Objective To obtain and identify dendritic cell (DC) from mouse bone marrow in vitro. Methods Recombinant mouse granulocyte and macrophage colony stimulus factor (GM-CSF) induction was used to induce mouse bone marrow cells to form dendritic cells in vitro. The morphological changes were observed with light inverted microscope and scanning electron microscope (SEM). CD11c, MHCⅡ, and CD86 were identified with flow cytometry. The biological function was studied with antigen phagocytosis test and mixed lymphocyte reaction (MLR). Result The cultured DC from mouse bone marrow displayed the typical morphological characteristics of DC. Immature DC, which had high expression in CD11c and low expression in MHCⅡ and CD86, could phagocytize antigen. Mature DC, which had high expression in CD11c, MHCⅡ and CD86, could stimulate allogenic mixed lymphocyte proliferation. Conclusion DC can be generated from mouse bone marrow cells through cytokine induction in vitro and be used for further study associated with DC.

Objective To explore the serum levels of growth differentiation factor-15 (GDF-15) in patients with acute exacerbation of chronic heart failure (CHF) and its correlation with other common indexes,to provide reference for its clinical diagnosis,treatment and prognosis.Methods Two hundred patients with acute exacerbation of CHF were selected as CHF group,and 100 matched healthy volunteers were selected as control group.Serum levels of GDF-15 and N-terminal pronatriuretic peptide (NT-proBNP) were detected,and left ventricular end diastolic diameter (LVESD),left ventricular end diastolic diameter (LVEDD),and left ventricular ejection fraction (LVEF) were measured by echocardiography within 2 hours after admitted to hospital,and after the symptoms improved of CHF group,and on health examination day of control group.Patients in CHF group were followed up to record CHF related adverse events.Correlations between GDF-15 and other indicators were analyzed by Spearman correlation analysis,and the clinical value of serum GDF-15 on diagnosing CHF was analyzed by the receiver operating characteristic curve (ROC) and the area under the curve (AUC).Results The serum levels of GDF-15 and NT-proBNP in each time-point of CHF group were all higher than those of control group (t =4.70 ～ 7.11,P ＜ 0.05 orP ＜ 0.01).The serum levels of GDF-15 and NT-proBNP had negative correlation with LVEF (r =-0.539,-0.572,P ＜ 0.01),and had positive correlation with LVESD,LVEDD,and NYHA cardiac functional grading (r =0.505 ～ 0.861,P ＜ 0.01).Serum GDF-15 had positive correlation with serum NT-proBNP (r =0.528,P ＜0.01).With the increase of serum GDF-15 level,CHF group's readmission (rate) and death (rate) were both increased (x2 =36.86,26.59,P ＜0.01).AUC of predicting readmission risk by serum GDF-15 was 0.822 (95％ CI:0.719 ～0.890,P ＜0.01),and the best predictive cutoff point was 2 876.30 ng/L (sensitivity was 91.86％,specificity was 73.27％).AUC of predicting mortality risk was 0.816 (95％ CI:0.715 ～ 0.885,P ＜ 0.01),and the best predictive cutoff point was 3 487.05 ng/L (sensitivity was 91.72％,specificity was 69.05％).Conclusions Serum GDF-15 level in patients with acute exacerbation of CHF is higher,decreases with symptoms improvement,has positive correlation with LVESD,LVEDD,and NYHA cardiac functional grading,and has negative correlation with LVEF,has higher sensitivity on predicting CHF-related adverse events,and the mechanism may be related to the activation of SMAD pathway.Therefore,it may be a promising biomarker for clinical diagnosis and prognosis of cardiovascular diseases.

Mitogen-activated protein kinases (MAPKs) signal is one of the important ways in eukaryotic cell,which adjusts and controls the structure and function of the cell. MAPKs in eukaryotes include p38, ERK, JNK and ERK5, etc. With the deepening research,we found that the activation of p38, ERK, JNK signal pathways were closely related with osteoarthritis (OA) cartilage injury. MAPKs are the key signaling systems involved in the production of matrix metalloproteinases and the regulation of cartilage cell proliferation, apoptosis and differentiation. Expecially the matrix metalloproteinases can accelerate the degradation of articular cartilage. So it has been the new spot in pathogenesis of osteoarthritis study.
Animals ; Cartilage, Articular ; pathology ; Humans ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinases ; metabolism ; Osteoarthritis ; etiology ; pathology

OBJECTIVETo investigate the immunological activity of Streptomyces polysaccharide (SMP) on normal and immunosuppressed mice.

METHODSThe effect of SMP on the proliferating activity of normal mouse splenocytes was tested in the mixed lymphocyte culture, and the changes of peripheral blood T lymphocytes were evaluated with acid a-naphthyl acetate esterase (ANAE) method. The ratio of Lyt2+ and L3T4+ T cell subsets was measured by flow cytometry.

RESULTSSMP stimulated obvious proliferation of mixed lymphocytes, showed protective effects on T lymphocyte and increased the ratio of Lyt2+ and L3T4+ cell subsets to nearly normal level in immunosuppressed mice.

CONCLUSIONSSMP can regulate the immune function in mice.

Animals ; CD4-Positive T-Lymphocytes ; immunology ; Female ; Immunocompromised Host ; immunology ; Lymphocyte Culture Test, Mixed ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Polysaccharides ; immunology ; Spleen ; cytology ; Streptomyces ; chemistry ; immunology ; T-Lymphocytes ; cytology ; immunology

Objective To investigate the effects of different CO2 pneumoperitoneum on cell cycle and cell cycle protein of a gastric cancer cell line. Methods Human gastric cancer cell line MNK-45 were exposed to 0,10,12 and 15 mm Hg CO2 pneumoperitoneum in vitro for 4 hrs. Cell cycle was measured by flowcytometry, the expression of CDK4 ,Cyclin D1、Rb and pRb was studied by Western-blot, and the binding ability of CDK4 and Cyclin D1 was evaluated by immunoprecipitation. Results The cell proliferation index in 15 mm Hg CO2 pneumoperitoneum group dropped significantly (27.4% ± 3. 7%) vs. (36. 4% ±3. 3%) ,P <0. 05, while that in other groups did not change significantly. The protein of CDK4、Cyclin D1and binding ability of Cyclin D1 and CDK4 dropped dramatically in the 15 mm Hg CO2 pneumoperitoneum group (0.71%±0.12%),(0.93% ±0.21%),(0.54%±0.11%),(0.18% ±0.02%) vs. (1.05% ±0.16%),(1.40% ±0.24%),(0.75% ±0.14%),(0.31% ±0.02%), all P<0.05. There were no changes of Rb in protein levels, while the phosphorylated Rb dropped obviously. Conclusion There was no obvious effects of clinical CO2 pneumoperitoneum on gastric cancer cells growth and proliferation.

Objective To investigate the effects of different CO2 pneumoperitoneum pressures on gastric cancer cells' growth and proliferation in nude mouse model of implanted tumor. Methods Human gastric cancer cell lines MNK-45 were exposed under 0、10、12 and 15 mm Hg CO2 pneumoperitoneum for 4 hrs respectively. 2 × 106 processed cells were inplanted into nude mice subcutaneously. Three weeks later, mice were sacrificed and the weight and bulk of the tumor measured. Then we observed the transplantation tumor by HE stain and Ki-67 stain. Results There was no significant difference in tumor's growing time, bulk and weight between 0, 10, 12 mm Hg CO2 pneumoperitoneum groups (7. 8 d, 7. 2 d, 7. 8 d; 1. 2 cm3, 1. 3 cm3, 1. 3 cm3; 1.5 g, 1. 9 g, 1. 6 g)and the control group (7. 3 d, 1. 2 cm3, 1.4 g) (P > 0. 05 ). The growing time of tumor in 15 mm Hg CO2 pneumoperitoneum (12. 5 d) was obviously longer than the control group ( P < 0.05 ) , the bulk and weight of tumor in 15 mm Hg CO2 pneumoperitoneum (0. 5 cm3, 0. 5 g) group significantly decreased compared with the control group (P <0.05). The positive rate of Ki-67 in 15 mm Hg CO2 pneumoperitoneum (27. 5% ) group was obviously lower than the control group (59.6%) (P<0.01). However, there were no significant differences between 0, 10, 12 mm Hg CO2 pneumoperitoneum groups (61.2%, 60.5%, 63.4%) and the control group (P > 0.05). Conclusion Clinically adopted CO2 pneumoperitoneum pressures have no significant effect on gastric cancer cells growth and proliferation.

Adult ; Arsenic ; pharmacology ; Chromosome Aberrations ; chemically induced ; Dose-Response Relationship, Drug ; Female ; Humans ; Lymphocytes ; drug effects ; metabolism ; Male ; Middle Aged

Objective Neonatal lupus erythematosus (NLE) is an uncommon" passive autoimmune disease, which is associated with transplacental passage of maternal antibodies. It is often misdiagnosed as intrauterine infection or sepsis. The main purpose of this retrospective study was to summarize its clinical manifestations related with pathogenesis.Methods Data of all the NLE neonates, including clinical manifestations, immunochemical evidence of serum antinuclear antibodies (ANA), antibody to Ro/Sjogren's syndrome A ( anti-Ro/SSA), antibody to La/Sjogren' s syndrome B (anti-La/SSB) and anti-dsDNA antibodies in both infants and mothers, and images from head ultrasound and CT scans were analyzed. Follow-up was performed until one and half years of age or when all the clinical abnormalities had been resolved.Results Totally 8 cases (3 males and 5 females ) seen between September 2003 and February 2006 met the diagnostic criteria of NLE, in whom 4 were small for gestational age and one was born prematurely. Mean gestational age was (38.1 ± 1.9 ) weeks, mean birth weight (2605 ± 420) g, mean admission age (22.4 ± 27.7 ) days (2 hours-72 days) and mean age of onset (9.4 ± 12. 1)days (0-28 days). The common clinical manifestations included cutaneous lupus lesions (8 infants ), neural system abnormalities (2 infants ) and congenital heart block (2 infants). Annular, erythematous or desquamative lesions were seen in skin and all disappeared before 6 months of age. One patient presented with third degree atrio-ventricular block and was delivered by cesarean section because of " fetal distress" He did not recover by the end of one and half years follow-up. One infant was hypotonic with delayed neuro-motor development initially and during follow-up with both abnormal neonatal behavioral neurological assessment (NBNA) and imaging findings. Brain CT scan showed generalized low density involving periventricular and deep white matter at one week of age. At the age of one and a half years, he presented with normal mental development index determined by Child Development Center of China (CDCC) infant intelligence mensuration. Other abnormal clinical findings such as hepatosplenomegaly, anemia, thrombocytopenia, cholestasis and elevated liver enzyme activities were all resolved before 6 months of age. Only 3 mothers of the NLE infants were diagnosed as systemic lupus erythematosus (SLE) before parturition and only one received partial therapy. At least anti-Ro/SSA antibody or anti-La/SSB antibody or ANA was found in the affected patients. Seven cases had circulating anti-Ro and/or anti-La antibodies in the mothers and in the newborns, while ANA was positive in seven newborns and in all mothers. All the clinical symptoms disappeared before 18 months ot age except for congenital heart block. No special intervention was applied.Conclusions Serum auto-antibodies should be investigated to rule out NLE when a newborn infant has congenital heart block or rashes or thrombocytopenia, although there is no maternal history of SLE. Central nervous system abnormalities in NLE are likely to be a transient phenomenon and whether it will cause long-term sequelae is uncertain.

Objective To investigate the protective effects and mecha-nism of tetramethylpyrazine ( TMP ) on isoproterenol ( ISO ) -induced acute myocardial ischemia.Methods BALB/c mice were randomly divided into 6 groups:normal group ( saline 10 mg· kg -1 · d-1 ) , model group ( ISO 3 mg· kg -1 · d-1 , three times for three days ) , control group ( propranolol 0.4 mg · kg -1 · d-1 ) , different dose TMP group (60, 30, 15 mg· kg -1· d-1), 10 mice in each group.All groups were administered drugs for 7 days.Then ISO was given intraperitoneally to establish acute myocardial ischemia model.Tetrazolium chloride ( TTC ) staining was used to determine focal myocardial ischemia and hematoxylin-eosin ( HE ) staining was performed in histopathological examination.MB isoenzyme of creatine kinase ( CK -MB ) , lactate dehydrogenase ( LDH) levels and hydrogen peroxide ( H2 O2 ) , total an-tioxidant capacity ( T -AOC ) of serum were detected.Results TMP obviously attenuated abnormal electrocardiogram with ST -segment elevation , myocardial damage and ischemia region area in model mice.Compared with model group [ CK -MB: ( 1562.46 ± 138.92 ) U · L-1; LDH:(6973.28 ±285.50 ) U· L-1; T-AOC: ( 2.99 ±1.25 ) U · mL-1;H2 O2:( 92.52 ±11.42 ) mmol · L-1 ] , TMP group significantly decreased the levels of serum [ CK -MB:( 605.81 ±117.72 ), ( 838.67 ±159.73 ), (1264.37 ±134.17) U· L-1);LDH:(2661.79 ±229.27),(5776.16 ±331.39), (6014.59 ±291.56) U· L-1);H2O2:(31.98 ±7.38), (67.06 ±18.43), (86.62 ±13.41) mmol· L-1] and increased T-AOC [(7.54 ±2.51), (5.80 ±2.53 ) , (4.93 ±2.21 ) U · mL-1 ] levels of myocardial ischemia mice ( P <0.05 or P <0.01 ) . Conclusion TMP would alleviate Iso-induced acute myocardial ischemia , which may be associated with the decrease of cardiac enzymes and its protective effects against oxidative stress.

To study the protective effect of Danshen Tongluo capsule on rat hearts in the myocardial infarction (MI) model. After being fed with high fat diets for one month, the SD rats were randomly divided into the sham group and the model group according to the left ventricular ejection fraction (EF). The MI model group was duplicated by ligating coronary artery and then divided into five groups: the model group, the positive control group (model + captopril), the small dose group (model + Danshen Tongluo), the medium dose group (model + Danshen Tongluo) and the high dose group (model + Danshen Tongluo). Four weeks later, changes in myocardium ultra-structure were observed by hemodynamics, cardiac ultrasound and electron microscope. The results showed: (1) All doses of Danshen Tongluo capsule could significantly reduce the left ventricular end diastolic pressure (LVEDP) (P <0.01), increase the maximum internal pressure of the left ventricle (+ dp/dtmax), and lower the drop rate of left ventricular pressure (- dp/dtmax), with statistical significances in medium and high dose groups; (2) The B ultrasound results showed increase in EF and left ventricular shortening fraction (FS) in all dose groups of Danshen Tongluo capsule; (3) The medium dose group showed significant decrease in myocardial infarction index (P <0.01) and injured and fractured myofilament and sarcomere of ischemic myocardium in myocardial ultra-structure; All of Danshen Tongluo capsule-treated groups revealed reduction in myocardial injury and myocardial infraction area. The study preliminarily proves that Danshen Tongluo capsule can improve hemodynamic function, and has a protective effect on myocardial ischemia.
Animals ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Hemodynamics ; drug effects ; Male ; Myocardial Infarction ; drug therapy ; pathology ; physiopathology ; Myocytes, Cardiac ; ultrastructure ; Rats ; Rats, Sprague-Dawley ; Ventricular Function, Left ; drug effects