Objective To investigate in vitro anti-tumor activity and the related mechanism of four alkaloid monomers from Sophora alopecuroides,and to evaluate their toxicity on normal cells,thereby to screen the drugs with high efficiency and low toxicity anti-tumor effects.Methods Methyl thiazolyl tetrazolium (MTT) assay was employed to determine the effects of four alkaloid monomers on the proliferation activity of human lung cancer cells and hamster lung cells.Morphology of apoptotic lung cancer cell nucleus was observed by laser scanning confocal microscopy with Hochest33342 staining.Cell apoptosis and necrosis were measured by Annexin Ⅴ-FITC/PI double staining.Results The four alkaloid monomers showed different degree of proliferation inhibition on lung cancer cells with dose dependent manner.Oxysophoridine had the strongest inhibition effect on lung cancer cells and the minimun toxicity on normal lung cells.Hochest33342 staining and flow cytometry analysis demonstrated that oxysophoridine could induce the apoptosis and necrosis of A549 cells.Conclusions All four alkaloid monomers have certain anti-tumor effect on lung cancer cells.Oxysophoridine is the most active one with high efficiency and low toxicity characteristics,which is suitable for use as anti-tumor drug candidate for further development.

BACKGROUND:Pancreatic stelate cels transforming growth factor β1/Smads signaling pathway activation is probably a main molecular mechanism of pancreatic fibrosis. If this pathway can be blocked, the progression of fibrosis of tissues with chronic pancreatitis wil be inhibited. OBJECTIVE:To study the inhibitory effect of colchicine on transforming growth factor β1/Smads pathway in chronic pancreatitis rat models. METHODS:Healthy male Sprague-Dawley rats were randomly divided into colchicines-treated group and chronic pancreatitis group. After successful establishment of rat models of chronic pancreatitis, the rats in the colchicines-treated group were intraperitonealy injected with colchicine 150 μg/kg daily. The rats in the chronic pancreatitis group were intraperitonealy injected with equal volume of physiological saline daily. Pancreatic tissues were colected after 3 months. Hematoxylin-eosin staining was used to observe histopathological changes of pancreatic tissue. Immunohistochemical staining was used to detect the expressions of transforming growth factor β1 in pancreatic tissue. Western blot assay was utilized to detect the expressions of P-Smad2, P-Smad3 and α-SMA protein in pancreatic stelate cels. RESULTS AND CONCLUSION: Hematoxylin-eosin staining results revealed that compared with the colchicines-treated group, glandular tissue had reduced, while fibrous connective tissue and inflammatory cels had increased obviously and replaced the pancreatic gland tissue in the chronic pancreatitis group. Immunohistochemical staining results demonstrated that the expression levels of transforming growth factor β1 and the index of positive cels were significantly lower in the colchicines-treated group than those in the chronic pancreatitis group (P < 0.05). Western blot assay results revealed that the results of P-Smad2/β-actin, P-Smad3/β-actin andα-SMA/β-actin in pancreatic stelate cels were significantly lower in the chronic pancreatitis group than those in the colchicines-treated group (P < 0.05). Results suggested that colchicine could inhibit the activity of transforming growth factor β1/Smads pathway and pancreatic tissue fibrosis in chronic pancreatitis rats. Therefore, colchicine can be used as a new candidate therapeutic scheme for chronic pancreatitis fibrosis.

Objective To quantize the synergistic force in movement of upper limbs between shoulders and elbows. Methods The trans-verse forces of elbows and shoulders during movement were recorded in a healthy adult with an upper-limb-force-measuring plate form which comprised of 2 three-dimensional force sensors, respectively. Then he performed shoulder abduction/adduction and elbow extension/flexion at 100%, 75%, 50%and 25%of the maximum contraction force, respectively. The ratio of the active action force and the joint action force (named assessment index) was used to assess the synergistic degree of the forearm and the upper arm. Results In the shoulder abduc-tion motion, the assessment index decreased as the strength of active action decreased, meant interference of joint action increased. Howev-er, it was almost stable in the shoulder adduction, increased in the elbow extensionas, and was irregular in the elbow flexion, as the active ac-tion strength decreased, respectively. Conclusion It may be more difficult to control upper arm than the forearm.

Objective To investigate the effect of bifidobacteria on dextran sulphate sodium(DSS)- induced acute ulcerative colitis in mice.Methods Thirty BALB/C mice were randomly divided into nor- mal control group (n=10),0501 strain group (n=10) and c122 strain group (n=10).Fifty BALB/ C mice received 5% dextran sulphate sodium(DSS) for 7 days to induce ulcerative colitis.The mice were then divided to model group,negative control group(perfused with 0.9 NaCl solution ),positive control group(perfused with SASP of 20 mg/ml),DSS + 0501 strain group(perfused with 1?10~9 CFU/ml bifidobacteria 0501 strain solution and DSS + c122 strain group (perfused with 1?10~9 CFU/ml bifidobacteria c122 strain solution).All mice were sacrificed 9 days later.The colon specimens were measure by histoehemical staining with H-E.The expressions of interleukin-10 (IL-10) and its protein were detected by RT-PCR and immunohistochemistry respectively.Results The degree of colon inflam- mation in mice both in DSS+ 0501 strain and DSS+ c122 strain groups were aggravated and expressions of IL-10 mRNA and protein were reduced compared to model group.No colon inflammation was found in 0501 strain and c122 strain groups.Conclusion Some strain of bifidobaeteria may aggravate colon in- flammation in mice when mucosal harrier is destroyed.

Objective To study the interrelation between highly pathogenic avian influenza viral pneumonia(HPAIVP) and cellular factors interleukin-1?(IL-1?),interleukin-6(IL-6),interleukin-8(IL-8)and tumor necrosis factor-?(TNF-?) in HPAIVP.Me-thods Sixty Kunming mice were divided into 2 groups randomly:experiment group and control group,each group consisted of 30 mice.The highly pathogenic avian influenza virus was used to establish HPAIDP models.The expressions of serum IL-1?,IL-6,IL-8 and TNF-? in experiment group of different moment and control group were detected by enzyme linked immunosorbent assay(ELISA).Result The levels of serum IL-1?,IL-6 and TNF-? in experiment group were significantly higher than those in control group(Pa

Objective To investigate whether epithelial-mesenchymal transition (EMT)is involved in the anti-invasion and anti-metastasis effects of curcumin on MDA-MB-2 3 1 breast cancer cell line and further analyze the underlying mechanisms.Methods MTT method was used to detect the anti-proliferative effect of curcumin on MDA-MB-2 3 1 cell line induced by lipopolysaccharide (LPS).The morphological changes were determined by optical and transmission electron microscopy,respectively.The expressions of Vimentin,E-cadherin,NF-κB and Snail were analyzed using RT-PCR and Western blotting.Results Curcumin could inhibit the occurrence of LPS-induced EMT of MDA-MB-2 3 1 breast cancer cell line, decrease the expression of LPS-induced EMT marker Vimentin and increase the expression of E-cadherin, resulting in the inhibition of in vitro cell motility and invasiveness.These actions were mediated by inactivating NF-κB-Snail signaling pathways.Conclusion Our data provide a new perspective of the anti-invasion mechanism of curcumin,indicating that the effect is in part due to its ability to inhibit the EMT process.

OBJECTIVETo construct primary cultured granulosa cells model of Zi Gooses tansfected by alpha-enolase (ENO1) overexpression adenovirus vector, and to detect the effect of ENO1 overexpression of granulose cells on progesterone secretion.

METHODSGranulosa cells were infected with Ad-CMV-ENO1 in gradient multiplicity of infection(MOI) levels:100, 250, 350 and 400 pfu/cell. Twenty four hours and 48 h after infection, green fluorescent protein (GDP) was respectively detected by fluorescence inverted microscopy. The effect of ENO1 overexpression of granulose cells on progesterone secretion was detected by the step double antibody sandwich enzyme-linked immunosorbent assay (ELISA).

RESULTSThe optimal infection rate (100%) was achieved when MOI was 800 pfu/cell,48h after infection. Real time RT-PCR and Western blot showed that the level of mRNA and protein expression ENO1 were increased significantly after infection (P < 0.01); The granulosa cells progesterone secretion of Ad-CMV-ENO1 group increased signigicantly (P < 0.01).

CONCLUSIONENO1 overexpression could make the primary culture follicle granulosa cells in vitro improve progesterone secretion.

Adenoviridae ; Biomarkers, Tumor ; genetics ; Cell Line ; Cells, Cultured ; DNA-Binding Proteins ; genetics ; Female ; Genetic Vectors ; Granulosa Cells ; metabolism ; Green Fluorescent Proteins ; genetics ; Humans ; Ovarian Follicle ; cytology ; Phosphopyruvate Hydratase ; genetics ; Progesterone ; secretion ; RNA, Messenger ; genetics ; Transfection ; Tumor Suppressor Proteins ; genetics

Objective To verify the reference interval for D-dimer assay and analyze the influence of age and gender on the ref-erence interval.Methods Inclusion criteria for reference individuals were established.60 healthy males and 63 females were enrolled and divided to three groups by age,including 20 to 39 years old group (20 males and 20 females),40 to 59 years old group (20 males and 23 females)and above 60 years old group (20 males and 20 females).Blood samples were drawn in cit-rate sodium anticoagulated tubes and D-dimer concentration was determined by three different coagulation analyzers using o-riginal reagents.According to CLSI guideline C-28-A3,the reference interval for each measurement system from reagent manufacturer was verified and the difference of D-dimer concentration between different age-group and sex-group was ana-lyzed using non-parameters tests.Results All reference intervals were verified for people under age 40,while one reference interval cannot be verified for people from 40 to 59 years old as same as one for people above 60 years old.D-dimer concen-tration increased with age and there was significantly different between 20~39 years old group and 40~59 years old group or above 60 years old group(P<0.05).There was only a significant difference between sex-group for people under age 60(P<0.05).Conclusion D-dimer concentration was associated with age and sex.For people under age 40,the reference inter-val from reagent manufacture can be verified and directly used in laboratory,while for people above age 60,the reference in-terval from reagent manufacture cannot be verified.The cause should be investigated and a new reference interval should be established separately when necessary.