OBJECTIVETo observe Smads protein expression in lung tissue of quartz exposed mice and to explore its association with pulmonary fibrosis in silicosis.

METHODSThe experimental mice were divided into control and quartz groups. 0.2 g/kg weight of quartz was injected intratracheally in quartz group. Samples were collected at the 1st, 3rd, 5th, 7th, 14th and 28th day after injection. Immunohistochemical methods with quantitative image analysis were used to assay the protein expression of transforming growth factor beta(1) (TGF-beta(1)), Smad 2/3, Smad 4, and Smad 7 protein levels. Protein expression level is presented by positive unit (PU).

RESULTSSmad 2/3 protein expression increased from day 3, reaching its peak level in day 14 [(42.2 +/- 2.4) PU], and decreased gradually. The elevation of Smad 4 protein level began from day 5, and the highest degree came into day 14 [(40.0 +/- 1.8) PU], decreased thereafter. The expression of Smad 7 presented a decreasing tendency at the beginning and reaching the lowest level in day 14 [(33.5 +/- 3.3) PU]. It seemed to elevate in day 28, but was still lower than the controls. There were positive correlation between Smad 2/3, Smad 4 and TGF-beta(1) (r = 0.91, r = 0.71, respectively, P < 0.05) and also between Smad 2/3 and hydroxyproline contents of lung tissue (r = 0.85, P < 0.05) except Smad 7.

CONCLUSIONSmad protein may have certain association with pulmonary fibrosis in silicosis.

Animals ; DNA-Binding Proteins ; immunology ; metabolism ; Lung ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Pulmonary Fibrosis ; chemically induced ; metabolism ; Quartz ; toxicity ; Smad2 Protein ; Smad3 Protein ; Smad4 Protein ; Smad7 Protein ; Trans-Activators ; immunology ; metabolism ; Transforming Growth Factor beta ; metabolism

Objective To investigate the efeects of microRNA(miR)-103a-3p regulates tumor protein 53-regulated inhibitor of apoptosis 1(TRIAP1)on osteoblast differentiation and bone mass in ovariectomized mice.Methods MC3T3-E1 cells were divided into normal group,miR-103a-3p-NC group,miR-103a-3p mimic group,miR-103a-3p mimic+TRIAP1-NC group,miR-103a-3p mimic+TRIAP1 mimic group.mRNA expression of miR-103a-3p,TRIAP1,P53 were detected by Real-time PCR;Cell proliferation and apoptosis were detected by MTT test and flow cytometry;cytoskeleton and mineralization of cells were detected by F-actin immunofluorescence staining and alizarin staining;alkaline phosphatase(ALP)activity was detected by ELISA.24 female mice were divided into sham group,osteoporosis(OP)group,miR-103a-3p antagonist-NC group,miR-103a-3p antagonist group(six in each group),extract bilateral ovaries to establish an OP model,sham group mice only isolated fat around ovarian tissue.mRNA expression of miR-103a-3p,TRIAP1,P53,ALP,osteocalcin(OCN),osteopontin(OPN)of bone tissue were detected;microCT detect bone mineral density(BMD),bone mineral content(BMC);haematoxylin eosin staining was used to observe pathological changes of bone tissue.Results After miR-103a-3p mimic was transfected into cells,the miR-103a-3p and P53 expression increased,TRIAP1 expression decreased,cell proliferation decreased,apoptosis increased,F-actin expression decreased,the number of calcium nodules decreased,and ALP enzyme activity decreased(P<0.01);however,after TRIAP1 mimic was additionally transfected into cells,the above result caused by miR-103a-3p mimics were significantly reversed(P<0.01).In OP group,the miR-103a-3p and P53 expression in bone tissue increased,the TRIAP1,ALP,OCN and OPN expression decreased,BMD and BMC were decreased,and bone tissue construct was damaged(P<0.05);in miR-103a-3p antagonist group,the miR-103a-3p and P53 expression in bone tissue decreased,TRIAP1,ALP,OCN,OPN expression increased,BMD and BMC increased,and bone tissue construct was improved(P<0.05).Conclusion MiRNA-103a-3p mediate TRIAP1/P53 to inhibit proliferation and mineralization of osteoblast,while miR-103a-3p antagonistic treatment reduce bone loss in OP mice.

OBJECTIVETo study the distribution of 5-FU in rat plasma and liver tissue following systemic or local 5-FU infusion.

METHODS5-FU was administered at the dose of 20 mg/kg systemically via bolus injection through the jugular vein or locally via infusion through the hepatic artery and portal vein of the rats. High-performance liquid chromatography was used to measure 5-FU concentration in the plasma and liver tissue, and the pharmacokinetic parameters, penetration rate and therapeutic dominance of 5-FU were calculated.

RESULTSSystemic administration of 5-FU resulted in the peak 5-FU concentration (Cmax) and area under curve (AUC) in the liver tissue of 13.79-/+4.56 microg/g and 342.20-/+108.20 microg.min(-1).g(-1)g-1, with the plasma Cmax and AUC of 36.85-/+5.96 microg/g and 842.00-/+158.00 microg.min(-1).ml(-1), respectively. Local 5-FU administration through the hepatic artery resulted in Cmax and AUC in the liver tissue of 29.58-/+4.30 microg/g and 794.60-/+115.40 microg.min(-1).g(-1) and Cmax and AUC in the plasma of 24.39-/+4.63 microg/g and 639.70-/+133.80 microg.min(-1).ml(-1), respectively. After administration through the portal vein, the Cmax and AUC of 5-FU was 28.21-/+4.46 microg/g and 733.60-/+180.3 microg.min(-1).g(-1) in the liver tissue, and 21.02-/+4.06 microg/ml and 529.80-/+111.50 microg.min(-1).ml(-1) in the plasma, respectively.

CONCLUSIONCompared with systemic venous bolus injection, administration through the hepatic artery and portal vein can significantly increase 5-FU concentration in the liver, and decrease its concentration in the peripheral blood.

Animals ; Area Under Curve ; Female ; Fluorouracil ; administration & dosage ; blood ; pharmacokinetics ; Hepatic Artery ; Immunosuppressive Agents ; administration & dosage ; blood ; pharmacokinetics ; Infusions, Intra-Arterial ; Infusions, Intravenous ; Liver ; metabolism ; pathology ; Liver Neoplasms, Experimental ; blood ; drug therapy ; Male ; Metabolic Clearance Rate ; Portal Vein ; Rats ; Rats, Wistar

A simple and quick method is described for the determination of ferulic acid, senkyunolide I, senkyunolide H, senkyunolide A and ligustilide in rhizomes of Ligusticum chuanxiong. The 5 active ingredients in the sample was extracted using 40% ethanol and analyzed by reversed-phase high performance liquid chromatography (HPLC). Chromatography separation was performed using Agilent 1100 series HPLC system with a Symmetry C18 column and gradient elution with a mixture of three solvents : solvent A, acetonitrile, solvent B, methanol and solvent C, 1% aqueous acetic acid, 0 min to 5 min A: B: C 20: 40: 40, 5 min to 30 min A: B: C 60 to 100 : 0 : 40 to 0. The effluent was monitored using a VWD detector set at 321 nm (0-4.3 min) and 275 nm (4.31-30 min). The flow rate was set at 1 mL x min(-1) and the injection volume was 10 microL. The column temperature was maintained at 35 degrees C. The calibration curve was linear (r > or = 0.99) over the tested ranges. The average recovery was 94.44%-103.1% (n = 6). The method has been successfully applied to the analysis in different harvest periods of L. chuanxiong samples. In this paper, single-factor randomized block design to study the 5 components content of L. chuanxiong on ten collecting stages. For the L. chuanxiong collected from April 15th to May 30rd, the content of 5 ingredients increased primarily, and then decreased. Determine the appropriate harvest time has important significance to the promotion of the quality of L. chuanxiong.
4-Butyrolactone ; analogs & derivatives ; analysis ; Acetic Acid ; chemistry ; Acetonitriles ; chemistry ; Benzofurans ; analysis ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; Ligusticum ; chemistry ; Methanol ; chemistry ; Solvents ; chemistry ; Time Factors

OBJECTIVETo investigate the gene expression of transforming growth factor-beta(1) (TGF-beta(1)) in lung tissues of silica-treated mice.

METHODSThe experimental mice were divided into control and silica group. 0.2 g/kg body weight of silica was injected intratracheally in silica group. Samples of lung tissue were collected 1, 3, 5, 7, 14 and 28 d after injection. RT-PCR method was used to analyze the gene expression of TGF-beta(1) in lung tissue of silica-treated mice.

RESULTSThe expression of TGF-beta(1) gene in lung tissue elevated from the 3rd day (1.20 +/- 0.15) and the peak value was on the 7th day (1.74 +/- 0.19). Then the expression decreased from the 14th to 28th day. But there was still higher than control until the 28th day.

CONCLUSIONTGF-beta(1) may play an important role in silica-induced pulmonary fibrosis.

Animals ; Gene Expression Regulation ; drug effects ; Lung ; drug effects ; metabolism ; pathology ; Male ; Mice ; Pulmonary Fibrosis ; etiology ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Silicon Dioxide ; pharmacology ; toxicity ; Time Factors ; Transforming Growth Factor beta ; genetics

OBJECTIVETo investigate the protein expression of transforming growth factor-beta(1) (TGF-beta(1)) in lung tissues of silica-treated mice.

METHODSThe experimental mice were divided into control and silica groups. 0.2 g/kg body weight of silica was injected intratracheally in mice of silica group. Samples of lung tissue were collected 1, 3, 5, 7, 14 and 28 d after injection. The immunohistochemical method was used to analyze the protein expression of TGF-beta(1).

RESULTSIn control mice, the expression of TGF-beta(1) in lung tissue was slightly positive while it was markedly increased in silica-treated mice. The expression was significantly elevated from the 7th day to 14th day. The expression in alveolar macrophages reached the peak on the 5th day [(93.4% +/- 2.8%) vs (42.2% +/- 12.0%), P < 0.01].

CONCLUSIONTGF-beta(1) may play an important role in early development of silicosis.

Animals ; Immunohistochemistry ; methods ; Lung ; chemistry ; drug effects ; pathology ; Male ; Mice ; Pulmonary Fibrosis ; etiology ; pathology ; Silicon Dioxide ; pharmacology ; toxicity ; Silicosis ; etiology ; pathology ; Time Factors ; Transforming Growth Factor beta ; analysis

OBJECTIVETo explore the correlation between the biological characteristics of volunteer donors and sperm parameters.

METHODSSperm parameters were analyzed for 778 primary volunteer donors by CASA system with standard methods recommended by WHO including sperm concentration, motility rate, VCL, VSL, ALH, LIN, STR and BCF, and their biological characteristics were recorded.

RESULTSOf all the volunteer donors, students accounted for more than a third, 89.2 percent were under the age of 35 years, 71.6 percent between 166 and 175 cm in height, 91.7 percent of normal weight and 77.1 percent with college education. The median sperm concentration and motility rate were 107.00 x 10(6)/ml and 75 percent respectively. Age was weakly correlated with such semen indexes as concentration (r = 0.210, P = 0.000) , motility rate (r = 0.213, P = 0.000), volume (r = 0.165, P = 0.002), VAP (r = 0.259, P = 0.000), VSL (r = 0.281, P = 0.000), VCL (r = 0.190, P = 0.000), BCF (r = 0.243, P = 0.000) and LIN (r = 0.192, P = 0.000). The semen indexes of the 26-35 age group were a little better than those of the 20-25. Height showed no correlation to semen indexes. Education and occupation were somewhat related with other sperm parameters (P < 0.01) than ALH (P = 0.695/0.886).

CONCLUSIONAge, occupation and education bear some correlation with sperm parameters except ALH, while height has none with any of them.

Adult ; Age Factors ; Humans ; Image Processing, Computer-Assisted ; methods ; Male ; Occupations ; Semen ; cytology ; Sperm Count ; Sperm Motility ; physiology ; Spermatozoa ; physiology ; Students ; Tissue Donors ; statistics & numerical data

AIMTo investigate the production of p-hydroxymethylphenol-beta-D-glucoside (gastrodin) through biotransformation by plant cell suspension cultures.

METHODSUsing cell suspension cultures of Datura stramonium to convert the exogenous p-hydroxybenzaldehyde into gastrodin was conducted and the converted compounds were separated with a combination of multi-chromatography. Their chemical structures were determined on the basis of spectral analysis and chemical evidence.

RESULTSThe conversion procedure of p-hydroxybenzaldehyde into gastrodin by Datura stramonium cell suspension cultures was established. The synthesized gastrodin (II) was isolated from the fermental liquor and identified by spectral analysis. At the same time, the p-hydroxybenzyl alcohol (I) converted through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of Datura stramonium was also isolated and identified. Two compounds were also isolated from the cell cultures and they were identified as beta-D-furanoallulose (III) and n-butyloxystyryl-beta-D-pyranoallulose (IV).

CONCLUSIONDatura stramonium grown in suspension cultures can convert exogenous p-hydroxybenzaldehyde into the corresponding gastrodin.

Benzaldehydes ; metabolism ; Benzyl Alcohols ; isolation & purification ; metabolism ; Biotransformation ; Cell Culture Techniques ; methods ; Cells, Cultured ; Datura stramonium ; cytology ; metabolism ; Glucosides ; biosynthesis ; isolation & purification ; Plant Stems ; cytology ; Plants, Medicinal ; cytology ; metabolism

The conversion of exogenous p-hydroxybenzaldehyde to p-hydroxy-methyl-phenol-beta-D-glucoside (gastrodin) was studied by using cell suspension culture of Datura tatula L. The chemical structure of this synthesized gastrodin was identified based on the spectral analysis and chemical evidence. The conversion procedure of p-hydroxybenzaldehyde into gastrodin by D. tatula L. cell suspension cultures was established. The synthesized gastrodin (II) was isolated from the ferment liquor and identified by spectral analysis. At the same time, the p-hydroxybenzyl alcohol (I) converted through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of D. tatula L. was also isolated and identified. The efficiency of glucosylation of p-hydroxybenzaldehyde was remarkably enhanced by adding salicylic acid (0.1 mg/L) and keeping the lower pressure (0.001MPa) in 25L airlift loop bioreactor. The biotransformation of exogenous p-hydroxybenzaldehyde to gastrodin by cell suspension culture of D. tatula L. is a promising approach.
Benzaldehydes ; metabolism ; Benzyl Alcohols ; chemistry ; Bioreactors ; Biotransformation ; Datura ; metabolism ; Glucosides ; biosynthesis ; chemistry ; Salicylic Acid ; pharmacology ; Suspensions

OBJECTIVETo study the BRCA1 mutations in patients with early-onset breast cancer and their affected relatives in Guangdong province and explore the relationship between BRCA1 mutation and the expressions of estrogen receptor(ER), progesterone receptor(PR), HER2 and ALN.

METHODSFrom 58 patients with early-onset breast cancer and their affected relatives, the genomic DNA was extracted from the peripheral blood mononuclear cells and the coding regions of the BRCA1 gene was amplified using polymerase chain reaction. BRCA1 gene mutations were screened by denaturing high performance liquid chromatography (DHPLC) and subsequent direct DNA sequencing. The expression of ER, PR, HER2 and ALN were detected with immunohistochemistry and their relations with the gene mutation were analyzed.

RESULTSDisease-related BRCA1 mutations were detected in 2 of the 58 patients, who were younger than 35 years old, including 1 with a novel splice-site mutation (IVS5-1 G-->A). No association was found between this novel mutation and the expressions of ER, PR, HER2 and ALN.

CONCLUSIONThe incidence of BRCA1 mutation is significantly lower in patients with early-onset breast cancer and their affected relatives in Guangdong province than in the Western populations. The novel mutation identified in BRCA1 gene may represent a mutation characteristic of the patients in Guangdong province. BRCA1 gene mutations may not have any relation with the expression of ER, PR, HER2 and ALN.

Adult ; Age of Onset ; Base Sequence ; Breast Neoplasms ; genetics ; China ; DNA Mutational Analysis ; Female ; Genes, BRCA1 ; Genotype ; Humans ; Molecular Sequence Data ; Mutation ; Receptor, ErbB-2 ; genetics ; Receptors, Estrogen ; genetics ; Receptors, Progesterone ; genetics