Quality indicator is defined as the measure used to access the degree of inherent characteristics meeting the requirements .It is a powerful tool to improve laboratory quality to monitor and evaluate performance throughout critical steps in the total testing process .Targeted quality improvement can be obtained by quantizing quality levels in each phase when the quality indicators applied .Establishing and monitoring the quality indicators enables laboratory to compare over time between providers , and evaluate the effectiveness of delivered services and improving patient safety .

Objective:To establish the RPB5-mediating protein (RMP)-silenced stable cell lines and study the inhibitory effects of small interfering RNA (siRNA) targeting RMP gene on the proliferation and migration of human hepatoma SMMC-7721 cells. Methods:Three RMPi siRNAs were designed and synthesized in vitro and transfected into SMMC-7721 cells. The inhibitory effect of siRNA on RMP gene expression was measured by RT-PCR to select the best siRNA. The expression vector pGPU6-Neo-RMP-484 was transfected into SMMC-7721 cells by the lipofectamine and the cells stably expressing the siRNA were selected by G418. RT-PCR was used to detect the interference efficacy against RMP gene. Cell proliferation and adhesion were measured by MTT assay. Wound healing test was used to observe the migration ability of cells. Results:The SMMC-7721 cell lines with down-regulated RMP expression were established by using RNA interference technology. Compared with the negative control cells, expression of RMP mRNA was down-regulated by(83.67±2.56)% .The proliferation of stable-transfected cells was inhibited by(74.33±0.58)% . The adhesion capability of stable-transfected cells was enhanced but the migration capacity was decreased compared with the negative control cells. Conclusion:The pGPU6-Neo-RMP-484 cell lines with stable transfection of RMP siRNA recombinant vector are successfully screened,which can be used as a cellular model for studying the molecular mechanism of RMP. Down-regulation of RMP gene expression can effectively inhibit the proliferation, enhance the adhesion, and decrease the migration of SMMC-7721 cells.

New strategies are needed for prevention of Pseudomonas aeruginosa (P. aeruginosa) infections, a widespread disease caused by P. aeruginosa with strong drug resistance. The immunoprotective capacity of the receptor of autoinducers protein LasR/RhlR was examined in the BALB/c mice. At first, specialized expression plasmids were developed to facilitate expression of LasR/RhlR proteins in Escherichia coli (E. coli). Then, biofilms were grown from clinical isolated P. aeruginosa PA0305 to investigate the relative contributions of cell signaling for biofilm formation. Morphological characters of biofilm were quantified using Image-Pro Plus software. Fluorescence analysis demonstrated that cell signal molecule LasR/RhlR significantly (P ＜ 0.05) influenced development of P. aeruginosa biofilm. Active immunization of mice with LasR/RhlR was found to provide significant protection against homologous challenge with P. aeruginosa in mice lungs. In 10 days after lungs inoculation, the bacterial clearance rate of the immunized mice was clearly higher than that of non-immunized groups on the basis of microbiological and histological assays. The protective effects of immunization with LasR and Rh1R together were the same as the result of LasR or Rh1R immunized mice alone. These data indicate that the manner ofLasR, Rh1R or both is an important determinant of immunoprotection in mice lungs infection.

BACKGROUND:Biliary stricture following liver transplantation is mainly focus on biliary stoma stricture; while, balloon dilatation temporarily keeps biliary tract open but not works out a solution at all.OBJECTIVE: To discuss the diagnosis and treatment of postoperative biliary stricture after orthotopie liver transplantation by the endoscope technique.DESIGN, TIME AND SETTING: A case analysis, which was performed at Dalian Liver and Gall Surgical Institute. Ten patients hospitalized from the Department of Liver and Gall Surgery of Dalian Friendship Hospital and four patients hospitalized from the Department of Organ Transplantation of Tianjin First Central Hospital were diagnosed as biliary stricture after orthotopic liver transplantation.PARTICIPANTS: Among 14 patients, 10 males and 4 females with mean age of 46 years provided end-to-end biliary anastomose.METHODS: Fourteen cases of postoperative biliary stricture after orthotopic liver transplantation were analyzed and diagnosed by endoscope technique. And by endoscope technique, the stricture was supported with tube after balloon dilatation.MAIN OUTCOME MEASURES: Bile duct mucous membrane under T-tube radiography and endoscope; calculary distribution and bile duct mucous membrane at stoma; healing of biliary stoma of donors and recipients; inflammatory edema and stricture; recheck of above-mentioned parameters after stricture expansion by endoscopic stone extraction technique.RESULTS: Thirteen cases of postoperative biliary stricture after orthotopic liver transplantation were analyzed and diagnosed by endoscope technique, including one was induced by calculus, and one non-stoma stricture. One case was treated with balloon dilation; biliary infection and jaundice occurred in 2 cases after endoscopic sphincterotomy (EST) + basket lithotripsy + endoscopic nasobiliary drainage (ENBD), so operations or fibrocholedochoscope treatments had to be carried out. By T tube radiography, in 1 case there was strip-like negative simulacrum or no stricture, well-healed anastomosis and good mucous membranel transition; poor or no intrahepatic visualization were found in 2 cases, so anastomosis dilation was processed after the calculi removal by fibrocholedochoscope, stricture disappeared in 3 or 4 months; in 8 cases there were blur extrahepatic or intrahepatic biliary visualization, cord-like, column or branch-like negative simulacrum in biliary ducts and sign of non-anastomosis stricture, after removal of calculi, anastomosis stricture and congestion, edema were found, all these disappeared after average 2.5 months of dilation; the other 1 case was found stricture by T the radiography, but no calculi was found with fibrocholedochoscope, finally the Ttube was removed after 2 months of stricture dilation.CONCLUSION: Endoscopy is significant to directly reflect and reliably diagnose postoperative biliary stricture and effectively treat biliary stricture by anastomosis dilation.

With the dramatically increasing detection rate of ground-glass nodules (GGN), exact understanding and treatment strategy of them has become the hottest issue currently. More and more studies have begun to explore the underlying mechanisms of their indolent characteristics and favorable prognosis from the perspectives of molecular evolution and immune microenvironment. GGN has different dominating gene mutations at different evolutional stages. The pure GGN has a lower tumor mutation burden and genomic instability, while a gradually evolutionary feature of genomic mutation along with the pathological progression can be observed. GGN has less infiltration of immune cells, and they are under the pressure of immune surveillance with weakened immune escape. With the increase of solid components, an inhibitory immune microenvironment is gradually established and immune escape is gradually enhanced, leading to rapid tumor growth. Further exploration of the molecular characteristics of GGN will help to more precisely distinguish these highly heterogeneous lesions, which could be helpful to make personalized treatment plans.

With the dramatically increasing detection rate of ground-glass nodules (GGN), exact understanding and treatment strategy of them has become the hottest issue currently. More and more studies have begun to explore the underlying mechanisms of their indolent characteristics and favorable prognosis from the perspectives of molecular evolution and immune microenvironment. GGN has different dominating gene mutations at different evolutional stages. The pure GGN has a lower tumor mutation burden and genomic instability, while a gradually evolutionary feature of genomic mutation along with the pathological progression can be observed. GGN has less infiltration of immune cells, and they are under the pressure of immune surveillance with weakened immune escape. With the increase of solid components, an inhibitory immune microenvironment is gradually established and immune escape is gradually enhanced, leading to rapid tumor growth. Further exploration of the molecular characteristics of GGN will help to more precisely distinguish these highly heterogeneous lesions, which could be helpful to make personalized treatment plans.

This is a study on the biodegradable polymers as gene controlled-released coatings for gene transfer. The PELA (poly (Dl-lactic acid)-co-poly (ethylene glycol), and PLGAE (poly (lactic acid)-co-poly (ethylene glycol)-co-poly (glycolic acid) random copolymer) were synthesized and prepared as the coatings of plasmid pCH110 in the transfection. All kinds of factors affecting the loading efficiency, cytotoxicity, transfection efficiency and the course of the degradation and release in vitro were discussed. The average diameters of microspheres of PELA and PLGAE were 1-3 microm and 0.72 microm respectively. The loading efficiency levels of them were 62% and 70% respectively. The transfection efficiency levels of two kinds of pCH110 delivery system for COS-1 cells were higher and two of them had few cytotoxicity. After transfection, the X-gal assay was performed and reported positive for 96 h. The biodegradable polymeric materials as gene carriers possess their potential superiority.
Biocompatible Materials ; DNA ; chemistry ; Drug Carriers ; chemical synthesis ; chemistry ; toxicity ; Gene Transfer Techniques ; Lactates ; chemical synthesis ; chemistry ; toxicity ; Lactic Acid ; chemical synthesis ; chemistry ; toxicity ; Polyethylene Glycols ; chemical synthesis ; chemistry ; toxicity ; Polyglycolic Acid ; chemical synthesis ; chemistry ; toxicity ; Polymers ; chemical synthesis ; chemistry ; toxicity ; Transfection

Salmonella Typhi capsular polysaccharide vaccines were encapsulated in the Micro-particles made from polyethylene glycol-poly-DL-lactide (PELA). BALB/c mouse were divided into three groups with 20 mice in each. Mouse were immunized respectively with controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines and Salmonella Typhi capsular polysaccharide vaccines by oral and subcutaneous administration. The mice blood and salvia were collected at the 2nd, 4th and 8th weeks respectively for the titrating of IgG and sIgA antibodies by RIA. At the 8th week, live typhoid bacteria were injected into the immunized mice for the calculation of the rate of immunization protection. The IgG titers of the controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines group were higher than those of the other groups(P < 0.05). The IgA titers of the low groups of controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines (oral and subcutaneous) were higher than those of the group of Salmonella Typhi capsular polysaccharide vaccines (P < 0.05). The immunization protection rates of the three groups were 40%, 100% and 60% respectively. The controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines possess the advantages of releasing slowly in vivo and persisting long time immunogenicity.
Administration, Oral ; Animals ; Delayed-Action Preparations ; Female ; Immunoglobulin A, Secretory ; analysis ; Immunoglobulin G ; blood ; Injections, Subcutaneous ; Mice ; Mice, Inbred BALB C ; Microspheres ; Polysaccharides, Bacterial ; administration & dosage ; immunology ; Typhoid-Paratyphoid Vaccines ; administration & dosage ; immunology ; Vaccination

A total of 50 clinical imipenem-resistant isolates of Pseudomonas aeruginosa and Acinetobacter baumannii were subjected to the ceftazidime-2- mercaptoethanol -double-disk synergy test and to the PCR assays with primers specific for bla(IMP-1). After the process of sequencing the positive one to identify the results, PCR analysis was conducted with primers specific for class 1 integrons. For synergy test, 28 isolates gave positive results, among which were 27 Pseudomonas aeruginosa and Acinetobacter baumannii. Only one Pseudomonas aeruginosa was found to carry bla(IMP-1), and bla(Int1) at the same time. This is the first ascertainment of IMP-1 producing Pseudomonas aeruginosa isolate carrying bla(IntI1) in West China, which is of significance to the research on the clinical spread of these drug-resisitant genes.
Acinetobacter baumannii ; drug effects ; genetics ; Anti-Bacterial Agents ; pharmacology ; Ceftazidime ; pharmacology ; Drug Resistance, Multiple, Bacterial ; genetics ; Fermentation ; Imipenem ; pharmacology ; Mercaptoethanol ; pharmacology ; Microbial Sensitivity Tests ; Pseudomonas aeruginosa ; drug effects ; genetics ; beta-Lactamases ; genetics

BACKGROUND:Overactive osteoclasts disrupt bone homeostasis and play a bad role in the pathological mechanisms of related skeletal diseases,such as osteoporosis,fragility fractures,and osteoarthritis.Studies have confirmed that ellagic acid and ellagtannin have the potential to inhibit osteoclast differentiation.As their natural metabolites,urolithin A has antioxidant,anti-inflammatory,anti-proliferative and anti-cancer effects,but its effect on osteoclast differentiation and its underlying molecular mechanisms remain unclear. OBJECTIVE:To explore the effect of urolithin A on osteoclast differentiation induced by receptor activator for nuclear factor-κB ligand and its mechanism. METHODS:Mouse mononuclear macrophage leukemia cells(RAW264.7)that grew stably were cultured in vitro.Toxicity of urolithin A(0,0.1,0.5,1.5,2.5 μmol/L)to RAW264.7 cells were detected by cytotoxic MTS assay to screen out the safe concentration.Different concentrations of urolithin A were used again to intervene with receptor activator for nuclear factor-κB ligand-induced differentiation of RAW264.7 cells in vitro.Then,tartrate-resistant acid phosphatase staining and F-actin ring and nucleus staining were performed to observe its effect on the formation and function of osteoclasts.Finally,the expressions of urolithin A on upstream and downstream genes and proteins in the MAPK signaling pathway were observed by western blot and RT-qPCR assays. RESULTS AND CONCLUSION:Urolithin A inhibited osteoclast differentiation and F-actin ring formation in a concentration-dependent manner and 2.5 μmol/L had the strongest inhibitory effect.Urolithin A inhibited the mRNA expression of Nfatc1,Ctsk,Mmp9 and Atp6v0d2 and the protein synthesis of Nfatc1 and Ctsk,related to osteoclast formation and bone resorption.Urolithin A inhibited the activity of osteoclasts by downregulating the phosphorylation of p38 protein to inhibit the mitogen-activated protein kinase signaling pathway.