OBJECTIVEIt has been known that the intrahepatic renin-angiotensin-aldosterone system (RAAS) plays a key role in the fibrogenesis in livers. Aldosterone (Aldo), the principal effector molecule of the RAAS, exerts local effects on cell growth and fibrogenesis. However, the signal transduction mechanisms underlying the effects of Aldo on hepatic fibrogenesis remain to be fully elucidated. The present study aims to investigate the signal transduction mechanism underlying the effects of Aldo on the signal passageway of active protein-1 (AP-1).

METHODSIn vitro, HSCs-T6 cell line was treated with Aldo for 10 min, 30 min, 60 min, 120 min and 180 min, and protein expression of Phospho-P42/44 was detected by Western blot. In addition, HSCs-T6 cell line was preincubated for 60 min or not with U0126 (an inhibitor of the MAPK/ERK kinase), and also with antioxidant-N-acetylcysteine (NAC) prior to exposure to Aldo for the indicated times. Protein expression of Phospho-P42/44 was measured by Western blot. DNA biding activity of AP-1 was analyzed by electrophoretic gel mobility shift assay (EMSA). By means of RT-PCR, expression of alpha1(1) procollagen mRNA was detected.

RESULTSAldo stimulated HSC via extracellular signal-regulated kinase (ERK1/2) pathway. Time course experiments showed that Aldo induced Phospho-P42/44 expression, which was abrogated by U0126, reaching a maximum at 10 minutes, and then declined progressively. NAC inhibited the Phospho-P42/44 expression. EMSA showed that stimulation of HSC by Aldo markedly increased AP-1 DNA binding activity. U0126 markedly reduced AP-1 DNA binding activity induced by Aldo; NAC partly decreased AP-1 activity induced by Aldo. Aldo up-regulated expression of alpha1(1) procollagen mRNA, which was attenuated by U0126 and NAC.

CONCLUSIONStimulation of HSC by Aldo results in activation of AP-1 via ERK1/2 pathway, leading to up-regulation of AP-1 target gene alpha1(1) procollagen mRNA expression.

Aldosterone ; pharmacology ; Cell Line ; Collagen Type I ; biosynthesis ; genetics ; Hepatocytes ; cytology ; metabolism ; Humans ; Mitogen-Activated Protein Kinase 3 ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Transcription Factor AP-1 ; metabolism

OBJECTIVETo compare therapeutic effects of electroacupuncture at Houxi (SI 3) and medicine on acute lumbar sprain.

METHODSThree hundred cases of acute lumbar sprain were randomly divided into two groups, a electroacupuncture (EA) group and a medication group, 150 cases in each group. The EA group were treated with EA at Houxi (SI 3), once each day, 3 sessions constituting one course, and the medication group with Mobike, once daily, 7. 5 mg each time. Their therapeutic effects were evaluated after treatment for 7 days and one month respectively.

RESULTSFor the short-term therapeutic effect, the effective rate was 97. 3% in the EA group and 89. 2% in the medication group with a very significant difference between the two groups (P<0. 01); for the long-term therapeutic effect, the effective rate was 99. 3% in the EA group and 93. 2% in the medication group with a very significant difference between the two groups (P<0. 01).

CONCLUSIONBoth the short-term and the long-term therapeutic effects of EA at Houxi (SI 3) on acute lumbar sprain are better than those in the medication group.

Acute Disease ; Adult ; Electroacupuncture ; Female ; Humans ; Lumbosacral Region ; injuries ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Sprains and Strains ; therapy

Objective It is rarely reported whether myricetin inhibits the activation and function of cardiac fibroblasts and thereby prevents myocardial fibrosis. This study was to investigate the effects of myricetin on the activation,proliferation and secretion of cardiac fibroblasts and its possible molecular mechanisms. Methods Fibroblasts isolated from 1-3 days old rats were cultured and their activation,proliferation and secretion were induced with the transforming growth factor (TGF). The fibroblasts were incubated with myricetin at different concentrations of 1,3,10,30 and 100 μmol/L for 24 hours followed by detection of their proliferation with the CKK8 kit,the transcription levels of fibrotic factors by RT-PCR and the expression levels of α-SMA and signal proteins by immunoflu-orescence staining and Western blot,respectively. Results The expression of α-SMA was significantly up-regulated in the cardiac fi- broblasts of the rats in the TGF-β,30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups as compared with that in the control group (P<0.05) but down-regulated in the 30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups in com-parison with that in the TGF-β group (P<0.05). At 48 hours,the transcription levels of collagenⅠ,collagenⅢ,fibronectin and con-nective tissue growth factor were markedly higher in the TGF-β,30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups than in the control group (P<0.05) but lower in the 30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups than in the TGF-β group (P<0.05). The phosphorylation levels of smad2 and smad3 were remarkably elevated in the TGF-β,30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups and the expression of smad4 reduced in the TGF-β group as com-pared with the control group (P<0.05). The levels of smad2,smad3 and smad4 were all significantly decreased in the 30 μmol/L myr-icetin+TGF-β and the 100 μmol/L myricetin+TGF-β groups in comparison with the TGF-β group (P<0.05). Conclusion Myricetin suppresses the activation,proliferation and secretion of cardiac fibroblasts induced by TGF-β via inhibiting the smad signaling pathway.

Objective To learn the willingness to engage in elderly care among nursing students in junior college in order to provide the basis for the training of geriatric nursing staff.Methods Totally 1 1 76 junior college nursing students in Ningbo were selected by stratified sampling method and were investigated with a questionnaire based on the theory of planned behavior.The willingness and plan to engage in elderly care among the students were analyzed.Results Fifty six percent of the nursing students would like to work in geriatric nursing.The average score of behavioral attitude,subjective norm,and perceived behavior control were (52.1 2 ±9.52),(1 6.42 ±6.21 ),and (31 .54 ±7.94).The regression analysis showed that the score of behavior attitude (β=0.1 8),subjective norm (β=0.35 ),and perceived behavior control (β=0.25 )were the factors influencing the willingness to work in geriatric nursing.Different major,grade, experience in elderly care and knowledge had different attitude and behavior in the willingness to engage in elderly care work(P <0.05).There were positive correlations among behavioral attitude,subjective norm,perceived behavior control and behavior intention (P <0.01 )Conclusion The investigation suggested that training nursing staff achievements previews in this area.Nursing students in higher vocational colleges have good professional intention in the work of geriatric nursing work.

OBJECTIVETo study the biomarkers of styrene and to provide theoretical basis for bio-monitoring of styrene.

METHODSUrinary mandalic acid (MA), phenylglyoxalic acid (PGA) and mercapturic acid (MUA) of styrene were examined by high performance liquid chromatography (HPLC).

RESULTSThe correlation regression equations between exposure dose and MA, PGA and MUA level in morning urinary samples were: ŷ = 2.58x + 70.82; ŷ = 1.66x + 37.42; ŷ = 0.05x + 0.55 respectively. The correlation regression equations between exposure dose and MA, PGA and MUA level in post-shift urinary samples were: ŷ = 1.85x + 89.02; ŷ = 1.33x + 4.32; ŷ = 0.04x + 0.68 respectively. All showed close dose-response relationship.

CONCLUSIONSThe level of MA, PGA and MUA in morning or post-shift urinary samples may be used as bio-monitoring indexes of styrene.

Acetylcysteine ; urine ; Adult ; Biomarkers ; Chromatography, High Pressure Liquid ; Environmental Monitoring ; Glyoxylates ; urine ; Humans ; Male ; Mandelic Acids ; urine ; Regression Analysis ; Styrene ; metabolism

BACKGROUNDNatural killer (NK) cells play critical roles in host immune defense, while the quantities and subset distributions may vary among different races. To address the difference, we compared these variables among Chinese Han, the Caucasians and the Blacks. The study may provide critical background information for both basic research and clinical investigation.

METHODSBlood samples collected from populations of different races were tested within 12 hours after collection and subsets of NK cells were characterized using flow cytometry.

RESULTSThe absolute NK count in the Chinese Han was significantly higher than that in the Caucasian. The Han and Caucasian groups showed higher percentages of cytotoxic subset compared to that of the Black group. The percentage of cytokine-producing subset of Chinese Han group was lower than that of Caucasian and Black groups. Black group had a higher percentage of function-unknown NK subset than that of the Han and Caucasian groups.

CONCLUSIONOur data indicated that NK cell count and the distribution of different subsets varied among different races, which should be taken into consideration in related investigations.

Adult ; African Continental Ancestry Group ; Asian Continental Ancestry Group ; European Continental Ancestry Group ; Female ; Humans ; Killer Cells, Natural ; cytology ; metabolism ; Male ; NK Cell Lectin-Like Receptor Subfamily C ; metabolism

OBJECTIVETo identify the candidate auto-antigen of rheumatic heart disease as a molecular marker for this disease.

METHODSThe total RNA of the heart tissue of patients with rheumatic heart disease was extracted and reverse-transcribed into long cDNA to construct the phage expression library. The library was screened using the serum from patients with active rheumatic fever, and the positive clone was identified and analyzed by bioinformatics and expressed in vitro. The expressed products were evaluated with Western blotting and its cross-reactivity was assessed.

RESULTSThe phage expression library of the heart tissue of patients with rheumatic heart disease was constructed, with the titer of the primary library of 3.3×10(6) pfu/ml, recombinant rate of 99%, and 81% of the inserted segments were larger than 1 kb. An auto-antigen RHDAG1 was identified by screening, which was homologous to keratin 18. RHDAG1 was detected in the serum of patients with active rheumatic fever and of those with rheumatic heart disease, but not in the serum of healthy subjects.

CONCLUSIONPhage display library can be an effective strategy to screen the auto-antigens of rheumatic heart disease. The auto-antigen RHDAG1 can be a candidate molecular biomarker of rheumatic heart disease and/or rheumatic fever.

Autoantibodies ; blood ; immunology ; Autoantigens ; immunology ; isolation & purification ; Autoimmune Diseases ; blood ; immunology ; Humans ; Peptide Library ; Rheumatic Heart Disease ; immunology

OBJECTIVETo study the methods and mechanisms of immune tolerance in cardiac transplantation.

METHODSMale DA rat hearts were transplanted to male Lewis rats using Ono's model and randomly divided into five groups: untreated, intravenous injection of 1 x 10(8) DA splenocytes to Lewis rat, intraperitoneal injection of cyclophosphamide (100 mg/kg) to Lewis rat, intravenous injection of 1 x 10(8) DA splenocytes combined with intraperitoneal injection of cyclophosphamide (100 mg/kg) to Lewis rat, multiple injection of DA rat splenocytes with intraperitoneal injection of cyclophosphamide, 11 days later heart transplantation was performed. Mean survival time (MST), histological changes, mixed lymphocyte reaction (MLR), the role of interleukin-2 (IL-2) to MLR and the role of tolerant rat splenocytes to MLR were measured after operation.

RESULTSThe survival time of heart allografts in the group of multiple injection of DA rat splenocytes with intraperitoneal injection of cyclophosphamide [MST: (85.3 +/- 7.5) d, t = 0, P < 0.01] was significantly longer than in the groups of untreated [MST: (7.3 +/- 1.0) d], intravenous injection of 1 x 10(8) DA splenocytes to Lewis rat [MST: (7.9 +/- 0.9) d], intraperitoneal injection of cyclophosphamide (100 mg/kg) to Lewis rat [MST: (8.1 +/- 1.2) d], intravenous injection of 1 x 10(8) DA splenocytes combined with intraperitoneal injection of cyclophosphamide (100 mg/kg) to Lewis rat [MST: (25.8 +/- 3.5) d]. Only a few inflammatory cells infiltrated in cardiac allografts in the group of multiple injection of DA rat splenocytes with intraperitoneal injection of cyclophosphamide. MLR in the group of multiple injection of DA rat splenocytes with intraperitoneal injection of cyclophosphamide were significantly decreased compared with those of normal control (t = 0, P < 0.01). IL-2 could partly reversed the hyporesponsiveness of MLR in tolerant rats, the tolerance could be transferred in vitro.

CONCLUSIONSMultiple injection of donor splenocytes combined with intraperitoneal injection of cyclophosphamide to recipients could induce immune tolerance to cardiac allografts.

Adoptive Transfer ; Animals ; Cell Transplantation ; Cyclophosphamide ; pharmacology ; Graft Enhancement, Immunologic ; methods ; Graft Survival ; Heart Transplantation ; immunology ; Injections, Intravenous ; Isoantigens ; administration & dosage ; immunology ; Male ; Rats ; Rats, Inbred BN ; Rats, Inbred Lew ; Rats, Inbred Strains ; Rats, Wistar ; Spleen ; cytology ; Transplantation Tolerance ; drug effects ; Transplantation, Heterologous ; immunology

Objective To study the risk factors on anemia among elderly women in rural areas of Xiuning county, Anhui province, China. Methods Xiuning county was selected as working field and elderly women aged 50-75y were selected as subjects. Finger hemoglobin (Hb) was measured and basic health survey was face-to-face interviewed. 220 elderly women with anemia entered into the ease group; and matched by age, another 220 women with normal Hb concentration entered the control group. Survey on diet, questionnaire regarding health and lifestyle and related blood indexes were studied and tested. Results When comparing the data from both case and control groups, weight was (49.4±7.3) kg vs. (52.5±8.4) kg (t= 3.97,P<0.01 ), waist circumference was (75.8±7.8) cm vs. (79.1±9.3) cm (t=3.85,P<0.01), BMI was (21.8±2.6) kg/m2 vs. ( 22.9±3.2) kg/m2 (t = 3.775, P< 0.01 ), respectively. The total protein was ( 76.4± 5.0) g/L vs. (78.4±5.6)g/L (t=3.83 ,P<0.01 ), albumin was (45.7±3.1 ) g/L vs. (47.3±2.9)g/L (t=5.24, P<0.01 ), serum iron was ( 10.3±4.1 ) μmol/L vs. ( 12.7±4.6) μmol/L (t=5.48, P<0.01 ), and saturation of transferrin was ( 19.0±7.6)% vs. (23.1±9.1 )% (t=4.90, P<0.01 ), respectively. Results from multifactor conditioned logistic regression analysis showed that the odd ratios (OR) for anemia with staple food, BMI and vitamin A were 1.54, 1.89, 1.69, and the OR for anemia with BMI, staple food, animal food, carbohydrate and vitamin A were 2.0, 1.6, 1.6, 1.4, 1.6, with their confidence intervals (CI) as 1.3-2.9, 1.1-2.3,1.0-2.3,1.0-2.1, 1.1-2.4, respectively. Conclusion The quality of diet, health status and related blood indexes on anemia among elderly women were lower than that in control group. Lower BMI, less staple food and animal food, less carbohydrate and vitamin A intake appeared to be risk factors of anemia.

This study was aimed to investigate whether mesenchymal stem cells (MSC) can be isolated from bone marrow filters which have always been discarded. The bone marrow (BM) particles from BM filters of 2 healthy donors were cultivated by primary explant culture. After expansion, the number of MSC was counted and their immunophenotype and differentiation potential were detected. The results indicated that many MSC were found in bone marrow particles from filters, and nearly 10(7) MSC were obtained at 3 passages of expansion. They not only possessed the characteristics of morphology and immunophenotype of MSC, but also could differentiate into osteoblasts, chondrocytes and adipocytes. In conclusion, a large amount of MSC can be obtained from BM filters if the BM particles were cultivated by primary explant culture.
Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; cytology ; Tissue Culture Techniques