Objective To prepare monoclonal antibody specifically against carcinoembryonic antigen glypican-3 (GPC3) and its preliminary application. Methods GPC3 was cloned with PCR to pET16b vector and expressed in E. co-liBL21. Spleen cells were obtained from Balb/c mice embedding immunized with purified antigen intraperitoneally, and fused with Sp2/0 cells. Hybridoma cells were screened by indirect ELISA, and identified by Western blot assay using puri-fied protein after the cell fusion. The indirect immunofluorescence method was used to detect the GPC3 expression in HepG2 cell line. Results The prokaryotic expression vector of GPC3 was successfully constructed, and GPC3 was stably expressed in E. coliBL21. A mouse hybridoma cell line secreting monoclonal antibody to GPC3 was obtained. Western blot analysis showed that monoclonal antibody specifically recognized recombinant protein. Monoclonal antibody could be used to detect GPC3 protein expression in HepG2 cell line by indirect immunofluorescence. Conclusion The monoclonal antibody against GPC3 was successfully obtained.

OBJECTIVETo observe the effect of Wenyang Yiqi Pingchuan Recipe (WYPR) on the pathomorphology of lung and its regulation on lung tissue contents of nitric oxide (NO) and endothelin-1 (ET-1) in rats with bronchial asthma.

METHODSSixty SD rats were randomly divided into 6 groups: the normal group, the model group, and the four treated groups treated with high dose WYPR, low dose WYPR, Guilong Kechuanning Capsule and aminophylline, respectively, 10 in each group. Except those in the normal group, all rats of bronchial asthma model were established by egg protein sensitization and provocated by inhalation. The treatments were given via gastrogavage every day starting from the first provocation (the 3rd week of modeling) to the execution. Rats were sacrificed after 4-week treatment, their lung was taken for determining the contents of NO and ET-1, and histopathological changes in lung were observed as well.

RESULTSCompared with the normal group, the contents of NO and ET-1 in the lung tissue, the thickness of bronchus wall and bronchus smooth muscle, the number of eosinophil granulocytes increased in the model group and the low dose WYPR group, showing statistical significance (P < 0. 01). As compared with the model group, all the above-mentioned indices were lower in all the 4 treated groups (P < 0.05 or P < 0.01), but the lowering in the WYPR treated groups (either high or low dose) was more significant than in the Guilong Kechuanning treated group (P < 0.05 or P < 0.01); while compared with the aminophylline treated group, the high dose WYPR group was superior in reducing eosinophile granulocytes (P < 0.01), but no significance between them was shown in NO and ET-1 levels (P > 0.05).

CONCLUSIONSWYPR could reduce the eosinophilic infiltration, decrease the contents of NO and ET-1 in the lung tissue, restrain the air passage inflammation and inhibit the pathological process as airway remodeling.

Animals ; Asthma ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Endothelin-1 ; metabolism ; Inflammation ; pathology ; Lung ; metabolism ; pathology ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVE To study clinical characteristics of liver failure with fungal infections in the elderly patients and risk factors associated with treatment failure.METHODS Eighty four elderly patients with liver failure followed by fungal infections since 1986 were divided into two groups: effective group and ineffective group.RESULTS The common pathogens were Candida albicans(58.33%),Aspergillus fumigatus(9.52%) and Candida tropicalis(8.33%).The lungs(43.88%),mouth(32.65%),intestinal tract(9.18%) and blood(5.10%) were the main sites of fungal infection.Among them after treatment,35 cases(41.67%) were effective compared with 49 cases(58.33%) ineffective.The risk factors for treatment failure included age,complication with multiple organ dysfunction syndrome(MODS) and aspergillosis.In multivariate analysis,we found MODS in patients was an independent factor in predicting the prognosis.CONCLUSIONS To improve the treatment outcome,important measures include preventing infection,enhancing the treatment of liver failure,monitoring and supporting multiple organs: heart,brain,lungs and kidneys,and promptly rational administration of antifungal agents in elderly patient with liver failure.

OBJECTIVE To study the feature of liver disease and liver failure complicated with fungous infection. METHODS The patients with liver disease complicated with fungous infection were collected form 1986 to 2005. The time was divided into four stages:from 1986 to 1990,from 1991 to 1995,from 1996 to 2000 and from 2001 to 2005. All patients with liver failure complicated with fungous infection in different stages were investigated for the incidence,the use of antibiotics and corticosteroids,the category and site of fungous infection and prognosis. RESULTS End-stage liver disease accounted for 82.6% and HBV infection was the main etiology in 475 cases of fungous infection. Fungous infection occurred mainly in hospital. Hospital acquired infection and community acquired infection were similar in different stages. The use of antibiotics and corticosteroids accounted for 88.8% and 48% in all patients before fungous infection,respectively. The use of antibiotics had no difference and the use of corticosteroids decreased in different stages. Candida were the main infection strains and the lungs and pharynx oralis were the main infection sites. The rate of healing and improvement of fungous infection and underlying diseases increased year by year. Healing and improvement rate of underlaying disease positively correlated with that of fungous infection. CONCLUSIONS End-stage liver disease patients are susceptible to fungous infection and Candida are the common infection strains. Lungs and pharynx oralis are the common infection sites. Anti-fungous therapy is important in the treatment of liver failure complicated with fungous infection.

Pigeon circovrius (PiCV) is a member of circovirus, which is usually regarded as an immunosuppression agent. There were reports that pigeons infected by PiCV showed symptoms of lethargy, weight loss, vomiting, diarrhea, respiratory distress, etc. In this study, we established a PCR method for the detection of PiCV DNA. Samples from 5 different farms in Zhejiang Province were examined and samples from a farm in Hangzhou were positive. Furthermore, the genomic segments of 2 strains of PiCV were amplified, cloned and sequenced using designed primers and the complete genomes of the strains were then assembled and named as PiCV-zj1 and PiCV-zj2, respectively. The sequences were deposited in GenBank under the GenBank Accession number of DQ090945 and DQ090944, respectively. Sequence Analysis had shown that the complete genomes of 2 strains of PiCV from Zhejiang Province had 2 039 nucleotides totally in length and common characters of circovirus such as a stem-loop structure and conserved motifs for Rep protein, which were supposed to be related to the replication of the virus. Pairwise comparisons showed that the nucleotide sequence of the genome of PiCV strains from Zhejiang Province had 86%-89.1% identities to that of 11 published PiCV strains, and that the amino acid identities of the replication-associated protein (Rep) and capsid protein (Cap) displayed 92.1%-94.7% and 76.6%-81.4%, respectively. A phylogenic tree was built using PHYLIP with bootstrap support for 1 000 replicates. The result showed that 10 strains from Europe and America formed one big branch and the others from Zhejiang Province and Australia formed the other two, respectively. This was the first report on the detection and full genome sequencing of PiCV in China.
Animals ; Base Sequence ; Circoviridae Infections ; virology ; Circovirus ; classification ; genetics ; Cloning, Molecular ; Columbidae ; virology ; Genome, Viral ; genetics ; Models, Genetic ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Viral Proteins ; genetics

To optimize the formulation and preparation method of multivesicular liposome of thymopentin and to investigate its pharmacokinetics in rats, the multivesicular liposome of thymopentin was prepared by double emulsification method and the formulation was optimized by orthogonal design. The release characteristics of thymopentin from multivesicular liposome in PBS (pH 7.4) and in plasma were investigated. The multivesicular liposome of thymopentin labeled with fluorescein isothiocyanate was prepared by double emulsification method. Its pharmacokinetics was evaluated following intramuscular injection in rats. The optimal formulation of multivesicular liposome of thymopentin were formulated with 7.5% glucose in aqueous phase and 2.25 mol x L(-1) triolein, 2.68 mol x L(-1) DPPG and 16.96 mol x L(-1) DOPC in organic phase. The entrapment efficiency of the multivesicular liposome of thymopentin was above 85% and the mean particle size was about 22 microm. The in vitro release of thymopentin from multivesicular liposome in PBS (pH 7.4) and in plasma was found to be in a sustained manner. The release curves were fitted to Higuchi equation. The pharmacokinetics following intramuscular injection of the multivesicular liposome of thymopentin labeled with fluorescein isothiocyanate in rats showed that the peak concentration of thymopentin was lower and elimination of it was slower significantly than that of thymopentin labeled with fluorescein isothiocyanate solution in the same dose. The plasma concentration of thymopentin maintained above quantitative limitation at 120 h after administration of multivesicular liposome of thymopentin. The optimized formulation and preparation technology of multivesicular liposome of thymopentin with higher entrapment efficiency are feasible with good reproducibility. Multivesicular liposome of thymopentin showed significant sustained-release property following intramuscular injection in rats.
Adjuvants, Immunologic ; administration & dosage ; pharmacokinetics ; Animals ; Area Under Curve ; Delayed-Action Preparations ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Glucose ; chemistry ; Liposomes ; chemistry ; Male ; Particle Size ; Phosphatidylcholines ; chemistry ; Phosphatidylglycerols ; chemistry ; Rats ; Rats, Sprague-Dawley ; Thymopentin ; administration & dosage ; pharmacokinetics ; Triolein ; chemistry

OBJECTIVETo assess the clinical significance of electrophysiological tests of bulbocavernosus reflex (BCR), ischiocavernosus reflex (ICR) and pudenda somatosensory evoked potential (PSEP) for patients with erectile dysfunction (ED).

METHODSElectrophysiological tests of BCR, ICR and PSEP were performed for 232 ED patients with IIEF-5 scores of 2-20 (10.39 +/- 5.52), another 21 normal volunteer controls underwent the same tests, and the results were compared.

RESULTSAbnormal results, such as prolonged and advanced latencies, were found in 94 (40.5%) of the patients, which suggested neurotic ED with different degrees of cauda equine nerve injuries.

CONCLUSIONElectrophysiological tests of BCR, ICR and PSEP can objectively reveal the functional state of the cauda equine nerve and offer an important support to the diagnosis of nerve-mediated ED.

Adult ; Aged ; Case-Control Studies ; Electrophysiology ; Erectile Dysfunction ; physiopathology ; Evoked Potentials, Somatosensory ; Humans ; Male ; Middle Aged ; Muscle, Smooth ; physiopathology ; Penis ; innervation ; physiopathology ; Reflex ; physiology

UNLABELLEDTo study the method for dose calculation and beam weight optimization of intensity-modulated radiation therapy (IMRT).

METHODSThe IMRT dose calculation model based on two-dimensional convolution was constructed, the program of dose calculation and beam weight optimization with genetic algorithm was written with Visual c#.Net, and the optimization results were analyzed.

RESULTSGenetic algorithm optimization of beam weights can produce highly conformal dose distributions within a clinically acceptable computation time.

CONCLUSIONGenetic algorithm is valid and efficient in IMRT beam weight optimization, which may facilitate IMRT treatment planning.

Algorithms ; Humans ; Models, Statistical ; Models, Theoretical ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; methods ; Radiotherapy, Conformal ; methods ; standards

OBJECTIVETo investigate the risk factors related to post-asphyxial multiple organ dysfunction (PA-MOD) in neonates.

METHODSA total of 397 neonates with birth asphyxia were enrolled from January 2009 to December 2010.The patients were divided into PA-MOD group (n=179) and non-PA-MOD group (n=218). The risk factors of PA-MOD were retrospectively studied.

RESULTSMultivariate logistic regression analysis showed that severe asphyxia, fetal distress, abnormal labor, and decreased amniotic fluid were the risk factors for PA-MOD among the neonates. Spearman rank correlation analysis showed that the number of the involved organs increased along with the increase of age at admission (P<0.05) and with the decrease of gestational age and birth weight (P<0.05).

CONCLUSIONSThe efforts should be made to enhance perinatal care for neonates, especially for preterm infants and low-birh-weight infants, to decrease the incidence of MOD.

Asphyxia Neonatorum ; complications ; Female ; Humans ; Infant, Newborn ; Logistic Models ; Male ; Multiple Organ Failure ; etiology ; prevention & control ; Risk Factors

Objective To determinate the prognostic value of etiology in patients with chronic systolic heart failure (CSHF).Methods Data of in-hospital patients with CSHF were investigated between 2000 and 2010 from 12 hospitals in Hubei province.All patients were followed up through telephone calls.Univariate and multivariate Cox proportional hazards analyses were then used to explore the differences in the all-cause mortality,heart failure (HF) mortality and sudden cardiac death (SCD) among patients caused by different etiologies.Kaplan-Meier curve were then constructed and Univariate and multivariate Cox regression analyses were used to select demographic and clinical variables in predicting the all-cause mortality,HF mortality and SCD in CSHF patients.Multivariate logistic models and ROC curve were developed with or without the cinfirmed etiology to assess the incremental additive information related to different etiologies.Results (1)Over the median 3 (2-4) years follow-up program,6453 (38.69％) patients died,including 5505 (33.00％) due to HF prognosis and 717 (4.30％) died of SCD.All-cause mortality rates accounted for 34.50％,54.30％,41.48％ and 15.76％,with HF mortality rates as 30.11％,44.95％,36.25％ and 13.10％.SCDs accounted 8.46％,8.45％,9.84％ and 1.05％ in patients with CHD,DCM,HHD and RHD,respectively.(2) Compared with RHD patients,the adjusted HRs for all-cause mortality were 1.554 (1.240 to 1.947;P＜0.001),1.405(1.119 to 1.764;P=0.003) and 1.315(1.147 to 1.467;P=0.005) while the adjusted HRs and 95％CIs for HF mortality were 1.458( 1.213-1.751 ;P＜0.001 ),1.763( 1.448-2.147;P＜0.001 ) and 1.281 ( 1.067-1.537; P=0.008),in patients with CHD,DCM and HHD,respectively.There were no significant differences in CHD (HR 3.345; 95％ CI,1.291 to 8.666; P=0.013 ) or HHD (HR 2.062; 95％CI,0.794 to 5.352; P=0.137 ),while only DCM ( HR 4.764; 95％CI,1.799 to 12.618;P=0.002) remained significant in SCD despite of the multivariate adjustment.(3) Etiology increased the sensitivity and specificity of predicting models for all-cause mortality(AUC 0.839,95％CI,0.832to 0.845 vs.0.776,95％CI,0.768 to 0.784) and HF mortality(AUC 0.814,95％CI,0.806 to 0.822 vs.0.796,95％CI,0.788 to 0.804) but not with SCD (AUC 0.777,95％CI,0.749 to 0.809 vs.0.747,95％CI,0.727 to 0.766).Conclusion CSHF due to CHD,DCM and HHD carried a worse prognosis than that of RHD.Different etiologies provided significant incremental prognostic information beyond readily available clinical variables for all-cause mortality and HF mortality.