Background Choroidal neovascularization(CNV)has been descrihed as a main reason of visual loss in a lot of ocular diseases.Researches showed that local hypoxia and retinal pigment epithelial(RPE) cells play an important role in the formation of CNV.A closely relationship of angiopoietin-2 (Ang-2) and angiogenesis has been proved.However,whether the expression of Aog-2 in hypoxic cultured human RPE cells is associated with the pathogenesis of CNV is still below understood.Objective This study was to investigate the effects of hypoxia on expression of Ang-2 in cultured human RPE cells in vitro,and discuss the possible effects of Ang-2 in the formation of CNV.Methods Human RPE cells were cultured and passaged,and 4-7 generation of cells were used in the experiment.The cells were incubated in cultural plate at the density of 5×107 cells/L.The culture medium containing 200 μmol/L CoCl2 was used to establish the hypoxia model of human RPE cells culturecd in vitro for 0.5,1,2,4,6,12and 24 hours,and the RPE cells cultured under normoxia were as controls.Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of Ang-2 mRNA in cultured human RPE cells,and enzymelinked immunosorbnent assay(ELISA) was used to assay the content of Ang-2 protein in supernatant of cultured human RPE cells.Results The survival rate of human RPE cells was 90％ after resoscitation.The fourth generation of cells showed the fusiform with the less pigment in them.The Ang-2 mRNA/β-actin mRNA value in human RPE cells was significantly different among various groups(F=1086.30,P=0.00),The Ang-2 mRN A/β-actin mRNA value in hypoxia cultured for 0.5 hours group began to increase and peaked in hypoxia culture for 4-6 hours group,with the significant differences in comparison with normoxia control group(P＜0.05).The Ang-2 mRNA/β-actin mRNA value decreased to the baseline level at hypoxia for 24 hours.The ELISA analysis showed that the concentration of Ang-2 protein in supernatant of RPE cells showed significant difference among groups(F=1034.00,P=0.00).The concentration of Ang-2 protein increased at hypoxia culture for 0.5 hours and peaked at 6 hours,showing significant differences in comparison with the control group (P＜0.05).Conclusions Hypoxia could significantly up-regulate the expression of Ang-2 in human RPE cells cultured in vitro.Ang-2 expresses highly in the early stage of hypoxia,implying that Ang-2 participates in the formation of CNV.

The genes of maltooligosyl trehalose synthase (MTSase) and maltooligosyl trehalose tetrahydrolase(MTHase) from Sulfolobus solfataricus ATCC 35092 were amplified using PCR. The expression plasmids, pTrc99a-MTSase and pTrc99a-MTHase, were constructed by inserting these two DNA fragments into E. coli expression vector pTrc99a. The specific activity of MTSase and MTHase in E. coli BL21(DE3) at optimal fermentation conditions reached 31.3U/g (wet cell) and 403U/g (wet cell), respectively. The biotransformation of partially hydrolyzed starch to trehalose catalyzed by MTSase and MTHase was carried out at 75℃ and pH 5.0. The highest yield of trehalose (ca. 53.6%) was gained when the original starch concentration was 15%(w/v) and the DE value was 10.

Objective To explore the association of chemokines and their receptors with immunologi- cal abnormality in newly diagnosed systemic lupus erythematosus(SLE) patients.Methods The serum con- centration of MIP-1?,MIP-1?,RANTES,IFN-?IL-4 were measured by enzyme-linked immunoabsorbent assay (ELISA) in 37 newly diagnosed.SLE patients and 20 normal controls.The expression rate of CCR1, CCR3,CCR5 on CD4~+T cells were detected by flow cytometry in 18 SLE patients and 10 normal controls.Re- suits Serum MIP-1?,MIP-1?concentrations were significantly higher in SLE patients than in normal control group (P＜0.01),the concentration of MIP-1?positively correlated with MIP-1?(r=0.609,P＜0.01);the per- centage of CD4~+CCR1~+ and CD4~+CCR5~+ cell were significantly lower in newly diagnosed SLE patients than in normal control group (both P＜0.01),the percentage of CD4~+CCRI~+ cells correlated negatively with the level of serum MIP-1?and IFN-?r=-0.525,P=-0.017;r=-0.442,P=0.045);the percentage of CD4~+CCR5~+ cell corre- lated negatively with the level of serum IFN-?(r=-0.645,P=0.001);the ratios of CD4~+CCR3~+/CD4~+CCR5~+ was significantly higher in newly diagnosed SLE patients than in the normal control group (P＜0.01).Conclusion Abnormal change and interaction of chemokines and their receptors with cytokines lead to immunologic dys- function and may participate in the initiation of SLE.

Ticks are obligate ectoparasites and vectors of arboviruses, vickettsiate, spirochetes and parasitil protozoa of humans and domestic animals. Immunological protection of mammalian hosts against tick infestation has been proposed as the most sustainable alternative tick control method to the current use of acaricides. The success of this method is dependent on the identification of key molecules for use as tick vaccine antigens. Proteolytic enzymes are involved in a wide range of cellular processes, thus they can be considered as good target antigens for a tick vaccine. In the present study, we used rapid amplification of cDNA ends protocol and primers that were designed based on the consensus amino acid motifs flanking present in all papain-like cysteine proteinases, to amplify, sequence and characterize two Rhipicephalus haemaphysaloides haemaphysaloides cathepsin L-like cysteine proteinases, named as cysA and cysB. The full length of cysA is 1168bp, encoding a 332 amino acid residue polypeptide with 36.33kD predicted molecular mass; the full length of cysB is 1153bp, encoding a 335 amino acid residue polypeptide with 37.56kD predicted molecular mass. The consensus amino acid motifs flanking presence in both deduced amino acid sequences. And both genes show high sequence homology to other tick cathepsin L-like cysteine proteinase, so they were identified as members of the cysteine proteinase gene family. Expression analysis by RT-PCR revealed that cysA and cysB were expressed differently in different periods of tick development.
Amino Acid Sequence ; Animals ; Cathepsin L ; Cathepsins ; genetics ; Cloning, Molecular ; Cysteine Endopeptidases ; genetics ; Female ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction ; Rhipicephalus ; enzymology ; Sequence Analysis

Objective To examine the relationship between hyperthyroidism and excessive iodine intake from drinking water through epidemiological studies in the iodine excess and the iodine normal villages. Methods Mengzhuang village of Pei county and Xingji village of Pizhou city in Jiangsu province, with median water iodine concentration of 1284.1 μg/L and 77.9 μ/L, respectively,were selected in 2006. Thyroid diseases of all local residents in the two villages were assessed clinically to compare the prevalence and the distribution of hyperthyroidism. Results A total of 17 471 residents were recruited from the iodine excess village, 26 of them were diagnosed with hyperthyroidism. The prevalence of hyperthyroidism was 1.49‰, 0.75‰ (7/9264) in male,2.32‰( 19/8207) in female, and the age-standardized prevalence was 1.48‰. A total of 12 765 residents were recruited from the iodine normal village, among them 27 residents were diagnosed with hyperthyroidism. The prevalence of hyperthyroidism was 2.12‰, 0.96‰(7/6823) in male, 3.26‰(20/5942) in female, and the agestandardized prevalence was 2.02‰. The prevalence and age-standardized prevalence was significantly lower in the iodine excess village than those in the iodine normal village (u = 2.88, 2.89; all P < 0.01). The prevalence of hyperthyroidism was lower among females in the iodine excess village (2.32‰) than that in the iodine normal one (3.37‰, u = 2.89, P < 0.01). Residents aged 20 - 50 years had higher prevalence of hyperthyroidism[(19.36 -38.96 )/10 000]in the two villages. The proportion of Graves diseases was 50.00% (13/26) in the iodine excessvillage, higher than that in the iodine normal village[29.41%(5/17) , χ2 = 5.853, P < 0.01]. Conclusions Chronic excessive iodine intake does not increase the chance of suffering from hyperthyroidism. On the contrary, the prevalence of hyperthyroidism in the iodine excess village decreases significantly compared with that of the iodine normal village. The prevalence is higher among females.

Human defensin is a family of cationic antimicrobial peptides in human being. During the last two decades a series of endogenous alpha-and beta-human defensins have been discovered. They are important components of the first barrier in human's body against the invasion of various microorganisms, and they are thought to play an important role in linking the innate and adaptive defense system of human being. The recent advances in the research of human defensins were reviewed, including their discovery, molecular and genetic properties, expression regulation, and mechanisms of antimicrobial activity. The possibility to produce human defensins via genetic engineering was also discussed. And the application outlook of human defensins in medicine and curing patients infected with antibiotics-resistant microbials was presented.
Amino Acid Sequence ; Defensins ; chemistry ; genetics ; metabolism ; physiology ; Genetic Engineering ; Humans ; Molecular Sequence Data ; Sequence Homology, Amino Acid

To study the chemical constituents of K. oblongifolia, silica gel column chromatography, MCI and Sephadex LH-20 were used to separate the 70% acetone extract of the stems of K. oblongifolia. The structures of the isolated compounds have been established on the basis of physicochemical and NMR spectroscopic evidence as well as ESI-MS in some cases. Twenty compounds were obtained and identified as heteroclitalignan A (1), kadsulignan F (2), kadoblongifolin C (3), schizanrin F (4), heteroclitalignan C (5), kadsurarin (6), kadsulignan O (7), eburicol (8), meso-dihydroguaiaretic acid (9), kadsufolin A (10), tiegusanin M (11), heteroclitin B (12), (7'S)-parabenzlactone (13), angeloylbinankadsurin B (14), propinquain H (15), quercetin (16), kadsulignan P (17), schizanrin G (18), micrandilactone C (19) and (-)-shikimic acid (20). Compouds 1, 5, 8, 11-15, 18 and 20 were isolated from this plant for the first time. Toxicity of compounds 1-10 were evaluated with zebrafish model to observe the effect on its embryonic development and heart function. The results showed that compounds 7, 9 and 10 caused edema of zebrafish embryo and decreased the heart rate of zebrafish, which exhibited interference effect on heart development of zebrafish.
Animals ; Embryo, Nonmammalian ; drug effects ; Guaiacol ; analogs & derivatives ; toxicity ; Kadsura ; chemistry ; Lignans ; toxicity ; Plant Extracts ; toxicity ; Quercetin ; toxicity ; Triterpenes ; toxicity ; Zebrafish ; embryology

OBJECTIVETo evaluate a new technique for breast reconstruction with multiple miniprostheses,

METHODSEach silicone miniprosthesis(10 approximately 15 ml) was implanted one by one beneath the prepared muscle pocket until the desired volume was achieved.

RESULTSSeven patients were treated with the above mentioned technique, including five after gland fibroadenoma excision, one hemangioma excision and one reduction mammoplasty. The results were satisfactory.

CONCLUSIONThe above mentioned technique with the miniprostheses may be another good way for breast augmentation and breast reconstruction with many advantages such as smaller incision, easy intraoperative assessment of the ideal size of the breast, easy handling the technique, perhaps low incidence of capsular contracture.

Adult ; Breast ; surgery ; Breast Implants ; Female ; Humans ; Patient Satisfaction ; Reconstructive Surgical Procedures ; methods ; Treatment Outcome

OBJECTIVETo study the effects of intestinal microflora alteration on specific and nonspecific immune function and hematopoietic function of mice.

METHODSSixty BALB/C mice were divided at random into two groups, experimental group and control group, with 30 mice in each. The mice in the experimental group were given kanamycin 50 mg while those in the control group were given distilled water intragastrically everyday for consecutive 10 days. After the 10 day treatment all the mice were sacrificed, and the cecal contents were collected for quantitative analysis of the intestinal bacterial flora. Certain indexes of immune function, including phagocytosis rate of macrophages, number of T lymphocytes positively stained by esterase and serum interleukin 2 (IL-2) content, and the weight of the spleen, granulocyte-macrophage colony stimulating factor etc. as indexes of hematopoietic function were determined.

RESULTSIn the group, the quantity of Enterobacteriaceae, Enterococcus, Bifidobacterium and Lactobacillus were significantly lower than that in the control group (P < 0.01). The number of PFC (plaque forming cells), the phagocytosis rate of macrophage, the number of T lymphocytes with positive NANE staining, the level of IL-2 significantly decreased when compared with that in the control group (P < 0.01). The weight of the spleen in the experimental group decreased when compared with that in the control group (P < 0.01). Levels of IL-3, GM-CSF, the total number of WBC and the proportion of neutrophil remarkably decreased as compared to that in the control group (P < 0.01). Analysis of the correlations between normal microflora, immunologic and hematopoietic indexes showed that marked positive correlations between the quantity of Bifidobacteria and each immune index including the levels of IL-3 and GM-CSF. There was a positive correlation between IL-2 and IL-3, IL-2 and GM-CSF as well.

CONCLUSIONThe application of antibiotics may cause changes in the structure and quantity of intestinal microflora. The dysbacteriosis may decrease the immune function of organism. The dysbacteriosis may decrease the hemopoietic function. The dysbacteriosis, the decrease in immune and hematopoietic function may affect one another. The balance in microecosystem should be emphasized and antibiotics should be applied rationally to reduce the side effects such as dysbacteriosis.

Animals ; Anti-Bacterial Agents ; pharmacology ; Esterases ; biosynthesis ; Feces ; microbiology ; Granulocyte-Macrophage Colony-Stimulating Factor ; analysis ; Interleukin-2 ; blood ; Intestines ; drug effects ; microbiology ; Kanamycin ; pharmacology ; Macrophages ; drug effects ; physiology ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Organ Size ; Phagocytosis ; drug effects ; Spleen ; drug effects ; pathology ; T-Lymphocytes ; drug effects ; metabolism

OBJECTIVETo identify the relationship between amino acid mutations in Neisseria gonorrhoeae isolates and their antibiotic resistance.

METHODSPI gene fragments of Neisseria gonorrhoeae from 17 clinical isolates were obtained with PCR amplification. They were cloned into the PCR cloning vector pBS-T to form pBS-T-PI and sequenced. The sequences of PI genes were analyzed. At the same time, minimum inhibitory concentration (MIC) of penicillin and tetracycline to these 17 isolates were measured and contrasted with the corresponding PI sequence.

RESULTSThe recombinants of PI gene from 17 clinical isolates of Neisseria gonorrhoeae were successfully constructed and sequenced. They were divided into PIA and PIB subtypes according to the results from blastn software by comparing the sequences with the GenBank. Mutations were found at the sites of 120 and 121. There were only some of the sequences having an aspartic acid (D) mutation on 120 and 121 sites, which was not the same as reported. On the other hand,there were two PI sequences,5-9 and 6-1, whose mutations on No. 120 were lysine, similar to those documented.

CONCLUSIONSome relationship between PI amino acids mutations at sites 120 and 121 in Neisseria gonorrhoeae isolates from Chengdu, China and their resistance to penicillin and tetracycline were found. However,further studies need to be done in the future to confirm this hypothesis.

Amino Acid Sequence ; Anti-Bacterial Agents ; pharmacology ; DNA Mutational Analysis ; DNA, Bacterial ; Drug Resistance, Bacterial ; Mutation ; Neisseria gonorrhoeae ; drug effects ; genetics ; isolation & purification ; Polymerase Chain Reaction