AIM: To investigate the variation and significance of mRNA and protein concentration of myeloid cell leukemin-1 (Mcl-1) in apoptotic HepG2 cells induced by apoptin. METHODS: The apoptin expression vector pCDNA3.0-VP3 was transfected into HepG2 cells via liposome. Mcl-1 mRNA was analyzed by real-time quantitative reverse transcriptase-polymerase chain reaction. The protein of apoptin, Mcl-1 and cytochrome C were detected by Western blotting. RESULTS: The VP3 gene was transfected into HepG2 cells successfully and expressed steadily. Compared to blank control, Mcl-1 mRNA and protein levels of VP3 positive cells were decreased (mRNA: 0.09%±0.00% vs 0.41%±0.14%, P<0.05; protein: 0.43%±0.01% vs 0.90%±0.04%, P<0.01). Released cytochrome C from mitochondrion was increased (0.98%±0.02% vs 0.62%±0.03%, P<0.01). CONCLUSION: In the course of the apoptosis of HepG2 cells induced by apoptin, the amount of Mcl-1 mRNA and protein is decreased, and released cytochrome C from mitochondrion is increased. The apoptosis induced by apoptin may be correlated with the down-regulation of Mcl-1 mRNA and protein.

Objective To investigate the diagnostic value of transgastric peritoneal endoscopy in diagnosis of ascites with unknown origin.Methods Endoscopy was introduced into peritoneal cavity through gastric wall in 23 patients with exudative ascites which was able to be diagnosed by routine methods and biopsy was made through endoscopy to get pathological diagnosis.Results Definite diagnosis was made in 22 patient (95.7%),of which 12 (54.6%) were malignant tumors,8 (36.4%) were tuberculosis peritonitis,1 (4.5%) was spontaneous peritonitis associated with liver cirrhosis and 1 (4.5%) was eosinophilic enteritis.Conclusion Natural orifice transluminal endoscopy combined with biopsy is an effective and accurate procedure for diagnosis of ascites of unknown canses.

BACKGROUND: It has been believed mesenchymal stem cells (MSCs) play a role in treatment through paracrine mechanism. Various side effects such as embolism, tumorigenesis and immunological reaction caused by direct injection of MSCs can be avoided by extracting MSC lysate. However, there is a larger difference in current collection methods and standards of MSC lysate. OBJECTIVE: To compare repeated freeze-thaw and ultrasonication for the collection of lysate of MSCs. METHODS: Adipose-derived mesenchymal stem cells (ADMSCs) were isolated from the abdominal subcutaneous fat of healthy individuals, and purified with adherence screening method, followed by in vitro amplification using fetal bovine serum medium. The common surface makers of these cells were tested by flow cytometry (1×109, 2×109, 4×109/L). Repeated freeze-thaw and ultrasonication were employed for cell cytoclasis at three different densities respectively in saline and double distilled water, and a comprehensive comparison was performed on cytoclasis rate and the content of protein in cell lysate between the two methods. RESULTS AND CONCLUSION: (1) ADMSCs obtained from in vitro isolated human adipose tissue grew in a swirl or radial pattern with a homogenous size and neat arrangement. CD44, CD90, CD105 and other commonly used surface markers were highly expressed. (2) The study for optimization of lysate collection revealed that the higher cell density implicated a longer time for cell wall disruption and cytoclasis, as well as significantly increased cytoclasis rate. (3) BCA protein assay showed that the highest content of protein was obtained in saline solvent using ultrasonication method. Comprehensive analysis on the results leads to a conclusion that ultrasonication method with saline as the solvent is the optimized method for extraction of ADMSCs lysate, and the cell concentration of less than 4×109/L is recommended.