Objective To detect the effect of cell density off both integrin αγβ6 expression and matrix metalloproteinase-9(MMP-9)secretion in colon cancer cells. Methods Flow cytometry was applied to analyze αγβ6 expression in human WiDr colon cancer cell lines and human HaCaT keratinocyte cells, respectively,at high-and low-cell density culture.The MMP-9 aetivity level for various coloIl cancer cell lines, WiDr and SW480 cells at high-and low-cell density culture was analyzed using Biotrak MMP-9 activity assay and Gelatin Zymography assay, respectively. Results High cell density significantly enhances integrin αγβ6 expression for WiDr cells expressing αγβ6 compared with low density,but no increase was observed for human keratinocyte HaCaT cells. Biotrak MMP-9 assay indicated that the amount of MMP-9 secreted per cell for WiDr and SW480 B6 cells at high cell density culture was(3.3±1.2)×10-7ng/cell and(27.2±3.0)× 10-7ng/cell respectively; However,at low cell density it was(1.8±0.7)× 10-7ng/cell and(10.9±2.0)×10-7 ng/cell,respectively. It was 2-3-fold higher for WiDr and SW480 β6 cells at high cell density compared with that at low cell density, but no density-dependent increase observed for SW480 wild cells lack αγβ6 expression(t=0.47,P＞0.05),MMP-9 secretion for SW480 wild cells was(3.9±1.7)× 10-7 ng/cell at hish cell density and(3.8 ±0.7)×10-7 ng/cell at low eell density(P＞0.05),respectively. Gelatin zymography assay also indicated that the level of MMP-9 in SW480 B6 cells expressing αγβ6 was evidently higher at high density than at low density, however no density-dependent increase observed for SW480 wild cells and HaCaT cells. Conclusions High cell density induces integrin αγβ6 expression and promotes MMP-9 secretion in colon cancer cells, which constitutes the basis for a self-perpetuating system of tumor infiltrating growth in colon cancer progression.

Objective To investigate the in vitro inhibitory effects of regulatory T cells ( Treg ) from unpregnant mice and pregnancy-induced regulatory T cells ( piTreg) on the proliferation of na?ve T cells and their differences .Methods The numbers of piTreg cells from allogeneic pregnant mice ( C57/B6 fe-male×BALB/c male) on day 12.5 (E12.5d) of gestation and Treg cells from unpregnant C57/B6 mice were detected respectively by flow cytometry .The percentages of piTreg cells and Treg cells in CD 4+T cells of age-matched female mice and their intracellular expression of Foxp 3 were analyzed .The in vitro inhibitory effects of piTreg cells and Treg cells on the CFSE-labeled na?ve T cells ( effector cells ) were compared in a one-way mixed lymphocyte culture system using mitomycin C-inactivated CD4-T cells as stimulator cells . Results The level of piTreg cells in splenic mononuclear cells was significantly higher than that of Treg cells (P<0.001) from normal mice.Foxp3 was highly expressed in both piTreg cells and Treg cells , howev-er slightly increased in piTreg cells .Moreover , piTreg cells had a significant stronger in vitro inhibitory effect on na?ve T cells proliferation than that of Tregs cells (P<0.006), which was in a cell-dependent manner. Conclusion The present study suggests that the piTreg cells have a stronger inhibitory effect on na ?ve T cell proliferation as compared with Terg cells from unpregnant mice , The differential activity of CD 4+CD25+Treg might be mediated by the paternal antigens during pregnancy .

Eleven compounds were isolated and purified from the barks extract of Nothopanax delavayi and their structures were identified as serratagenic acid-3-O-alpha-L-arabinopyranosyl-28-O-beta-D-glucopyranosyl ester (1), serratagenic acid-3-0-alpha-L-arabi-nopyranosyl-28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester (2), serratagenic acid (3), serratagenic acid-3-O-alpha-L-arabinopyranoside (4), serratagenic acid-beta-O-beta-(2', 4'-O-diacetyl) -D-xylopyranosyl-28-O-[alpha-L-rhamnopy-ranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->46)-beta-D-glucopyranosyl] ester (5), serratagenic acid-3-O-alpha-(4'-O-acetyl)-L-arabino pyrano-syl-28-0- [-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester(6), serratagenic acid-3-O-alpha-(2'-O-acetyl)-L-arabinopyranosyl-28-O-[-alpha-L-rhamnopyranosyl- (1-->4) -beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester(7), serratagenic acid-3-0-beta-D-xylopyranosyl-28-O-[-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester (8), protocatechuic acid (9), ethyl caffeate (10) and caffeic anhydride (11) by physicochemical properties and spectroscopic data analysis. Among them, compounds 3-4 and 9-11 were firstly isolated from the genus Nothopanax, and compounds 5-8 were isolated from this plant for the first time.
Araliaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Bark ; chemistry

OBJECTIVETo investigate the species of Ligularia distributed in the northwestern China and their medicinal uses in the local area.

METHODField investigation, specimen collection, taxonomic study and datum check were adopted.

RESULTThere are 29 species and 1 varieties of Ligularia distributed in the northwestern China, and 18 species of them had been used as folk medicines with the function of resolving phlegm, relieving cough, clearing heat and toxins.

CONCLUSIONThe northwestern China is abundant in medicinal resource of Ligularia.

Anti-Asthmatic Agents ; isolation & purification ; pharmacology ; Antitussive Agents ; isolation & purification ; pharmacology ; Asteraceae ; chemistry ; classification ; China ; Conservation of Natural Resources ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ecosystem ; Humans ; Hypnotics and Sedatives ; isolation & purification ; pharmacology ; Pharmacognosy ; Plants, Medicinal ; chemistry ; classification

To study the protective effect of astragalus saponin extracts (AS) on kidneys of diabetic rats. Totally 32 diabetic rats induced by streptozotocin (STZ) were divided into AS high and low dose groups, the positive control group and the model group (DM group) and orally administered with 50 mg x- kg(-1) x d(-1) AS 200, 25 mg x kg(-1) x d(-1) valsartan, 10 mL x kg(-1) x d(1) physiological saline, respectively. Another 8 healthy rats were collected in the normal control group (NC group, physiological saline 10 mL x kg(-1). d(-1)). All rats were treated for consecutively 6 weeks. After the administration, the body weight was measured every week, the concentration of blood glucose was monitored on week 2, 4 and 6. The total urine and total urinary protein (U-TP) in 24 h were measured by the metabolic cage method on week 6; At the end of week 6, blood samples were collected from hearts to detect blood urea nitrogen (BUN), serum creatinine (Scr), uric acid (UA) , total cholesterol (CH) triglyceride (TG) by biochemical methods. Kidneys were collect to calculate the kidney hypertrophy index and observe the pathological sections. The laboratory results show that in the DM group, the blood glucose, metabolic cost in 24 h, kidney hypertrophy index, U-TP, BUN, Scr, UA, TG were significantly higher than that in the NC group (P < 0.01, P < 0.05) , with significant pathological changes; After the intervention with AS, the metabolic value in 24 h, kidney hypertrophy index, U-TP, BUN, Scr, UA, TG were significantly lower in the high dose group (P < 0.01, P < 0.05), and the kidney hypertrophy index, BUN, Scr, UA, TG in the low dose group were also significantly lower (P < 0.05), with slight reduction in renal pathological changes in both groups. In conclusion, Astragalus saponin extracts have a certain protective effect on kidneys of diabetic rats.
Animals ; Astragalus Plant ; chemistry ; Blood Glucose ; metabolism ; Blood Urea Nitrogen ; Diabetic Nephropathies ; metabolism ; prevention & control ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Kidney ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Saponins ; administration & dosage ; Uric Acid ; metabolism

Metabolic engineering is the analysis of metabolic pathway and designing rational genetic modification to optimize cellular properties by using principle of molecular biology. Aromatic metabolites such as tryptophan, phenylalanine, and tyrosine are essential amino acids for human and animals. In addition, phenylalanine is used in aspartame production. Escherichia coli and many other microoganism synthesize aromatic amino acids through the condensation reaction between phospho-enolpyruvate (PEP) and erythrose-4-phosphate(E4P) to form 3-deoxy-D-arabinoheptulosonate 7-phosphate(DAHP). But many enzymes compete for intracellular PEP, especially the phosphotransferase system which is responsible for glucose transport in E. coli. This system uses PEP as a phosphate donor and converts it to pyruvate, which is less likely to recycle back to PEP. To channel more carbon flux into the aromatic pathway, one has to overcome pathways competing for PEP. ppsA and tktA are the key genes in central metabolism of aromatic amino acids biosynthesis. ppsA encoding phosphoenolpyrucate synthetase A (PpsA) which catalyzes pyruvate into PEP; tktA encoding transketolase A which plays a major role in erythrose-4-phosphate (E4P) production of pentose pathway. We amplified ppsA and tktA from E. coli K-12 by PCR and constructed recombinant plasmids of them in pBV220 vector containing P(R)P(L) promoter. Because of each gene carrying P(L) promoter, four productions of ligation were obtained. The monoclonal host containing recombinant plasmids was routinely grown in Luria-Bertani (LB) medium added Ampicillin at 37 degrees C overnight, and then inoculated in LB (Apr) medium by 3%-5% in flasks on a rotary shaker at 30 degres C, induced at 42 degrees C for 4.5 hours when OD600 = 0.4, cells were obtained by centrifugation at 10,000 r/min at 4 degrees C. The results of SDS-PAGE demonstrated that the bands at 84kD and 73kD were more intensive than the same ones of the controls. The specific activity of PpsA in crude extracts was increased by 10.8-fold, and TktA, by 3.9-fold. When both genes were co-expressed in E. coli, the activity of PpsA varied from 2.1-9.1 fold comparing to control, but the activity of TktA was relatively stable(3.9-4.5 fold). Whatever the two genes were expressed respectively or cooperatively, both could promote the production of DAHP, the first intermediate of the common aromatic pathway, but co-expression was more effective on forming DAHP. The results demonstrate that co-expression of ppsA and tktA can improve the production of DAHP to near theoretical yield. This report details a different strategy based on co-expression of two genes in one vector in vivo to release the burden and paves the way for construction of genetic engineering bacteria for further research.
Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; enzymology ; genetics ; metabolism ; Escherichia coli Proteins ; genetics ; metabolism ; Plasmids ; genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics ; Pyruvate Synthase ; genetics ; metabolism ; Transketolase ; genetics ; metabolism

OBJECTIVETo clarify the resource of medicinal plants of genus Polygonum s. lat. distributed in Anhui Province.

METHODConducting field investigation and consulting related specimens and data.

RESULT AND CONCLUSIONThe distribution, growing environment and medicinal use of 32 taxa have been clarified. A scientific basis for further study for these medicinal plants has been provided.

Analgesics, Non-Narcotic ; pharmacology ; Antidiuretic Agents ; pharmacology ; China ; Conservation of Natural Resources ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ecosystem ; Pharmacognosy ; Plants, Medicinal ; anatomy & histology ; chemistry ; classification ; Polygonum ; anatomy & histology ; chemistry ; classification

Adult ; Bone Plates ; Female ; Humans ; Male ; Middle Aged ; Pubic Symphysis Diastasis ; surgery

OBJECTIVE: To determine the effect of CagA(+) Helicobacter pylori(H.pylori)strain and anti-H.pylori drugs on the expression of connexin 43(Cx43) and cell proliferation of BGC-823 cells in vitro,and to investigate the relation between the changes of Cx43 expression, cell proliferation of BGC-823 cells and CagA(+)H.pylori. METHODS: BGC-823 cells were co-cultured with CagA(+) H.pylori strain(NCTC J99) or CagA(-) H.pylori strain(NCTC 12908)at bacteria/cells ratio of 20:1,100:1 and 500:1 for 24 hours and 48 hours respectively. anti-H.pylori drugs was given in the group co-cultured at bacteria/cells ratio of 100:1 after 16 hours. In the control group, BGC-823 cells were cultured for 24 hours and 48 hours respectively,but without H.pylori or antij H.pylori drugs. Immunocytochemical SABC method and the image analysis of the computer were applied to detect the changes of Cx43 expression in BGC-823 cells. The cell proliferation was examined by methyl tetrazolium (MTT) method. RESULTS: (1)The expression of Cx43 in the control group after cultivation for 48 hours was higher than that for 24 hours (P< 0.05). The expression of Cx43 in the groups co-cultured with CagA(+) H.pylori strain after cultivation for 48 hours was lower than that co-cultured for only 24 hours, and that of the groups co-cultured with CagA(+) H.pylori strain was lower than that of the control group for both 24 hours and 48 hours (P< 0.05). The expression of Cx43 in the groups at bacteria/cells ratio of 500:1 was lower than that at bacteria/cells ratio of 20:1 and 100:1 for both 24 and 48 hours (P< 0.05),and that at bacteria/cells ratio of 100:1 was lower than that at bacteria/cells ratio of 20:1 for 48 hours (P< 0.05).However, there was no significant difference in Cx43 expression between 24 and 48 hours in the groups co-cultured with CagA(-) H.pylori strain (P>0.05). Cx43 expression in the groups co-cultured with CagA(-) H.pylori strain at the ratio of 100:1 and 500:1 was lower than that in the control group, and Cx43 expression at the ratio of 500:1 was lower than that at the ratio of 20:1 for 24 hours and 48 hours. Cx43 expression increased after the intervention with anti-H.pylori drugs for 48 hours. (2) In the groups co-cultured with CagA(+)H.pylori strain, the optical density value of MTT indicated that the cell proliferation at the bacteria/cells ratio of 100:1 was higher than that in the control group, but no significant difference was found in other two groups co-cultured for 24 hours. After co-culturing for 48 hours, the cell proliferation at the bacteria/cells ratio of 20:1 and 100:1 was significantly accelerated, while the cell proliferation at 500:1 was inhibited. In the groups co-cultured with CagA(-) H.pylori strain,there was no change in the cell proliferation. Intervention with anti-H.pylori drugs could suppress the cell proliferation. CONCLUSION CagA(+) H.pylori can down-regulate the expression of Cx43 in BGC-823 cells,which is related to the reaction time and the density of H.pylori. Low density of CagA(+)H.pylori suspensions can accelerate the proliferation of BGC-823 cells, while high density can suppress the cell proliferation. The CagA(-) H.pylori has no effect on the cell proliferation. Intervention with anti-H.pylori drugs can up-regulate the expression of Cx43,and suppress the cell proliferation of BGC-823 cells.
Anti-Bacterial Agents ; pharmacology ; Antigens, Bacterial ; genetics ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Connexin 43 ; biosynthesis ; Helicobacter pylori ; drug effects ; genetics ; metabolism ; Humans ; Immunohistochemistry ; Stomach Neoplasms ; metabolism ; microbiology ; pathology

OBJECTIVETo find a procedure for facial rejuvenation which is simple, safe with lasting aesthetic results based on facial anatomic study.

METHODSAnatomy study was performed on 12 sides from 6 head specimens. Observe the range and thickness of fat lateral to the nasolabial grooves. Observed the location of the skin retaining ligaments and reappraised their functions combining with clinical observations.

RESULTSSkin and subcutis and SMAS (including mimic muscles) become slackening with aging, but the loosening degrees are different, especially in the region lateral to the nasolabial groove. So they should be handled respectively. The fat lateral to the nasolabial groove is thick and is mobile with aging . So the subcutaneous detachment need not beyond the anterior border of the masseter. In the past two years, we performed rhytidectomy on 100 patients by limited subcutaneous detachment and SMAS double-plication. Satisfactory results were obtained. There are no serious complications observed.

CONCLUSIONSRhytidectomy by limited subcutaneous detachment and SMAS double-plication is a simple and safe procedure with lasting aesthetic results.

Face ; anatomy & histology ; Head ; anatomy & histology ; Humans ; Male ; Middle Aged ; Neck ; anatomy & histology ; Rhytidoplasty ; methods