Objective To investigate the effects of low molecular weight polypeptide(LMP) extracted from chicken embryo on learning and memory in senile mice induced by D-galactose(Gal). Method The 16 d old chicken embryo was acid and alkali hydrolyzed,enzymolyzed and ultrafiltered to formulate LMP. 48 mice were divided randomly into four groups: control,aging model ,aging+low dose LMP,aging+high dose LMP. The aging model and two LMP groups were treated with Gal 80 mg/ (kg bw?d) by nape subcutaneous injection,while the control group with normal saline 8ml/(kg bw?d). Control and model groups were given i.g. normal saline 20 ml/(kg bw?d),and two LMP groups 10 ml or 20 ml/(kg bw?d) LMP respectively. Learning and memory of mice were tested with Morris water maze. Results The escape latency of model group was longer and the percentage of swimming distance in platform region higher than that of control group (P

Objective To investigate the effect of low molecular weight substance extracted from chick embryo on mitochondrial DNA(mtDNA)deletion in senile mice induced bv D-galactose.Methods Senile mice induced by D-galactose were treated with low molecular weight substance extracted from chick embryo.The deleted fragment of mtDNA was examined by using polymerase chain reaction technique and agarose gel electrophoresis.A relative quantitation of band densities was performed by using densitometry scanning techniques.The deletion was identified by using direct sequencing analysis. Results The 4239 bD mtDNA deletion were present in liver,cerebral cortex and hippocampus tissues in all of the mice.and it was significantly higher in senile model mice than in control mice(all P＜0.01).Low molecular weight substance extracted from chick embryo reduced the mtDNA deletion in various tissues in senile model mice(all P＜0.05).Deletion was more abundant in liver than in cerebral cortex and hippoeampus. Conclusions The 4239 bp mtDNA deletion is a common deletion,and low molecular weight substance extracted from chick embryo could decrease the incidence of 4239 bD mtDNA deletion.

Objective To develop an LC-MS/MS method for determining the concentration of wogonin in dog plasma and investigate the pharmacokinetics and bioavailability by different administrations of wogonin in Beagle's dogs. Methods LC-MS/MS was employed in determining the concentration of wogonin with the selected ion monitoring model after liquid-liquid extraction with ethyl acetate of dog plasma samples. The lower limit of quantification was 0.105 μg/L. Target ions were at m/z 285.0→270.0 for wogonin and 373.3→305.3 for finasteride. In a randomized, self-control, and cross-over study, six male Beagle's dogs were treated with different administration methods in three test periods. Pharmacokinetic parameters were calculated with DAS software (Ver. 2.0). Results The calibration curve was linear in the range of 0.105-107.36 μg/L for wogonin in dog plasma samples. The main pharmacokinetic parameters of ig administration (native drug of 15 mg/kg and solution preparation of 5 mg/kg) and iv route were as follows: Cmax (2.5 ± 1.1), (7.9 ± 3.3), and (6838.7 ± 1322.1) μg/L, tmax (0.7 ± 0.3) and (0.3 ± 0.2) h for the both former, AUC0-1 (7.1 ± 2.0), (21.0 ± 3.2), and (629.7 ±111.8) μg·h/L. The absolute bioavailability of native and solution of wogonin were (0.59 ± 0.35)% and (3.65 ± 2.00)%, respectively. Conclusion The validated method is convenient, sensitive, and specific, and the improvement of wogonin solubility could remarkably increase the absolute bioavailability.

Objective To explore the application of cardiac operation under direct vision through Lower median ministernotomy,and to search the new methods to improve the orthodox cardiac operation under direct vision.Methods The method of the cardiac operation under direct vision through lower median ministernotomy was used to 296 cases with heart disease who needed cardiac surgery,including 250 cases with congenital heart disease,40 cases with rheumatic heart disease,5 cases with left atrial myxom and 1 case with coronary heart disease.Results All cases performed the cardiac operation under direct vision through lower Median ministernotomy successfully without death and any residual.Conclusion The cardiac operation under direct vision through lower median ministernotomy can be used in majority cardiac operation under direct vision,it has well surgical and cosmetic result,but cannot be used in treating some patients with complex cardiac deformity or large heart.

Objective To evaluate the performance of peripheral blood T-SPOT.TB and cerebrospinal fluid (CSF) interferon (IFN)-γ detection in the diagnosis of tuberculous meningitis (TBM).Methods Among the 182 consecutive cases with suspected TBM in Huashan Hospital from March 2011 to March 2013,30 patients were included in the case group according to the latest diagnostic criteria of TBM.Thirty-nine patients diagnosed with non-tuberculous meningitis were included in the control group.T-SPOT.TB was employed to detect tuberculosis-specific IFN-γ-secreting T cells in the peripheral blood.And IFN-γ in CSF was detected simultaneously by enzyme-linked immunosorbent assay (ELISA) without antigen stimulation.The CSF was collected from 10 patients of TBM group after anti tuberculosis treatment for 4 weeks to observe the dynamic changes.The t-test was used for analysis of continuous variables with normal distribution and Kruskal-Wallis test was used for analysis of variables with abnormal distribution.Results ()f the 30 TBM cases,6 were confirmed cases and 24 were highly suspected cases.The control group was comprised of 12 viral encephalitis,16 suppurative meningitis and 11 cryptococcal meningitis.The positive rate of T-SPOT.TB was significantly higher in TBM group compared with control group (70％ vs 13％,x2 =12.15,P＜0.01).The mean concentration of CSF IFN γ of TBM group was 244.35 pg/mL,which was significantly higher than that of control group 9.48 pg/mL (Z=-4.646,P＜0.01).The CSF IFN-γ was significantly decreased after 4 weeks of treatment (271.02 pg/mL vs 81.36 pg/mL,Z=-3.099,P=0.002).The sensitivity and specificity of peripheral blood T-SPOT.TB in the diagnosis of TBM were 70％ and 87％,respectively.The area under the receiver operating characteristic (ROC) curve of CSF IFN-γ for TBM diagnosis was 0.819; the optimal cut-off point was 81.36 pg/mL; the corresponding sensitivity and specificity were 83 ％ and 85 ％,respectively.Conclusion Both the detection of peripheral blood T-SPOT.TB and CSF IFN-γ are of great importance for the diagnosis of TBM.Dynamic observation of CSF IFN-γ is important for disease monitoring.

Objective:To evaluate the outcome and prognostic factors of conventional radiotherapy followed by intensity-modulated radiation therapy (IMRT) as a boost combined with chemotherapy in the treatment of high-grade gliomas (HGG). Methods:From Nov. 2004 to Nov. 2006, 112 HGG patients were treated with conventional radiotherapy followed by IMRT as a boost combined with chemotherapy. The radiotherapy regimen included first-course conventional radiotherapy and latE-course IMRT as a boost with the total radiation dosage of 57.5- 62.5Gy (27-29 fractions in 37-45 days). All the patients received chemotherapy. Sixty-three patients received temozolomidE-based chemotherapy and 49 patients received semustine plus teniposide chemotherapy. The treatment reaction was recorded. The overall survival rate and local progression-free survival rate were calculated. The prognostic factors were analyzed by using multivariate COX regression model. Results:The acute treatment toxicity was the most at grade 1 to grade 2 and no grade 4 adverse reaction was observed. Three cases had late radiation-induced brain necrosis. The overall 1-, 2-, and 3-year survival rates were 78.9%, 54.7%, and 30.8%, respectively. Multivariate analysis revealed that tumor location (P=0.001) and KPS score (P=0.011) were independent prognostic factors. The 1-, 2-, 3-year local progression-free survival rates were 63.8%, 38.9%, and 10.5%, respectively. Multivariate analysis revealed that tumor location (P=0.001), KPS score (P=0.001), and histologic type (P=0.005) were independent prognostic factors. Multivariate analysis did not reveal the significant correlation of temozolomide chemotherapy with the prognosis of the patients. Conclusion:Postoperative conventional radiotherapy followed by IMRT as a boost combined with chemotherapy has better short-term efficacy in the treatment of HGG and the adverse reaction is tolerable. KPS score, tumor location, and histologic type are important prognostic factors. Temozolomide chemotherapy is not associated with the prognosis of patients.

BACKGROUND: It is a hot investigation to many scholars that how to cure and prevent renal ischemic reperfusion injury in a utility way, but the mechanism is unclear at present. The investigation indicates that aquaporiin-1 plays an important role during this process. OBJECTIVE: To research the correlation between aquaporin-1 expression and renal function change following renal ischemic reperfusion injury. DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at Histology and Embryology Laboratory of Dalian Medical University from June 2007 to December 2008. MATERIALS: A total of 80 healthy female adult Wister rats were randomly divided into control group and ischemia-reperfusion group. Rats in each group were observed at days 1, 2, 3, 5, and 7 after operation, with 8 rats for each group. The ischemia-reperfusion injury was established on the left kidney. METHODS: Right kidney was removed. The left renal pedicle was freed and occlused to establish ischemia-reperfusion injury model. After 40 minutes, the blood was re-flowed. If the kidney colored from dark red to bright red within 2-5 minutes, the ischemia-reperfusion injury models were successfully established, and the thrombus was not formed in the kidney vessels. If the kidney was still dark red after 5 minutes, the thrombus was formed, and the rats were excluded from the ischemia-reperfusion group. The abdomen was sutured after 40 minutes.MAIN OUTCOME MEASURES: Rats were sacrificed at days 1, 2, 3, 5, and 7 after ischemia-reperfusion injury. Samples of urine, serum, and kidney were performed with the examinations of urine, renal function, renal pathology and morphology, immunohistological assay of aquaporiin-1, and RT-PCR assay. RESULTS: After ischemia-reperfusion injury, the rats had hydrouria, urine osmotic pressure depress, symptoms of carnine and urea nitrogen increasing. HE staining demonstrated that renal tubular epithelial cells were swelling, necrosis, and desquamate. Aquaporin-1 expression and its mRNA level was decreased; in particular, the expression and level were the lowest at day 1 after ischemia-reperfusion injury and recovered to normal value at day 5 after ischemia-reperfusion injury. CONCLUNSION: The down expression of aquaporin-1 maybe one of the important indicators to reflect renal functional changes of acute renal failure following renal ischemia-reperfusion injury.

BACKGROUND: Mesenchymal stem cells are the stem cells that possess the capability for self-renewal and multi-directional differentiation. Umbilical cord is the tissue outside the embryos and would be fallen off after parturition. In addition, it has wide source and no ethical restriction, so it is promising to be the first choice for mesenchymal stem cells. OBJECTIVE: To detect the surface markers CD29, CD44, CD49e, CD73, CD90, CD34, CD45, and CD271 of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) prior to and after cryopreservation and resuscitation. METHODS: After isolation and culture, morphology of the primary, P4 and P8 hUCMSCs was observed prior to cryopreservation and after resuscitation. Surface markers CD29, CD44, CD49e, CD73, CD90, CD34, CD45, and CD271 of primary, P4, and P8 hUCMSCs were detected through the use of flow cytometry prior to cryopreservation and after resuscitation RESULTS AND CONCLUSION: hUCMSCs prior to cryopreservation and hUCMSCs of different passages after resuscitation present the same phenotype, i.e., positive for CD29, CD44, CD49e, CD73, and CD90, and negative for CD34, CD45, CD271. These findings suggest that primary hUCMSCs do not present changes in surface markers after cryopreservation and resuscitation.

Urokinase plasminogen activator receptor (uPAR) is a membrane protein which is attached to the cellular external membrane. The uPAR expression can be observed both in tumor cells and in tumor-associated stromal cells. Thus, in the present study, the human amino-terminal fragment (hATF), as a targeting element to uPAR, is used to conjugate to the surface of superparamagnetic iron nanoparticle (SPIO). Flowcytometry was used to examine the uPAR expression in different tumor cell lines. The specificity of hATF-SPIO was verified by Prussian blue stain and cell phantom test. The imaging properties of hATF-SPIO were confirmed in vivo magnetic resonance imaging (MRI) of uPAR-elevated colon tumor. Finally, the distribution of hATF-SPIO in tumor tissue was confirmed by pathological staining. Results showed that the three cells in which we screened, presented different expression characteristics, i. e., Hela cells strongly expressed uPAR, HT29 cells moderately expressed uPAR, but Lovo cells didn't express uPAR. In vitro, after incubating with Hela cells, hATF-SPIO could specifically combined to and be subsequently internalized by uPAR positive cells, which could be observed via Prussian blue staining. Meanwhile T2WI signal intensity of Hela cells, after incubation with targeted probe, significantly decreased, and otherwise no obvious changes in Lovo cells both by Prussian blue staining and MRI scans. In vivo, hATF-SPIO could be systematically delivered to HT29 xenograft and accumulated in the tumor tissue which was confirmed by Prussian Blue stain compared to Lovo xenografts. Twenty-four hours after injection of targeting probe, the signal intensity of HT29 xenografts was lower than Lovo ones which was statistically significant. This targeting nanoparticles enabled not only in vitro specifically combining to uPAR positive cells but also in vivo imaging of uPAR moderately elevated colon cancer lesions.
Cell Line, Tumor ; Colonic Neoplasms ; diagnosis ; Ferric Compounds ; Humans ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; chemistry ; Molecular Imaging ; methods ; Receptors, Urokinase Plasminogen Activator ; chemistry ; Staining and Labeling

Objective To investigate the effects of early nasal intermittent positive pressure ventilation (NIPPV) compared with early continuous positive airway pressure (NCPAP) in low birth weight preterm infants with respiratory distress syndrome (RDS).Methods We performed a prospective,randomized controlled trial involving 364 low birth weight preterm infants with respiratory distress syndrome within 6 hours of birth.The infants were randomly assigned to NIPPV (n=188) or NCPAP (n=176) groups.Non invasive ventilation was initiated in the neonatal intensive care unit (NICU).The rate of mechanical intubation (within 3 days or 7 days),the use of pulmonary surfactant (PS),the rate of complications and mortality were compared between the two groups.Mann Whitney U,t and Chi-square tests were used for statistical analysis.Results The average time of invasive mechanical ventilation in NIPPV group were lower than that in NCPAP group[2.0 (1.0-4.0) d vs 7.0 (3.0-8.5) d,U=-3.457,P=0.001].The need for intubation and mechanical ventilation by day 3 and day 7 in the NIPPV group were less than those in the NCPAP group [day 3:4.8％ (9/188) vs 10.8％ (19/176),x2=4.621,P=0.032; day 7:9.0％ (17/188) vs 16.5％ (29/176),x2=4.551,P=0.033].In the NIPPV group,infants who got PS therapy was less than that in the NCPAP group [3.2％ (6/188) vs 8.5％ (15/176),x2=4.752,P=0.029].There was no significant difference in the fatality rate between the NIPPV and the NCPAP group [12.8％ (24/188) vs 10.8％ (19/176),P ＞ 0.05].There were no significant difference in the incidence of air leak,intracranial hemorrhage,periventricular leukomalacia,retinopathy of prematurity,necrotizing enterocolitis,patent ductus arteriosus,and bronchopulmonary dysplasia between the NIPPV group and the NCPAP group.Conclusion Among low birth weight prcterm infants with RDS,the early use of NIPPV reduces the need for PS,intubation and invasive ventilation compared with NCPAP.