Liver cancer stem cell plays an important role in hepatocarcinoma occurrence and metastasis.Presently, the two theories about the source of liver cancer stem cells are mature hepatocytes' dedifferentiation and hepatic stem cells' blocked differentiation.Side population cell sorting and different surface antigen labeling are general sorting methods of hepatocarcinoma stem cells.Targeting the liver cancer stem cells population provides an effective way to the treatments of liver neoplasms.

Establishing medical service payment system reform organization of diagnosis related groups and prospective payment system(DRGs-PPS) based on the theory of matrix management and learning organization.Firstly joint committee mechanism hasbeen established to guide the implementation of DRGs-PPS with the strong support of Beijing local government; Secondly series of related technical standards and organizational systems including AP-DRGs,AR-DRGs Group,Data Dictionary,ICD Group,International Classification of Surgical Operations Group,Pricing Classification Group,Training Group,Supervision Group,Support Group,Data Validation Group,BJ-DRGs Group have been created to support standard maintenance,project inspection and data analysis; Furthermore,learning organization has been realized by setting up various project groups and the establishment of DRGs-PPS platform help ensure performance evaluation of healtheare institutions and payment system reform.

Objective To observe the effects of betulinic acid (BA) on proliferation of human hepatoma stem cell;To discuss its anti-cancer mechanism from the aspects of cell cycle and cell apoptosis. Methods HepG2 stem cells were cultivated in vitro and testified the self-renewal capacity. The effects of BA in concentration of 40, 20, 10, 5, 2.5, 1.25μmol/L on the cell vitality of cultured human liver cancer stem cells for 24 and 48 hours were measured with CCK-8 method. The human hepatoma stem cell line HepG2 was administrated by BA at concentrations of 40, 20, 10, 5μmol/L for 48 hours, and cell cycle and apoptosis rate were measured by flow cytometry. Results BA could inhibit HepG2 stem cell proliferation obviously with dose-effect relationship. BA influenced cell cycle, and induced tumor stem cell apoptosis. 40μmol/L BA blocked cell cycle in S phase, and cell apoptosis rate reached 10.86%. Conclusion BA has obvious inhibitory effects on proliferation of HepG2 liver cancer stem cell, which probably plays a part in anti-cancer by influencing cell cycle and inducing cell apoptosis.

Objective To investigate the therapeutic effect of kansui root on patients with severe acute pancreatitis(SAP). Methods Clinical data of 54 cases of severe acute pancreatitis treated with kansui root(kansui root group) were analyzed and compared with 54 cases of severe acute pancreatitis treated without kansui root (control group).Results The releivng time of abdominal pain was significantly shorter than that in control group( P

Objective To study the skills and effects of endoscopic retrograde appendicitis therapy (ERAT) in treating patients with uncomplicated acute appendicitis .Methods We enrolled 21 patients with suspected acute appendicitis who then underwent emergent ERAT between October 2014 and January 2015 .The data of treatment were collected and the operative skills and effects of ERAT were analyzed . Results ERAT was completed successfully in all the patients ,resulting in a success rate of 100% .Mean operation time of ERAT was (49 .7 ± 18 .2) min and mean hospital stay was (3 .3 ± 1 .6)d .Cannulation of the appendix lumen was the most critical step of ERAT ,and cannulation time [(5 .7 ± 4 .9)min , P< 0 .05] was shortened significantly by the use of LoopTip guidewire . Fourteen patients with intraluminal appendicoliths (7 of massive appendicoliths , 4 of sand‐like appendicoliths and 3 of sand‐like appendicoliths with luminal stenosis ) underwent endoscopic lithotomy successfully with balloon or basket ,with the success rate of 100% .One patient who presented perforation after appendicolith removal by basket was cured with conservative treatment .Appendix stent was inserted ,then pulled out after 1 week in 9 patients ,while no complaint or complication of the stent was observed .Operation time of ERAT shortened with the increase of case number .Conclusion ERAT is an effective and safe therapy for treating patients with uncomplicated acute appendicitis .The high success rate and safety of ERAT will be achieved by selecting suitable instruments for cannulation and appendicolith removal ,deciding suitable indications for stenting ,and accumulating of operative cases .

Objective To study the effect and safety of endoscopic retrograde appendicitis therapy (ERAT) in treating patients with uncomplicated acute appendicitis. Methods Patients with uncomplicated acute appendicitis were enrolled and divided into ERAT group and LA group received laparoscopic appendectomy. Then compare treat-ment condition, complications and follow-up of the two groups. Results ERAT were completed successfully in all the patients in ERAT group, while one patient underwent a reversion to open appendectomy for technical difficulties in LA group. Mean operative time was (49.7 ± 18.2) min for ERAT group and (68.9 ± 25.9) min for LA group (P <0.05). Fever relief time (1.3 ± 0.5) d, WBC normalization time (2.0 ± 0.9) d, mean bed time (0.1 ± 0.2) d and mean hospital stay (3.3 ± 1.6) d for ERAT group were significantly lower than LA group (P <0.05). 14 patients with intra-luminal appendicoliths (7 of massive appendicoliths, 4 of broken appendicoliths and 3 of broken appendicoliths with luminal stenosis) underwent endoscopic lithotomy successfully in ERAT group, resulting in a success rate of 100.00%. One patient presented perforation after ERAT was cured with conservative treatment. During the follow-up of at least 1/2 year, the rate of recurrence was 10.00% in ERAT group. 1 patient (5.00%) underwent LA at the 5th month after ERAT during the follow-up. Conclusion ERAT is an effective and safe therapy in treating patients with uncomplicated acute appendicitis with advantages of minimal invasiveness and quick recovery. Uncomplicated acute appendicitis with appendicoliths and/or luminal stenosis are the most suitable indications for ERAT.

Objective To study the expressions of myeloid cell leukemia-1 (Mcl-1 ) gene in mouse macrophages Raw264.7 and human macrophages THP-1,to screen out the cell lines with high levels of expression as the experimental cells,and based on the screening results to construct the short hairpin RNA(shRNA)eukaryotic expression plasmid targeting mice Mcl-1 gene for transfection and further screen out the shRNA expression plasmid with the most obvious effect in silencing Mcl-1 gene.Methods Semi-quantitative PCR method was used to detect the expression of Mcl-1 mRNA in the two kinds of macrophages.Western blotting was adopted to detect the expressions of Mcl-1 proteins in the two kinds of macrophages.Three different gene loci for Mcl-1 shRNA fragments were designed with small molecules interfering RNA (siRNA)software.Eukaryotic expression plasmid Mcl-1 shRNA 1-3 carrying the shRNA fragments was constructed by a company.And the eukaryotic expression plasmid vector was transfected into scavenger cells Raw264.7 mice via through the liposome.The transfection results 24 h and 48 h after transfection were observed under the inverted fluorescence microscope;Mcl-1 mRNA and protein expression were detected by real time quantitative PCR and Western blot,respectively.Results The relative expression levels of Mcl-1 mRNA and protein in mouse macrophages Raw264.7 were significantly higher than those of human macrophages (P <0.05).The shRNA expression vector constructed within 24 h and 48 h could decrease Mcl-1 mRNA and protein levels in Raw264.7 cells,especially with the most obvious silencing effect at 48 h.The 48-h transfection group differed significantly from normal group,liposome group and negative control group (P <0.05).Compared with Mcl-1shRNA1and Mcl-1shRNA2,Mcl-1shRNA3 had the strongest effect in silencing Mcl-1mRNA and protein.Conclusion We have successfully screened out experimental Raw264.7 cells and Mcl-1 shRNA eukaryotic expression plasmid which has an obvious silencing effect targeting on Mcl-1 in mice macrophages Raw264.7.

Objective:To transfect Mcl-1shRNA into Raw264.7 cells,and screen out specific shRNA eukaryotic expression plasmids with the most significant effect of silent Mcl-1 gene to figure out the effect of shRNA on Mcl-1 expression in murine macrophage cell line Raw 264.7.Methods: Specific shRNA was transfected into murine macrophage cell line Raw 264.7 via lipofectamine.Semi-quantitative RT-PCR and Western blot were respectively employed to test the changes in Mcl-1 mRNA level and Mcl-1 protein expressions 24 h and 48 h after the transfection ,and the silencing effects of the three pairs of specific shRNA fragments corresponding to different sites were analyzed.Results: Specific shRNA fragments at 24 h and 48 h could effectively reduce Mcl-1 mRNA and protein level ,with higher silencing effects than those of the normal group ,the lipofectamine group and the negative control group.There were statistically significant differences among them ( P<0.05 ).Among the three pairs of specific shRNA fragments corre-sponding to different sites ,Mcl-1 shRNA3 showed the most significant inhibiting effect on Mcl-1 mRNA and proteins.Conclusion:RNA interference can downregulate the level of Mcl-1 mRNA in murine macrophage cell line Raw 264.7 and greatly downregulate the expression of Mcl-1protein.Specific shRNA eukaryotic expression plasmids with the most significant effect of silent Mcl-1 gene have been screened out successfully.

The aim of this study is to investigate the immune effects of BCG primary immunization and IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine booster immunization on mice .We randomly divided the mice into 7 groups ,namely PBS negative controls ,BCG controls ,pcAg85A controls ,BCG primary immunization combined with Ag85A booster immunization controls ,BCG primary immunization combined with Ag85A and IL-12 booster immunization controls , BCG primary immunization combined with IL-12 booster immunization controls ,and BCG primary immunization combined with pcDNA3 .1 booster immunization controls .Implementing the immune in procedure of BCG primary immunization and cytokine IL-12 booster immunization combined with mycobacterium tuberculosis Ag85A DNA ,we observed the immune effect on mice by detecting the mice serum total IgG , specific lymphocyte proliferation and cytokine levels in 4 ,6 ,8 weeks after the last immunization .Comparing the mice immunized in the strategy of BCG primary immunization and cytokine IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine strengthening immunization to the mice in other groups by other immune ways ,we found that ,in BCG/Ag85A+ IL-12 groups ,IgG in-creased significantly (P＜ 0 .05) ,specific lymphocyte prolifera-ted significantly ,and after strengthening immunization IFN-γlevels ,IL-2 levels and IL-4 levels in the three periods were 128 .2 ±20.4,190.2±16.51,244.2±39.14 ;146.2±17.29,271.6±16.36and16.36±28.12 ;68.6±6.62,96.6±5.5and5.5± 10 .71 ,respectively ,which were higher than those in other groups (P＜0 .05) .It’s suggested that the immunization way of BCG primary immunization and cytokine IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine booster immu-nization could significantly enhance the humoral and cellular immunity of the bodies ,and provide the basis for further study on protective effect test in animals .

AIM:To investigate the effect of inhibiting Mcl-1 gene expression on apoptosis of mouse peritoneal macrophages infected with different virulence of Mycobacterium tuberculosis using a technique of RNA interference .METH-ODS:The BALB/c mice were infected with prepared bacterium of the virulence strains of Xinjiang , H37Rv, H37Ra and BCG.Mcl-1-shRNA was applied to the mouse model of infection , and the control groups were set up .On 1 d, 3 d, 5 d and 7 d, the mouse peritoneal macrophages were collected .The expression of Mcl-1 at mRNA and protein levels was deter-mined by real-time PCR and Western blot .The apoptotic rate of peritoneal macrophages was analyzed by flow cytometry . RESULTS:The expression of Mcl-1 at mRNA and protein levels was up-regulated in the peritoneal macrophages from the mice infected with different virulence of Mycobacterium tuberculosis, and the cells from the mice infected with virulence strains of Xinjiang and H37Rv expressed higher level of Mcl-1 than the uninfected control cells (P<0.05).The expres-sion of Mcl-1 at mRNA and protein levels was reduced by RNA interference as compared with control group ( P<0.05 ) . Inhibition of Mcl-1 expression induced apoptosis of peritoneal macrophages in the mice .CONCLUSION: The Mcl-1 ex-pression at mRNA and protein levels in mouse peritoneal macrophages infected with different virulence of Mycobacterium tu-berculosis was effectively suppressed by Mcl-1-shRNA, which can induce macrophage apoptosis .