Objective To study the effect of Numb gene overexpression on invasion and migration in human renal cell carcinoma cells and its related mechanism. Methods The Numb-ORF plasmid was transfected into renal cell carcinoma cells 786-O, set the negative control and blank control. The expression of Numb and matrix metalloproteinase-9 (MMP-9) was detected by real-time PCR and western blot. Then abilities of cells invasion and migration were measured respectively by transwell assay and scratch-wound-healing assay. Results Compared to negative control (2.51±0.27) and blank control (2.87±0.21), Numb expression in Numb-ORF group (36.13 ±8.33) was significantly higher (P= 0.00). Meanwhile, MMP-9 level in Numb-ORF group was decreased: Numb-ORF group was 2.36±0.29, negative control was 8.73±0.91, blank control was 8.99±0.78 (P=0.00). In scratch-wound-healing assay,the repair ratio of 12 h was as follow: Numb-ORF group was 0.53 ±0.06, negative control was 0.73 ±0.09, blank control was 0.75 ±0.08 (P= 0.02). In transwell assay, the number of invasion cells in Numb-ORF group was 31.40±5.96, less than negative control (126.93±13.61) and blank control (131.87 ±15.42) (P= 0.00). Conclusion Overexpression of Numb significantly decreases abilities of invasion and migration in 786-O cells, and the suppressive effect may be due to the down-regulation of MMP-9 expression in human renal cell carcinoma.

Objective To establish a high performance liquid chromatography(HPLC) method for the simultaneous determination of eight components in Gynostemma pentaphyllum(Thunb.) Makino (ginsenosides Rb1, Rb3, Rd, Re, F2, and gypenosides A, XLIX and XVII). Methods HPLC was performed on Agilent Eclipse XDB-C18(4.6 mm × 250 mm, 5 μm) chromatographic column with 0.3％ formic acid solution (A)- acetonitrile (B) as the mobile phase by gradient elution, flow rate was 1.5 mL·min-1, detection wavelength was 203 nm, and column temperature was set at 50 ℃. Results There was a good linear relationship between peak area and concentration of eight components (r ≥ 0.999) , the recovery was in the range of 97％ to 100％. Conclusion The established method is simple, rapid, accurate, reliable, and reproducible, and can be used for the quality control of Gynostemma pentaphyllum(Thunb.) Makino.

OBJECTIVETo observe the morphological characteristics of Plasmodium yoelii schizogony in their ghost erythrocytes.

METHODSSaponify, hypotonic shock, and electron microscopy were used to observe the different fashions of erythrocytic parasites and their characteristic organellae in ghost erythrocytes.

RESULTSThe malarial parasites and their fine structures were dramatically well preserved in the ghost erythrocytes, such as the ring-like early trophozoites, the brassiere-like early schizonts, the emerging buds on the surface of late schizonts, and the grape-cluster like late schizonts. The cytostome, food vacuole, and crystallized malarial pigments were found in the early trophozoites. The proliferations of nucleoplasma and nuclear membrane as well as and the clot-like nuclear division were followed by the budding during the schizogony.

CONCLUSIONThe saponify technique that makes the erythrocytic malaria parasites and their fine organellae to be dramatically revealed in their ghost erythrocytes, may be a useful method in the Plasmodium biological research and anti-malaria immunological researches.

Animals ; Erythrocyte Membrane ; parasitology ; ultrastructure ; Female ; Mice ; Mice, Inbred BALB C ; Microscopy, Electron ; Plasmodium yoelii ; ultrastructure

OBJECTIVETo determine the therapeutic effectiveness and safety of polyethylene glycol 4000 (forlax) in the treatment of constipation in children over 8 years old.

METHODSThis study was designed as a randomized, positive medicine (lactulose) controlled multicenter trial. A total of 216 children with constipation from 8-18 years old from 7 hospitals across China who were matched with a uniform entry criteria were enrolled in this study. The 216 patients were randomized to receive either oral forlax (20 g/d, n=105) or lactulose (15 mL/d, n=111) for 2 weeks. The therapeutic effects, including bowel movement frequency, stool consistency, clinical complete remission rate of constipation and abdominal symptoms, and the safety of forlax and lactulose were evaluated at 1 and 2 weeks of treatment.

RESULTSThe median weekly frequency of bowel movement in the forlax group increased by 4 and 5 times respectively after 1 and 2 weeks of treatment, and increased by 3 and 4 times in the lactulose group (P < 0.05). The stool consistency of the two groups was both improved significantly after treatment. The Bristol score of stool consistency of the forlax and lactulose groups were 3.41+/-1.11 and 3.64+/-1.33 respectively (P < 0.05) after 1 week of treatment, and were 4.26+/-0.89 and 3.63+/-1.33 respectively (P < 0.05) after 2 weeks of treatment. The clinical complete remission rate of constipation in the forlax and lactulose groups was 70% and 40% respectively (P < 0.05) by week 1 of treatment, and that was 72% and 41% respectively (P < 0.05) by week 2 of treatment. Abdominal pain disappeared in 75% of patients in the forlax group but in only 57% in the lactulose group by week 2 of treatment (P < 0.05). No serious adverse events happened and no abnormalities were found in laboratory tests and physical examinations in the two groups after medication.

CONCLUSIONSForlax is safe and effective in the treatment of constipation in children over 8 years old.

Adolescent ; Cathartics ; adverse effects ; therapeutic use ; Child ; Constipation ; therapy ; Female ; Humans ; Lactulose ; therapeutic use ; Male ; Polyethylene Glycols ; adverse effects ; therapeutic use

OBJECTIVETo evaluate the effects of simvastatin combined with omega-3 fatty acids on high sensitive C-reactive protein (HsCRP), lipidemia, and fibrinolysis in coronary heart disease (CHD) and CHD risk equivalent patients with mixed dyslipidemia.

METHODSA randomized, double-blind placebo controlled and parallel group trial was conducted. Patients with CHD and CHD risk equivalents with mixed dyslipidemia were treated with 10 or 20 mg simvastatin for 6-12 weeks. Following with the treatment of patients whose low-density lipoprotein cholesterol (LDL-ch) reaching goal level (< 100 mg/dL) or close to the goal (< 130 mg/dL), while triglyceride (TG) > or = 200 mg/dL and < 500 mg/dL, was combined with omega-3 fatty acids (3 g/d) or a placebo for 2 months. The effects of the treatment on HsCRP, total cholesterol (TC), LDL-ch, high-density lipoprotein cholesterol (HDL-ch), TG, lipoprotein (a) [LP (a)], apolipoprotein A1 (apoA1), apolipoprotein B (apoB), plasminogen activator inhibitor-1 (PAI-1), and tissue plasminogen activator (tPA) were investigated. Forty patients finished the study with each group consisting of twenty patients.

RESULTS(1) There were significant reductions of HsCRP, TG, TC, and TC/HDL-ch, which decreased by 2.16 +/- 2.77 mg/L (38.5%), 94.0 +/- 65.4 mg/dL (31.1%), 13.3 +/- 22.3 mg/dL (6.3%), 0.78 +/- 1.60 respectively in the omega-3 fatty acids group (P < 0.01, < 0.001, < 0.05, < 0.05) compared to the baseline. HsCRP and triglyceride reduction were more significant in omega-3 fatty acids group compared to the placebo group (P = 0.021 and 0.011 respectively). (2) In the omega-3 fatty acids group, the values and percentage of TG reduction had a significantly positive relation with HsCRP reduction (r = 0.51 and 0.45, P = 0.021 and 0.047 respectively).

CONCLUSIONIn CHD and CHD risk equivalent patients with mixed dyslipidemia, dyslipidemia's therapeutic effect using simvastatin and omega-3 fatty acids may result from not only the combination of lipid adjustment, but also enhancement of their own nonlipid influences.

Aged ; C-Reactive Protein ; metabolism ; Coronary Disease ; blood ; Double-Blind Method ; Drug Therapy, Combination ; Fatty Acids, Omega-3 ; therapeutic use ; Fibrinolysis ; drug effects ; Humans ; Hypolipidemic Agents ; therapeutic use ; Lipids ; blood ; Male ; Middle Aged ; Simvastatin ; therapeutic use ; Triglycerides ; blood

OBJECTIVETo analyze the clinical manifestations, laboratory findings, treatment and prognosis of patients with hemophagocytic syndrome (HPS).

METHODSA retrospective study was carried out to analyze the underlying disease, clinical characteristics, laboratory findings and outcomes of 46 patients with HPS.

RESULTSThis cohort included 19 cases of HPS secondary to cancer, 11 cases of HPS secondary to infection, 10 cases of suspected malignant lymphoma based on PET-CT findings (without biopsy), and 6 cases of unknown etiology. The coincidence rate of the clinical characteristics of the patients with the indices listed in HPS-2004 criteria were: fever (100%), elevated serum ferritin (100%), cytopenias (93.48%), splenomegaly (91.30%), hemophagocytosis in the bone marrow, spleen or lymph nodes (84.78%), hypofibrinogenemia (67.39%), and hypertriglyceridemia (54.05%). The cases of cancer, infections and unknown etiology showed significant differences in serum levels of ferritin and β2MG (P<0.05), and significant differences were found in triglycerides, LDH, and fibrinogenemia between the nonfatal and fatal cases (P<0.05).

CONCLUSIONHPS can be secondary to various underlying diseases, many associated with Epstein-Barr virus infection. Cancer, especially NK/T-cell lymphoma, is the main cause of HPS. Persistent fever, elevated serum ferritin level and cytopenias are the most sensitive indicators for diagnosis of HPS, and early diagnosis and treatment are critical to lower the mortality rate of this disease.

Adult ; Aged ; Anthracosis ; diagnostic imaging ; therapy ; Bronchoalveolar Lavage ; Humans ; Lung ; diagnostic imaging ; Male ; Middle Aged ; Radiography

OBJECTIVETo develop a new DNA chip with coloration, which can be used for rapid and economical detection of the genotyping of hepatitis C virus (HCV).

METHODSProbes and primers were designed according to the sequence of HCV 5' non-coding region (5' NCR) to fabricate DNA chip. Experimental group consisted of 60 positive serum samples and control group consisted of 20 negative serum samples. To obtain the aimed gene, then they were hybridized with DNA chip. Finally, the results showed in a nylon film. The results of DNA sequencing of samples were used as the control in double blind experimental.

RESULTSUsing DNA chip, HCV was detected in positive of all serum specimens of experimental group and negative in control group. The determination of HCV genotype by DNA chip showed corresponding rate of 96.7% with those by sequence assay.

CONCLUSIONIt showed higher specialty and sensitivity using DNA chip to detect the genotype of HCV. It would be valuable for the clinical genotyping of HCV

5' Untranslated Regions ; genetics ; Base Sequence ; Genotype ; Hepacivirus ; classification ; genetics ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Sequence Analysis, DNA

OBJECTIVETo investigate the antibacterial effect of a particular antimicrobial peptide Cecropin B(CB) on Pseudomonas aeruginosa infection of wound in mice.

METHODSThirty ICR mice were enrolled in the study, and the Pseudomonas aeruginosa infection model was reproduced by excision of the full layer of dorsal skin with an area of 1 cm x 1 cm. Then they were randomly divided into C ( control, n = 10, with wet compress of isotonic saline at 3 postinjury hour( PIH) ) , M (with hydropathic compress of 100 g/L mafenide at 3 PIH), A (with wet compress of 1 000 mg/L Cecropin B at 3 PIH) groups. The changes in body temperature and hemogram in each group were determined before and 4 days after injury. Quantitative examination were used to detect the quantity of bacteria in muscular tissue of the wounds, and the survival of the mice were observed on 4 post-injury day( PID).

RESULTSThe wounds were moist with more exudation in C group,while that in other groups were dry without obvious exudation. The body temperature of the majority of the mice in each group were elevated, but the number of leucocytes in each group was lowered after operation. The quantity of bacteria in muscle in A group[ (42 +/- 50) CFU/g] was obviously lower than that in M group [(886+/-804) CFU/g, P <0.05] , and it was all obviously lower than that in C group[ (41 +/-28) x 10(5) CFU/g, P <0.01]. The number of surviving mice after 4 PID in C group was evidently smaller than that in A and M groups( P <0. 05).

CONCLUSIONThe cecropin B possesses obvious anti-bacterial effect on the Pseudomonas Aeruginosa infected wounds of ICR mice, and it can reduce the mortality.

Animals ; Antimicrobial Cationic Peptides ; therapeutic use ; Disease Models, Animal ; Insect Proteins ; therapeutic use ; Male ; Mice ; Mice, Inbred ICR ; Pseudomonas Infections ; drug therapy ; Wound Infection ; drug therapy ; microbiology

OBJECTIVE: To localize the gene of autosomal dominant familial dilated cardiomyopathy with conduction defect. METHODS: A Chinese family which was diagnosed as dilated cardiomyopathy with conduction defect was studied. Venous blood (3 - 5 mL) from some family members was collected, and genomic DNA was extracted from the blood. Then whole genome wide scan was performed after excluding the known markers on the candidate loci (CMD1A, CMD1 E, CMD1F, and CMD1H) by two-point linkage analysis. RESULTS: No significant evidence for linkage was found in the two point linkage analyses to the known markers in the analyzed family. And the whole genome wide scan showed the maximum LOD score reached 2.68 at marker D3S1614 ( at recombination fraction theta = 0). CONCLUSION The related gene in this kindred is located on 3q26 other than on CMD1A, CMD1H, CMD1E, and CMD1F.
Adult ; Arrhythmias, Cardiac ; etiology ; genetics ; Cardiomyopathy, Dilated ; genetics ; Chromosomes, Human, Pair 3 ; genetics ; Female ; Genetic Linkage ; Humans ; Male ; Microsatellite Repeats ; Middle Aged ; Pedigree