Bone Morphogenetic Protein 1 ; metabolism ; Carcinoma ; metabolism ; pathology ; Carcinoma, Papillary ; metabolism ; pathology ; Cell Adhesion Molecules ; metabolism ; Epithelial-Mesenchymal Transition ; Humans ; Survival Rate ; Thyroid Neoplasms ; metabolism ; pathology

Objective To investigate the feasibility of low-frequency ultrasound combined with microbubbles improving transfection of enhanced green lfuorescent protein plasmid (pEGFP) to subcutaneously transplanted prostate cancer in nude mice, and to optimize the parameters of intensity. Methods The model of nude mice of subcutaneously transplanted prostate cancer were built. Then they were divided into 2 groups including low-frequency ultrasound group and low-frequency ultrasound combined with microbubbles group, each group with 15 mice. Then 250μl of mixture of saline and pEGFP (50μg) were co-injected in low-frequency ultrasound group and 250μl of mixture of SonVue and pEGFP (50μg) were co-injected in low-frequency ultrasound combined with microbubbles group through tail vein. According to intensity, they were divided into 3 subgroups respectively, including group 1 W/cm2, group 2 W/cm2 and group 3 W/cm2, each group with 5 mice. Ultrasound was applied at 80 kHz input frequency with 50%duty cycle for 10 minutes after pEGFP injection. The tumors were collected on the third day after treatment. The expression of pEGFP in tumor was examined by laser scanning confocal microscope (ISCM) and the average lfuorescence intensity were estimated. At the same time, routine pathological examination was performed. The mean lfuorescence intensity of low frequency ultrasound group and low frequency ultrasound combined with microbubble group were compared with one-way ANOVA and those of any two groups were compared by SNQ-q test. Results TThe mean lfuorescent light in low-frequency ultrasound combined with microbubbles group were 23.75±2.54, 30.25±1.67 and 59.60±2.03, which were obviously stronger than those of low-frequency ultrasound treatment group (14.04±1.35, 14.66±0.98 and 14.32±1.20), the difference was statistically signiifcant (the value of q were 18.26, 14.12 and 13.88, P<0.05). The rate of gene transfection increased along with the enhancement of the ultrasound intensity (the value of q were 15.33, 17.81 and 13.79, P<0.05). Histology analyses performed by HE staining showed that there was no damage to the tumor tissues in all groups, tumor tissues were intact and without infection. Conclusions Low-frequency ultrasound combination with microbubbles can significantly enhance pEGFP transfecting into subcutaneously transplanted prostate cancer in nude mice. In certain range, improving the ultrasound intensity can increase the rate of gene transfection.

Objective To investigate the effect of 20 kHz low-intensity ultrasound combined with microbubbles on autophagy of both PC3 cells and DU145 cells. Methods Ultrasound with a frequency of 20 kHz and intensity of 80 mW in continuous wave mode was used. Both PC3 cells and DU145 cells were divided into four groups, including control group (A), microbubbles group (B), ultrasound group (C) and ultrasound combined with microbubbles group (D). Twenty-four hours after treatment, the acid vesicular organelles were detected by acridine orange lfuorescence staining, and transmission electron microscopy (TEM) was used to observe autophagosomes. Results Acridine orange lfuorescence staining showed formation of acid vesicular organelles (AVOs) in the cytoplasm with normal nucleus in both PC3 cells and DU145 cells in group D, while in group A cells were basically normal. Lots of autophagosomes with double-membrane structure were detected by transmission electron microscope in group D. Conclusions Low-frequency and low-intensity ultrasound combined with microbubbles can induced autophagy in both PC3 cells and DU145 cells.

Actins ; metabolism ; Adenomyoepithelioma ; metabolism ; pathology ; surgery ; Aged ; Axilla ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Female ; Humans ; Keratin-7 ; metabolism ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Mastectomy, Modified Radical ; S100 Proteins ; metabolism

Animals ; Cadherins ; metabolism ; Cell Cycle Proteins ; genetics ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Early Growth Response Protein 1 ; genetics ; metabolism ; Estradiol ; physiology ; Eukaryotic Initiation Factor-3 ; metabolism ; Genes, Tumor Suppressor ; physiology ; HSP90 Heat-Shock Proteins ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Iron ; metabolism ; Neoplasms ; metabolism ; pathology

Bone Neoplasms ; diagnostic imaging ; pathology ; secondary ; surgery ; Chondrosarcoma ; diagnostic imaging ; pathology ; secondary ; surgery ; Fingers ; Humans ; Male ; Middle Aged ; Osteochondroma ; pathology ; Radiography

Objective To screen differentially expressed genes in placentas with hepatitis B virus (HBV)infection and to discuss the molecular mechanism of HBV intrauterine infection.Methods Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group.The suppression subtractive hybridization(SSH)technique was used.Total RNAs of placenta tissue of the study group were mixed as the tester,and total RNAs of placenta tissue of the control group were mixed as the driver.A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems.Amplifications of the library were carried out with E.coil strain DH5? by reverse spot hybridization.RT-PCR confirmed that phosphatidylinositol 3-kinase(PI3K)was up-regulated in placenta tissue with HBV infection.Results Colony PCR showed that the clones contained 200-1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics.Thirty three known genes and 2 genes with unknown function were obtained.RT-PCR preliminarily confirmed that PI3K gene was up-regulated in HBV infected placenta.Conclusions The differentially expressed genes in placentas with hepatitis B virus(HBV)infection using SSH technique has been screened out successfully.These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation,and signal conduction-antiapoptosis pathway.This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.

Abdominal Pain ; etiology ; Acute Pain ; etiology ; Child ; Humans ; Lymphoma ; complications

Objective To study the clinical significance of fecal smear examination on diagnosing intestinal flora imbalance in infantile diarrhea.Methods A sterile cotton swab was used to spread a layer of fresh feces quantum satis from a sterile container on a clean slide;the smear was fixed and stained with Gram′s methods after it was air-dried,then the specimen was observed with a microscopy(field lens 100 ? eye lens 10) and recorded.Results In the acute diarrhea group(40 cases),the distribution feature of 3 floras on the fecal smears:≥50% Gram-negative bacilli in 6 cases(15%),≥50% Gram-positive cocci in 30 cases(75%) and ≥68% Gram-positive bacilli in 4 cases(10%).In the delayed and chronic diarrhea group(62 cases),the distribution feature of 3 floras on the fecal smears:≥50% Gram-negative bacilli in 7 cases(11.29%),≥50% Gram-positive cocci in 44 cases(70.97%) and ≥68% Gram-positive bacilli in 6 cases(9.68%).In the normal control group(32 cases),the distribution feature of 3 floras on the fecal smears:≥50% Gram-negative bacilli in 1 case(3.13%),≥50% Gram-positive cocci in 1 case(3.13%),and ≥68% Gram-positive bacilli in 17 cases(53.13%).For the distribution of 3 floras in the 3 groups,chi-squared test was performed,and the results showed that the difference was significant in Gram-positive cocci(?~2=47.76 P0.05).Conclusions Acute,delayed or chronic diarrhea can lead to flora imbalance.In order to timely and rapidly know the flora imbalance in children with diarrhea,clinically the simple,easily operated and practical smear staining method shall be widely applied.

Full-length NS5A gene of the hepatitis C virus was amplified by PCR using plasmid pBAC25 containing HCV nonstructural gene as template. The amplified fragment (about 1.34 kb) was cloned into plasmid pQE32, and the recombinant plasmid pQENS5A was expressed in JM109 strain. The NS5A protein was purified by NiSO4 metal chelating resin, and characterized by Western-blot. Its antigenecity was determined by ELISA. The positive detection rate of anti-NS5A was 75% (69/92) in ninety-two clinic sera. The positive rate of anti-NS5A was 82.5% (33/40) in fourty positive standand sera, and the negative rate of anti-NS5A was 100% (40/40) in fourty negative standand sera. The results showed that the Full-length NS5A proteinn had the higher sensitivity and specificity in the detection of HCV antibody in sera, we suggested that NS5A protein was a useful antigen for blood screening.