The anti-tumour mechanism of Shuanghuang Qizhen Keliu Prescription based on network pharmacology / 中国药理学通报

Aim To explore the anti-tumour mechanism of Shuanghuang Qizhen Keliu Prescription based on network pharmacology. Methods The active components and targets of Shuanghuang Qizhen Keliu Prescription were searched and selected through the TCM-SP database. Related targets of tumor were screened and retrieved by OMIM. The key anti-tumor targets of Shuanghuang Qizhen Keliu Prescription were obtained by using Venn diagram analysis. The key target PPI network was constructed and the key targets were obtained by Cytoscape. Metascape platform was used for GO and KEGG enrichment analysis of key targets. Results There were many active components such as quercetin, kaempferol, and beta-sitosterol in Shuanghuang Qizhen Keliu Prescription, and their targets might be TP53, AKT1, JUN, MAPK1, TNF, RELA, IL-6 and so on. The anti-tumour mechanism might be proliferation inhibition, migration inhibition and apoptosis promotion. Conclusions This study preliminarily discusses the potential anti-tumour mechanism of Shuanghuang Qizhen Keliu Prescription, providing ideas and basis for subsequent experimental research.

Effect of extracorporeal membrane oxygenation measures on prognosis of children with fulminant myocarditis / 中国小儿急救医学

Objective To investigate the clinical features and the clinical effects of extracorporeal membrane oxygenation(ECMO) in children with fulminant myocarditis. Methods The data of one hundred and sixty one pediatric patients with fulminant myocarditis at Beijing Children′s Hospital from March 1,2007 to May 31,2018 were analyzed retrospectively,including clinical manifestations,electrocardiogram,echocar￣diography,myocardial enzyme,the application of ECMO,curative effect and prognosis. Results Among 161 cases of children with fulminant myocarditis,74 cases (46. 0%) had respiratory tract symptoms,54 cases (33. 5%) had digestive tract symptoms,24 cases (14. 9%) had nervous system symptoms,only 33 cases (20. 5%) had cardiovascular symptoms. One hundred and thirty￣two cases(82. 0%) had elevation of crea￣tine kinase isoenzymes( CK￣MB) and/or cardiac troponin I( cTnI),and 107 cases(66. 5%) had abnormal echocardiography. One hundred and fifty￣two cases ( 94. 4%) showed abnormal electrocardiogram, and the changes of ST￣T and atrioventricular block were most common. Five cases survived in 8 children treated with ECMO. They underwent ECMO with an median supporting time of 101. 5 (52. 75,142. 8) hours through a venous￣arterial ECMO model. After the application of ECMO,blood pressure,the heart rate and oxygen satura￣tion were obviously improved in the 8 children,and the level of blood lactic acid was statistically reduced com￣pared with that before the application of ECMO(P<0. 001). During the follow￣up period of 1 months,the 5 surviving children recovered with normal cardiac function,without abnormal function of other organs. There were a total of 17 cases(10. 6%) died,4 children(8. 9%) in 45 patients died since the application of ECMO and the other 13 children(11. 6%) in 116 patients died before that time. This proportion of death was reduced after the application of ECMO,whereas the difference was not statistically significant(P>0. 05). Conclusion Our study indicates the diverse symptoms in children with fulminant myocarditis. Comprehensive application of electrocardiogram,echocardiography,and myocardial enzymology markers may be of benefit to the timely diag￣nosis of fulminant myocarditis. The application of ECMO in fulminant myocarditis remains an effective approach in reduction of mortality rate,and can effectively support recovery of cardiopulmonary functions.

Role of hippocampal nNOS-PSD95 coupling in short-term memory retrieval disorder induced by sevoflurane in mice / 中华麻醉学杂志

Objective To evaluate the role of hippocampal neuronal nitric oxide synthase(nNOS)-postsynaptic dense protein 95(PSD95)coupling in short-term memory retrieval disorder induced by sevoflu-rane in mice.Methods Sixteen clean-grade healthy Kunming mice of both sexes,aged 2-3 months,weighing 30-35 g,were divided into 2 groups(n＝8 each)according to the random number table method:sevoflurane group(S group)and nNOS-PSD95 uncoupling agent ZL006 group(Z group).After successful establishment of dark avoidance memory,3.3％sevoflurane and 40％O2 were inhaled for 2 h in both groups,and normal saline 1.5 ml was intraperitoneally injected in group S and ZL006 1 mg/kg in group Z at 30 min before anesthesia.The step-through latency and error times were recorded before anesthesia and at 12 h after the end of anesthesia.The mice were then sacrificed,and hippocampal tissues were taken for de-termination of the expression of nNOS and PSD95(by Western blot)and co-expression of nNOS and PSD95(by immunoprecipitation).Results Compared with that before anesthesia,the step-through latency was significantly shortened,and the error times were increased at 12 h after anesthesia in group S(P＜0.05),and no significant change was found in the above indicators in group Z(P>0.05).Compared with group S,the step-through latency was significantly prolonged,error times were decreased,the co-expression of nNOS and PSD95 was down-regulated(P＜0.05),and no significant change was found in the expression of nNOS and PSD95 in group Z(P>0.05).Conclusion The mechanism by which sevoflurane induces short-term memory retrieval disorder may be related to promoting the coupling of nNOS to PSD95 in the hippocam-pus of mice.

Transiciones a la adultez: exploración sobre las relaciones románticas y normativas de masculinidad de los adolescentes en el Bañado Sur, Asunción / Transitions to adulthood: exploration of romantic relationships and norms of masculinity adolescents in Bañado Sur, Asunción

Introducción: muchas veces los hombres jóvenes se definen a sí mismos y a su identidad masculina a través de relaciones románticas y sexuales; sus decisiones sexuales pueden afectar su transición hacia la madurez, así como la adquisición de infecciones de transmisión sexual, VIH y tasas de embarazos. Objetivo: Este artículo analiza cómo influyen en el modo en que actúan en las relaciones sexuales y románticas y la formación de su identidad masculina, los grupos de amigos de hombres jóvenes paraguayos, sus familias. Metodología: En el 2010 llevamos a cabo cinco charlasen grupo en Asunción, Paraguay, en las que se examinaron las normas comunitarias con grupos de adolescentes entre 14 y 19 años de edad. Luego entrevistamos a la mitad de los miembros de cada grupo para examinar sus relaciones con amigos, la familia, mujeres jóvenes y sus creencias sobre las normas de género existentes. Resultados: Los jóvenes describieron dos tipos de normas masculinas (“pareja/proveedor” y “macho”) y dos tipos de relaciones románticas (“casual” y “formal”). Muchas veces se encontró una concordancia en el lenguaje utilizado para describir cada abanico de conductas, lo que pone de relieve la relación existente entre las normas masculinas y las relaciones románticas.

Introduction: Many times young men define themselves and their male identity through romantic and sexual relationships; their sexual decisions can affect their transition to adulthood as well as the acquisition of sexually transmitted infections, HIV and pregnancy rates. Objective:This article discusses how to influence the waythey act in sexual and romantic relationshipsand the formation of their male identity, groupsof young men Paraguay an friends, their families. Methodology: In 2010 we conducted five talks group in Asuncion, Paraguay, in which Community rules were examined with groups of teenagers between 14 and 19 years old. Then we interviewed half of the members of each group to examine their relationships with friends, family, young women and their beliefs about existing gender norms. Results: Young described two types of male norms(“partner / supplier” and “male”) and two types of romantic relationships (“casual” and “formal”).Many times a match was found in the languageused to describe each range of behaviors, which highlights the relationship between male normsand romantic relationships. The rules received bythe neighborhood as models that were more machobehavior by young people themselves informedcharacteristics. The rules can not be changed unlessyoung people talk about their behavior clearly notnormative. This demonstrates that further studieson the formation, meaning and transformation of male standards are needed.

Population pharmacokinetics and inter-laboratory variability of sildenafil and its metabolite after oral administration in Korean healthy male volunteers

This study was to clarify population pharmacokinetics (PK) of sildenafil and its metabolite, N-desmethyl sildenafil (NDS) in Korean healthy male population using a pooled data from multiple clinical trials in consideration of inter-institution and inter-laboratory difference. A population PK analysis was performed with data of 243 healthy volunteers from five single-center (4 centers) comparative PK trials. The dataset included 7,376 sildenafil and NDS concentration (3,688 for each analyte) observed during 24 hours after the single dose of original sildenafil (either 50 mg or 100 mg of Viagra®). The plasma concentration was assayed in two laboratories. Various model structure was tested and the final model was evaluated using visual predictive checks. Demographic and clinical variables were assessed as potential covariates for PK parameters. A one-compartment first-order elimination model with proportional error was selected for the dispositional characteristics of sildenafil, and two-compartment model was chosen for NDS. Three transit compartments with Erlang-type absorption for fast absorption pathway and one compartment for slow absorption pathway constructed overall absorption model. The first-pass effect was rejected since it does not improve the model. The difference of NDS level by the bioanalysis laboratory was selected as the only covariate. Even though a direct comparison was difficult, the general trend in PK of sildenafil and NDS for Korean healthy male was considered similar to that of the other populations reported previously. It is recommended that the laboratory effect should be explored and evaluated when dataset is built using results from several laboratories.

Effect of cetuximab combined with adriamycin on proliferation and apoptosis of triple negative breast cancer cells / 中国药理学通报

Aim To detect the effects of cetuximab combined with adriamycin on the proliferation and ap-optosis of triple-negative breast cancer cells.Methods Cell viability was evaluated by MTT assay.The cell apoptosis was analyzed by flow cytometry with propidi-um iodide staining.JC-1 staining was used to deter-mine mitochondrial membrane potential.The expres-sions of glucose regulated protein78 (GRP-78),Bcl-2 and Caspase-3 were measured with Western blot.Re-sults MTT assay showed that cetuximab had inhibi-tion effect on the breast cancer cell MDA-MB-231 growth,and the effect was related to concentration of drug.The inhibition effect of adriamycin on MDA-MB-231 had remarkabe relationship with time and concen-tration.When combined with each other,they could re-markably increase inhibition effect.The viability of cells in combination group for 1 2 h,24 h,48 h,sig-nificantly lower than that in cetuximab or adriamycin group (P group (P <0.01 ).JC-1 staining indicated that cetux-imab or adriamycin could reduce the mitochondrial membrane potential,but the reduction effect was more remarkable in the combination group.Western blot re-vealed that cetuximab could reduce the expression of GRP-78 and Bcl-2,and increased the expression of Caspase-3 and its activity.The expressions of Bcl-2, Caspase-3 had no significant change in adriamycin group,but GRP-78 was increased.In combination group,the expression of GRP-78 and Bcl-2 was signifi-cantly decreased,but Caspase-3 was increased nota-blely compared to adriamycin group.Conclusions The combination of cetuximab and adriamycin enhances the inhibition effect on the triple-negative breast cancer MDA-MB-231 cells,and increases cell apoptosis.The mechanism may be that cetuximab reduces the endo-plasmic reticulum stress level,then activates the mito-chondrial pathways by decreasing the expression of Bcl-2,reducing the mitochondrial membrane potential,and promoting cell apoptosis.

Anti-proliferative effect of salinomycin on doxorubicin-resistant human breast cancer MCF-7/DOX cells / 中国药理学通报

Aim To investigate the anti-proliferative effect of salinomycin on doxorubicin-resistant human breast cancer MCF-7 /DOX cells.Methods MCF-7 and MCF-7 /DOX cells were treated or untreated with salinomycin.Cell viability was detected by MTS assay. Cell apoptosis was detected by Annexin V-FITC /PI as-say.Reactive oxygen species (ROS)was measured by DCFH-DA staining.Mitochondrial membrane potential was measured by JC-1 assay.The expression of apopto-sis related proteins BAX, BCL-2, caspase-3, and caspase-9 were evaluated by Western blot analysis. Results The cell viability was significantly reduced by salinomycin treatment in a dose-dependent manner. The flow cytometry results showed that salinomycin in-duced MCF-7 /DOX cell apoptosis,increased ROS pro-duction,and decreased mitochondrial membrane poten-tial.Furthermore,salinomycin decreased the expres-sion of BCL-2,and increased the expression of BAX, cleaved caspase-3,and cleaved caspase-9.Moreover, the antioxidant N-acetylcysteine (NAC ) markedly blocked the above effects.Conclusions Our results suggest that salinomycin-induced apoptosis in MCF-7 /DOX is associated with induction of ROS production, and activation of mitochondria apoptosis pathway, which may become a potential chemotherapeutic agent for the therapy of doxorubicin resistant breast cancer.

Detection and characterization of metallo-β-lactamases in Pseudomonas aeruginosa by phenotypic and molecular methods from clinical samples in a tertiary care hospital / Detección y caracterización de la metallo-betalactamasa en la Pseudomonas aeruginosa mediante métodos fenotípicos y moleculares a partir de muestras clínicas en un hospital de atención terciaria

AIMS: The aim of this study was to detect and characterize the presence of metallo-β-lactamase (MBL) production in multidrug resistant (MDR) P aeruginosa collected from clinical samples in a tertiary care hospital. METHODS AND MATERIALS: A total of 67 non-repetitive isolates of MDR P aeruginosa recovered from various clinical specimens were screened for MBL production by IPM/MEM-EDTA combined disc test. Polymerase chain reaction was performed on all isolates using blaIMP and blaVIM consensus primers to characterize them genotypically. RESULTS: Among 67 P aeruginosa isolates, 62.7% (42/67) and 70.1% (47/67) were resistant to imipenem and meropenem respectively and 47 (70.1%) were found to be MBL producers. Among this 47 MBL-producing isolates, 41 (61.1%) strains carried the blaVIM gene and 2 (3%) strains carried the blaIMP gene. Three strains were phenotypically negative but positive genotypically for blaVIM gene. One strain was resistant to both imipenem and meropenem but did not show phenotypic positivity. CONCLUSION: This study confirms the dissemination of blaVIM genes among MDR Pseudomonas aeruginosa and hence it is indispensible to identify and aptly control the threat of horizontal and vertical transfer.

OBJETIVO: El objetivo de este estudio es descubrir y caracterizar la presencia de producción de metallo-betalactamasa (MBL) en P aeruginosa resistente a los multifármacos (RMF), recogida de muestras clínicas de un hospital de atención terciaria. MÉTODO: Un total de 67 aislados no repetitivos de P aeruginosa RMF obtenidos de varios specímenes clínicos, fueron tamizados en busca de producción de MBL, mediante una prueba de disco combinado IPM/MEM-EDTA. Se efectuó una reacción en cadena de la polimerasa sobre todos los aislados, usando iniciadores de consenso blaIMP y blaVIM para la caracterización genotípica. RESULTADOS: Entre los aislados de P aeruginosa, 62.7% (42/67) y 70.1% (47/67) fueron resistentes al Imipenem y al Meropenem respectivamente, mientras que se halló que 47 (70.1%) eran productores de MBL. De los 47 aislados productores de MBL, 41 (61.1%) cepas eran portadoras del gen blaVIM en tanto que 2 (3%) cepas eran portadoras del gen blaIMP. Tres cepas fueron fenotípicamente negativas, pero genotípicamente positivas con respecto al gen blaVIM. Una cepa fue resistente tanto al Imipenem como al Meropenem, pero no mostró positividad fenotípicamente. CONCLUSIÓN: El presente estudio confirma la diseminación de los genes blaVIM entre las Pseudomonas aeruginosa RMF. Es importante identificar así como controlar adecuadamente la amenaza de la transferencia horizontal y vertical.

Evaluación de la reproducibilidad de la retinoscopia dinámica monocular de Merchán / Evaluation of the reproducibility of the Merchán monocular dynamic retinoscopy

Objetivo: Evaluar la reproducibilidad de la retinoscopia dinámica monocular y su nivel de acuerdo con la retinoscopia estática binocular y monocular, retinoscopia de Nott y Método Estimado Monocular (MEM). Métodos: Se determinó la reproducibilidad entre los evaluadores y entre los métodos por medio del coeficiente de correlación intraclase (CCI) y se establecieron los límites de acuerdo de Bland y Altman.Resultados: Se evaluaron 126 personas entre 5 y 39 años y se encontró una baja reproducibilidad interexaminador de la retinoscopia dinámica monocular en ambos ojos CCI ojo derecho: 0.49 (IC95% 0.36; 0.51); ojo izquierdo 0.51 (IC95% 0.38; 0.59). El límite de acuerdo entre evaluadores fue ±1.25 D. Al evaluar la reproducibilidad entre la retinoscopia dinámica monocular y la estática se observó que la mayor reproducibilidad se obtuvo con la estática binocular y monocular y, en visión próxima, entre el método estimado monocular y la retinoscopia de Nott. Conclusiones: La retinoscopia dinámica monocular no es una prueba reproducible y presenta diferencias clínicas significativas para determinar el estado refractivo, en cuanto a poder dióptrico y tipo de ametropía, por tanto, no se puede considerar dentro de la batería de exámenes aplicados para determinar diagnósticos y correcciones refractivas tanto en la visión lejana como en la visión próxima.

Objective: To assess the reproducibility of monocular dynamic retinoscopy and its level of agreement with binocular static retinoscopy, the Nott retinoscopy, and the Monocular Estimated Method (MEM). Materials and methods: The reproducibility was determined among evaluators and among the methods through the intraclass coefficient of correlation (ICC) and set the limits of agreement of Bland and Altman. Results: A total of 126 subjects between 5 and 39 years of age were evaluated and low inter-rater reproducibility of the monocular dynamic retinoscopy was found in both eyes: ICC right eye: 0.49 (IC 95% 0.36; 0.61) left eye 0.51 (IC 95% 0.38; 0.59). The limit of agreement between evaluators was ±1.25D. In assessing reproducibility between the monocular dynamic retinoscopy and the static retinoscopy, it was observed that the greatest reproducibility was obtained with the binocular and monocular static retinoscopy and in near vision between the estimated method monocular and Nott retinoscopy. Conclusions: Monocular dynamic retinoscopy is not a reproducible test and presents clinical differences meaningful to determine the refractive state, regarding power dial and type of ametropy; therefore, it could not be considered within the battery of tests used to determine diagnoses and refractive corrections both in distant vision and in near vision.

Construction of human interleukin-10 eukaryotic expression vector pcDNA4/HisMaxA-hIL-10 and its expression in rabbit synovial cells / 中华风湿病学杂志

Objective To construct an efficient eukaryotic expression recombinant vector of human interleukin-1O(hIL-lO),and observe its expression in rabbit synoviocytes(RSCs).Methods Total RNA was extracted from peripheral blood mononuclear cells(PBMCs)of a patient with drug allergy.Specific Drimers for full-length open reading frames(ORFs)of hIL-10 were designed according to GeneBank(NM 000572).Withtotal RNA as the template,full-length ORFs of hIL-10 were amplified by reverse transcription Dolymerase chain reaction(RT-PCR).RT-PCR products were digested by restrictive endonucleotidase.then inserted into plasmid pcDNA4/HisMaxA.Both restrictive endonucleotidase analysis and DNA sequencing Were carried out for inserts verification.RSCs were transfected with recombinant plasmid expression vector PcDNA4 HisMaxA-hiL10 by liposome-mediated gene transfer methods,then cultured in vitro.The supernatants were collected af-ter transfection for 12 hours,24 hours,48 hours,72 hours,7 days,14 days respectively for IL-10 measure-ment by enzyme linked immunosorbent assay(ELISA).Results Full-length ORFs of hIL-1o(0.54 kb)had been successfully cloned from PBMCs through RT-PCR.The inserts and insert location of pcDNA4 HisMaxA were in a fight way verified by enzyme analysis and DNA sequencing.ELISA results showed that exogenous hIL-10 gene had expressed in the transfected RSCs from 12 hours to 7 days after transfection,and hIL-10level of transfection group significantly higher than that of the control group.Conclusion pcDNA4 HisMaxA-hiL10,the hIL-10 efficient eukaryotic expression vectors,has been suecessfully constructed.

Display of heterologous proteins on the surface of E. coli by Red recombineering / 中华微生物学和免疫学杂志

Objective To display of heterologous proteins on the surface of E. coli . Methods The 1653 bp luciferase report gene was knocked in Ipp and ompA genes of chromosome by Red recombine system and selection-counter selection kan/sacB. Results The quantitative analysis results of exogenous lu-ciferase expression displayed that it could be expressed as fusion with the outer membrane proteins on the cell surface. The fusion protein of foreign protein and outer membrane protein Lpp-OmpA-Luc could be high-effi-ciently displayed on cell surface. Conclusion The stable expression of exogenous gene on the surface of E. coli had no effect on the bacterial growth and propagation.

Effect of Earle's solution and MEM on activities of neurons in culture / 中国康复理论与实践

@# ObjectiveTo observe the activities of cultured neurons incubated with Earle's solution, new minimal essential medium (MEM) or former MEM.MethodsActivity changes of cultured neurons were measured by determining the leakage of lactate dehydrogenase (LDH) and metabolic rate of 3-(4,5-dimethylthiazol)2,5 diphenyltetrazolium bromide (MTT).ResultsActivities of neurons incubated with different culture medium for 12 h or 24 h were significantly different (P<0.01).ConclusionDifferent culture medium can influence neuronal activities.

Comparison between Cuprophane and Polysulfone Membrane in Biocompatibility / 대한신장학회잡지

OBJECTIVE: It has been proposed that the contact between blood and dialysis membrane during hemodialysis therapy induces harmful reactions to patients. Membrane biocompatibility is the characteristic that makes less adverse reaction. We tried to compare the biocompatibility between Cuprophane and Polysulfone membranes. MRTHODS: Nine chronic renal failure patients who have undergone maintenance hemrodialysis therapy with Hemophan membrane three times per week were included in this study. Cuprophane membranes were used in the first week: Hemophan membranes in the second week; Polysulfone membranes in the third week. Each membrane was used three times a week. On the day of third dialysis with the Cuprophane membrane(first week) and Polysulfone membrane(third week), serial blood sampling was obtained from the afferent line at hemodialysis initiation, 15 minutes, 30 minutes, 60 minutes, 120 minutes and 30 minutes to measure serum complement activity(C3a, C5a), blood polymorphonuclear leukocyte and platelet count, and arterial oxygen pressure which have been regarded as biocompatibility parameters. The parameters measured during the first week(Cuprophane) and the third week(Polysulfone) were compared to evaluate the difference in biocompatibility of both membranes. RESULTS: 1) C3a desArg In both groups, the level of C3a desArg increased significantly from basal level and the Cuprophane group showed significantly higher level of C3a desArg than that of Polysulfone group. 2) C5a desArg : In both groups, the level of C5a desArg did not increased sigpificantly from basal level. Only at 30 minute after hemodialysis, Cuprophane group showed significantly higher level of C5a desArg than that of Polysulfone group(p=0.037). 3) Polymorphonuclear leukocyte : In both groups, the polymorphonuclear leukocyte counts decreased significantly at 15 minutes and 30 minutes from basal level. The polymorphonuclear leukocyte count was lower in Cuprophane group than that of Polysulfone group at 15 minute after hemodialysis(p=0.001). 4) Platelet: In Cuprophane group, the platelet count was decreased significantly at 15 minute(p=0.004) but there were no difference in platelet counts between the two groups. 5) Arterial oxygen pressure Both group showed no consistent pattern of variation of oxygen pressure and there was no significant difference between the two groups. CONCLUSION: The biocompatibility of Polysulfone membrane was superior to the Cuprophane membrane with respect to the increased activation of complement C3a. decrease of polymorphonuclear leukocyte and decrease of platelet count.

Effects of High Temperature and Noise on Erythrocyte Me mbrane ATPase Activity in Pilots During Flight / 航天医学与医学工程

Objective To determine the effect of heat and noise on erythrocyte memb rane ATPase activities in pilots during flying. Method Twenty- four pilots per forming bombing for 3h (45～53℃, 122～97dB in the cabin) served as the su bjects . 21 ground personnel served as control (27℃ in the room). Blood samples were t aken from both groups before flying (6:00 a.m), and immediately (12:00 a.m.) and 8h (8:00 p.m.) after flying. Na+-K+ATPase, and Ca2+-Mg2+ATPase activities in erythrocyte membran e were determined with colorimetry. Result The Na+-K+ATPase activity in eryth rocyte membrane at 6:00 a.m.in pilots was higher than that in control group at the s ame time (P<0.01). The Ca2+-Mg2+ATPase activities in erythrocyte membrane at 12:00 a .m. and 8:00 p.m. in pilots were significantly higher, compared with those in co ntrol group at the same time (P<0.01). Conclusion The ATPase values obtained in our study were all within normal range, and the daytim e variation of both groups are the same. Exposure of human body to heat and noi se for long time may be harmful, the higher ATPase activity is, the more catabol ism of ATP will be. ATP exhaustion will lead to Ca2+ overload in erythrocy te thus stiffen the red cell membrane.

Endothelial cells from the rat aorta in culture / 대한해부학회지

Endothelial cells were isolated from the intima of Sprague-Dawley rat aorta with 0.2% collagenase solution and cultured in MCDB 131 medium. All endothelial cells regardless of passages and the freezing-thawing were cultured in M-199, alpha-MEM, and MCDB 131 media. Thereafter, the morphological findings of the cells were observed under light and electron microscopes. The activities of nitric oxide synthetase of endothelial cells were investigated with NADPH-diphorase staining. Production of von Willebrand factor, cytoskeletal proteins, and extracellular matrix proteins in endothelial cells was identified with PAP. By treating with 0.2% collagenase solution for 30 minutes, and then incubating the cells in MCDB 131 medium for 3 days, only endothelial cells could be separated. Endothelial cells formed a monolayer with typical cobblestone pattern (polygonal-shaped morphology) 7-10 days after seeding. The growth patterns of endothelial cells were similar regardless of their cultured states (primary cultured cells, subcultured or thawed). The existence of nitric oxide synthetase, von Willebrand factor, beta-actin, fibronectin, and laminin in endothelial cells was confirmed. However, above-mentioned materials were quantitatively less in cells grown with MCDB 131 medium compared to those cultured with M-199 and alpha-MEM. Weibel-Palade bodies were observed by transmission electron microscopy. Cultured cells in MCDB 131 medium compared to those cultured with M-199 and alpha-MEM were relatively fewer in number of rough endoplasmic reticulum, mitochondria, free ribosome, vesicle, and liposome and especially showed the absence of basement membrane. Taken together, the cultivation of endothelial cells from the intima of rat aorta was possible. Endothelial cells in vitro synthesized various materials such as von Willebrand factor etc. and had characteristic organelles (Weibel-Palade body etc.). It suggested that it would be more advantageous to culture the endothelial cells in M-199 with 20% fetal bovine serum and alpha-MEM with 10% bovine calf serum rather than in MCDB 131 endothelial cell culture medium only with 0.7% dialyzed serum to maintain characteristics of endothelial cells in vivo

Effects of the Combination of Shenmai Injection and Aminophylline on Diaphragmatic Function in Rabbits / 中药新药与临床药理

Objective: Effects of Shenmai injection and aminophylline alone or in combination on transdiaphragmatic pressure and electrical activity of diaphragm were observed, so as to evaluate their effecfs on diaphragmatic fatigue. Methods: The diaphramatic fatique (DiF) model was estab- lished by stimulating bilateral phrenic nerves for 40 minutes. The animals were allocated to three groups: Shenmai group (2mL/kg, N=7), aminophylline group (20mg/kg, N=7) and combi- nation group (Shenmai injection 2mL/kg+aminophylline 20mg/kg, N=7) The transdi- aphragmatic pressure (Pdi) and diaphragmatic electromyogram (EMGdi) were recorded before and after treatment. EMGdi was analysed by computer and then the high/low frequency ratio (H/L) and the central frequency (Fc) were caculated. The diaphragm evoked potential (DEP) was record- ed simutaneously. All data was analysed by analysis of variance and q test. Results: Pdi, H/L, Fc and amplitude of DEP were markedly increased after the treatment with aminophyline and Shen- mai injection alone (Compared with DiF. P