Objective: To study the effect of light quality on the growth as well as triterpenic acid content and component of Ganoderma lucidum mycelium. Methods: Light-emitting diode (LED) was used as sources of light, the growth index, content and component of triterpenic acids in G. lucidum mycelium cultured under different LED conditions were investigated along with culture time. Results: The growth rate, morphology, biomass, total contents and components of triterpenic acid in G. lucidum mycelium varied under different lights. The mycelium cultured under blue light maintained stable growth during the whole period; Under red light, both growth rate and biomass of mycelium were the highest in early time, subsequently, decreased sharply and grew slowly. Under green light, both growth rate and biomass of mycelium were the lowest in early and medium phases, while increased significantly later, especially dry matter. Triterpenic acids were highly produced on the day 7 of cultivation, while decreased in content and increased in types. Green light and blue light had a distinct advantage of improving the type and content triterpenic acids in G. lucidum. Conclusion: Light quality affects the growth as well as the content and component of triterpenic acids in G. lucidum mycelium, and blue light irradiation is the best for the steady growth and triterpenic acids accumulation.

Objective:To study the influence of morphine on the expression of nestin in the ependymal epithelia,central gray and hippocampal formations in mice.Methods:Twenty health mice were evenly randomized into control group and experiment group.Mice in the control group were injected with normal saline(0.1 ml daily)and those in the experimental group were injected with morphine (0.1 ml,1 mg daily).Thirty days later,the mice brain samples were harvested and made into paraffin sections.Immumohistochemical ABC technique was used to observe the expression of nestin under light microscope.The images were analyzed with the image analytical system.Results:In the control group,the ependymal epithelia,the central gray,the periventricular gray substances and the hippocampal formations had weak expression of the nestin,with a mean gray scale of 150.98?13.31;there were 5 kinds of nestin-positive cells:(1) the basal cells of ependymal epithelium,(2)cells distributed in the periventricular gray substance and the deep lamella of central gray, (3)cells distributed in the superficial lamella of central gray,the subiculum,the parahippocampal gyrus and the cortex inⅡ,Ⅲlayers of the entorhinal area,(4)cells frequently seen in the rectum of midbrain and the subiculum,and(5)cells distributed in the tectum of midhrain,the hippocampus,gyrus dentatus,parahippocampal gyrus and the cortex in V layer of the entorhinal area;the density of nestin in the subiculum and entorhinal area was(7.20?1.23)mm~2.In the experiment group,the ependymal epithelia,the central gray,the periventricular gray substances and the hippocampal formations had positive expression of the nestin,with the mean gray scale being 133.03?22.28;the density of the above-mentioned 5 kinds of cells increased;the density of nestin in the subiculum and entorhinal area was(10.50?1.43)mm~2.The mean value of gray scale and nestin-positive neurons were significantly different between the 2 groups (P

In the current study, nine flavonoids were isolated and purified from 75% ethanol extract of Selaginella uncinata (Desv.) Spring by column chromatographic techniques over macroporous resin, polyamide, silica gel, Sephadex LH-20 and pre-HPLC. On the basis of their physico-chemical properties and spectroscopic data analyses, these compounds were elucidated as cirsimarin (1), nepitrin (2), apigenin-6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (3), apigenin-6-C-β-D-glucopyranosyl-8-C-α-L-arabinopyranoside (4), apigenin-7-O-β-D-glucopyranoside (5), 2,3-dihydroamentoflavone (6), 4'-O-methylamentoflavone (7), 2,3-dihydro-4'-O-methyl-amentoflavone (8), and 2,3,2",3"-tetrahydron-4'-O-methyl-robustaflavone (9). Compounds 1-5 belong to flavonoid glycosides and were isolated from the genus Selaginella for the first time.
Flavonoids ; analysis ; Selaginellaceae ; chemistry

OBJECTIVETo observe the effects of deguelin on the apoptosis and proliferation of human esophageal cancer cell Ec-109, and to explore its possible mechanisms.

METHODSHuman esophageal cancer cells Ec-109 were in vitro cultured. They were divided into the blank control group, and 5, 10, 20, and 40 nmol/L deguelin groups. The inhibition on the proliferation was detected at 24, 48, and 72 h using CCK-8 assay. The early apoptosis rate at 24 h was detected by flow cytometry. The expressions of apoptosis-related proteins Bcl-2 and Bax were detected at 24 and 48 h respectively.

RESULTSCompared with the blank control group at the same point, the growth inhibition rate in all deguelin groups increased at 24, 48, and 72 h, showing statistical difference (P <0.05). The early apoptosis rate was 4.37% +/- 0.35%, 6.71% +/-0.14%, 15.62% +/- 0.21%, and 19.78% +/- 0.15% in 5, 10, 20, and 40 nmol/L deguelin groups, respectively, showing statistical difference when compared with that of the blank control group (1.10% +/- 0.08%, P < 0.05). Compared with the blank control group, Bcl-2 protein expression obviously decreased, and Bax protein expression obviously increased in 10, 20, and 40 nmol/L deguelin groups, showing statistical difference (P <0.05). The aforesaid indices were in time- and dose-dependent manners.

CONCLUSIONDeguelin showed obvious effects on inhibiting the proliferation of Ec-109 cells and promoting their apoptosis, which was correlated with up-regulating Bax protein expression and down-regulating Bcl-2 protein expression.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Esophageal Neoplasms ; pathology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rotenone ; analogs & derivatives ; pharmacology ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo observe the effect and mechanism of curcumin derivative B06 on kidney from rats with hyperlipidemia and type 2 diabetes.

METHODSThirty five male SD rats were randomly divided into five groups(n = 7): the normal control group, high-fat group, high-fat + B06-treatd group, diabetic group, diabetic + B06-treated group. After fed with high-fat diet for 4 weeks, the later two groups were in- jected with streptozotocin intraperitoneally to induce type 2 diabetes mellitus. B06-treated groups were given B06 by gavage at a dosage of 0.2 mg/kg . d for 8 weeks. After the treatment, the serum creatinine, blood urea nitrogen and uric acid were detected biochemically, the morphology of kidney was observed with light and transmission electron microscopy, the expression of collagen fibers was observed with Masson staining, the protein expression of collogen IV and fibronectin in kidney were determined by Immunohistochemistry.

RESULTSIt was showed that the levels of the serum creatinine and blood urea nitrogen elevated significantly in diabetic group. In high-fat and diabetic groups, increased glomerular mesangial matrix and collagen fiber and thicken glomerular basal membrane were observed under light microscopy, swelling and fusion of foot process were found under electron microscope; increased green matrix within glomeruli was observed under Masson staining. collogen IV and fibronectin protein expression were significantly enhanced in high-fat group and diabetic group. After B06's intervention, the levels of serum creatinine and blood urea nitrogen were decreased in diabetic groups, the morphological change of kidney was obviously relieved, Collogen IV and fibronectin protein expression reduced.

CONCLUSIONCurcumin derivative B06 exerts a protective effect on kidney in type 2 diabetic rats, reduced expressions of collogen IV and fibronectin, inhibition of the accumulation of extracellular matrix and glomerular mesangial proliferation, and then prevention of renal fibrosis may be the mechanism.

Animals ; Blood Urea Nitrogen ; Collagen Type IV ; metabolism ; Creatinine ; blood ; Curcumin ; pharmacology ; Diabetes Mellitus, Experimental ; complications ; drug therapy ; Diabetes Mellitus, Type 2 ; Drugs, Chinese Herbal ; Fibronectins ; metabolism ; Kidney ; metabolism ; physiopathology ; Kidney Diseases ; drug therapy ; Male ; Rats ; Rats, Sprague-Dawley ; Streptozocin ; Uric Acid ; blood

By using a yeast two-hybrid system, a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins, and its effects on the growth of yeast cells and the activation of reporter genes were investigated. Total mRNA extracted from Hela cells was reversely transcribed into cDNA. Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector. The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector were transformed into the yeast cell AH109, respectively. After they were cultured respectively in YPDA liquid medium and nutrition-deficient culture medium, their toxicity and transcriptional activation were tested by both the phenotype assay and the color assay. The bait plasmid HPV18 E6 was successfully obtained. After being cultured in YPDA liquid medium for 16h, the A (600 nm) values of two yeast fluids were 0.98+/-0.03 and 0.99+/-0.02, respectively. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector could grow to white colonies on SD/-Trp/X-alpha-gal plates, while no colony could survive on SD/-His/-Trp/X-alpha-gal, SD/-Ade/-Trp/X-alpha-gal plates, indicating that the bait plasmid pGBKT7-HPV18 E6 was constructed successfully and expressed correctly, and could not activate the transcription of reporter gene alone. The yeast two-hybrid GAL4 system 3 can be utilized to find HPV18 E6 interacting proteins.

Objective To compare the similarities and the differences of nursing care on patients with idiopathic thrombocytopenic purpura. Underwent laparoscope splenectomy and open splenectomy. Methods A total of 122 cases admitted from October 1996 to September 2008 were selected and investigated prospectively.Among them, 72 cases were underwent laparoscope spleneetomy, the other 50 cases were accepted open splenectomy. Indexes of the recovery after operation were compared. Results The mean operation time was longer in the laparoscopic splenectomy group than that in the open group ( 135.3 min vs 108.5 min,P ＜0. 05).The laparoscopic group decreased more significantly than the open group in blood loss ( 110 ml vs 185 ml),abdominal drainage volume ( 100 ml vs 230 ml), the off-bed ambulation time(26. 2 h vs 46.9 h), the anal aerofluxus time(28.9 h vs 68. 1 h) ,food intake time(32. 2 h vs 72.3 h), and post operative hospitalization (8.5 d vs 15. 1 d). Postoperative pain was significantly less in LS group ( P ＜ 0. 05 ). There were no differences in postoperative complication, treatment effectiveness and temperature between two groups (P ＞ 0. 05 ).Conclusions Laparoscope splenectomy, whereas of less traumatic and low morbidity, results in comparable effects as open splenectomy for the treatment of idiopathic thrombocytopenic purpura.. It has important significance to know both similarities and differences of clinical nursing care for patients undergoing the two ways of splenectomy, in order to enhance the nursing quality for peri-operative patients with splenectomy via laparoscope and promote their postoperative rehabilitation.

To investigate the immune regulatory effects of human bone marrow mesenchymal stem cells on alloantigen T lymphocyte in vitro, human MSCs were isolated and expanded from bone marrow cells, and identified with cell morphology, and the phenotypes were assessed by immunohistochemistry and flow cytometry. As the stimulation factor of T lymphocytes proliferation, either PHA or dendritic cells isolated from cord blood were cocultured with CD2(+) T lymphocytes from peripheral blood mononuclear cells by magnetic beads with or without MSC in 96-well plats for seven days. T cell proliferation was assessed by [(3)H]-thymidine incorporation using a liquid scintillation counter. T cell subsets, Th1, Th2, Tc1 and Tc2 were analyzed by flow cytometry after co-culture of CD2(+) T cells with MSCs for 10 days. The results showed that a significant decrease of CD2(+) T cell proliferation was evident when MSC were added back to T cells stimulated by DC or PHA, and an increase of Th2 and Tc2 subsets were observed after co-culture of MSC with T lymphocytes. It is suggested that allogeneic MSC can suppress T cell proliferation in vitro and the cause of that was partly depend on interaction of cells and the alteration of T cell subsets.
Bone Marrow Cells ; cytology ; immunology ; CD2 Antigens ; immunology ; Cell Communication ; immunology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Flow Cytometry ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; cytology ; immunology ; T-Lymphocyte Subsets ; cytology ; immunology ; T-Lymphocytes ; cytology ; immunology

Objective To observe the clinical efficacy of warm needling plus Chinese medication for external application in treating post-stroke shoulder pain. Method Two hundred patients with post-stroke shoulder pain were randomized into a treatment group and a control group, 100 cases each. The two groups both received rehabilitation training for shoulder joint. In addition, the treatment group was given warm needling plus Chinese medication for external application, while the control group was given warm needling. Visual Analogue Scale (VAS) for pain, upper-limb Fugl-Mayer Assessment (FMA) and Modified Barthel Index (MBI) were adopted to evaluate the two groups before and after the treatment. The clinical efficacies of the two groups were also compared. Result The total effective rate was 100.0％ in the treatment group versus 87.0％ in the control group, and the between-group difference was statistically significant (P<0.01). The VAS, FMA and MBI scores were significantly changed after the treatment in both groups (P<0.01). After the treatment, the VAS, FMA and MBI scores of the treatment group were significantly different from those of the control group (P<0.05, P<0.01). Conclusion Warm needling plus Chinese medication for external application and rehabilitation training can obviously reduce post-stroke shoulder pain, and enhance the upper-limb motor function and activities of daily living.

AIMTo study the effects of cryopreservation length on the proliferative potential of hematopoietic cells derived from cord blood.

METHODSUsing Dextran-40 and 10% DMSO as cryoprotectants, separated nuclear cells were stored in liquid nitrogen after they were freezed according programme. One month or 4 months later, they were thawed and expanded in serum-free medium for culture and expansion of hematopoietic cell (SFEM) for 5 weeks. Dynamic results were detected every week.

RESULTSAt the 5th week of expanding, TNC were expanded for 1499.0 +/- 115.6-folds and 1513.0 +/- 110.4-folds, respectively. CD34+ cells and CFCs reached to their highest level at the 2nd week and at the 3rd week. CD34+ cells were expanded for 63.8 +/- 6.1-folds and 62.4 +/- 5.7-folds, respectively. CFCs were expanded for 53.8 +/- 6.3-folds and 54.8 +/- 6.7-folds, respectively. Between the two kinds of cells, statistical significant difference in proliferative potential wasn't detected.

CONCLUSIONIn ideal cryopreservative condition, the cryopreservation length would do not affect the proliferative potential of cord blood hematopoietic cells.

Cell Proliferation ; Cell Survival ; Cells, Cultured ; Cryopreservation ; methods ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Time Factors