Objective To investigate the expression of p21-activated kinase4 (PAK4) in human gastric cancer, and to evaluate the correlation of PAK4 protein level with clinical pathologic parameters. Methods Immunohistochemical method was employed for detecting the expression of PAK4 in tissue samples obtained from 95 gastric cancer patients who underwent surgical treatment. Western blot was also used to measure the expression of PAK4 in gastric cancer tissues and adjacent non-cancerous gastric mucosa in 40 patients and also in 7 human gastric cancer cell lines. Results Immunohistochemical staining results showed that positive PAK4 expression in the gastric cancer tissues was 67.4%, and 96.9% of those (62/64) was located in the cytoplasm, while only 3.1% (2/64) was on the cell membrane. Expression of PAK4 was revealed to be significantly and positively correlated with both TNM stage (X2=10. 426, P<0.01) and lymph node metastasis (X2= 4.912, P<0.05 ). Western blot showed that PAK4 expression of gastric cancer tissues was significantly higher than that of adjacent non-cancerous gastric mucosa (t=-5. 978, P<0.01). Expression of PAK4 could also be detected in 7 human gastric cancer cell lines. Compared with GES-1, expression of PAK4 is higher in MKN45, AGS, N87 and BGC823, but it was lower in MGC803, MKN1 and SGC7901. Conclusion The overexpression of p21-activated kinase4 (PAK4) may be correlated with the carcinogenesis and prognosis of gastric cancer.

Objective To investigate the expression of p21-activated kinase4 (PAK4) in human gastric cancer, and to evaluate the correlation of PAK4 protein level with clinical pathologic parameters. Methods Immunohistochemical method was employed for detecting the expression of PAK4 in tissue samples obtained from 95 gastric cancer patients who underwent surgical treatment. Western blot was also used to measure the expression of PAK4 in gastric cancer tissues and adjacent non-cancerous gastric mucosa in 40 patients and also in 7 human gastric cancer cell lines. Results Immunohistochemical staining results showed that positive PAK4 expression in the gastric cancer tissues was 67.4%, and 96.9% of those (62/64) was located in the cytoplasm, while only 3.1% (2/64) was on the cell membrane. Expression of PAK4 was revealed to be significantly and positively correlated with both TNM stage (X2=10. 426, P<0.01) and lymph node metastasis (X2= 4.912, P<0.05 ). Western blot showed that PAK4 expression of gastric cancer tissues was significantly higher than that of adjacent non-cancerous gastric mucosa (t=-5. 978, P<0.01). Expression of PAK4 could also be detected in 7 human gastric cancer cell lines. Compared with GES-1, expression of PAK4 is higher in MKN45, AGS, N87 and BGC823, but it was lower in MGC803, MKN1 and SGC7901. Conclusion The overexpression of p21-activated kinase4 (PAK4) may be correlated with the carcinogenesis and prognosis of gastric cancer.

Adult ; Biopsy ; DNA ; genetics ; Female ; Gene Expression Profiling ; Glomerulonephritis, IGA ; genetics ; pathology ; Humans ; Kidney ; pathology ; Male ; Middle Aged ; Young Adult

This article introduces the definition, classification, premarket admission and other administering specialities about In-Vitro Diagnostic Reagents in the U.S.A. and China. And by analyzing manufacture and administration of In-Vitro Diagnostic Reagents in our country, It is pointed out that a suitable administering model in accordance with the characteristics of In-Vitro Diagnostic Reagents should be adopted to perfect the administration.
China ; Device Approval ; Indicators and Reagents ; classification ; standards ; Quality Control ; Reagent Kits, Diagnostic ; classification ; standards ; United States ; United States Food and Drug Administration

This article makes a pilot study on the key points of the quality management system of in-vitro diagnostic reagents by analyzing the technical characteristics and production methods of these products as well as the status in quo, and problems the in-vitro diagnostic reagent industry in China is facing nowadays. It can serve as a reference to the supervision departments and the manufacturers in this field which are establishing and running the quality management system.
China ; Equipment and Supplies ; standards ; Humans ; Indicators and Reagents ; chemistry ; standards ; Pilot Projects ; Quality Assurance, Health Care ; organization & administration ; Quality Control ; Reagent Kits, Diagnostic ; standards ; Safety Management ; Technology, Pharmaceutical ; organization & administration ; standards ; Total Quality Management

OBJECTIVE: To investigate the influence of commercial peritoneal dialysate (CDS) on function of macrophage. METHODS: Cultured peritoneal macrophages were divided into two experimental groups and their controls.(1) Macrophages were cultured in conditioned culture medium containing 50%CDS (0.139 mol/L glucose) for 24 h. (2)Macrophage were exposed to CDS containing 0.139mol/L glucose for 10, 30 and 60 min respectively, and then cultured in CDS-free medium for 24 h. The nitric oxide (NO) production and MTT in two groups were measured. In each control group, CDS was replaced by same amount of culture medium. RESULTS: NO production and MTT reduction ability (related to cell viability) of experimental groups were remarkably lower than those of controls and the NO production and MTT reduction in 60min CDS-exposed group were lower than that of 10 min and 30 min CDS-exposed group. CONCLUSION: Dialysate may have detrimental effects on viability and other function of macrophage and these effects may related to time length of CDS exposure.

OBJECTIVETo investigate the prevalence of viral infections and putative association of viral infection with illness severity in young children with severe lower respiratory tract infection (LRTI) in Chongqing.

METHODRespiratory secretion specimens were collected from 119 hospitalized patients with severe pneumonia from December 2006 to March 2008.After being processed, the samples were detected for respiratory viruses including respiratory syncytial virus (RSV), adenovirus (ADV), human metapneumovirus (hMPV), human bocavirus (HBoV), parainfluenza virus 1, 2, 3 (PIV 1, 2, 3), influenza virus A and B (IVA and IVB) either by PCR or RT-PCR. Clinical data were analyzed along with virological data by using appropriate statistical methods.

RESULTViral pathogens were identified in specimens of 86 (72.3%) cases, among which RSV was detected in 49 (41.2%) patients. More than one virus was detected in 23 individual (26.7%) samples, of which 19 were dual positive for RSV and another virus. Bacterial cultures were performed for 69 patients. Both bacterial and viral pathogens were identified in 53 (76.8%) patients. Bacterial and viral coinfection was demonstrated in samples from 41 (59.4%) cases.

CONCLUSIONViral pathogens are the main etiology of severe pneumonia in young children in Chongqing area during the study period. RSV was the most frequent viral pathogens, followed by ADV and hMPV. Coinfection with respiratory common viruses was relatively common, though co-infection with viruses did not appear to aggravate the patients' condition.

Adenoviridae ; isolation & purification ; Child, Preschool ; China ; epidemiology ; Human bocavirus ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; Influenza A virus ; isolation & purification ; Metapneumovirus ; isolation & purification ; Pneumonia, Bacterial ; microbiology ; virology ; Pneumonia, Viral ; microbiology ; virology ; Respiratory Syncytial Viruses ; isolation & purification ; Virus Diseases ; virology

AIMTo establish the methods for determining the activity of sphingosine kinase(SPK) and the content of sphingosine 1-phosphate (S1P) in biological samples.

METHODSThe ECV304 cells were transfected with pcDNA3 vector encoding Flag-labeled SPK gene. The expression of SPK was measured by Western blot assay and the activity of SPK was determined by enzymatic reaction, isotope incorporation and thin-layer chromatography methods. The S1P in biological samples was extracted, digested by alkaline phosphatase and then catalyzed by SPK. The S1P contents were determined according to the amounts of products.

RESULTSSPK gene transfection could enhance the expression and activity of SPK in cells markedly, and the cellular S1P was also increased obviously. HGF stimulation could increase the activity of SPK and cellular S1P in ECV304 cells.

CONCLUSIONMethods for determining the activity of SPK and the content of SPK in biological samples were established.

Cell Line ; Cytophotometry ; Humans ; Isotope Labeling ; Lysophospholipids ; metabolism ; Phosphotransferases (Alcohol Group Acceptor) ; metabolism ; Sphingosine ; analogs & derivatives ; metabolism

AIM:To understand the clinic effect of intravitreal anti-vascular endothelial growth factor(VEGF) drugs injection in the treatment of fundus disease in the real-world study (RWS).METHODS:The clinical cases treated with anti-VEGF drugs in our department from September 2012 to June 2015 were enrolled in this study.Retrospective investigation was reviewed to the kinds of diseases, frequency, usage, efficacy, adverse reaction, and the effects on visual acuity, fundus and macular thickness which were treated with intravitreal anti-VEGF drugs injection.RESULTS:In 305 patients (340 eyes) treated with anti-VEGF drugs, 53 patients (60 eyes, 17.6%) were wet age-related macular degeneration (AMD), polypoidal choroidal vasculopathy (PCV) 16 cases (18 eyes, 5.3%), diabetic macular edema (DME) 120 cases (134 eyes, 39.4%), branch retinal vein occlusion (BRVO) secondary macular edema 61 cases (68 eyes, 20.0%), central retinal vein occlusion (CRVO) secondary macular edema 29 cases (32 eyes, 9.4%), idiopathic choroidal neovascularization (ICNV) 16 cases (18 eyes, 5.3%), high myopia with choroid neovascularization 4 cases (4 eyes, 1.2%), neovascular glaucoma 4 cases (4 eyes, 1.2%), retinal angiomatous proliferation (RAP) 1 cases (1 eyes, 0.2%) and optic papillary neovascularization 1 cases (1 eyes, 0.2%).The minimum age was 16 years old, and the maximum age 90 years old.There were 247 cases (275 eyes, 80.9%) were treated with intravitreal ranibizumab injection, 58 cases (65 eyes, 19.1%) intravitreal conbercept injection.The time number of all patients accepted anti-VEGF drugs treatment was 465, with an average of 1.7 times per eye.Which, the 3 + PRN treatment method in 98 patients (109 eyes, 32.1%), 1 + PRN treatment in 207 patients (231 eyes, 67.9%).69 cases (77 eyes, 22.6%) were used alone to receive anti-VEGF drugs therapy, 10 cases (11 eyes, 3.2%) combined with intravitreal triamcinolone injection(TA), 35 cases (39 eyes, 11.5%) combined with vitrectomy, 26 cases (29 eyes, 8.5%) combined with photodynamic treatment (PDT), 165 cases (184 eyes, 54.1%) combined with simple laser treatment.After anti-VEGF drug treatment, majority of patients' the best corrected visual acuity (BCVA), fundus and central macular thickness(CMT) were significantly improved, compared with the pre-treatment, the difference is significant (P<0.05).So that anti-VEGF drugs can effectively improve visual function and ocular fundus for fundus diseses.There were no serious adverse reactions except 3 patients appearling skin redness, itching, rash, 1 patient low low-grade fever and 1 patient acute cerebral infarction during the treatment.CONCLUSION:Intravitreal anti-VEGF drugs injection can significantly improve the visual function and ocular fundus for patients with fundus diseases, but there are still some adverse events, which should be attached great importance to medical workers.

OBJECTIVESTo evaluate the inhibitory effect mediated by combination of small interfering RNAs (siRNAs) targeting different sites of hepatitis B virus (HBV) transcripts on the viral replication and antigen expression in vitro.

METHODS(1) Seven siRNAs targeting surface (S), polymerase (P) or precore (PreC) region of HBV genome were designed and chemically synthesized. (2) HBV-producing HepG2.2.15 cells were treated with or without siRNAs for 72 h. (3) HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay. (4) Intracellular viral DNA was quantified by real-time PCR (Polymerase Chain Reaction). (5) HBV viral mRNA was reverse transcribed and quantified by real-time PCR. (6) The change of cell cycle and apoptosis was determined by flow cytometry.

RESULTSOur data demonstrated that synthetic small interfering RNAs (siRNAs) targeting S and PreC gene could efficiently and specifically inhibit HBV replication and antigen expression. The expression of HBsAg and HBeAg and the replication of HBV could be specifically inhibited in a dose-dependent manner by siRNAs. Furthermore, our results showed that the combination of siRNAs targeting various regions could inhibit HBV replication and antigen expression in a more efficient way than the use of single siRNA at the same final concentration. No apoptotic change was observed in the cell after siRNA treatment.

CONCLUSIONOur results demonstrated that siRNAs exerted robust and specific inhibition on HBV replication and antigen expression in a cell culture system and combination of siRNAs targeting different regions exhibited more potency.

Apoptosis ; Cell Cycle ; Cell Line, Tumor ; DNA, Viral ; biosynthesis ; Flow Cytometry ; Gene Expression Regulation, Viral ; genetics ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B e Antigens ; metabolism ; Hepatitis B virus ; genetics ; physiology ; Humans ; RNA, Small Interfering ; genetics ; metabolism ; Virus Replication ; genetics