Child ; Encephalitis, Viral ; pathology ; Enterovirus A, Human ; Enterovirus Infections ; complications ; Heart Failure ; etiology ; Humans ; Respiratory Insufficiency ; etiology

Based on target cycling amplification and 3 ,4 ,9 ,10-perylenetetracarboxylic dianhydride derivative functionalized singal probe, an ultrasensitive electrochemiluminescence ( ECL) sensor was designed for the detection of lead ions. The hairpin substrate DNA was immobilized on the electrode through molecular self-assembly. In the presence of Pb2+and DNAzyme, the substrate was cleaved with single strand DNA fragments left on the electrode surface. Meanwhile, the target and DNAzyme was released for another cleaving circularly. As a result, the single strand DNA fragments hybridized with the assist hairpin probe H1, which leaded to the fabrication of H2 labeled with the 3 , 4 , 9 , 10-perylenetetracarboxylic dianhydride derivative functionalized hollow gold nanoparticles. With the increasing concentration of Pb2+, much more signal probe was been captured and the ECL signal of the biosensor in peroxydisulfate ( S2 O2-8 ) solution would increase. An ECL assay demonstrates that the sensor has a good linear response to Pb2+ concentration in the range of 1í10-12 mol/L-1í10-6 mol/L, with a detection limit of 1í10-12 mol/L. The fabricated sensor shows good selectivity toward Pb2+against other common metal ions.

Acrylonitrile ; poisoning ; Adult ; Female ; Humans ; Male ; Middle Aged ; Young Adult

Objective To evaluate the inhibitory effect of the small interfering RNA(siRNA) on hepatitis B virus(HBV)in vivo which targets HBV S gene region.Methods An animal model of HBV infection was developed hydrodynamically by injecting pcDNA3.1-HBV together with siRNA through the tail vein of Balb/c.HBsAg was analyzed by time resolved immunofluorometric assay, HBV DNA was analyzed by fluorogenic quantitative PCR(FQ-PCR),HBV S-mRNA was detected by semi-quantitative RT-PCR,and viral specific proteins(HBsAg and HBcAg)in the liver were assayed by immunohistochemical staining.Results In the mice,the siRNA could effectively inhibit the secre- tion of HBsAg,reduce the titers of HBV DNA,and immunohistochemical results also indicated that the number of HBsAg and HBcAg positive cells was reduced.The inhibitory effect of siRNA on HBV lasted 3 clays at least.Conclusion These results demonstrate that the siRNA targeting HBV S gene region can substantially and specifically inhibit HBV replication and expression in vivo.

OBJECTIVETo evaluate the objectivity and comprehensiveness of Response Evaluation Criteria of Traditional Chinese Medicine for Solid Tumor (Draft, REC-TCM-ST) in application of Chinese medicine therapeutic effect in patients with advanced non-small cell lung cancer (NSCLC).

METHODSA retrospective clinical research was used in 104 NSCLC patients in stages of III-IV, 53 cases were in Chinese medicine (CM) group and 51 cases were in Western medicine (WM) group. The therapeutic effect of the two groups was evaluated with both REC-TCM-ST and Response Evaluation Criteria in Solid Tumor (RECIST). Kaplan-Meier method was used to analyze the survival time. Kappa test method was used to test the consistency of the two kinds of evaluation results.

RESULTSAccording to REC-TCM-ST, the effective rate on relieving tumor mass in the CM group was significantly lower than that in the WM group (P<0.05), but there was no significant difference in tumor-mass stable rate (P>0.05); the symptom of weakness in the CM group was improved significantly, indicating better therapeutic effect than that in the WM group (P<0.01). Karnofsky score in the CM group was significantly better than that in the WM group (P<0.01). In terms of survival conditions, the median survival time and the survival rate of 6 months, 1 year and 2 years of the CM group were higher than the WM group. The total effective rate was 9.62%, and the total stable rate was 72.12% for 104 cases according to RECIST; while the total effective rate was 34.62%, and the total stable rate was 84.62% according to REC-TCM-ST, thus there were significant differences between the results of the two criteria (P<0.01), and there was also some consistency between them, but not satisfactory.

CONCLUSIONSREC-TCM-ST was used to evaluate the therapeutic effect of CM in the treatment of advanced NSCLC, which shows that its evaluation results can better reflect the advantages and disadvantages of CM, and the effectiveness of CM is more objective and comprehensive than RECIST, so REC-TCM-ST is worthy of further improvement and clinical expansion.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Female ; Humans ; Karnofsky Performance Status ; Lung Neoplasms ; drug therapy ; pathology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Neoplasm Staging ; Response Evaluation Criteria in Solid Tumors ; Survival Analysis

OBJECTIVETo isolate CD133(+)/CD44(+)/ESA(+) subsets cells from SW480 colon cancer cells, and to observe the tumor formation.

METHODCD133(+)/CD44(+)/ESA(+) subsets cells, CD133(-)/CD44(+)/ESA(+) subsets cells and CD133(-)/CD44(-)/ESA(-) subsets cell were sorted by flow cytometry from SW480 colon cancer cells, then three subsets were separately inoculated in five NOD/SCID mice and the growth rates were calculated.

RESULTThe proportion of CD133(-)/CD44(-)/ESA(-), CD133(-)/CD44(+)/ESA(+) and CD133(+)/CD44(+)/ESA(+) subsets cells in SW480 cells were (86.38±10.23)%,(1.26±0.28)% and(0.38±0.07)%. After inoculation, tumor nodules could be formed three days later in CD133(+)/CD44(+)/ESA(+) group, and they could be formed 9 days later in CD133(-)/CD44(+)/ESA(+) group, while they could be formed 15 days later in CD133(-)/CD44(-)/ESA(-) group. Eighteen days later, tumor sizes in three groups were(13.82±5.04) mm(3), (9.25±4.57) mm(3) and (4.76±3.92) mm(3) respectively, and the differences were statistically significant(P<0.05).

CONCLUSIONESA(+)-CD44(+) is one of the surface markers for colonic cancer stem cells, and CD133(+)-CD44(+)-ESA(+) cells are SW480-like cancer stem cells.

Animals ; Biomarkers, Tumor ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; Flow Cytometry ; Humans ; Hyaluronan Receptors ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Neoplastic Stem Cells ; cytology ; Sialic Acid Binding Ig-like Lectin 3

OBJECTIVETo investigate the feasibility and clinical application value of selecting viable spermatozoa by noncontact diode laser.

METHODSWe obtained immotile spermatozoa from 2 infertile men with obstructive azoospermia or severe asthenospermia and selected viable spermatozoa using a single laser shot at the sperm tail. Those that responded to the laser shot by a curling reaction of the tail were regarded as presumably viable and used for intracytoplasmic sperm injection (ICSI).

RESULTSThe mean fertilization rate was 88.89% after ICSI with the laser-selected viable spermatozoa. Both of the embryo transfers resulted in a single pregnancy.

CONCLUSIONNoncontact diode laser is a useful alternative for the assessment of sperm viability, which may help to achieve successful pregnancy.

Embryo Transfer ; Female ; Fertilization ; Humans ; Infertility, Male ; therapy ; Male ; Pregnancy ; Pregnancy Outcome ; Sperm Injections, Intracytoplasmic ; Sperm Motility ; Sperm Tail ; physiology

OBJECTIVE Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited disease with a high morbidity around 1/1000-1/400, characterized by progressive enlargement of fluid-filled cysts derived from renal tubular epithelial cells. Massive cysts gradually compress renal parenchyma destroying normal renal structures and compromising renal functions. Unfortunately, it will cause end-stage renal disease in most of the patients but without effective therapy now, who have to live on hemodialysis or kidney transplantation. Based on this present situation, it is of great significance to find early intervention to inhibit renal cyst development. The projective of this study was to investigate whether Ganoderma triterpenes (GT) can inhibit renal cyst development and study the related mechanism. METHODS and RESULTS First, we used MDCK cyst model, cultivated MDCK cells in vitro to form fluid-filled cysts surrounded by monolayer cells. GT inhibited MDCK cyst formation significantly, and inhibited cyst enlargement dose-dependently proving GT cyst inhibition in vitro. Then we used an embryonic kidney cyst model, wile-type mice kidneys were taken out on embryonic day 13.5 to form renal cysts stimulated with 8-Br-cAMP. GT inhibited embryonic kidney cyst development significantly in a dose-dependent and reversible manner proving GT cyst inhibition at organ level. Furthermore, we used two ADPKD mouse models with severe cystic kidney disease phenotypes. GT dramatically inhibited renal cyst development, decreased ADPKD mouse kidney volume and the cyst index inside proving GT cyst inhibition in vivo. By Western blot, we proved GT down-regulated Ras/MAPK signal pathway without detectable effect on mTOR signal pathway both in MDCK cells and two ADPKD mouse kidneys. CONCLUSION GT retard renal cyst development both in vitro and in vivo significantly. The related mechanisms were involved in GT promoting renal tubular epithelial cell differentiation, down-regulating intracellular cAMP level and Ras/MAPK signal pathway.

OBJECTIVETo design pSilencer3.1-H1hygro plasmid expressing short interfering RNAs (siRNA) that targets HBV core gene region, and to evaluate inhibitory effect of this siRNA on HBV in vitro.

METHODSHepG2 2.2.15 was used as target cells. The plasmid and liposome metafectene were cotransfected into the cultured cells, HBV DNA were analyzed by fluorogenic quantitative PCR (FQ-PCR), HBV C-mRNA was detected by semi-quantitative RT-PCR.

RESULTSThe plasmid expressing siRNA was successfully constructed. The two constructed siRNAs could effectively inhibit HBV replication, and their inhibitive effect on HBV was dose-dependent.

CONCLUSIONThese results showed that siRNA could substantially inhibit HBV replication in the infected cells

Hepatitis B virus ; genetics ; physiology ; Humans ; Liver Neoplasms ; virology ; RNA Interference ; Tumor Cells, Cultured ; Virus Replication ; genetics

OBJECTIVETo evaluate the inhibitory effect of small interfering RNA (siRNA) targeting HBV C gene region on hepatitis B virus (HBV) in vivo.

METHODSAn animal model of HBV infection was developed hydrodynamically, and pcDNA3.1-HBV and siRNA were together injected into the tail vein of the BALB/c mice. HBsAg was analyzed by time-resolved immunofluorometric assay, HBV DNA was analyzed by fluorogenic quantitative PCR (FQ-PCR), HBV C-mRNA was detected by semi-quantitative RT-PCR, and viral specific proteins (HBsAg and HBcAg) in the mice livers were assayed using immunohistochemical staining.

RESULTSIn the mice, the siRNA effectively inhibited HBV replication and expression compared with the controls. The inhibitive effect of siRNA on HBV lasted at least 3 days.

CONCLUSIONThese results demonstrate that RNAi can substantially inhibit HBV replication and expression in vivo.

Animals ; Female ; Hepatitis B ; therapy ; Hepatitis B virus ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; RNA, Small Interfering ; physiology ; RNA-Induced Silencing Complex ; Random Allocation ; Virus Replication ; genetics