cells could hardly invade the matrigel membrane; the coculture supernatant did not induce the invasion of SHI-1 cells. When contacting directly with BMSCs, SHI-1 cells invaded to the lower chamber of millicell were significantly increased. The functional blocking peptide of CXCR4 and EMMPRIN could significantly inhibit the invasion triggered by BMSCs. When co-culturing with BMSCs, the expression of CXCR4, MMP-2, MMP-9 and TIMP-2 mRNA in SHI-1 cells were significantly elevated in company with a significantly higher level of SDF-1 in the co-cultured serum-free supernatant.

CONCLUSIONThe interactions of leukemic cells and BMSCs play important roles in leukemic cell infiltration.

Basigin ; physiology ; Cell Communication ; Cell Line, Tumor ; Coculture Techniques ; Humans ; Leukemia, Monocytic, Acute ; pathology ; Mesenchymal Stromal Cells ; physiology ; Neoplasm Invasiveness ; Receptors, CXCR4 ; physiology

OBJECTIVETo compare the effects of the cryoprotectant containing glucose and that containing sucrose on the motility of post-thaw human sperm.

METHODSThe cryoprotectant containing glucose and that containing sucrose were applied to 50 semen samples and the motility of the post-thaw human sperm was compared before and after cryopreservation and between the study groups.

RESULTSThe forward motility and total motility of the sperm were (58.4 +/- 5.7)% and (63.4 +/- 6.1)% before cryopreservation, (43.8 +/- 7.6)% and (48.4 +/- 7.6)% after thawing with the cryoprotectant containing glucose, and(42.6 +/- 8.9)% and (48.0 +/- 8.5)% after thawing with the cryoprotectant containing sucrose. Decreased sperm motility was observed after cryopreservation, with statistic significance (P < 0.01). There was no significant difference in the forward and total motility of the post-thaw sperm between the two cryoprotectants.

CONCLUSIONCryopreservation inflicts obvious damage on sperm. Sucrose is a feasible sperm cryoprotectant.

Adult ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Glucose ; pharmacology ; Humans ; Male ; Semen Preservation ; methods ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects ; physiology ; Sucrose ; pharmacology

OBJECTIVETo investigate the influence of tissue inhibitor of metalloproteinase 2 (TIMP-2) on the infiltrative patterns of human monocytic leukemic cell line SHI-1 in nude mice.

METHODS1) 1 x 10(7) TIMP-2 gene transduced SHI-1 (SHI-1-TIMP-2) and SHI-1 transduced MSCV gene (SHI-1-MSCV) cells were inoculated via tail vein into 6-week nude mice, which pretreated by splenectomy, cytoxan intraperitoneal injection, and sublethal irradiation(referred as SCI nude mice). 30 days after inoculation, half of the mice were sacrificed, and the infiltration patterns were investigated by histological exam and human CD45 immunohistochemistry, other mice were observed for survival time. 2) Leukemic cells inoculated subcutaneously into the axillary area of mice without any pre-treatment. On day 23 and 30, mice were sacrificed to measure the volume of neoplasm. TIMP-2 protein expression and the micro vein density were detected by immunohistochemistry.

RESULTSIn SCI nude mice inoculated via caudal vein with SHI-1-TIMP-2 cells, the survival time was shorter and infiltration (including in central nervous system) was higher than that in those inoculated with SHI-1-MSCV cells. However, in inoculated subcutaneously group, the neoplasm though grew rapidly at first, over expression of TIMP-2 limited the tumor growth and angiogenesis.

CONCLUSIONThe functions of TIMP-2 are diversity; the role of TIMP-2 in tumor infiltration and metastasis was worthy of further investigation.

Animals ; Cell Line, Tumor ; DNA, Complementary ; genetics ; Genetic Vectors ; Humans ; Leukemia, Experimental ; genetics ; pathology ; Leukemic Infiltration ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; Transfection