Adult ; Aged ; Female ; Humans ; Lung Neoplasms ; blood ; genetics ; Male ; Middle Aged ; Mutation ; Tumor Suppressor Protein p53 ; blood ; genetics

Objective:To explore the relationship between the severity of cardiovascular disease with the expression of apolipoprotein(a) [apo(a)] and apolipoprotein B(apoB) in peripheral blood and their location in peripheral blood cells.Methods: In this report,we selected 4 patients with angiography which indicated that three coronary arteries were narrowed and 5 control patients with normal angiography.Arterial blood was collected and analyzed for lipid parameters in plasma.The mRNA expression of apo(a) and apoB in peripheral white blood cells and platelets were determined by RT-PCR and their protein expression by western blot.Moreover,the expression and location of apo(a) and apoB in white blood cells were determined by confocal microscopy and computer 3D analysis.Results: In plasma,levels of high density lipo-protein-cholesterol(HDL-C) and apolipoprotein A-Ⅰ(apoA-Ⅰ) in cardiovascular disease(CVD) patients were significantly less than those in the control patients[(0.62?0.05) mmol/L,(0.78?0.08) mmol/L vs(0.81?0.15) mmol/L,(0.9?0.07) mmol/L,P0.05).Studies with confocal microscopy indicated that proteins of apo(a) and apoB were co-expressed by a few cells of leukocytes and the ratio of apoB/apo(a) in cardiovascular disease patients was significantly less than that in the control patients(optical density value 1.60?0.12 vs 4.40?0.35,P

Cigarette smoke extract (CSE) contains abundant oxidants and free radicals. Oxidative stress caused by cigarette smoking results in the destruction of the alveolar cell walls and emphysema. However, there exists discrepancy about how CSE works in the process. In the present study, we observed the effect of CSE on the cell growth of type II alveolar epithelial cell-derived A549 cell line, and provided molecular understanding of this effect. The MTT assay results showed that CSE decreased the cell viability of A549 cells in a dose- and time-dependent manner, and cell cycle was arrested in G(1)/S phase. Furthermore, CSE-induced apoptosis of A549 cells was verified by Hoechst 33258 staining, electron microscopy in morphology, and the appearance of DNA fragmentation and annexin V-FITC/propidium iodide (PI) staining assay at molecular level. It was found that CSE treatment resulted in the upregulation of Fas/APO-1 receptor and activation of caspase-3. CSE also initiated accumulation of intracellular reactive oxygen species, which was detected by laser confocal microscopy. Taken together, CSE could inhibit the cell growth and induce apoptosis of A549 cells through Fas receptor pathway. Oxidative stress caused by CSE may be the radical factor leading to apoptosis as well as cell growth inhibition in alveolar epithelial cells.
Apoptosis ; drug effects ; Cell Cycle Checkpoints ; drug effects ; Cell Line, Tumor ; Epithelial Cells ; drug effects ; Humans ; Lung Neoplasms ; pathology ; Pulmonary Alveoli ; cytology ; drug effects ; pathology ; Smoke ; adverse effects ; Tobacco ; toxicity

Animals ; Cell Hypoxia ; Cells, Cultured ; Chemokine CXCL12 ; genetics ; metabolism ; Endothelial Cells ; cytology ; metabolism ; Female ; Gene Expression Regulation ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Immunohistochemistry ; Male ; Pulmonary Artery ; cytology ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Swine

OBJECTIVETo investigate the expression of fatty acid synthase (FAS) in adenosis, atypical ductal epithelial hyperplasia, ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) of breast, and the correlation of FAS expression with HER2 gene amplification in IDC.

METHODSImmunohistochemical EnVision method staining for FAS was performed in 100 cases of breast lesions and 10 normal breast tissues. HER2 gene amplification was detected with FISH in 60 cases of IDC.

RESULTSThe cohort included 10 cases of adenosis, 10 atypical ductal epithelial hyperplasia, 20 DCIS (8 high-grade, 9 intermediated-grade and 3 low-grade), and 60 cases of IDC (5 grade 1, 40 grade 2 and 15 grade 3). FAS expression was negative in all 10 normal breast tissues; in the 10 cases of adenosis, strongly positive FAS expression was detected in one case, positive in 2, weakly positive in 4, and negative in 3; in the 10 cases of atypical ductal epithelial hyperplasia, FAS immunohistochemistry showed that 1 was strongly positive, 4 positive, 4 weakly positive, and 1 negative; in the 20 cases of DCIS, FAS immunostaining showed that 12 were strongly positive, 5 positive, 1 weakly positive, and 2 negative; FAS expression showed a clear increasing trend from normal breast tissue, atypical ductal epithelial hyperplasia to DCIS (χ(2) = 42.02, P < 0.01). Likewise, the increasing trend was also demonstrated from adenosis to DCIS (χ(2) = 34.69, P < 0.01). There was also a positive correlation between FAS expression and extent of lesion among normal breast tissue, adenosis, atypical ductal epithelial hyperplasia and DCIS (χ(2) = 86.02, P < 0.01; r = 0.568, P < 0.01). FAS expression was not correlated with the grade of DCIS (χ(2) = 9.12, P = 0.16). In the five cases of grade 1 IDC, FAS immunostaining showed that 4 cases were strongly positive and 1 positive; in the 40 cases of grade 2 IDC, FAS immunostaining showed that 27 strongly positive, 12 positive, and 1 negative; in the 15 cases of grade 3 IDC, FAS immunostaining showed that 6 were strongly positive, 5 positive, 3 weakly positive, and 1 negative; FAS expression was stronger and more extensive in DCIS, IDC grades 1 and 2 than that in other groups. However, FAS expression was weaker in the IDC grade 3 (χ(2) = 11.26, P = 0.01). The positive expression rate of FAS in IDC was generally higher than that in benign breast lesions (χ(2) = 47.19, P < 0.01). In the 60 cases of IDC, FISH showed HER2 gene amplification in 22 cases, but not in the remaining 38 cases. FAS expression in IDC was highly correlated with HER2 gene amplification (r = 0.44, P < 0.01). The expression of FAS had significant correlation with status of ER and PR and tumor size (P < 0.05). There was no significant correlation with age, immunohistochemical HER2 expression, lymph node metastasis and clinical stage (P > 0.05).

CONCLUSIONSFAS may be closely related to the carcinogenesis of breast IDC. FAS expression is closely associated with HER2 gene amplification in IDC.

Breast ; metabolism ; pathology ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; genetics ; metabolism ; pathology ; Fatty Acid Synthases ; metabolism ; Female ; Fibrocystic Breast Disease ; metabolism ; Gene Amplification ; Genes, erbB-2 ; Humans ; Hyperplasia ; Lymphatic Metastasis ; Middle Aged ; Receptor, ErbB-2 ; metabolism

OBJECTIVETo explore the effect of hypoxia and hypoxia-inducible factor-1alpha (HIF-1alpha) on the expression of multidrug resistance gene-1 (mdr-1) and its coded p-glyeoprotein (P-gp) as well as the chemotherapeutic sensitivity of human ovarian cancer cells to paclitaxel and its mechanism.

METHODSThe mRNA and protein levels of HIF-1alpha, mdr-1 and p-gp were studied by immunocytochemistry, semiquantitative reverse transcription-ploymerase chain reaction (RT-PCR) and Western blot analysis in human ovarian cancer cells (A2780) in 5% CO2 + 1% O2 hypoxic culture and 21% O2 normoxic culture, respectively. Methyl thiazolyl tetrazolium (MIT) was used to evaluate the chemotherapeutic sensitivity of A2780 cells to paclitaxel by inhibition rate. RNA interference technique was used and small hairpin RNAs (shRNAs) eukaryotic expression vector targeting HIF-1alpha was constructed as pSiHIF-1alpha, and transfected into A2780 cells. RT-PCR and Western blot were used to detect gene silencing effect on HIF-1alpha, the expressions of mdr-1 and p-gp. The inhibition rate was observed after HIF-1alpha gene silence.

RESULTSHIF-1alpha at both mRNA and protein levels was induced significantly under hypoxia. The HIF-1alpha expression at mRNA level was oxygen gradient-independent, while HIF-1alpha expression at protein level was oxygen gradient-dependent. The inhibition rate of paclitaxel to hypoxic A2780 cells in 5% CO2 + 1% O2 was significantly lower than that in normoxic A2780 cells (P <0.05). The shRNAs plasmid targeting HIF-1alpha was constructed successfully and HIF-1alpha gene was silenced in A2780 cells efficiently followed by mdr-1 and p-gp down-regulation. The inhibition rate was greatly increased in hypoxic A2780/siHIF-1alpha cells.

CONCLUSIONHypoxia can decrease the chemotherapeutic sensitivity of human ovarian cancer A2780 cells to paclitaxel through HIF-1alpha regulating the expression of mdr-1 and p-gp.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; Antineoplastic Agents, Phytogenic ; pharmacology ; Blotting, Western ; Cell Hypoxia ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Down-Regulation ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; biosynthesis ; genetics ; Ovarian Neoplasms ; genetics ; metabolism ; pathology ; Paclitaxel ; pharmacology ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Adolescent ; Adult ; Child ; Child, Preschool ; Exons ; Factor VIII ; genetics ; Hemophilia A ; genetics ; Humans ; Infant ; Male ; Middle Aged ; Mutation

OBJECTIVETo study the effects of irrigation volume on the growth and quality of Lonicera japonica.

METHODDifferent volume of irrigation water was applied to the potted L. japonica before the first florescence of honeysuckle in order to keep the relative moisture content of the soil as 100%, 80%, 50%, 30%, and observe the growth and development of shoot and leaves, yield and quality of honeysuckle.

RESULTAs the of irrigation volume was reduced, the internodal elongation of shoot and the increase of the knot were restrained, and the yield of honeysuckle dropped. But the content of chlorogen acid in the buds treated by 80% irrigation volume was not affected, while that of the other treatment all decreased. The moisture contents of the leaves treated by 30% irrigation volume was significantly lower than that of the CK, while the specific leaf weight (SLW) increased significantly comparing to the CK. The content of chlorophyl in the leaves of 50% and 30% increased, while the ratio of chlorophyl A and B fell significantly. The content of dissoluble suger in the leaves of 80% and 50% irrigation volume was much higher than that in the CK. The results also showed that, the content of dissoluble protein in the leaves of 30% irrigation volume increased comparing with that in 50%, and the content of proline in leaveas of 30% irrigation volume increased significantly.

CONCLUSIONUsing less irrigation volume (keeping 80% of relative moisture content of the soil) does not affect the quality of honeysuckle, but decreased dry weight of honeysuckle. It is necessary to take irrigation management during cultivation of honeysuckle.

Biomass ; Chlorogenic Acid ; metabolism ; Flowers ; drug effects ; growth & development ; metabolism ; Lonicera ; drug effects ; growth & development ; metabolism ; Plant Leaves ; drug effects ; growth & development ; metabolism ; Plant Proteins ; metabolism ; Plant Stems ; drug effects ; growth & development ; metabolism ; Plants, Medicinal ; drug effects ; growth & development ; metabolism ; Proline ; metabolism ; Soil ; Water ; metabolism ; pharmacology

Objective To explore the dosage and injection method of concanavalin A(Con A) for inducing Wistar rats into the acute hepatic injury model. Methods (1)According to the dosage of Con A, 42 Wistar rats were randomly divided into groups A, B, C, D, E, N, 7 rats in each group. Group N was given tail intravenous injection of normal saline as normal control group. Groups A, B, C, D, E were given intravenous injection of 4, 8, 16, 30, 40 mg/kg of Con A respectively. At the 8th hour after modeling, the levels of alanine transaminase(ALT), aspartate aminotransferase(AST), albumin(ALB), interleukin(IL)-2 , IL-10, interferon (IFN)-γ, and tumor necrosis factor(TNF)-αwere detected. And HE staining was used to observe the pathological feature of hepatic tissue. (2)According to the injection method of Con A, 21 Wistar rats were randomly divided into normal control group, intraperitoneal injection group and tail intravenous injection group, 7 rats in each group. The dosage of Con A for the rats in intraperitoneal injection group and tail intravenous injection group was 16 mg/kg. At the 8th hour after modeling, the levels of serum ALT, AST, and ALB were determined. Results The number of abnormal deaths in various dose Con A groups at the end of each experiment was 0 in groups A, B, C, and 2 in group D, and 7 in group E. A small amount of spotty necrosis, inflammatory cell infiltration, and hepatic lobule with almost integrity of structure were found in groups A, B, while obvious bridging-like necrosis was seen in groups C, D. Serum ALT, AST, and ALB levels in intraperitoneal injection group had no statistically significant difference as compared with the normal control group. Conclusion Tail intravenous injection of 16 mg/kg of Con A can be used to induce an acute immunological liver injury rat model successfully.

In order to detect coagulation factor VIII (FVIII) inhibitor in patients with severe hemophilia A (HA) and preliminarily study the genetic mutation in patients with inhibitor positive. Totally 58 patients with HA (FVIII: C < 1%) were enrolled. FVIII: C activity was measured by one-stage coagulation assay. FVIII inhibitor was screened by using APTT method and FVIII inhibitor in screened positive patients with HA was quantitatively analyzed by using Bethesda method. Using genomic DNA as template, 12, 14, 16 exons of FVIII in screened positive patients were amplified, and the mutations of amplified products were detected by direct sequencing. The results indicated that the FVIII inhibitor could be detected in 4 patients (6.9%) from 58 HA patients, no gene mutations in 12, 14, 16 exons of FVIII were found. It is concluded that the positive rate of FVIII inhibitor in HA patients is lower than that reported in literature. The causes of inhibitor production needs to further investigate.
Adolescent ; Adult ; Blood Coagulation Factor Inhibitors ; isolation & purification ; Blood Coagulation Tests ; Child ; Child, Preschool ; Exons ; Factor VIII ; antagonists & inhibitors ; genetics ; Genetic Testing ; Hemophilia A ; diagnosis ; genetics ; Humans ; Infant ; Middle Aged ; Mutation ; Young Adult