Aged ; Diagnosis, Differential ; Fibronectins ; metabolism ; Glomerulonephritis, Membranoproliferative ; pathology ; Humans ; Immunohistochemistry ; Kidney Diseases ; metabolism ; pathology ; Kidney Glomerulus ; metabolism ; pathology ; ultrastructure ; Male ; Microscopy, Electron

Background Doxycycline is a broad spectrum antibiotic,and it is frequently used in the treatment of ocular surface diseases.Objective The purpose of the present study was to investigate the effect of doxycycline on the inhibition of cell proliferation in bovine corneal myofibroblasts in vitro and assess its contribution to ocular surface repairing mechanism.Methods Six fresh bovine corneas were collected.The corneal stromal layer was isolated by two-step method of 1.0 g/L and 2.0 g/L collegenase-1.Isolated cells were plated at mantaryay culture flask in 10％ FBS of RPMI-1640.Vimentin and alpha-smooth muscle actin (α-SMA) organization were evaluated by immunocytochemistry,and the cells with influoresccence staining for vimentin and α-SMA were identified as the corneal myofibroblasts.Doxycycline at the concentrations of 10,20,40,60,80 mg/L was added to the medium,respectively,in different concentrations of doxycycline groups.Dexamethasone (120 mg/L)was used in the same way in the positive control group,and no drug was used in the negative control group.Cell proliferation was evaluated by MTT and the cell cycle was analyzed by BD FACScan flow cytometer assay 24 hours and 48 hours after addition of any drug.Results The cells grew well and showed the positive response for vimentin and α-SMA.MTT assay showed that the A570values of bovine corneal myofibroblasts were gradually declined with the increase of the concentration of doxycycline and lapse of active time,showing statistically significant difference (Fconcentration =1233.778,P＜0.001 ; Ftime =227.564,P ＜ 0.001).And the difference between the two factors was also statistically significant (Ftime*concentration =51.656,P＜0.001).Flow cytometry cell cycle analysis showed that 24 hours after 10,20,40,60,80 mg/L doxycycline treated,the perentage of of corneal myofibroblast cell in G0-G1 phase was 82.85％,84.36％,85.18％,87.12 ％ and 89.31％,showing significant increase in comparison with 63.89％ of the negative control group (all P＜0.05),and that of 40 mg/L doxycycline group was near the positive control group.Forty-eight hours after 10,20,40,60,80 mg/L doxycycline treated,the perentage of of corneal myofibroblast cell in G0-G1 phase was 82.78％,86.15％,88.23％,89.57％,93.00％,with significant increase in comparison with 70.17％ of the negative control group (all P ＜ 0.01),and that of 40 mg/L doxycycline group was near the positive control group.Conclusions The growth of the bovine corneal myofibroblasts is inhibited by doxycycline in time-and dosedependent manner in the range from 10 mg/L to 80 mg/L,and 40 mg/L of doxycycline has an obviously inhibitory action as 120 mg/L dexamethasone.

Objective To understand the situations of home establishments and the related domestic behaviors that causing injuries. Methods A total of 9760 families with regular residents in a community in Shanghai were investigated. Questionnaire was designed bascd on the Guidelines for Conducting Community Surveys on Injuries and Violence and International Classification of External Causes of Injuries. Results Inside the home settings, relative factors were found on issues as fire protection and using of electricity. In terms of household settings, 14.85% of the families had smoke alarm systems in the kitchen; 40.75% had no windows set for emergence. 50% to 70% of the residents had the idea of safe behaviors, including 35.93% of the families stored cleaning products,other chemicals or medical substances in alternative containers, such as used drinking bottle. Only 1.81% of the people being investigated thought that home was also a high risk place where injury might occur and it was placed number 9 in a multiple choice questionnaire. Data from the multiple level model analysis showed that factors as number of family members, space, education, profession etc. were closely related to the situation of home settings and their resident's behaviors. Conclusion Many injury related factors were found related to home establishments and people' s daily behaviors at home which called for interventions to be taken.

Objective To investigate whether recombinant human endostatin can create a time window of vascular normalization prior to vascular pruning to alleviate hypoxia in Lewis lung carcinoma in mice. Methods Kinetic changes in morphology of tumor vasculature in response to recombinant human endostatin were detected under a confocal microscope with immunofluorescent staining in Lewis lung carcinomas in mice. The hypoxic cell fraction of different time was assessed with immunohistochemical staining . Effects on tumor growth were monitored as indicated in the growth curve of tumors . Results Compared with the control group vascularity of the tumors was reduced over time by recombinant human endostatin treatment and significantly regressed for 9 days. During the treatment, pericyte coverage increased at day 3, increased markedly at day 5, and fell again at day 7. The vascular basement membrane was thin and closely associated with endothelial cells after recombinant human endostatin treatment, but appeared thickened, loosely associated with endothelial cells in control tumors. The decrease in hypoxic cell fraction at day 5 after treatment was also found. Tumor growth was not accelerated 5 days after recombinant human endostatin treatment. Conclusions Recombinant human endostatin can normalize tumor vasculature within day 3 to 7, leading to improved tumor oxygenation. The results provide important experimental basis for combining recombinant human endostatin with radiation therapy in human tumors.

A high activity isoflavone-glucosidase, which hydrolysis glycosides, was obtainde using liquid fermentation from Absidia sp. R strain. The isoflavone-glucosidase was purified 11 folds with yielding rate of 10.9% after ammonium sulfate precipitation and DEAE-Cellocuse (DE-52) ion exchange chromatography. SDS-PAGE results showed that the molecular weight is 53kD. And the optimum temperature, the optimum pH, Km and pI of the enzyme are 50 deegrees C, 5.0, 1.3 x 10(-2) mol/L and 3.2, respectively. The isoflavone-glucosidase is also rather stable under 60 degrees C and in pH range from 5.0 to 7.0. The enzyme can be activated by Co2+ and Ca2+, and be inhibited by Ag+ and Cu2+.
Absidia ; enzymology ; Glucosidases ; isolation & purification ; metabolism ; Hydrogen-Ion Concentration ; Isoflavones ; metabolism ; Temperature

OBJECTIVETo explore the roles of granulocyte colony-stimulating factor in the pathogenesis of moyamoya disease.

METHODSSerum G-CSF concentrations were measured using enzyme linked immunosorbent assay (ELISA) in 20 children with moyamoya disease and 20 healthy children.

RESULTSSerum G-CSF concentration (35.7+/-10.3 pg/mL) in children with moyamoya disease was significantly higher than that in healthy controls (23.5+/-3.8 pg/mL) (p<0.01).

CONCLUSIONSThe elevated serum G-CSF concentration in children with moyamoya disease suggests that G-CSF may play an important role in the pathogenesis of moyamoya disease.

Child ; Child, Preschool ; Female ; Granulocyte Colony-Stimulating Factor ; blood ; physiology ; Humans ; Male ; Moyamoya Disease ; blood ; etiology ; Vascular Endothelial Growth Factor A ; analysis ; physiology

Objective To design radioactive biliary stents and to evaluate the feasibility and safety of the stents.Methods Plastic stents with inserted iodine-125 seeds were designed and tested in sixteen normal pigs. In the brachytherapy group,the pigs were implanted radioactive stents in the common bile duct (CBD) and then divided into three groups on the basis of radiation dose of the reference point,50 Gy group (n=4),100Gy group (n=4),and 150 Gy group (n=4).In the control group,the same plastic stents with non-radioactive seeds were implanted (n=2),whilst in the blank control group,no stents were implanted (n=2).Blood routine,serum amylase,liver and kidney function were tested before and 1,7,14,30,60 days after the implantation of stents. Animals were sacrificed on the 14,30 and 60 days after stem implantation.Some relevant index such as perito- neal bleeding and inflammation,ascites,injury of adjacent organs,as well as perforation,stricture and dilatation of bile duct were investigated.Bile duct tissues were stained with H-E,and observed under microscopy. Results The radioactive plastic biliary stents were successfully prepared and implanted.There was no effusion, hemorrhage or necrosis in the adjacent organs of radioactive biliary stent implanted group.Perforation of the CBD wall was not observed in the brachytherapy group.By pathological examination in the CBD,obvious hyperplasia of the mucosa and mucosal glands were seen in the control group.Necrosis of mucosal layer existed in brachy- therapy groups.In 50 Gy group,mucosal layer was incomplete and mild hyperplasia of mucosal glands was seen. In 100 Gy group,mucosal layer disappeared and almost no hyperplasia of the mucosal glands could be found.In 150 Gy group,mucosal layer disappeared and mucosal glands obviously decreased.There were no obvious abnormalities noted in blood tests after implantation in each group.Conclusions The design of radioactive biliary stents are feasible and safe.The radioactive stents have obvious radiation effect besides the mechanical effect on the mucosal layer of CBD.

AIM: To investigate the antitumor activity and mechanism of Paecilomyces tenuipes polysaccharide(PTPS).METHODS: PTPS-I was obtained by water extraction and alcohol precipitation,and purified by DEAE-cellulose and Sephadex G-100 chromatography.Human erythroleukemia cell line K562,laryngocarcinoma cell line Hep2 and hepatic carcinoma cell line SMMC-7721were co-cultured with PTPS-I or the conditioned medium which prepared with PTPS-I-stimulated human mononuclear cells(PTPS-I-MNC-CM),and the proliferation of tumor cells was determined.The cell counting kit-8(CCK-8) was used to determine the proliferation of MNCs.The FQ-RT-PCR was applied to investigate the expression of TNF-? and IL-6 mRNA in MNCs.RESULTS: PTPS-I-MNC-CM inhibited the proliferation of K562,Hep2 and SMMC-7721 cells in vitro(P

To study the expression of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) in lung cancer cells, and to testify if the PPAR-gamma agonists can inhibit human lung cancer cell growth through induction of apoptosis, PPAR-gamma was detected in two lung cancer cell lines by RT-PCR and immunohistochemistry, the inhibition of human lung cancer cell growth was investigated by MTT and cell counts, and the apoptosis was assessed by TUNEL. The results showed that: (1) PPAR-gamma expressed on two lung cancer cell lines; (2) PPAR-gamma activated by ligands could inhibit human lung cancer cell growth remarkably; (3) PPAR-gamma agonists could induce apoptosis to inhibit lung cancer cell growth. It was concluded that PPAR-gamma expressed in lung cancer cell can be activated by ligands and can inhibit lung cancer cell growth through induction of apoptosis.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; pathology ; Carcinoma, Small Cell ; pathology ; Cell Division ; drug effects ; Gene Expression Regulation, Neoplastic ; Humans ; Ligands ; Lung Neoplasms ; pathology ; Prostaglandin D2 ; analogs & derivatives ; pharmacology ; Receptors, Cytoplasmic and Nuclear ; agonists ; metabolism ; Transcription Factors ; agonists ; metabolism ; Tumor Cells, Cultured

Adolescent ; Adult ; Aged ; Chronic Disease ; Female ; Hepatitis B e Antigens ; analysis ; Hepatitis B virus ; genetics ; Humans ; Liver Diseases ; virology ; Male ; Middle Aged ; Mutation ; Oligonucleotide Array Sequence Analysis ; Promoter Regions, Genetic