Objective To observe the effects of ursolic acid (UA) on the activation of nicotinamide adenine dinucleotide phosphate oxidase (NOX) and the downstream signaling pathways in platelet derived growth factor (PDGF) activated rat hepatic stellate cell (HSC-T6).Methods Rat HSC-T6 cells were divided into blank control group (no treatment),UA control group (50 μmol/L UA),PDGF group (10 μg/L PDGF),UA intervention group (50 μmol/L UA + 10 μg/L PDGF),diphenyleneiodonium intervention(DPI) group (20 μmol/L DPI+ 10 μg/L PDGF),SB203580 (p38 mitogen-activated protein kirase(p38MAPK) inhibitor) intervention group (10 μmol/L SB203580 + 10 μg/LPDGF),LY294002 (phosphatidylinositop 3 kinase(PI3K) inhibitor) intervention group (10 μmol/L LY294002 + 10 μg/L PDGF) and rosup positive control group (5 μg/mL rosup).Except rosup positive control group,the expression of type Ⅰ collagen at mRNA level of each group was detected by fluorescence quantitavepolymerase chain reaction (RT-PCR).The expression of membrane protein p47phox (except rosup positive control group),PI3K(except rosup positive control group and SB203580 intervention group),p-protein kinase B (p-AKT,except rosup positive control group and SB203580 intervention group) and phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK,except rosup positive control group and LY294002 intervention group) were tested by Western blot.Except SB203580 intervention group and LY294002 intervention group,the fluorescence intensity in the cells of each group was analyzed with active oxygen detection kit and fluorescence microplate reader.Single factor analysis of variance and LSD test were performed for comparison between groups.Results Type Ⅰ collagen at the mRNA level of PDGF group (3.74±0.32) was higher than that of blank control group (1.00±0.00) ; Type Ⅰ collagen at the mRNA level of UA group (0.21 ±0.02) was lower than that of blank control group,UA intervention group (1.02 ± 0.12),DPI intervention group (1.09±0.21),SB203580 intervention group (1.18± 0.27),and LY294002 intervention group (1.15 ± 0.26) were all lower than PDGF group,and the differences were statistically significant (t =15.667,-4.501,-15.553,-15.154,-14.642 and -14.813,all P＜0.05).p47phox at the protein expression level of PDGF group (1.98±0.53) was higher than that of blank control group (1.00±0.00) ; that of UA group (0.48±0.10) was lower than blank control group; those of UA intervention group (0.95 ± 0.26),DPI intervention group (0.99 ± 0.28),SB203580 intervention group (0.93±0.31),and LY294002 intervention group (1.07±0.19) were all lower than PDGF group (t=4.209,-2.234,4.424,-4.252,-4.510 and-3.909,all P＜0.05).The protein expression level of PI3K of PDGF group (2.27±0.46) was higher than that of blank control group (1.00±0.00); that of UA intervention group (0.14 ± 0.07) was lower than PDGF group and blank control group; that of UA group (0.14±0.07) was lower than blank control group; those of DPI intervention group (0.53±0.25) and LY294002 intervention group (0.35±0.14) were all lower than PDGFgroup (t 6.205,8.208,-2.003,4.202,-8.502 and-9.831,all P＜0.05).The protein expression level of p-Akt of PDGF group (2.54±0.49) was higher than that of blank control group (1.00± 0.00); those of UA intervention group (0.74± 0.20),DPI intervention group (0.94 ± 0.37) and LY294002 intervention group (1.17±0.41) were all lower than PDGF group; that of UA group (0.59± 0.15) was lower than blank control group (t=5.927,-6.928,-6.158,-5.273 and-1.578,all P＜ 0.05).The protein expression level of p-p38MAPK of PDGF group (1.98±0.35) was higher than that of blank control group (1.00±0.00); those of UA intervention group (0.68±0.28),DPI intervention group (0.63±0.27) and SB203580 intervention group (0.67 ± 0.29) was all lower than PDGF group; that of UAgroup (0.28±0.13) was lower than blank control group (t=4.897,-6.479,-6.727,-6.529 and-3.561,all P＜0.05).The level of active oxygen of PDGF group (105.57±7.51) was higher than that of blank control group (69.60±8.63) ; those of UA intervention group (64.56±9.11),DPI intervention group (65.75 ± 6.62) was lower than PDGF group,UA group (29.84 ±3.19) was lower than blank control group (t=6.368,-7.288,-7.071 and-7.255,all P＜0.05).Conclusion UA could inhibit membrane displacement of NOX subunit p47phox and reduce active oxygen production in PDGF induced rat HSC-T6 cells,and then block phosphorylation of PI3K Akt,p 38MAPK signal pathways and inhibited the expression of type Ⅰ collagen at mRNA level.

The murine bone marrow endothelial cell line (mBMEC) has been maintained by means of subculture and cryopreservation for over 10 years since it was established in our laboratory. This study was aimed to newly identify biological characteristics of this cell line for further study. The cultured mBMEC cells were observed by inverted microscopy and transmission electron microscopy (TEM). PECAM-1 (CD31) and von Willebrand factor (vWF) were detected by immunofluorescent staining. The phagocytotic activity of the cells in culture was tested by using fluorescent acetylated low-density lipoprotein (Dil-Ac-LDL). The cell growth kinetics analysis and karyotype analysis were performed. The results showed that the adherent cells were mostly elliptical, rounded and spindle-shaped, and some of them connected to each other to form cord- and network-like arrangements in mBMEC cultures at subconfluence. The adherent cells grew up to confluence as a cobblestone-like monolayer. Several ultrastructural features of the endothelial cells could be observed in TEM sections of the cultured cells. More than 94% of mBMEC cells were positive for either CD31 or vWF. The phagocytotic ingestion of Dil-Ac-LDL occurred in 98.5% of cells. In normal culture conditions, the cells grew with a mean population doubling time of 54.6 hours and the maximal mitotic index was 38 per thousand in the rapid growth period. The colony yields were 4.33% to 7.40% depending on the plating density of cells. Karyotypes of all the cells were aneuploidy with a greater percentage of hyperdiploid. It is concluded that mBMEC cells retain the fundamental properties of endothelial cells, but the growth kinetics and biological behaviors are slightly different from those in the early days after the establishment of this cell line.
Animals ; Bone Marrow Cells ; cytology ; Cell Line ; Endothelial Cells ; cytology ; physiology ; Karyotyping ; Mice ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; von Willebrand Factor ; metabolism

Carcinoma, Hepatocellular ; genetics ; metabolism ; Cell Proliferation ; drug effects ; Humans ; Liver Neoplasms ; genetics ; metabolism ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Tumor Cells, Cultured ; beta Catenin ; biosynthesis ; genetics

OBJECTIVETo explore the effects of rearing patterns on diet and temperament traits among infants in urban areas.

METHODSA total of 480 25-30-month-old infants were randomly selected from the birth cohort in Hefei Maternal and Child Health Care Center in 2008. A household survey was conducted using China Toddler Temperament Scale (CTTS), Dietary Characteristics Questionnaire and Family Environment Questionnaire.

RESULTSOf the 430 surveyed households, there were three main rearing patterns including parents rearing pattern (Group A), grandparents rearing pattern (Group B) and joint rearing pattern (Group C), accounting for 33.0%, 21.2% and 45.8%, respectively. Infants in Group A tended to adopt processed food pattern, with poor rhythmicity and adaptability; infants in Group B tended to adopt fruit, vegetable, and cereals-based food pattern, with relatively poor rhythmicity; infants in Group C tended to adopt aquatic products and fruit/vegetable-based food pattern, with good rhythmicity and adaptability. Linear regression model showed that infants who consumed more aquatic products, high-protein food, and fruits/vegetables had more positive temperamental traits, whereas infants who consumed more processed foods had more negative temperamental traits.

CONCLUSIONSA joint rearing pattern may be a favorable rearing style for infants aged 25-30 months in urban areas in terms of diet and temperament traits.

Child Rearing ; Child, Preschool ; Diet ; Humans ; Infant ; Infant Nutritional Physiological Phenomena ; Temperament

OBJECTIVETo evaluate the impacts of maternal weight gain during pregnancy on offspring weight and obesity from birth to 24 months of age.

METHODSThe information on maternal pre-pregnancy body mass index (BMI), gestational weight gain and demographic characteristics were collected from 317 pregnant women. The information on offspring weight, BMI and breastfeeding data was obtained from various follow-up examinations from 0 to 24 months of age.

RESULTSThe logistic regression analysis showed that excessive gestational weight gain resulted in an increased risk of obesity in children at age of 6 months (adjusted RR=3.56, 95% CI:1.31-8.35) and 9 months (adjusted RR=2.87, 95% CI: 1.04-3.28) after adjustment for potential confounding factors. The linear regression model showed that there were significant correlations between gestational weight gain and Z score of weight in offsprings at birth (β=0.032, 95% CI: 0.008-0.057), 3 months (β=0.037, 95% CI: 0.013-0.062), 6 months (β=0.043, 95% CI: 0.017-0.068), 9 months (β=0.038, 95% CI: 0.013-0.063) and 12 months (β=0.034, 95% CI: 0.009-0.059), but not at 18 months and 24 months.

CONCLUSIONSExcessive gestational weight gain may affect offspring weight and increase the risk of obesity in children from birth to 12 months of age. During their second year of life, this effect will temporarily disappear.

Body Mass Index ; Body Weight ; Female ; Follow-Up Studies ; Humans ; Linear Models ; Logistic Models ; Obesity ; etiology ; Pregnancy ; Weight Gain

OBJECTIVECurrent study was conducted to investigate and compare the impact of temperature and pH on the activities of amylase, protease and lipase in alimentary tract of Whitmania pigra.

METHODThe responses of amylase, protease, and lipase activities were determined over a wide range of temperatures (7-52 degrees C) and pH gradient (2.2-11.2).

RESULTThe highest lipase activity was found under 37 degrees C, pH 8.2, and the highest amylase activity was detected under 37 degrees C, pH 5.2, while protease activity peaked at 42 degrees C, pH 3.2 or pH 9.2.

CONCLUSIONThe optimal temperature in alimentary tract of Wh. pigra for lipase and amylase was 37 degrees C, and the responding temperature for protease was 42 degrees C. The optimal pH value in alimentary tract of Wh. pigra for lipase and amylase was pH 8.2 and pH 5.2, respectively. While pH 3.2 or 9.2 seems to be both favorable for high protease activity.

Amylases ; chemistry ; metabolism ; Animals ; Digestive System ; chemistry ; enzymology ; Enzyme Stability ; Hydrogen-Ion Concentration ; Leeches ; chemistry ; enzymology ; Lipase ; chemistry ; metabolism ; Peptide Hydrolases ; chemistry ; metabolism ; Temperature

Objective To explore the transmission chain of COVID-19 by serum antibody detection, and to provide scientific evidence for the prevention and control of the epidemic. Methods Field epidemiological investigation was used to determine the COVID-19 cases and their close contacts. The 2019-nCoV nucleic acid in throat swabs and anal swabs were examined by RT-PCR. Serum specimens were collected for anti-2019-nCoV IgM antibody detection and combined IgM/IgG detection. Results Case A had no confirmed exposure to COVID-19. However, case C and D had dinner and lived together with case A; they reported contact history and dinner history with other confirmed COVID-19 cases(H, L). Case A tested positive for 2019-nCoV nucleic acid, whereas case C and D were negative. Moreover, case A and C were IgM antibody positive, while case D was negative. Case A, C and D were all positive for combined IgM/IgG. In addition, case D had clinical symptom, while case C did not. Conclusion Serum antibody detection can be used as an effective supplement to the inference of transmission chain of COVID-19, which may facilitate determining the source of infection and improving the evidence.

OBJECTIVETo evaluate the anticoagulant and antineoplastic activities of chemically modified low-molecular-weight heparin (LMWH).

METHODSLMWH obtained by splitting unfractionated heparin (UFH) with sodium periodate oxidation and sodium borohydride reduction was subjected to acetylation catalyzed by DCC and DMAP to produce acetylated LMWH (ALMWH). The anticoagulant activity of ALMWH was determined in mice, and its antiproliferative and anti-invasion activities was assessed in human breast cancer cells MDA-MB-231 and MFC-7.

RESULTSThe anticoagulant activity of LMWH was decreased significantly after acetylation. The concentrations of commercial LMWH* and ALMWH for doubling the coagulation time (CT) were 33.04 µmol/L and 223.56 µmol/L, respectively, and the IC(50) of ALMWH for doubling CT was 6 times of that of LMWH*. ALMWH and LMWH at 0.1, 0.3, 0.9, 2.7 and 8.1 mmol/L both significantly inhibited the proliferation of MDA-MB-231 and MCF-7 cells in a concentration-dependent manner, but ALMWH produced stronger inhibitory effects. The IC(50) of LMWH and ALMWH for inhibiting cell proliferation was 3168.4 µmol/L and 152.6 µmol/L in MCF-7 cells, and 12299.6 µmol/L and 22.2 µmol/L in MDA-MB-231 cells, respectively. ALMWH and LMWH all markedly suppressed the invasion of MDA-MB-231 cells with comparable effects.

CONCLUSIONChemical modification of structure can endow LMWH with a low anticoagulant and high antiproliferative activities.

Animals ; Anticoagulants ; chemistry ; pharmacology ; Antineoplastic Agents ; chemistry ; pharmacology ; Blood Coagulation ; Blood Coagulation Tests ; Cell Line, Tumor ; Heparin ; chemistry ; Heparin, Low-Molecular-Weight ; chemistry ; pharmacology ; Humans ; Mice

OBJECTIVETo study the molecular mechanisms of nm23-H1 for regulating PKC signal pathway before and after transfection with nm23-H1 gene.

METHODSUsing Western-blot, Boyden-chamber, MTT and laser scanning confocal microscopy (LSCM) techniques to detect the distribution of PKC in cytosol and plasma membrane, changes of invasion and proliferation activity, PKC translocation status and changes of intracellular Ca(2+) concentration among different human pulmonary carcinoma cells with transfected or untransfected nm23-H1 gene, and changes of the three cell lines after treatment with Calphostin C, a PKC inhibitor.

RESULTS(1) The expression of PKCalpha, PKCbeta II on L9981 and L9981-pLXSN cell membrane, which was in activated status, was remarkably higher than those in L9981-nm23-H1 cell line (P < 0.001). The expression of PKCalpha, PKCbeta II in cytosol in L9981 and L9981-pLXSN cell lines, which was in inactivated status, was lower than those in L9981-nm23-H1 cell line (P < 0.001). It means that the PKC signal pathway was activated in L9981 and L9981-pLXSN cell lines. (2) PKCalpha and PKCbeta II mainly located in nuclei and perinuclear area in L9981 and L9981-pLXSN cells, which were in active status, and the Ca(2+) concentration in these cells was obviously higher than that in L9981-nm23-H1 cell line (P < 0.01). In L9981-nm23-H1 cell line, which was transfected with nm23-H1 gene, PKCalpha and PKCbeta II mainly located in soluble cytosolic section, in an inactive status. (3) The invasion and proliferation ability of L9981 and L9981-pLXSN lung cancer cells was higher than that of L9981-nm23-H1 cell line (P < 0.001). There was no statistically significant difference between L9981 and L9981-pLXSN cell lines (P > 0.05). (4) After treated with PKC inhibitor Calphstin C, the expression of PKC and PKCbeta II in membrane in L9981 and L9981-pLXSN cell lines was down-regulated (P < 0.001), PKCalpha and PKCbeta II were mainly located in cytosolic area, mainly in an inactive status, and the Ca(2+) concentration was found to be decreased in all the three cell lines. The invasion and proliferation ability of the three lung cancer cell lines were obviously decreasing (P < 0.001). However, the invasion and proliferation ability of L9981-nm23-H1 lung cancer cell line was still lower than that of L9981 and L9981-pLXSN lung cancer cell lines (P < 0.001). There was also no significant difference between L9981 and L9981-pLXSN cell lines (P > 0.05).

CONCLUSIONThe results of this study suggest that nm23-H1 gene might inhibit the invasion and metastasis of lung cancer cells by down-regulating PKC signaling pathway. The Ca(2+) in cells might be involved in this process.

Calcium ; metabolism ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Cell Proliferation ; drug effects ; Cytosol ; metabolism ; Down-Regulation ; Humans ; Lung Neoplasms ; enzymology ; metabolism ; pathology ; NM23 Nucleoside Diphosphate Kinases ; genetics ; Naphthalenes ; pharmacology ; Neoplasm Invasiveness ; Protein Kinase C ; antagonists & inhibitors ; metabolism ; Protein Kinase C beta ; Protein Kinase C-alpha ; metabolism ; Signal Transduction ; Transfection

OBJECTIVETo investigate the clinical characteristics and prognosis of patients with different subtypes of breast cancer: basaloid, HER-2 and luminal types, and try to find the evidence of individualized treatment for the patients.

METHODS1280 histologically and immunohistochemically proven patients with resectable breast cancer were treated, and the clinical data including characteristics, relapse and survival of the patients with different subtypes of breast cancer were analyzed retrospectively.

RESULTSOf the 1280 breast cancer patients, basaloid, HER-2 and luminal types accounted for 20.9%, 23.2% and 55.9%, respectively. Basaloid type was more likely to be found in younger patients frequently with a family history of breast cancer. HER-2 type usually had a tumor of larger size with more advanced stage disease and more metastatic lymph nodes. Luminal type was likely to occur in aged patients with an earlier stage disease. The recurrence rates in basaloid, HER-2 and luminal types were 25.0%, 27.9% and 11.7%, respectively. Patients with basaloid or HER-2 type were found to have a significantly higher recurrence rate than the patients with luminal type breast cancer (P < 0.001), but no significant difference was observed between the basaloid and HER-2 types. However, patients with basaloid type breast cancer were more likely to develop lung metastasis than HER-2 type (13.4% vs. 7.1%, P = 0.017). Up to December 2006, the 5-year disease-free survival (DFS) rates for patients with basaloid, HER-2 and luminal types were 72.2%, 68.2% and 86.2% (P < 0.001), respectively. The overall 5-yr survival (OS) rates of the three groups were 88.6%, 83.8% and 95.8% (P < 0.001) , respectively. Of the patients with luminal type breast cancer, HER2-negative patients had a higher DFS (86.2% vs 57.0%, P < 0.001) and OS (95.8% vs 87.7%, P = 0.0001) compared with those with HER2-positive. The results of Multivariate Cox Regression showed that tumor size and lymph node state were the most important factors influencing the prognosis.

CONCLUSIONEach subtype of breast cancer has somewhat its own specific clinical features in terms of recurrence pattern and prognosis, therefore, individualized treatment regimen may be required.

Adult ; Aged ; Breast Neoplasms ; classification ; metabolism ; pathology ; therapy ; Chemotherapy, Adjuvant ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; secondary ; Lymphatic Metastasis ; Mastectomy ; methods ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Proportional Hazards Models ; Radiotherapy, Adjuvant ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Retrospective Studies ; Survival Rate ; Young Adult