Objective: To study the effects of extremely low frequency (ELF) magnetic field with fixed parameters on human liver cancer cells (SK-HEP-1) at different aspects. Methods: SK-HEP-1 cells were exposed to 50Hz, 20mT magnetic field during the whole culture process, and then proliferation activity, growth kinetics, metabolic profile and cell cycle were analyzed. Results: 50Hz, 20mT magnetic field inhibits the growth and metabolism of SK-HEP-1 cells, and hampers their mitotic division. Conclusion: 50Hz, 20mT magnetic field could be a potential therapy in the treatment of human malignant tumors.

Objective To investigate the influence of medicated cerebrospinal fluid of modified Sanjia San on inflammatory reactions of BV2 microglia cells induced by lipopolysaccharide (LPS).Methods BV2 microglia cells were divided into blank control group, model group, three modified Sanjia San groups and huperzine group.After the cells cultured and adhered to wall, modified Sanjia San groups and huperzine group were, respectively, given medicated cerebrospinal fluid (10.0%, 5.0%, 2.5%, 10.0%) and incubated at 37 ℃ for 2 h.Except of blank control group, all groups were given LPS (1 mg/L as final concentration) and incubated continuously for 24 h, and then supernatant was collected for extracting protein samples.The content of nitric oxide (NO) in supernatant was detected by using nitrate reductase method in all groups.The content of inducible nitric oxide synthase (NOS), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in supernatant were detected by using enzyme-linked immunosorbent assay (ELISA) in all groups.The expressions of c-Jun N-terminal kinase (JNK) and phospho-c-Jun N-terminal kinase (p-JNK) were detected by using Western blot test in all groups.Results The content of supernatant NO, iNOS, IL-1β and TNF-α increased significantly, and expression of p-JNK was up-regulated in model group.The content of supernatant NO, iNOS, IL-1β and TNF-α decreased significantly, and expression of p-JNK was down-regulated in 10.0% and 5.0% modified Sanjia San groups.Conclusion The medicated cerebrospinal fluid of modified Sanjia San has the regulative effect on inflammatory reactions of BV2 microglia cells induced by LPS, and the mechanism may be related to the controlling of p-JNK expression and reducing of generation of inflammatory media.

China ; epidemiology ; Female ; Humans ; Liver Diseases, Alcoholic ; epidemiology ; etiology ; Male ; Obesity ; complications ; Risk Factors ; Sex Factors

OBJECTIVETo explore the effect of hypoxia on the fibrosis of corpus cavernosum smooth muscle cells (CCSMC) in SD rats.

METHODSCCSMCs were cultured in vitro, identified by immunohistochemistry, and then exposed to hypoxia at the concentration of 1% O2 for 12, 24, 48 and 72 hours. Those exposed to normal oxygen concentration for the corresponding lengths of time were used as the control. The relative expressions of TGF-131, type I collagen and type DI collagen were determined by RT-PCR.

RESULTSThe in vitro cultured CCSMCs grew well, and the anti-a-smooth muscle actin monoclonal antibodies were positive on immunohistochemical staining. The relative expression levels of TGF-beta1, type I collagen and type mI collagen were positively correlated with the time of hypoxia interference within 48 hours, and did not increase further with prolonged exposure.

CONCLUSIONWhen exposed to hypoxia, the relative expressions of TGF-beta1, type I collagen and type mI collagen in the CCSMCs of SD rats increased with the length of time, and reached the peak at 48 hours. Hypoxia can cause fibrosis of CCSMCs in SD rats.

Animals ; Cell Hypoxia ; Cells, Cultured ; Collagen Type I ; metabolism ; Extracellular Matrix ; Fibrosis ; Male ; Muscle, Smooth ; cytology ; pathology ; Oxygen ; metabolism ; Penis ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism

In this study, 17 kinds of Dendrobium species of Fengdous including 39 individuals were collected from 4 provinces. Mitochondrial gene sequences co I, nad 5, nad 1-intron 2 and chloroplast gene sequences rbcL, matK amd psbA-trnH were amplified from these materials, as well as nrDNA ITS. Furthermore, suitable sequences for identification of Dendrobium species of Fengdous were screened by K-2-P and P-distance. The results showed that during the mentioned 7 sequences, nrDNA ITS, nad 1-intron 2 and psbA-trnH which had a high degree of variability could be used to identify Dendrobium species of Fengdous. However, single fragment could not be used to distinguish D. moniliforme and D. huoshanense. Moreover, compared to other combined fragments, new type combined fragments nrDNA ITS+nad 1-intron 2 was more effective in identifying the original plants of Dendrobium species and could be used to identify D. huoshanense and D. moniliforme. Besides, according to the UPGMA tree constructed with nrDNA ITS+nad 1-intron 2, 3 inspected Dendrobium plants were identified as D. huoshanense, D. moniliforme and D. officinale, respectively. This study identified Dendrobium species of Fengdous by combined fragments nrDNA ITS+nad 1-intron 2 for the first time, which provided a more effective basis for identification of Dendrobium species. And this study will be helpful for regulating the market of Fengdous.
DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; classification ; genetics ; Genes, Chloroplast ; Genes, Plant ; Introns ; Plants, Medicinal ; classification ; genetics

OBJECTIVETo investigate the initial changes in the gut microenvironment that accompany intestinal endotoxemia related to alcoholic fatty liver disease (ALD) in order to explore the potential initiating factors and to observe the effect of probiotic therapy on these factors.

METHODSFifty Sprague-Dawley male rats were randomly divided into an ALD model group (alcoholic intragastric administration), an intervention group (ALD with probiotic intragastric administration), and a control group (physiological saline intragastric administration). Histological changes of the liver were evaluated using hematoxylin-eosin staining and light microscopy. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and triglycerides (TG), and plasma endotoxin and coli bacillus were determined. The structural integrity of intestinal mucosa and tight junctions were observed by transmission electron microscopy. Occludin protein expression in intestinal epithelial cells was detected by immunohistochemistry.

RESULTSAfter four weeks, the three groups showed significant differences in the plasma endotoxin levels [control: (0.67+/-0.14) pg/ml, model: (4.42+/-1.28) pg/ml, and intervention: (2.88+/-0.83) pg/ml; F = 27.288, P = 0.000] and numbers of Escherichia coli [control: (2.31+/-0.39) lg3/ml, model: (3.23+/-0.41) lg3/ml, and intervention: (2.24+/-0.44) lg3/ml; F = 10.692, P = 0.001]. The plasma endotoxin level and E. coli number were significantly higher in the model group than in the control group and the intervention group (all P less than 0.05). The three groups showed no significant differences in the levels of ALT, AST, and TG at four weeks. After eight weeks, however, all three serum markers were significantly different between the three groups [ALT: control: (62.33+/-7.12) U/L, model: (95.50+/-8.73) U/L, and intervention: (81.33+/-6.19) U/L; F = 18.051, P = 0.000]; [AST: control: (90.50+/-10.67) U/L, model: (130.00+/-14.91) U/L, and intervention: (110.33+/-7.26) U/L; F = 30.170, P = 0.000]; [TG: control: (0.84+/-0.84) mmol/L, model: (1.40+/-0.17) mmol/L, and intervention: (1.10+/-0.17) mmol/L; F = 10.592, P = 0.001]. In addition, the three groups showed significant differences in E. coli number [control: (2.23+/-0.46) lg3/ml, model: (4.81+/-0.29) lg3/ml, and intervention: (3.61+/-0.50) lg3/ml; F = 23.579, P = 0.000] and plasma endotoxin level [control: (0.52+/-0.21) pg/ml, model: (12.46+/-2.61) pg/ml, intervention: (6.83+/-1.74) pg/ml; F = 30.731, P = 0.000]. The levels of ALT, AST, TG and endotoxin, and the number of E. coli were all significantly higher in the model group than in the control group and the intervention group (all P less than 0.05). Small intestinal epithelial cell structural failure was more apparent and intercellular gaps more broad after eight weeks than after four weeks for all three groups. However, the intervention group showed clearer cell connection structures and less extensive cell gap broadening than the model group at eight weeks. After eight weeks, the occludin protein had become significantly down-regulated and distributed in a non-continuous pattern in the model group, as compared with the control group. However, the occludin protein expression was higher in intervention group than in the model group.

CONCLUSIONIntestinal endotoxemia related to perturbations in the microenvironment occurs in the early phase of ALD, and the increased intestinal permeability appears to be the initial factor of elevated plasma endotoxin, which may lead to liver damage. Probiotic therapy can reduced plasma endotoxin levels and postpone ALD progression by altering the composition of the gut microbiota and up-regulating expression of the occludin protein in intestinal epithelial cells.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Endotoxins ; blood ; Escherichia coli ; isolation & purification ; Fatty Liver, Alcoholic ; microbiology ; therapy ; Intestinal Mucosa ; metabolism ; microbiology ; Intestine, Small ; metabolism ; microbiology ; Male ; Occludin ; metabolism ; Probiotics ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Triglycerides ; blood

OBJECTIVETo investigate the changes in circulating bone marrow stem cells in pregnant rabbits after AMI (AMI) and their relationship with estradiol.

METHODSThree groups of rabbits were used, namely pregnancy and AMI group, AMI group without pregnancy, and sham operation group with pregnancy. The ratio of CD90(+) cells in the peripheral blood was determined with flow cytometry in all the rabbits, and serum estradiol level measured. Four weeks after AMI, hemodynamic measurements were carried out. The morphological changes of the myocardial tissues were examined with ImageJ 1.31.

RESULTS AND CONCLUSIONFour weeks after AMI, the two pregnancy groups showed a higher Left ventricular end systolic pressure(LVESP) and+dp/dtmax, lower left ventricular end-diastolic pressure (LVEDP) and -dp/dtmax and high levels of CD90(+) cells in peripheral blood than AMI group without pregnancy (P<0.01). The ratio of circulating CD90(+) cells increased gradually with gestational age and peaked at the end stage of pregnancy. After delivery the circulating CD90+ cell ratio decreased sharply, showing a significant correlation with serum estradiol level (r=0.725, P<0.01). Four weeks after AMI, the pregnancy group had smaller myocardial infarction (MI) volume than the non-pregnant group (22.17+/-6.34% vs 38.86+/-5.97%, P<0.05). Circulating bone marrow stem cells increased during pregnancy with gestational age and peaked at the end stage of pregnancy. Ten days after delivery, the stem cells resumed basically the normal level. The proportion of circulating bone marrow stem cells was significantly correlated with the level of serum estradiol during pregnancy, and mobilization of the bone marrow stem cells induced by acute ischemic event in pregnant rabbits was advanced. 4 weeks after AMI, the pregnant rabbits showed better heart contraction and diastolic function than the non-pregnant ones.

Animals ; Bone Marrow Cells ; cytology ; Estradiol ; blood ; Female ; Hematopoietic Stem Cells ; cytology ; Myocardial Contraction ; Myocardial Infarction ; blood ; physiopathology ; Pregnancy ; Pregnancy Complications, Cardiovascular ; blood ; Rabbits ; Thy-1 Antigens ; blood ; Time Factors

Bletilla striata has been used as traditional Chinese medicine for several centuries. In recent years, the quality and quantity of wild B. striata plants have declined sharply due to habitat deterioration and human over-exploitation. Therefore, it is of great urgency to evaluate and protect B. striata wild plant resource. In this study, sequence-related amplified polymorphism (SRAP) markers were applied to assess the level and pattern of genetic diversity in twelve populations of B. striata. The results showed a high level of genetic diversity (PPB = 90.48%, H = 0.349 4, I = 0.509 6) and moderate genetic differentiation among populations (G(st) = 0.260 9). Based on the unweighted pair-group method with arithmetic average (UPGMA), twelve populations gathered in three clusters. The cluster 1 included four populations. There are Nanjing, Zhenjiang, Xuancheng and Hangzhou. The seven populations which come from Hubei Province, Hunan Province, Jiangxi Province and Guizhou Province belonged to the cluster 2. The cluster 3 only contained Wenshan population. Moreover, Mantel test revealed significant positive correlation between genetic distances and geographic distances (r = 0.632 9; P < 0.000 1). According to the results, we proposed a series of conservation consideration for B. striata.
China ; Genetic Markers ; Genetic Variation ; Genetics, Population ; Orchidaceae ; genetics ; Phylogeny ; Plants, Medicinal ; genetics

p53 (encoded by TP53) is undoubtedly one of the most extensively studied genes and proteins. It is a highly potent transcription factor which, under normal circumstances, is maintained at low level. Both genotoxic and non-genotoxic stresses can induce p53 stabilized leading to changes in the expression of p53-responsive genes. The biological outcome inducing this pathway can be either growth arrest and apoptosis or senescence to maintain the integrity of the genome or to delete the damaged cells. The biochemical activity of p53 itself and the cellular environment govern the choice between these outcomes in a cell type- and stress-specific manner. So, p53 is a pivotal tumour suppressor and a mainstay of our body's natural anticancer defence. This review could provide some useful information for further study on the mechanisms of tumorigenesis and its progression, and also could contribute to the discovery of antitumor agents.
Animals ; Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; DNA Damage ; DNA Repair ; Genes, p53 ; Humans ; Proto-Oncogene Proteins c-mdm2 ; metabolism ; Signal Transduction ; Tumor Suppressor Protein p53 ; genetics ; physiology

Objective To observe the effects of Langchuangqing granule on the Forssman cutaneous vasculitis reaction in guinea pigs , the antibo-dies, complements and immunoglobulin levels in MRL/lpr mice and the antibodies levels in the spleen lymphocytes.Methods In the forssman cutaneous vasculitis reaction , the immune serums were intradermally in-jected to the back skin of guinea pigs followed by tail vein injection of 0.5%evans blue.The skins dyed evans blue showed the model estab-lished.The guinea pigs were divided into five groups:model group , con-trol group and high-dose, middle-dose, low-dose ( Langchuangqing granule,18.27, 9.13, 4.56 g· kg -1) of test groups.The optical densi-ty( OD) of the skin was tested after continuously intragastric administra-tion for 8 days.In the lupus experiment in MRL/lpr mice, the models were established naturally with the mice growing up.The mice were divided into six groups: blank group , model group , control group and high-dose, middle-dose, low-dose(Langchuangqing granule,45.67, 22.83, 11.42 g· kg -1) of test groups.The blood was taken in all groups after intragastric administration for 6 weeks.Then the anti -double stranded DNA anti-body(dsDNA), antinuclear antibody (ANA), complement 3(C3), complement 4(C4), immunoglobulin M(IgM), immunoglobulin G(IgG) levels were measured by enzyme -linked immunosorbent assay (ELISA)method.In the spleen lymphocytes experiment , the dsDNA and ANA levels were measured in the lymphocyte supernatant after the effects of serum containing Langchuangqing granule on spleen lymphocytes.Results In the forssman cutaneous vasculitis reac-tion, compared with model group , the OD of the skin was significantly reduced in high dose test group in guinea pigs ( P<0.05 ).In the lupus experiment in MRL/lpr mice , compared with model group , high dose of langchuangqing granule significantly reduced dsDNA[(47.25 ±6.05) vs (37.51 ±3.26) ng· L-1] and IgM levels, improved C3 lev-els(P<0.05, P<0.01).Also in the middle dose group, IgM levels notably reduced and C3 increased(P<0.05,P<0.01 ).In the spleen lymphocytes experiment , the dsDNA levels in lymphocyte supernatant significantly reduced af-ter treated with the 20%drug-containing serum ( P<0.05 ).Conclusion The effects of Langchuangqing granule on the systemic lupus erythematosus may be related to the immunological functions of Autoantibodies , complement , immunoglobulin.