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.

Objective To explore the influencing factors of blood concentration of tacrolimus in pediatric living donor liver transplant recipients and provide rationales for individualized administration of tacrolimus .Methods Trough concentrations (C0 ) , doses of tacrolimus , recipient age , gender , body weight ,donor and recipient CYP3A5 genotypes ,hematocrit (HCT ) and liver/kidney function related indicators at 3 ,5 ,7 ,14 days ,1 month , 2 months and 3 months post living donor liver transplantation were collected from a total of 100 pediatric recipients .Taking ratio of concentration to dose (C0 /D) as a dependent variable ,the influencing factors of blood concentration of tacrolimus were analyzed by multivariate stepwise regression .Results The influencing factors of blood tacrolimus concentration at 3d post-transplantation were recipient CYP3A5 genotyp , donor CYP3A5 genotype and weight of recipients . The major influencing factors at 5d post-transplantation were recipient & donor CYP3A5 genotypes , recipient weight and HCT . The major relevant factors at 7d posttransplantation were CYP3A5 of recipients ,age and HCT .The influencing factors at 14 days were the same as those at 2 months ,i .e .CYP3A5 genotype and weight of recipients .At 1 month the major influencing factors were weight of recipients ,CYP3A5 of recipients and alkaline phosphatase (ALP) ; CYP3A5 genotype and weight of recipients at 3 months . Further study on CYP3A5 genotype of donors and recipients , the C0 /D ratio of CYP3A5 genotype non-expression group was significantly higher than that of expression group in recipients and C0 /D ratio of donor CYP3A5 genotype nonexpression group was significantly higher than that of expression group .Conclusions The influencing factors of concentration of tacrolimusvary at different timepoints after liver transplantation . Paying close attention to the changes of CYP3A5 genotype , weight of recipients and related biochemical indexes and considering various influencing factors facilitate individualized dosing for improving the prognosis of pediatric recipients .

Objective To explore application of nursing intervention of children's fun mode m anesthesia induction of preschool children.Methods A total of 180 preschool children undergoing elective surgery in our hospital were randomly divided into control group and observation group according to the random number table method,with 90 cases in each group,the control group was given routine nursing,and the observation group with children's fun mode nursing intervention,the curative effect is compared between the two groups.Results Before operation,there was no significant difference between the two groups in the improvement of the perioperative anxiety scale (mYPAS) score in the groups (P ＞ 0.05).During induction of anesthesia,mYPAS score of the two groups before operation were significantly higher than one day before surgery,and the differences were statistically significant (P ＜ 0.05),and anxiety score in the observation group was significantly lower than the control group (P ＜ 0.05).Before one day of operation,there was no significant difference in systolic blood pressure (SBP) and diastolic blood pressure (DBP) between the two groups (P ＞0.05).During induction of anesthesia,the SBP,DBP of two groups were significantly higher than one day before operation,and the differences were statistically significant (P ＜ 0.05),and the observation group had lower SBP and DBP than the control group,the difference was statistically significant (P ＜ 0.05).Before one day of surgery,there was no significant difference in heart rate (HR) between two groups (P ＞ 0.0 5).During induction of anesthesia,the two groups had higher HR than one day before operation (P ＜ 0.05),and the observation group had lower HR than the control group (P ＜ 0.05).The observation group during anesthesia induction had lower cooperation scale (ICC) score than the control group(P ＜ 0.05).Conclusion Nursing intervention of children's fun mode for preschool children can improve children's blood pressure,and has less influence on HR so as to improve cooperation of anesthesia induction.

Objective To explore application of nursing intervention of children's fun mode m anesthesia induction of preschool children.Methods A total of 180 preschool children undergoing elective surgery in our hospital were randomly divided into control group and observation group according to the random number table method,with 90 cases in each group,the control group was given routine nursing,and the observation group with children's fun mode nursing intervention,the curative effect is compared between the two groups.Results Before operation,there was no significant difference between the two groups in the improvement of the perioperative anxiety scale (mYPAS) score in the groups (P ＞ 0.05).During induction of anesthesia,mYPAS score of the two groups before operation were significantly higher than one day before surgery,and the differences were statistically significant (P ＜ 0.05),and anxiety score in the observation group was significantly lower than the control group (P ＜ 0.05).Before one day of operation,there was no significant difference in systolic blood pressure (SBP) and diastolic blood pressure (DBP) between the two groups (P ＞0.05).During induction of anesthesia,the SBP,DBP of two groups were significantly higher than one day before operation,and the differences were statistically significant (P ＜ 0.05),and the observation group had lower SBP and DBP than the control group,the difference was statistically significant (P ＜ 0.05).Before one day of surgery,there was no significant difference in heart rate (HR) between two groups (P ＞ 0.0 5).During induction of anesthesia,the two groups had higher HR than one day before operation (P ＜ 0.05),and the observation group had lower HR than the control group (P ＜ 0.05).The observation group during anesthesia induction had lower cooperation scale (ICC) score than the control group(P ＜ 0.05).Conclusion Nursing intervention of children's fun mode for preschool children can improve children's blood pressure,and has less influence on HR so as to improve cooperation of anesthesia induction.