Objective To explore the relationship between galactose-induced apoptosis of lens epithelial cells and cataractogenesis.Methods Galactose was injected retrobulbarly into Wistar rats.The opacity of lens was examined by slit lamp.The expression of C-MYC was measured by flow cytometer.The apoptosis of lens epithelial cells was observed by TUNEL techniques.Results More than 50 percent of lenses were in the Ⅱ stage of cataractogenesis on the(7 d) after galactose induction;Eighty-seven percent of lenses were in the Ⅳ-Ⅴ stage on the(14 d),and 100 percent of lenses were in the Ⅳ-Ⅴ stage on the(24 d).The rate of apoptotic cell on the(7 d) and on the(14 d) after galactose induction was respectively 5.6%～8.4% and 30.2%～41.8%,and 60% apoptotic cells were observed on the(24 d).The levels of C-MYC expression in lens epithelial cells induced by galactose on the(7 d) and(14 d) were higher than that of normal lenses,but the level of C-MYC expression backed to normal level on the(24 d).Conclusion It was suggested that there are some apoptosis of the lens epithelial cells during galactose-induced cataractogenesis and that the apoptosis of the lens epithelial cells might be caused by higher expression of C-MYC.

Objective To evaluate the strictness and self-consistency of the charge nurses in the nurses examination using venipunctnre trocar operation as research object.Methods FACETS,the polyhedral Rasch software was used in the examination.Results The strictness and self-consistency of charge nurses were evidently different.Conclusions The variable strictness and self-consistency of the charge nurses in the examination leads to unreliability of the results.Developing a scientific and reliable evaluation system is essential to improve the ability of the teaching nurses and the student nurses as well as the hospital nursing quality.

Objective To partially purify the toxic factor secreted by A. corymbifera and to analyze the mechanism of A. corymbifera-induced human umbilical vein endothelial cell (HUVEC) apoptosis. Methods Glycoprotein secreted by A. corymbifera was purified by Con A Lectin chromatography. The influence of different protein fractions on HUVEC apoptosis was determined by flow eytometer. Both denaturing and nondenaturing deglycosylation of purified glycoprotein was performed and the ability of the protein moiety and carbohydrate moiety to induce HUVEC apoptosis was evaluated respectively. Activation of related caspases during A. corymbifera-induced apoptosis was analyzed by Western blot. The role of caspase-8 and -9 in HUVEC apoptosis was investigated using caspase inhibitors. Caspase inhibitors were used to stop the suppression of HUVEC viability by XTT assay. Results Flow cytometric analysis shows the total protein as well as the glycoprotein fraction of A. corymbifera may induce HUVEC apoptosis in a dose dependent manner. In contrast, similar activity was not observed in the non-glycoprotein fraction. Neither deglycosylated protein nor carbohydrate moiety is able to induce HUVEC apoptosis alone. In the apoptotic signaling pathway, caspase9, caspase-3 and cytochrome C were activated significantly, except caspase-8. Moreover, caspase-9 inhibitor, instead of caspase-8 inhibitor, completely abrogates A. corymbifera-induced HUVEC apoptosis. Caspase9 and caspase-3 inhibitors completely waived the suppression of HUVEC viability by A. corymbifera. Conclusion Glycoprotein secreted by A. corymbifera is associated with HUVEC apoptosis. Intact glycoprotein is essential for the apoptotic progress. Intrinsic apoptotic signaling pathway mediates A. corymbifera-induced HUVEC apoptosis.

Objective To analyze safety,efficacy and resection methods of video-assisted thoracic surgery(VATS) for the treatment of intralobar pulmonary sequestration(IPS).Methods Data of 17 patients who were diagnosed as IPS and received VATS from December 2006 to September 2011 were retrospectively analyzed.The patients were 7 males and 10 females with the mean age of 40.3 (14-61) years.Diagnosis was confirmed in 9 patients by enhanced CT and unconfirmed in 8 patients.Three ports were used for surgery.After the aberrant artery was confirmed,liner stapler was used in 16 patients to cut it and Hem-o-lok was used in 1 patient because the aberrant artery was about 3 mm in diameter and long enough.If the diameter of the aberrant artery was longer than 10 mm,a stapling device without knife was used to occlude it centrally and a second stapling device was used to cut it peripherally.Wedge resection or lobectomy was performed due to the different conditions.When the lesion was small with linited range in CT image and the lesion was easily distinguished from normal lung tissue during operation,wedge resection was preferred.Results Seventeen patients underwent VATS successfully without any conversion to thoracotomy or any serious complications.Five patients were planned to receive wedge resection and one was converted to lobectomy.Another 12 patients were planned to receive lobectomy and all succeeded.The mean operating time was 128 (80-170)min.The mean blood loss was 80 (5-200) ml.The mean days of chest tube maintained were 4.0 (2-6) days.The mean postoperative hospitalization days were 7.6 (4-11) days.All patients were diagnosed as IPS according to operating in-sight and postoperative pathology.There was no patient suffering from chronic cough,bloody sputum or recurrent pneumonia during the follow-up.Conclusion VATS for the treatment of IPS is safe and feasible.If conditions permit,wedge resection or segmentectomy may be preferred.

Background Various studies demonstrated that the apoptosis of lens epithelial cells(LECs) is associated with the overexpression of the c-myc gene in LECs induced by galactose.Inhibiting the abnormal expression of the c-myc gene in LECs is an effective approach to mitigate the pathogenesis and development of cataract.Objective The goal of this study is to investigate the inhibitory effects of c-myc antisense oligodeoxynucleotide(c-myc ASODN) on the apoptosis of LECs in the eye with galactose-induced cataract.Methods Galactose-induced cataract models were established by the retrobulbar injection of 0.2 mL of 20% galactose once per day.Lipo-antisense oligodeoxynucleotide(Lipo-ASODN,0.2 mL) was retrobulbarly injected 4 hours after the injection of galactose at one-day intervals.The animals were sacrificed and lenses were obtained to evaluate the apoptosis of LECs and the effect of c-myc ASODN on LECs apoptosis induced by galactose was examined by TUNEL assay after 7,14 and 24 days.The ultrastructural changes of LECs were examined under the transmission electron microscopy(TEM).Results A significant difference in the apoptotic rate of LECs was found among the 7 day,14 day and 24 day groups(F_(7 days)=3 418.495,P＜0.01;F_(14 days)=1137.555,P＜0.01;F_(24 days)=2198.871,P＜0.01).The apoptotic rate of LECs in the galactose group was markedly higher than that in the normal saline solution group 7 days,14 days and 24 days after the experiment(P＜0.01).The apoptotic rate of LECs in the galactose+lipo+ ASODN group significantly declined in comparison to the galactose group after 7 days,14 days and 24 days(P＜0.05).TUNEL assay showed the condensation,breakage and irregularity of the nuclei of apoptotic cells in the galactose group.The destruction of the ultrastructure of the cells and organelles were observed under the transmission electron microscope.Conclusion Galactose induces apoptosis of LECs in cataractogenesis.C-myc ASODN inhibits apoptosis of LECs induced by galactose.

Objectives To investigate the expression and diagnostic value of 14-3-3 protein in brains of patients with sporadic Creutzfeldt-Jakob disease(sCJD).Methods 14-3-3 protein was immunohistochemically analyzed in tissue from the frontal lobe of 5 patients with sCJD and 4 non-CJD eases Using 14-3-3 ?and ?antibodies with reference to the results of KB,GFAP and PrP detection.Results The expressions of 14-3-3 protein in five brains of sCJD were more obviously,mostly in gray matters and astrocytes in three cases.The concentration was related to PrP deposition type,but not related to prion protein genotype.Except few expression of 14-3-3 protein in neurous of two cases of acute contusion,there were no expression in the other two cases in control group.Conclusions The expression of 14-3-3 protein in brain is useful to pathological diagnosis of CJD.

Objective To analyze the influence of Absidia corymbifera on cell activity of human umbilical vein endothelial cells (HUVEC) as well as the related mechanism. Methods Time course analy sis of the influence of A. corymbifera on cell viability of HUVEC was determined by cell counting after Trypan blue staining. Apoptosis of HUVEC induced by A. corymbifera was observed under fluorescence microscope after treatment with apoptosis detection kit. Time course analysis of HUVEC apoptosis induced by A. corymbifera was detected by flow cytometry quantitatively. Effect of caspase-3 inhibitor on A. corymbifera associated apoptosis was also evaluated at the same time. Activation of caspase-3 inside HUVEC was detected by Western blot. Results A. corymbifera inhibited cell viability of HUVEC in a time-dependent manner by Trypan blue staining. After 12 hours' co-culture, A. corymbifera began to show suppression on cell viability (P =0. 001 ). Fluorescence microscope observation revealed A. corymbifera induced apoptosis of HUVEC instead of necrosis. Flow cytometry analysis showed A. corymbifera induced apoptosis of HUVEC in a time-dependent manner. A. corymbifera began to show obvious effect on apoptosis after 12 h co-culture (P =0.0036). Moreover, A. corymbifera-associated apoptosis was almost abrogated completely by caspase-3 inhibitor. Western blot analysis demonstrated that A. corymbifera triggered the activation of caspase-3 inside HUVEC in a timedependent fashion. Conclusion A. corymbifera induces apoptosis of HUVEC in vitro. Such apoptotic signal is transmitted through caspase cascade reaction.

BACKGROUND: Adipose-derived stem cells (ADSCs) represent one of the promising cell sources for myocardial regeneration due to their easy accessibility and efficacy in the improvement of cardiac function following myocardial infarction. However, previously reported studies on the underlying mechanism of ADSCs-mediated cardioprotective effect mainly focused on the ADSCs cultured at room air. OBJECTIVE: To test the paracrine actions and anti-apoptotic effect of ADSCs under hypoxic conditions. METHODS: After isolation and culture, neonatal rat myocardial cells were injured by hydrogen peroxide and co-cultured with rat ADSCs under normoxia and hypoxia (10% O2) conditions. Ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) in the cell pellet and levels of vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1), and basic fibroblast growth factor (bFGF) were tested by ELISA. Expression of apoptotic proteins Bax and Bcl-2 were determined by western blot. RESULTS AND CONCLUSION: GSH/GSSG, VEGF, IGF-1, and bFGF were decreased in neonatal rat myocardial cells injured by hydrogen peroxide. ADSCs significantly attenuated hydrogen peroxide-induced myocardial apoptosis by increasing the ratio of GSH/GSSG and the secretion of VEGF, IGF-1 and bFGF. ADSCs also down-regulated Bax expression and up-regulated Bcl-2 expression. To conclude, hypoxic conditions can enhance the anti-apoptosis and cardioprotective effects of ADSCs through the paracrine mechanism.

Mytilin-derived-peptide-1 (MDP-1) and mytilin-derived-peptide-2 (MDP-2) are two truncated decapeptides with reversed sequence synthesized corresponding to the residues 20-29 of mytilin-1 (GenBank Accession No. FJ973154) from M. coruscus. The objective of this study is to characterize the structural basis of these two peptides for their antimicrobial activities and functional differences, and to investigate the inhibitory mechanism of MDPs on Escherichia coli and Sarcina lutea. The structures of MDP-1 and MDP-2 in solution were determined by 1H 2D NMR methods; the antibactericidal effects of MDPs on E. coli and S. lutea were observed by transmitted electron microscopy (TEM). Both MDP-1 and MDP-2 have a well-defined loop structure stabilized by two additional disulfide bridges, which resemble the-hairpin structure of mytilin-1 model. The surface profile of MDPs' structures was characterized by protruding charged residues surrounded by hydrophobic residues. TEM analysis showed that MDPs destroyed cytoplasmic membrane and cell wall of bacteria and the interface between the cell wall and membrane was blurred. Furthermore, some holes were observed in treated bacteria, which resulted in cell death. Structural comparison between MDP-1 and MDP-2 shows that the distribution of positively charged amino acids on the loop of MDPs is topologically different significantly, which might be the reason why MDP-2 has higher activity than MDP-1. Furthermore, TEM results suggested that the bactericidal mechanisms of MDPs against E. coli and S. lutea were similar. Both MDP-1 and MDP-2 could attach to the negatively charged bacterial wall by positively charged amino acid residues and destroy the bacteria membrane in a pore-forming manner, thus cause the contents of the cells to release and eventually cell death.
Animals ; Anti-Infective Agents ; chemical synthesis ; pharmacology ; Antimicrobial Cationic Peptides ; chemical synthesis ; chemistry ; pharmacology ; Cell Wall ; drug effects ; Escherichia coli ; drug effects ; Mytilus ; chemistry ; Sarcina ; drug effects

OBJECTIVETo explore the noninvasive indicators for predicting the occurrence of esophageal varices (EV) in patients with liver cirrhosis.

METHODSA total of 202 patients with liver cirrhosis caused by hepatitis B or C or alcoholic hepatic disease were enrolled in this study. EV and high risk esophageal varices (HREV) were confirmed in these patients by gastroscopy. The hematological, serum biochemical and ultrasonic parameters of the patients were analyzed, and a model for predicting EV was established by stepwise logistic regression analysis.

RESULTSThe areas under receiver operating characteristics curve (AUROC) of splenic thickness (SPT) for detecting EV and HREV were 0.827 and 0.766, respectively. The combined index USWA (SPT, US, WBC and albumin [ALB]) showed an AUROC of 0.873 for detecting EV, and the index SPA (combining SPT and ALB) showed an AUROC of 0.777 for detecting HREV. The direct combination of SPT with USWA or with platelet/splenic thickness ratio (PSA) was capable of completely excluding a definite diagnosis of EV, while the sequential combination of SPT with USWA or with platelet was capable of a diagnosis of EV and clarifying the condition of EV in approximately half of the cirrhotic patients in the absence of gastroscopic findings. The combination of SPT and SPA allowed for a definite diagnosis of the condition of HREV in 10% of the cirrhotic patients.

CONCLUSIONSPT combined with SPT derived indexes or platelet status allows for a definite diagnosis of EV in patients with liver cirrhosis to offer a noninvasive option for diagnosis.