To observe the expressions of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in rats with pulmonary fibrosis (PF) induced by bleomycin, and to explore the mechanisms of Shenlong Decoction in preventing and treating PF.

Fosmidomycin (100 micromol x L(-1)) which is the effective inhibitor of DXR, key enzyme in terpenoid MEP pathway, was used to treat with hairy roots of Salvia miltiorrhiza. The treated roots were harvested at 2, 4, 6, 8, 10, 16 and 21 d, mRNA level of SmDXR and tanshinone content in treated and negative control groups were detected. Results found that, after treated with fosmidomycin, color of S. miltiorrhiza hairy roots grew pale gradually comparing with controls; mRNA level of SmDXR in hairy roots varied as a shape of parabolic and the highest value achieved at the sixth day after treatment, then it decreased gradually; Content of four kinds of tanshinones were detected. Among of the four kinds of tanshinones, Tanshinone I content changed relatively little, while content of dihydrotanshinone I, cryptotanshinone and tanshinone II (A) decreased gradually in 21 days. The content of total tanshinones in NC groups was 5, 63 times more than FOS-treated roots in the 21th day. The previous results showed that SmDXR played an important role in the accumulation of tanshinone content in MEP pathway. Once the mRNA level of SmDXR was suppressed, the accumulation of secondary metabolites will be significantly affected.
Aldose-Ketose Isomerases ; genetics ; Diterpenes, Abietane ; metabolism ; Fosfomycin ; analogs & derivatives ; pharmacology ; Gene Expression Regulation, Plant ; drug effects ; Plant Roots ; drug effects ; growth & development ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Salvia miltiorrhiza ; drug effects ; genetics ; growth & development ; metabolism ; Time Factors

Objective To investigate the applicability of clinical grade and a non-contact measurement method in evaluation of underneath eye wrinkles and to compare two methods.Methods A lot of 46 healthy Chinese women were recruited for this study.Underneath eye wrinkles severity was evaluated using clinical grade and a non-contact measurement method.The correlations were calculated for clinical grade and non-contact measurement parameters and age.The non-contact measurement parameters were classified by factor analysis.The correlations between age,clinical grade and factors were analyzed.Results The correlation coefficient between clinical grade in comparison to subject's age was 0.818.The parameters getting from non contact measurement were obviously correlated with age and clinical grade except SEr and SEsc;the correlation coefficients between parameters and age were-0.601 to 0.605;the correlation cofficients between parameters and clinical grade were-0.630 to 0.570.The non-contact measurement parameters could be classified into two factors;one represented wrinkle depth and roughness;the other represented wrinkle width and counts.These two factors were also obviously correlated with age and clinical grade.Conclusions Clinical grade and non contact measurement methods are both applicable in evaluation of underneath eye wrinkles.The parameters getting from two methods are obviously correlated with each other.

Lonicerae Japonicae Flos was one of the most widely used traditional Chinese medicine for its special biological activities. The content of luteoloside, one of its major compounds, was an important standard for the quantity control of Lonicerae Japonicae Flos. The major method used for the detection of luteoloside was instrumental analysis. Compared with the ELISA method, instrumental analysis was time-consuming, complex pretreatment and low-throughout. Thus, it was significantly important to develop an enzyme-linked immunosorbent assay (ELISA) for luteoloside analysis. Here, the conjugates of luteoloside-bovine (LG-BSA) and luteoloside-ovalbumin (LG-OVA) were produced as the immunogen and coating antigen by the carbodiimide ( CDI) method, respectively. The conjugation ratio of carrier protein and the hapten in the conjugate were determined by UV-Vis spectrophotometry (UV). LG-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The titer and specificity of antiserum were detected by ELISA. The conjugation ratio of hapten and carries protein were 3. 7: 1 (LG-BSA) and 1. 0: 1 (LG-OVA). The antiserum titer was higher than 2 000 with the linear range of 18.4-4 852.4 μg x L(-1), R2 = 0.988 4 and IC50 = 298.7 μg x L(-1). The result showed that the conjugate antigen LG-BSA was synthesized successfully and the mice can produce specific antiserum injected with artificial antigen.
Animals ; Antibodies ; analysis ; immunology ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Immunization ; Lonicera ; chemistry ; immunology ; Mice ; Mice, Inbred BALB C

Objective To assess the clinical values of MR angiography(MRA)in the detection of deep vein thrombosis of the lower limbs.Methods Two-dimensional time of flight(2D TOF)MRA was performed in thirty patients who were suspected of having deep vein thrombosis in the lower limbs.The findings of MRA were compared to that of digital subtraction angiography(DSA).Results twenty-five cases showed deep vein thrombosis in the lower limbs,the MRA findings included venous filling defect (14 cases),occlusions and interruptions of veins(8 cases),venous recanalizations(3 cases),collateral veins(25 cases).Taking the results of DSA as a golden standard,MRA detected all of the affected cases with only one case as the false positive.Conclusion 2D TOF MRA is a method of choice in the diagnosis of deep vein thrombosis of the lower limbs.

OBJECTIVETo investigate association of mutation in WNK4 gene with essential hypertension and to analyze the expression of WNK4 gene.

METHODScSNP in WNK4 gene in a small samples was detected by sequencing, then PCR-RFLP was performed in 98 patients with essential hypertension and 95 control subjects. The expression profile of WNK4 gene was tested by RT-PCR.

RESULTSA cSNP was detected in WNK4 gene exon7 G1662A, and there were significant differences in the distribution of allele frequency of G1662A between essential hypertension group and control group. WNK4 gene were expressed in the tissues of kidney, brain, lung, heart, spleen and intestine of fetus.

CONCLUSIONWNK4 gene is well correlated with essential hypertension.

Adult ; Brain ; metabolism ; Female ; Gene Expression ; Gene Frequency ; Genes ; Genetic Predisposition to Disease ; Humans ; Hypertension ; genetics ; Kidney ; metabolism ; Lysine ; genetics ; Male ; Middle Aged ; Mutation ; Phenotype ; Protein-Serine-Threonine Kinases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics

AIMTo explore sodium selenite-induced oxidative stress and apoptosis in human promyelocytic leukemia NB4 cells.

METHODSThe growth inhibition of NB4 cells was measured by MTT test. Apoptosis was determined morphologically by Giemsa stain and by DNA ladder formation in electrophoresis. Quantitation of apoptosis was determined by percentage of PI stained cells containing subdiploid amount of DNA measured by flow cytometry. Generation of reactive oxidative species (ROS) in NB4 cells was determined by lucigenin dependent chemoluminescent (CL) test. Spectrophotometer was used to measure the level of reduced glutathione, superoxide dismutase (SOD) and glutathione peroxidase in the cell.

RESULTSSodium selenite was shown to inhibit the growth of NB4 cells. Sodium selenite induced apoptosis with dose and time dependency: the ratio of subdiploid cells in control group was 1.3% +/- 0.7%. The 5 mumol.L-1 group was 10.4% +/- 1.4%, 10 mumol.L-1 group was 16% +/- 1%, and the 20 mumol.L-1 group was 27.3% +/- 0.8%. Sodium selenite (> or = 5 mumol.L-1) enhanced the ROS level markedly in NB4 cells (in 20 mumol.L-1 group ROS level was increased by 17 times, compared with control group), accompanied with decrease of reduced intracellular glutathione. These effects were time and dose dependent. N-acytlcysteine as an antioxidant was found to inhibit sodium selenite-induced oxidative stress and apoptosis in NB4 cells.

CONCLUSIONSodium selenite can induce apoptosis of NB4 cells and would possibly be used as an agent for the treatment of malignancy. The main mechanism of action might be related to oxidative stress induced by sodium selenite, thereby, leading to apoptosis as shown in NB4 cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; Cell Division ; drug effects ; Humans ; Leukemia, Promyelocytic, Acute ; pathology ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Sodium Selenite ; pharmacology ; Tumor Cells, Cultured

OBJECTIVETo explore in vitro expansion of CD34+CD59+ cells from patients with PNH, and compare the capabilities of survival, proliferation and expansion between CD34+CD59+ cells from patients with PNH and CD34+ cells from normal control.

METHODSCD34+CD59+ cells from patients with PNH and CD34+ cells from normal control were selected from the bone marrow mononuclear cells by means of two-step sorting method with immunomagnetic microbead-flow cytometry, then underwent in vitro expansion for two weeks and semi-solid culture in vitro before and after expansion.

RESULTS(1) CD34+CD59+ cells from patients with PNH can be expanded effectively in vitro, and the biggest expansion of CD34+CD59+ cells was about 23.49 fold on the 7th day. (2) There were some similar characteristics between CD34+CD59+ cells from patients with PNH and CD34+ cells from normal control, such as: the best combination of hematopoietic factors for in vitro expansion was SCF+ IL-3 + IL-6 + FL + Tpo + Epo, and the seventh day was the most suitable in course of 4-14 days for in vitro expansion, and after in vitro expansion, the cells remained CD59 positive and strong capability of performing colony-forming. (3) CD34+ cells from normal control had better proliferation, expansion and stronger potential to survive than CD34+CD59+ cells from patients with PNH.

CONCLUSIONS(1) In vitro expansion of CD34+CD59+ cells from patients with PNH can be performed. The present study showed the possibility of performing ABMT or APBSCT clinically for patients with PNH. (2) There were some similar characteristics between CD34+CD59+ cells from patients with PNH and CD34+ cells from normal control, but the latter had better proliferation, expansion and stronger potential to survive than the former. CD34+CD59+ cells from patients with PNH were not completely normal cells.

Antigens, CD34 ; analysis ; Bone Marrow Cells ; cytology ; immunology ; CD59 Antigens ; analysis ; Cell Differentiation ; Cell Division ; Cell Survival ; Cells, Cultured ; Hemoglobinuria, Paroxysmal ; immunology ; pathology ; Humans ; Immunophenotyping

OBJECTIVEFor heart functional parameters, we commonly used normal range. The reference values and predict formulas of heart functional parameters and their relationships with individual characteristics are still lack.

METHODSLeft ventricular (LV) volumes (end-diastolic volume and end-systolic volume), stroke volume (SV), ejection fraction (EF) and cardiac output (CO) were measured by cardiac CT angiography (CAT) in 1 200 healthy Caucasian volunteers, men 807 and women 393, and age 20-90yr. The results are analyzed by high-accuracy three-dimensional imaging technology, and then measured the dynamic changes of the volumes of each atriam and ventricule during their contractions and relaxations. The gender, age, height and weight were analyzed by multiple linear regression to predict LV functional parameters.

RESULTSExcept the LVEF was lower in man than in women (P < 0.001), all other LV functional parameters of EDV, ESV, SV, FE and CO were higher in man (P < 0.001). Multiple linear regression indicated that age, gender, height and weight are all independent factors of EDV, ESV and SV (P < 0.001). CO could be significantly predicted by age, gender and weight (P < 0.001), but not height (P > 0.05). The predict equation for CO (L x min(-1)) = 6.963+0.446 (Male) -0.037 x age (yr) +0.013 x weight (kg).

CONCLUSIONAge, gender, height and weight are predictors of heart functions. The reference values and predict equations are important for noninvasive and accurate evaluation of cardiovascular disease and individualized treatment.

Adult ; Age Factors ; Aged ; Aged, 80 and over ; Body Height ; Body Weight ; Cardiac Output ; Female ; Heart ; physiology ; Humans ; Male ; Middle Aged ; Reference Values ; Sex Factors ; Stroke Volume ; Ventricular Function, Left ; Young Adult

An expression system is described for high-yield production of recombinant soluble human FasL (shFasL) in Dictyostelium discoideum cells. DNA encoding amino acids 141 - 281 of hFasL was PCR amplified from cDNA derived from activated human neutrophils. The resulting product was fused with a DNA fragment encoding hCG-beta signal peptide and cloned in the expression vector pMB12neo. Dictyostelium strain AX3 was transfected with this plasmid, yielding a recombinant strain called AX3-pCESFL95-H3. In order to improve the shFasL expression level, pMB12neo was optimized by replacing its transcriptional terminator/ polyadenylation segment of the 2H3 gene with an actin8 terminator/polyadenylation segment, yielding derived expression vector pMB74. The recombinant Dictyostelium strain called AX3-pLu8 was generated with this new plasmid. When the recombinant cells were cultivated in a complex HL-5C medium, a cell density of (1.5 - 2) x 10(7)/mL was reached, and the shFasL level expressed by strains AX3-pCESFL95-H3 and AX3-pLu8 was 23.5 microg/L and 206 microg/L, respectively. By using a newly developed synthetic medium called SIH as culture medium, higher cell density of (4 - 5) x 10(7)/mL was achieved. Correspondently, 111 microg/L and 420 microg/L shFasL were secreted by recombinant strains AX3-pCESFL95-H3 and AX3-pLu8, respectively.
Animals ; Chorionic Gonadotropin, beta Subunit, Human ; genetics ; Culture Media ; Dictyostelium ; genetics ; growth & development ; metabolism ; Fas Ligand Protein ; biosynthesis ; genetics ; Humans ; Neutrophils ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics