Objective To evaluate the feasibility to detect Prototheca in a mouse model of Prototheca zopfii cutaneous infection by using fluorescence in situ hybridization (FISH).Methods The model of Prototheca zopfii cutaneous infection was established by abdominal subcutaneous inoculation of Prototheca zopfii suspensions into 20 male BALB/c mice.Seven days after the inoculation,the mice were sacrificed,and tissue specimens were obtained from abdominal skin and subjected to microscopic examination,fungal culture and paraffin embedding.A PZ-probe was artificially synthesized and used to detect Prototheca in paraffin-embedded sections by using FISH.Moreover,both periodic acid-Schiff (PAS) and hematoxylin-eosin (HE) staining were performed to examine the paraffin-embedded sections.Skin specimens obtained from normal mice and Candida albicans-or Cryptococcus neoformans-infected mice served as the negative control.Results Clinical presentations,pathological examination and fungal culture results all confirmed the successful establishment of Prototheca zopfii skin infection model in mice.Prototheca was identified by FISH with the PZ-probe in the paraffin-embedded skin tissue sections from the murine model of Prototheca zopfii cutaneous infection,but not detected in the negative control tissue specimens,which was consistent with the results of PAS and HE staining.Conclusion FISH can be used to detect Prototheca in paraffin-embedded skin sections from the mouse model of Prototheca zopfii cutaneous infection.

OBJECTIVETo investigate the correlation of G487A polymorphism of aldehyde dehydrogenase-2 (ALDH2) gene with hypertension in patients with coronary heart disease complicated by type 2 diabetes.

METHODSThis study was conducted among 167 patients with coronary heart disease complicated by diabetes mellitus. The polymorphisms of gene G487A ALDH2 were determined using polymerase chain reaction-restricted fragments length polymorphism technique (PCR-RFLP). According to the genotypes, the patients were divided into GG group (n=105) and GA/AA group (n=62), and the incidence of hypertension, risk factors of hypertension, systolic and diastolic pressures, and pulse pressure indexes were compared between the two groups. Multivariate logistic regression analysis was performed to adjust the effects of the confounding factors.

RESULTSThe incidence of hypertension in GA/AA group was significantly higher than that in GG group (P<0.05), and the former group showed a significantly greater differences between systolic and pulse pressure; the diastolic pressure was comparable between the two groups. Multivariate logistic regression analysis showed that GA/AA was associated with an increased risk of hypertension in synergy with high insulin level and insulin resistance.

CONCLUSIONALDH2 gene G487A polymorphism may be associated with hypertension in patients with coronary heart disease complicated by type 2 diabetes, and the patients with an A allele have a greater risk of developing hypertension.

Aged ; Alcohol Dehydrogenase ; genetics ; Coronary Disease ; complications ; genetics ; Diabetes Mellitus, Type 2 ; complications ; genetics ; Female ; Humans ; Hypertension ; etiology ; Male ; Middle Aged ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Risk Factors

OBJECTIVETo study the inhibitory effect of CD133 monoclonal antibody labeled with ¹³¹I (¹³¹I-CD133mAb) on Huh-7 human liver cancer cell line overexpressing CD133 antigen in vitro and in mouse models bearing the tumor cell xenograft.

METHODS¹³¹I-CD133mAb was prepared by chloramines-T method and evaluated for its stability. Flow cytometry and immunohistochemistry were used to detect the expression of CD133 in Huh-7 cells and in Huh-7 cell-derived tumors, respectively. Huh-7 cells treated with ¹³¹I-CD133mAb plus cisplatin (DDP), ¹³¹I -CD133mAb, DDP, or no treatment (blank control) were examined for cell proliferation suppression by MTT assay with the IC₅₀ calculated. BALB/c mice bearing subcutaneous Huh-7 cell xenograft in the right forelegs were treated with ¹³¹I -CD133mAb, DDP, or both every two days for two weeks. The tumor size and volume were measured twice a week, and pathological examination of the tumor was carried out after the treatments. The tumor inhibition rate was calculated and tumor cell apoptosis observed with HE staining.

RESULTSThe labeling ratio of ¹³¹I-CD133mAb was 90.25% and the radiochemical purity was 97.78%. Huh-7 cells showed obviously higher CD133 expression than HepG2 cells. ¹³¹I-CD133mAb combined with DDP group resulted in a significantly higher tumor inhibition rate than other treatments in the tumor-bearing mice.

CONCLUSION¹³¹I-CD133mAb can inhibit the growth of liver cancer cells with a high CD133 expression both in vivo and in vitro.

AC133 Antigen ; Animals ; Antibodies, Monoclonal ; pharmacology ; Antigens, CD ; immunology ; Apoptosis ; Carcinoma, Hepatocellular ; drug therapy ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cisplatin ; pharmacology ; Glycoproteins ; immunology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Peptides ; immunology ; Xenograft Model Antitumor Assays

OBJECTIVETo investigate the effect of amphotericinB (AmB) on migration and invasion of esophageal carcinoma Eca109 cells exposed to hypoxia and explore the molecular mechanisms.

METHODSRoutinely cultured esophageal carcinoma Eca109 cells were treated with 0, 1.25, 2.5, or 5 µg/ml AmB in hypoxic condition (3% O2, 5% CO2, and 92% N2) for 24 h. The cell migration and invasion were assessed by cell scratch test and Transwell chamber assay, respectively. Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expressions of hypoxia-inducible factor-1α (HIF-1α), matrix metalloproteinase-2 (MMP-2), and E-cadherin in the cells, respectively.

RESULTSCompared with the control cells, the cells treated with different doses of AmB showed attenuated ability of migration and invasion (P<0.05). AmB treatment resulted in significantly lowered mRNA and protein expressions of MMP-2 (P<0.05) and increased expressions of E-cadherin (P<0.05); the protein expression of HIF-1α decreased significantly in cells after AmB treatment (P<0.05) but its mRNA levels showed no significant changes (P>0.05).

CONCLUSIONAmB can suppress the migration and invasion of esophageal carcinoma Eca109 cells in hypoxic microenvironment possibly by regulating the expressions of HIF-1α, MMP-2 and E-cadherin.

Amphotericin B ; pharmacology ; Cadherins ; metabolism ; Cell Hypoxia ; Cell Line, Tumor ; drug effects ; Cell Movement ; drug effects ; Down-Regulation ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; RNA, Messenger