Objective To investigate the inhibitory effect and the effect on cell proliferation and apoptosis of artemether(ART) on human gastric adenocarcinoma cell lines and pancreatic cancer cell lines in vitro. Methods The inhibitory effect of ART on human gastric cancer cell lines(SGC-7901 and MKN-45) and human pancreatic cancer cell lines(SW-1990 and BXPC-3) were detected by MTT assay,and the effect on cell cycle and apoptosis were evaluated by flow cytometry. Results It was indicated by MTT assay that the killing effect of ART on the cancer cell lines were positively correlated to time and dosage(P

Animals ; Chicory ; Fibronectins ; metabolism ; Insulin-Like Growth Factor Binding Proteins ; metabolism ; Liver ; drug effects ; metabolism ; Liver Cirrhosis, Experimental ; metabolism ; Male ; Plant Extracts ; pharmacology ; Rats ; Rats, Wistar ; Smad3 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo observe the effect of naringin of Drynaria Rhizome, a Chinese medical component of Zhuanggu Jianxi Recipe (ZJR) containing serum on caveolin-p38MAPK signal factors (such as caveolin-1, p-p38, p-ATF-2, IL-1β, and TNF-α) in IL-1β induced rabbit degenerated chondrocytes, and further to explore its mechanism for protecting articular cartilages.

METHODSNaringin of Drynaria Rhizome was obtained and analyzed by HPLC-TOF/MS. Four weeks old New Zealand rabbits were killed and their bilateral knee joints were isolated aseptically. CDs were isolated and then cultured in vitro. The second generation of CDs were used for later experiment. The effect of naringin on CDs proliferation was detected by MTT assay. The effect of naringin on the expression of IL-1β-induced collagen II in CDs was detected by immunohistochemical method. The effect of naringin on caveolin-1, p-p38, and p-ATF-2 protein in IL-1β-induced CDs was detected by Western blot. The effect of naringin on mRNA expression of IL-1β and TNF-α in IL-1β-induced CDs was detected by RT-PCR.

RESULTSThe appearance time of naringin in flow graphs of naringin standard solution and ZJR containing serum was 23.5 min, and the molecular weight ranged between 581.0 and 581.5 m/z. Naringin could promote the proliferation of CDs, and inhibit the effect of IL-1β on collagen II in CDs. Compared with the model group, naringin could reduce the expression of caveolin-1, p-p38, p-ATF-2, IL-1β, and TNF-α in IL-1β induced CDs (P < 0.05), which was approximate to the level of the normal group.

CONCLUSIONSNaringin could not only promote the proliferation of CDs, but also protect IL-1β-induced CDs. Its mechanism might be associated with decreasing the expression of caveolin-1, p-p38, and p-ATF-2 proteins, inhibiting caveolin-p38MAPK signal pathway, and further reducing mRNA expression of IL-1β and TNF-α in the downstream of caveolin-p38MAPK signal pathway.

Animals ; Blotting, Western ; Cartilage, Articular ; Caveolins ; Chondrocytes ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Flavanones ; pharmacology ; Interleukin-1beta ; metabolism ; Polypodiaceae ; Rabbits ; Rhizome ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Adult ; Female ; Humans ; Middle Aged ; Myxoma ; pathology ; surgery ; Vulvar Neoplasms ; pathology ; surgery

OBJECTIVETo explore the mechanism of three kinds extracts (saponins, volatile components, polysaccharide components) of Qingxin Kaiqiao Recipe (QKR) in improving learning and memory capabilities of Alzheimer's disease (AD) rats.

METHODSA controlled comparison method was used. Totally 56 male SD rats were randomly divided into seven groups, i.e., the normal control group, the sham-operation group, the model group, the Aricept group, the saponin group, the benzene group, and the polysaccharide group, 8 in each group. AD rat model was established by bilateral hippocampus injection of Aβ1-40 (2 µL, 2.5 µg/µL). The next day after modeling rats in the saponin group, the benzene group, and the polysaccharide group, the saponin group, the Aricept group were intragastrically administered with saponin (at the daily dose of 9 mL/kg, 2.1 g/mL) , benzene (at the daily dose of 3.33 mL/kg, 5.7 g/mL) , polysaccharide (at the daily dose of 8.33 mL/kg, 2.28 g/mL), Aricept (at the daily dose of 1.67 mg/kg), respectively, once a day for 2 consecutive weeks from 10 am every day. Equal volume of normal saline was intragastrically administered to rats in the normal control group and the model group. Learning and memory capabilities were detected using water maze 2 weeks later. Expression levels of synaptotagmin-1 (Syt-1), interleukin-1β (IL-1β), glia fibrillary acidic protein (GFAP), and β-amyloid precursor protein (βAPP) in the cortex and hippocampus of AD rats were detected using immunohistochemistry.

RESULTSLearning and memory capabilities could be improved by three kinds extracts of QKR. There was no statistical difference in the escape latency between the polysaccharide group and the model group (P >0. 05). The escape lacency was shortened in the rest treatment groups (P < 0.05). The escape latency was obviously prolonged in three kinds extracts of QKR groups, when compared with the Aricept group (P < 0.05, P < 0.01). Compared with the model group, times for crossing platforms were significantly increased in the saponin group and the Aricept group (P < 0.05). Compared with the Aricept group, average times for crossing platforms were significantly lessened in three kinds extracts of QKR groups (P < 0.01). Compared with the sham-operation group, expression levels of Syt-1, IL-1β, GFAP, and βAPP in the cortex and hippocampus were increased in the model group (P < 0.01). Compared with the model group, the expression of cortical Syt-1 increased in the saponin group and the benzene group; the expression of cortical IL-1β increased in the benzene group and the polysaccharide group; the expression of hippocampal GFAP increased in the three kinds extracts of QKR groups; expression levels of Syt-1, IL-1β, GFAP, and β-APP in the cortex and hippocampus decreased in the rest treatment groups (all P < 0.05, P < 0.01). Compared with the Aricept group, expression levels of Syt-1, IL-1β, GFAP, and βAPP in the cortex and hippocampus were significantly increased in three kinds extracts of QKR groups (P < 0.05, P < 0.01).

CONCLUSIONThree kinds extracts of QKR might play roles in anti-AD possibly by decreasing expression levels of Syt-1, IL-1β, GFAP, and βAPP in the cortex and hippocampus.

Alzheimer Disease ; Amyloid beta-Protein Precursor ; Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Glial Fibrillary Acidic Protein ; Hippocampus ; Interleukin-1beta ; Learning ; drug effects ; Male ; Memory ; Rats ; Rats, Sprague-Dawley ; Saponins

OBJECTIVETo compare the incidence of emotional disorder in patients with acute or stable coronary heart disease.

METHODSA total of 298 patients with suspected coronary heart disease (CHD) were designed into three groups based on of coronary angiography results: acute coronary syndrome (ACS, n = 128), stable angina pectoris (SAP, n = 108) and non-CHD (n = 62). All patients were evaluated by Zung Self-rating Depression Scale (SDS), Zung Self-rating Anxiety Scale(SAS) and Hamilton Depression Rating Scale (HRSD) for depression and anxiety before coronary angiography (CAG), 3 days after CAG, and 1 day before discharge.

RESULTSIncidences of depression and anxiety were significantly higher the ACS group (65.6%and 78.9% before CAG; 60.9% and 70.3% 3 days post CAG; 45.3%and 64.8% before discharge) compared patients with SAP (18.5% and 26.9% before CAG; 17.6% and 28.7% 3 days post CAG; 15.7% and 26.9% before discharge, all P < 0.05 vs. ACS) and non-CHD patients (32.3% and 25.8% before CAG; 27.4% and 24.2% 3 days post CAG; 29.0% and 30.6% before discharge, all P < 0.05 vs. ACS) while the depression and anxiety incidences were similar between patients with SAP and non-CHD in this cohort (P > 0.05).

CONCLUSIONEmotional disorder is common in patients with suspected heart diseases, especially in patients with ACS. Psychological distress of patients with suspected heart disease should be evaluated and treated.

Acute Coronary Syndrome ; psychology ; Adult ; Aged ; Anxiety Disorders ; etiology ; Coronary Disease ; psychology ; Depressive Disorder ; etiology ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Psychiatric Status Rating Scales ; Risk Factors

The present study aims to explore the possible mechanisms that trichostatin A (TSA), a histone deacetylases inhibitor (HDACi), affects the inflammatory signaling pathways of lipopolysaccharide/toll-like receptor 4/nuclear factor-κB (LPS/TLR4/NF-κB). Murine macrophage cell line RAW264.7 cells were employed. MTT assay was used to assess cell viability. The contents of TNF-α, IL-1β and IL-6 in culture supernatant were assayed by enzyme-linked immunosorbent assay (ELISA). TLR4 expression and NF-κB/p65 (Lys310) acetylation were examined by Western blotting. DNA binding activity of NF-κB/p65 was detected by using TransAM(TM) NF-κB/p65 activity assay kit. The results showed that, compared with control group, which was treated by DMSO, the cells treated with TSA (20, 40, 80 ng/mL) showed decreased percentages of cell survival (P < 0.05). The contents of TNF-α, IL-1β and IL-6 in culture supernatant were all increased by LPS (100 ng/mL), whereas reduced by 40 ng/mL TSA pretreatment (P < 0.05). TSA pretreatment inhibited LPS-induced up-regulation of TLR4 protein expression. Acetylation of NF-κB/p65(Lys310), which was already increased by LPS, was further enhanced by TSA (P < 0.05). On the contrary, LPS-increased DNA binding activity of NF-κB/p65 was decreased by pretreatment with TSA (P < 0.05). The results suggest that TSA-induced anti-inflammation may be attributed to decreases in the expression of TLR4 and DNA binding activity of NF-κB/p65.
Acetylation ; Animals ; Cell Line ; Hydroxamic Acids ; pharmacology ; Inflammation ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Lipopolysaccharides ; Macrophages ; drug effects ; metabolism ; Mice ; Signal Transduction ; Toll-Like Receptor 4 ; metabolism ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Up-Regulation

In this study, the current status for breast diseases in a region with high-incidence of cervical cancer were epidemiologically investigated. From March to August, 2009, 17618 women, from Wufeng area of Hubei province, China, were recruited to screen breast diseases by using breast infrared diagnostic apparatus. Other diagnostic methods, such as B-mode ultrasound, X-ray mammography, needle biopsy and pathological examination were, if necessary, used to further confirm the diagnosis. The screening showed that 5990 of 17618 cases (34.00%) had breast diseases, 5843 (33.16%) had mammary gland hyperplasia, 48 (0.27%) had breast fibroadenoma, 11 (0.06%) had breast carcinoma, and 88 (0.50%) had other breast diseases. The peak morbidity of breast cancer was found in the women aged 50-60 ages. The morbidity of breast cancer was significantly increased in women elder than or equal to 50 years old (n=8, 0.157%) in comparison with that in the subjects younger than 50 years old (n=3, 0.024%) (u=2.327, P<0.05). It was shown that the occurrence of breast diseases was concentrated in women aged 20-40 years, while the total morbidity reached its peak at the age of 30 years and then decreased sharply after age of 40. Compared with the patients elder than or equal to 40 years old (n=3289, 27.46%), the morbidity rate of breast diseases was significantly increased in women less than 40 years old (2648 cases, 47.18%; P<0.001). However, there was no significant difference in the morbidity of breast diseases between the age group of 20-29 years and that of 30-39 years (P=0.453), and both of them were high. There was no significant association between the morbidity of breast diseases and cervical cancer. Since the morbidity of breast diseases was higher among young women, more attention should be paid to the screening of breast diseases among young women for early diagnosis.

The ubiquitin-proteasome pathway is the principal mechanism for the degradation of short-lived proteins in eukaryotic cells. Recently, proteasome inhibitors have been shown to induce apoptosis in many kinds of human malignant cells. In this study, the mechanism of apoptosis induced by proteasome inhibitor in leukemic cells was examined. Evaluated by MTT assay, treatment of leukemic cells with Z-LLL-CHO, a reversible proteasome inhibitor, induced cell death in a dose-dependent manner. Appearance of the sub G(0)/G(1) fraction of cell cycle observed in flow cytometry assay suggested the induction of apoptosis, which was further proved by typical DNA ladder and morphological study. Western blot displayed the cleavage of bcl-2 into a shortened 22 kD fragment and the decrease in the levels of caspase-3 precursor. A highly sensitive colorimetric assay was employed and the elevation of caspase-3 activity was detected in both cell lines after treatment with Z-LLL-CHO. By comparison, these results showed that the leukemic cell line M-07e and KG-1a, which both express bcl-2 at a relative high level, had different susceptibility to undergo apoptosis induced by Z-LLL-CHO, which possibly due to their different levels of expression and activation of caspase-3 precursor, as well as their different degree of bcl-2 cleavage after treated by Z-LLL-CHO.

Background: It is well-established that serum testosterone in men decreases with age, yet the underlying mechanism of this change remains elusive. Methods: The expression patterns of Fancd2 opposite-strand (Fancd2os) in BALB/c male mice and testicular tissue derived cell lines (GC-1, GC-2, TM3, and TM4) were assessed using real-time polymerase chain reaction (RT-PCR), Western blot and immunofluorescence. The Fancd2os-overexpressing or knockdown TM3 cells were constructed by infecting them with lentivirus particles and were used to evaluated the function of Fancd2os. The testosterone production was measured using enzyme linked immunosorbent assay (ELISA) and the steroidogenic enzymes such as steroidogenic acute regulatory protein (StAR), P450 cholesterol side-chain cleavage (P450scc), and 3β-hydroxysteroid dehydrogenase (3β-HSD) were analysed using RT-PCR. The apoptosis of TM3 cells induced by ultraviolet light or testicular tissues was detected using flow cytometry, Western blot or dUTP-biotin nick end labeling (TUNEL) assays. Pearson correlation analysis was used to assess the correlation between the Fancd2os expression and TUNEL-positive staining in mouse testicular Leydig cells. Results: The Fancd2os protein was predominantly expressed in mouse testicular Leydig cells and its expression increased with age. Fancd2os overexpression inhibited testosterone levels in TM3 Leydig cells, whereas knockdown of Fancd2os elevated testosterone production. Fancd2os overexpression downregulated the levels of StAR, P450scc and 3β-HSD, while Fancd2os knockdown reversed this effect. Fancd2os overexpression promoted ultraviolet light-induced apoptosis of TM3 cells. In contrast, Fancd2os knockdown restrained apoptosis in TM3 cells. In vivo assays revealed that higher Fancd2os levels and mouse age were associated with increased apoptosis in Leydig cells and decreased serum testosterone levels. Pearson correlation analysis exhibited a strong positive correlation between the expression of Fancd2os and TUNEL-positive staining in mouse testicular Leydig cells. Conclusion Our findings suggest that Fancd2os regulates testosterone synthesis via both steroidogenic enzymes and the apoptotic pathway.