Objective To study the association between house dust endotoxin levels and the airway responsiveness as well as allergen sensitization in adolescence in the urban and rural areas of Guangdong province.Methods Middle school students,aged 13-14,188 (male 109,female 79) from Guangzhou,129(male 70,female 59) from Conghua with positive responses,and 231 (male 125,female 106) and 307(male 145,female 162) of their respective reference students are selected from questionnaire prescreening.A detailed questionnaire,the lung function test,histamine bronchial provocation test,skin prick test and peripheral blood eosinophil count are undertaken.Seventy-six students from Guangzhou and 80 from Conghua were selected randomly for mattress dust sample collection to detect the levels of endotoxin.Results The prevalence of wheeze symptom,diagnosed asthma,nasal symptoms and allergic rhinitis of Guangzhou was significantly higher than that of Conghua (P＜0.0001),with 28.6％,27.7％,66.0％,46.4％ for Guangzhou respectively,and 5.0％,2.5％,31.0％,6.2％ for Conghua respectively.The positive rates of bronchial provocation test and skin sensitization against Dermatophagoides pteronyssinus(Der.p),D.farinae (Der.f),cat and dog allergen in the students of Guangzhou were significantly higher than that of Conghua (P＜0.0001).The positive rate of cockroach of Conghua was significantly higher than that of Guangzhou (P＜0.0001).The levels of endotoxin (amount,concentration and load) of samples from Conghua were significantly higher than that from Guangzhou (Conghua:6452 EU,10.95 EU/mg,1794 EU/m2 ;Guangzhou:1591 EU,6.45 EU/mg,508.80 EU/m2,respectively).PD20-FEV1 was correlated positively with endotoxin amount(r=0.174,P＜0.05).The diameters of the skin wheal size for Der.p and Der.f were correlated inversely with endotoxin amount and load (P＜0.001).Conclusion The prevalence of wheeze symptom,diagnosed asthma,nasal symptoms and allergic rhinitis in 13-14 year-old adolescent of Guangzhou was significantly higher than that of Conghua.Cockroach was the second major allergen of Conghua followed by house dust mite.The level of endotoxin was correlated positively with PD20-FEV1,but correlated inversely with skin sensitization to Def.p and Der.f.The study suggests that environmental endotoxin may have effect to protect children from developing allergic diseases.

AIMTo study the transfection and anti-hepatitis B virus (HBV) effect of the co-modified hepatocytes-targeting cationic liposomes encapsulating anti-HBV antisense oligonucleotides (asON) , and to investigate the transfection mechanisms of the liposomes.

METHODSDipalmitoylphosphatidylcholine (DPPC) and 3beta-[N-(N',N'-dimethylaminoethane)-carbamoyl] cholesterol (DC-Chol) were used as the lipids, beta-sitosterol-beta-D-glucoside (sito-G) and Brij 35 were used to modify the liposomes. Flow cytometry (FCM), fluorescence microscopy and enzyme-linked immunosorbent assay (ELISA) were utilized to evaluate the transfection improvement of the asON encapsulated in the liposomes in primary rat hepatocytes and the antigens inhibition activity in HepG 2.2.15 cells. The transfection mechanisms were evaluated based on the influence of wortmannin, nigericin, and asialofetuin on the antigens inhibition in HepG 2.2.15 cells by ELISA.

RESULTSThe co-modification with sito-G and Brij 35 significantly improved the transfection of the liposomes in primary rat hepatocytes and antigens inhibition effect in HepG 2.2.15 cells. Both transfection efficiency and antigens inhibition effect showed to be concentration-dependent with the asON-encapsulating liposomes. In fluorescence microscopy, the transfected cells showed strong fluorescence in primary rat hepatocytes, especially in the nuclei. Wortmannin, nigericin and asialofetuin decreased the antigens inhibition of the asON-encapsulating liposomes to different levels. Cationic liposomes modification with sito-G and Brij 35 could improve the transfection and antigens inhibition effect of the asON. The transfection mechanisms of the co-modified liposomes included endocytosis and membrane fusion. The ligand sito-G was confirmed to be able to enhance asialoglycoprotein receptor (ASGPR)-mediated endocytosis.

CONCLUSIONCo-modified hepatocytes-targeting cationic liposomes would be a specific and effective carrier to transfer asON into hepatocytes.

Androstadienes ; pharmacology ; Animals ; Asialoglycoproteins ; pharmacology ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Cell Survival ; Cells, Cultured ; Endocytosis ; drug effects ; Female ; Fetuins ; Flow Cytometry ; Hepatitis B Antigens ; metabolism ; Hepatitis B virus ; genetics ; immunology ; Hepatocytes ; cytology ; drug effects ; metabolism ; Humans ; Liposomes ; Microscopy, Fluorescence ; Nigericin ; pharmacology ; Oligonucleotides, Antisense ; chemistry ; genetics ; Polyethylene Glycols ; chemistry ; Rats ; Rats, Wistar ; Sitosterols ; chemistry ; Transfection ; methods ; alpha-Fetoproteins ; pharmacology

OBJECTIVETo study the effect of combining the injection of beta-sheet breaker H102 with human umbilical cord mesenchymal stem cell (hUCMSC) on APP transgenic mice behavior, P-tau, apoptosis and the expression of relevant enzymes in the brain.

METHODSAPP transgenic mice were randomly divided into model group, hUCMSC group, H102 group, H102 with hUCMSC group and a group of C57BL/6J mice with the same age and background was set as normal. After two weeks and four weeks, the ability of spatial reference memory was tested by Morris Water Maze. After four weeks, immunohistochemical stain and Western blot were done to detect the content of Bad, Bax, Bcl-2, Bcl-xl, P-tau, GSK-3beta, PP-2A and PP-1 in mice brain.

RESULTSThe ability of memory of hUCMSC in 2 weeks group was slightly improved than that in the model group. hUCMSC in four weeks group, H102 group and H102 with hUCMSC group significantly improved the ability of and memory, and reduced the phosphorylation of tau and brain cell's apoptosis of the Alzheimer disease (AD) mice.

CONCLUSIONBeta-sheet breaker H102 together with transplanting hUCMSC is an effective therapeutic strategy for AD.

Alzheimer Disease ; therapy ; Amyloid beta-Protein Precursor ; genetics ; Animals ; Disease Models, Animal ; Humans ; Maze Learning ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Peptides ; therapeutic use ; Umbilical Cord ; cytology

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnostic imaging ; pathology ; Female ; Humans ; Liver Neoplasms ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Ultrasonography

OBJECTIVETo study mechanism of the apoptosis of rat pancreas islet β cell strain (INS-1 cells) induced by sodium arsenite.

METHODSINS-1 cells were exposed to sodium arsenite at the different concentrations. MTT assay was used to detect the viability of INS-1 cells. The potentials on mitochondrial membrane and lysosome membrane of INS-1 cells were determined with the fluorescence spectrophotometer. The apoptotic levels of INS-1 cells exposed to sodium arsenite were observed by a fluorescence microscope and flow cytometry.

RESULTSAfter exposure to sodium arsenite, the viability of INS-1 cells significantly decreased with the doses of sodium arsenite. At 24 h after exposure, the OD values of the mitochondrial membrane potentials declined observably with the doses of sodium arsenite (P < 0.01). At 48 h after exposure, the OD values of the lysosome membrane potentials significantly increased with the doses of sodium arsenite (P < 0.01). At 72 h after exposure, the apoptotic cells were observed under a fluorescence microscope and enhanced with the doses of sodium arsenite. The apoptosis cells with light blue, karyopyknosis, karyorrhexis, apoptotic body and chromatin concentration appeared. The results detected with flow cytometry indicated that after exposure, the apoptotic INS-1E cells significantly increased with the doses of sodium arsenite.

CONCLUSIONSThe sodium arsenite can induce the apoptosis of INS-1 cells through the mitochondria-lysosome pathway.

Animals ; Apoptosis ; drug effects ; Arsenites ; toxicity ; Cells, Cultured ; Insulin-Secreting Cells ; drug effects ; Lysosomes ; metabolism ; Membrane Potentials ; drug effects ; Mitochondria ; metabolism ; Rats ; Sodium Compounds ; toxicity

OBJECTIVETo investigate the clinicopathological significance of positive CK7 expression in human colorectal carcinoma (HCC).

METHODSImmunohistochemistry was used to detect CK7 and CK20 protein expressions in 68 cases of HCC, 20 cases of canalicular adenoma (CA), 5 cases of serrated adenoma (SA) and 20 cases of hyperplastic polyps (HP).

RESULTSThe positivity rate of CK20 expression was 89.7% in HCC, and 100% in CA, SA and HP. In HCC, the expression rate of CK7 (39.7%) was not correlated with Dukes' classification, differentiation and tumor location. CK7 positivity rate in colon cancer was 35.7% (15/52) and 42.3% (11/26) in rectal cancer. CK7 expression was negative in CA. CK7 positivity rate in SA was 49% and 30% in HP.

CONCLUSIONCK7 is a possible marker for colorectal carcinogenesis from HP to SA, and ultimately to HCC, and examination of the colorectal polypoid lesions for CK7 expression can be significant for estimating the colorectal polypous cancerization.

Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Cytoplasm ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Intestinal Mucosa ; metabolism ; pathology ; Keratin-7 ; metabolism ; Male ; Middle Aged

AIMTo prepare fluorescein sodium (FS) cationic liposomes and investigate the influence of cationic lipid (DC-chol) and polyethylene glycol (PEG) with different molecule weight (MW) on cationic liposome incorporation efficiency, cellular delivery and fluidity of liposome membrane.

METHODSUsing FS as a model material for encapsulation, the liposomes were prepared and separated (by sephadex G-50 1 cm x 20 cm column), and the liposome incorporation efficiencies was measured. The interaction between the FS and cationic liposomes was investigated by measuring the change of fluorescent spectrum. The cellular uptake of different liposome forms by choosing HepG2 2.2.15 as an in vitro cell culture assay model, and the influence of PEG on the fluidity of liposome membrane with the technique of fluorescence polarization were investigated.

RESULTSCationic lipid and different PEGs showed great effects on increasing liposome incorporation efficiency (from 0.64% to 86.57%), cellular uptake (from 2.18% to 48.46%) and fluidity of liposome membrane. The effect of PEG was MW dependent, and with the increase of MW, the incorporation efficiency and transfection was improved, and the fluidity of liposome membrane increased.

CONCLUSIONAddition of cationic lipid and high MW PEG into cationic liposomes can enhance the cellular delivery and fluidity of cationic liposomes. Also, they can improve the incorporation efficiency of cationic liposomes.

Cholesterol ; analogs & derivatives ; pharmacology ; Drug Delivery Systems ; Fluorescein ; Hepatoblastoma ; pathology ; Humans ; Liposomes ; Liver Neoplasms ; pathology ; Membrane Fluidity ; drug effects ; Molecular Weight ; Polyethylene Glycols ; chemistry ; pharmacology ; Transfection ; methods ; Tumor Cells, Cultured

OBJECTIVETo explore hepatitis C virus (HCV) non-structural protein 5A (NS5A)'s influence on inhibition of AFP expression executed by p53 protein and its possible molecular mechanism.

METHODSPlasmid transfection and MEIA were employed to observe p53's inhibitive effect on AFP expression of Huh7 cells and the HCV NS5A's influence on p53 function. Western blot was employed to find out if HCV NS5A affects p53 protein expression and GST pull down assay was applied to examine the interaction between HCV NS5A and p53.

RESULTSThe AFP concentration in the supernatant of the culture of the Huh7 cells transfected with pRc/CMV was (14322+/-2412) ng/ml, and that of the Huh7 cells transfected with pCNS5A was (13843+/-3218) ng/ml; no significant difference existed between these two groups (t = 1.42, P > 0.05). After transfection with pC53-NS3, the AFP level was decreased to (10 241+/-1326) ng/ml, and in comparison to the above two groups it had a statistically significant difference (t values were 2.41 and 2.38, P < 0.05). When co-transfected with pCNS5A and pC53-NS3, the AFP expression (14582+/-1238) ng/ml returned to the level of pRc/CMV transfected, and there was a remarkably significant difference between this and that of the pC53-NS3 transfected cells (t = 3.12, P < 0.01). HCV NS5A had no function on the p53 protein expression with Western blot experiment. In the GST pull down assay, an HCV NS5A protein band was found after GST-p53 was added, but not detected with GST only.

CONCLUSIONWe found that p53 has an inhibitive function on the AFP expression in Huh7 cells and HCV NS5A minimized this p53 function. HCV NS5A did not affect p53 protein expression, but was able to form a complex with p53, by which HCV NS5A inactivated this p53 function.

Carcinoma, Hepatocellular ; metabolism ; virology ; Hepacivirus ; genetics ; Humans ; Liver Neoplasms ; metabolism ; virology ; Tumor Cells, Cultured ; Tumor Suppressor Protein p53 ; genetics ; pharmacology ; Viral Nonstructural Proteins ; genetics ; alpha-Fetoproteins ; biosynthesis ; genetics

BACKGROUNDTrichophyton rubrum represents the most common infectious fungus responsible for dermatophytosis in human, but the mechanism involved is still not completely understood. An appropriate model constructed to simulate host infection is the prerequisite to study the pathogenesis of dermatophytosis caused by T. rubrum. In this study, we intended to develop a new T. rubrum infection model in vitro, using the three-dimensional reconstructed epidermis - EpiSkin ®, and to pave the way for further investigation of the mechanisms involved in T. rubrum infection.

METHODSThe reconstructed human epidermis (RHE) was infected by inoculating low-dose (400 conidia) and high-dose (4000 conidia) T. rubrum conidia to optimize the infection dose. During the various periods after infection, the samples were processed for pathological examination and scanning electron microscopy (SEM) observation.

RESULTSThe histological analysis of RHE revealed a fully differentiated epidermis with a functional stratum corneum, which was analogous to the normal human epidermis. The results of hematoxylin and eosin staining and the periodic acid-Schiff staining showed that the infection dose of 400 conidia was in accord with the pathological characteristics of host dermatophytosis caused by T. rubrum. SEM observations further exhibited the process of T. rubrum infection in an intuitionistic way.

CONCLUSIONSWe established the T. rubrum infection model on RHE in vitro successfully. It is a promising model for further investigation of the mechanisms involved in T. rubrum infection.

Animals ; Disease Models, Animal ; Epidermis ; microbiology ; Humans ; Keratinocytes ; cytology ; Tissue Culture Techniques ; Trichophyton ; pathogenicity

OBJECTIVETo evaluate the effect of a phospholipid-coated microbubble contrast agent for myocardium opacification in comparison with a albumin-coated microbubble contrast agent (Quanfuxian).

METHODSIn 10 dogs with single coronary artery stenosis involving the anterior descending branch or circumflex branch randomly received infusion of the two contrast agents through the femoral vein. The myocardial blood flow, heart rate and blood pressure were analyzed qualitatively and quantitatively. The concentration and the particle diameter of the two contrast agents were determined.

RESULTSThe concentration of the phospholipid-coated microbubbles was (1.06-/+0.22) x10(9)/ml, with a diameter of 3.04-/+0.34 microm, similar to the concentration and diameter of Quanfuxian ((1.31-/+0.33)x10(9)/ml and 2.88-/+0.58 microm, respectively, P>0.05). Both of the agents achieved grade three myocardium opacification, and produced no obvious effect on the heart rate and blood pressure. Quantitative analysis of myocardial opacification in terms of myocardial blood volume (A), blood velocity (beta), and blood flow (A x beta) revealed no significant difference between the two agents (P>0.05), and the parameters derived from the two agents showed good correlations (P<0.05, rA=0.809, r beta=0.932, rA.beta=0.925).

CONCLUSIONThe phospholipid-coated microbubble contrast agent shows good effect for myocardial opacification without significant difference from Quanfuxian. Both of the agents are good ultrasound contrast agents for quantitative analysis of myocardium blood flow.

Albumins ; chemistry ; Animals ; Contrast Media ; administration & dosage ; chemistry ; Coronary Stenosis ; diagnostic imaging ; Dogs ; Echocardiography ; methods ; Female ; Male ; Microbubbles ; Phospholipids ; chemistry