Hong, Hai Xia;Zhang, Yan Ming;Xu, Hao;Su, Zheng Yuan;Sun, Pei
Molecules and Cells
/
v.24
no.3
/
pp.358-363
/
2007
Swine endothelial cells are commonly used as an in vitro model for studying features of the blood-brain barrier and some hemorrhagic diseases. However, primary cultures of swine cells have finite lifespans. To establish immortalized swine umbilical vein endothelial cells (SUVECs) using human telomerase reverse transcriptase (hTERT), the plasmid pCI-neo-hTERT was transfected into SUVECs by lipofection. Clones were selected for G418 resistance, and positive clones were amplified. One of the clones was cultured for up to 50 passages. Factor VIII-related antigen and CD34 were detected. The immortalized cells shared the properties of normal cells, such as contact inhibition, serum requirement and anchorage dependence. Karyotype analysis revealed that the immortalized cells were in the diploid range. In addition, both in vivo and in vitro assays of tumorigenicity showed no neoplastic transformation. Furthermore, NO, $PGI_2$, and ET-1 concentrations in the transfected cells were normal. These results suggest that the SUVECs immortalized by hTERT retain their original characteristics.
Liu, Bruce Yao Wen;Chen, Cheng-Lung;Lee, Shin-Yu;Chang, Fu-Hsiung;Lin, Win-Li;Chia, Chih-Ta;Chen, Yang-Yuan
Biomaterials and Biomechanics in Bioengineering
/
v.1
no.1
/
pp.41-50
/
2014
Gold nanorod's exceptional light to heat transduction is a robust phonomenon that has been extensively verified. The phenomenon is a trait from which many novel applications across disciplines have been proposed. In this investigation, the feasibility of utilizing heat harvested from such photothermal method to combat cancer is presented. Using non-invasive laser methods, an in vivo study is conducted on mouse ear tumors administered with gold nanorods (Au NRs). An emphasis is placed on monitoring the tumor developments after photothermal treatments, over time. The findings reveal significant tumor growth surpression at a threshold laser power of $0.6W/cm^2$ lasting 2 minutes; this energy also brought about dramatic size reduction in treated tumors. Furthermore, the apparent formation of an eschar over the laser treated region indicates extensive hemorrhagic necrosis of the tumor tissue; a phenomenon implicative to the inhibition of angiogenesis.
Background: The dysfunction of multiple organs is found to be caused by reactive oxygen species as a major modulator of microvascular injury after hemorrhagic shock. Hemorrhagic shock, one of many causes inducing acute lung injury, is associated with increase in alveolocapillary permeability and characterized by edema, neutrophil infiltration, and hemorrhage in the interstitial and alveolar space. Aggressive and rapid fluid resuscitation potentially might increased the risk of pulmonary dysfunction by the interstitial edema. Therefore, in order to improve the pulmonary dysfunction induced by hemorrhagic shock, the present study was attempted to investigate how to reduce the inflammatory responses and edema in lung. Material and Method: Male Sprague-Dawley rats, weight 300 to 350 gm were anesthetized with ketamine(7 mg/kg) intramuscular Hemorrhagic Shock(HS) was induced by withdrawal of 3 mL/100 g over 10 min. through right jugular vein. Mean arterial pressure was then maintained at $35{\sim}40$ mmHg by further blood withdrawal. At 60 min. after HS, the shed blood and Ringer's solution or 5% albumin was infused to restore mean carotid arterial pressure over 80 mmHg. Rats were divided into three groups according to rectal temperature level($37^{\circ}C$[normothermia] vs $33^{\circ}C$[mild hypothermia]) and resuscitation fluid(lactate Ringer's solution vs 5% albumin solution). Group I consisted of rats with the normothermia and lactate Ringer's solution infusion. Group II consisted of rats with the systemic hypothermia and lactate Ringer's solution infusion. Group III consisted of rats with the systemic hypothermia and 5% albumin solution infusion. Hemodynamic parameters(heart rate, mean carotid arterial pressure), metabolism, and pulmonary tissue damage were observed for 4 hours. Result: In all experimental groups including 6 rats in group I, totally 26 rats were alive in 3rd stage. However, bleeding volume of group I in first stage was $3.2{\pm}0.5$ mL/100 g less than those of group II($3.9{\pm}0.8$ mL/100 g) and group III($4.1{\pm}0.7$ mL/100 g). Fluid volume infused in 2nd stage was $28.6{\pm}6.0$ mL(group I), $20.6{\pm}4.0$ mL(group II) and $14.7{\pm}2.7$ mL(group III), retrospectively in which there was statistically a significance between all groups(p<0.05). Plasma potassium level was markedly elevated in comparison with other groups(II and III), whereas glucose level was obviously reduced in 2nd stage of group I. Level of interleukine-8 in group I was obviously higher than that of group II or III(p<0.05). They were $1.834{\pm}437$ pg/mL(group I), $1,006{\pm}532$ pg/mL(group II), and $764{\pm}302$ pg/mL(group III), retrospectively. In histologic score, the score of group III($1.6{\pm}0.6$) was significantly lower than that of group I($2.8{\pm}1.2$)(p<0.05). Conclusion: In pressure-controlled hemorrhagic shock model, it is suggested that hypothermia might inhibit the direct damage of ischemic tissue through reduction of basic metabolic rate in shock state compared to normothermia. It seems that hypothermia should be benefit to recovery pulmonary function by reducing replaced fluid volume, inhibiting anti-inflammatory agent(IL-8) and leukocyte infiltration in state of ischemia-reperfusion injury. However, if is considered that other changes in pulmonary damage and inflammatory responses might induce by not only kinds of fluid solutions but also hypothermia, and that the detailed evaluation should be study.
Background: The purpose of this study was to develop risk-adjustment models for acute stroke mortality that were based on data from Health Insurance Review and Assessment Service (HIRA) dataset and to evaluate the validity of these models for comparing hospital performance. Methods: We identified prognostic factors of acute stroke mortality through literature review. On the basis of the avaliable data, the following factors was included in risk adjustment models: age, sex, stroke subtype, stroke severity, and comorbid conditions. Survey data in 2014 was used for development and 2012 dataset was analysed for validation. Prediction models of acute stroke mortality by stroke type were developed using logistic regression. Model performance was evaluated using C-statistics, $R^2$ values, and Hosmer-Lemeshow goodness-of-fit statistics. Results: We excluded some of the clinical factors such as mental status, vital sign, and lab finding from risk adjustment model because there is no avaliable data. The ischemic stroke model with age, sex, and stroke severity (categorical) showed good performance (C-statistic=0.881, Hosmer-Lemeshow test p=0.371). The hemorrhagic stroke model with age, sex, stroke subtype, and stroke severity (categorical) also showed good performance (C-statistic=0.867, Hosmer-Lemeshow test p=0.850). Conclusion: Among risk adjustment models we recommend the model including age, sex, stroke severity, and stroke subtype for HIRA assessment. However, this model may be inappropriate for comparing hospital performance due to several methodological weaknesses such as lack of clinical information, variations across hospitals in the coding of comorbidities, inability to discriminate between comorbidity and complication, missing of stroke severity, and small case number of hospitals. Therefore, further studies are needed to enhance the validity of the risk adjustment model of acute stroke mortality.
Shiga toxins (Stxs) are the main virulence factors expressed by the pathogenic Stx-producing bacteria, namely, Shigella dysenteriae serotype 1 and certain Escherichia coli strains. These bacteria cause widespread outbreaks of bloody diarrhea (hemorrhagic colitis) that in severe cases can progress to life-threatening systemic complications, including hemolytic uremic syndrome (HUS) characterized by the acute onset of microangiopathic hemolytic anemia and kidney dysfunction. Shiga toxicosis has a distinct pathogenesis and animal models of Stx-associated HUS have allowed us to investigate this. Since these models will also be useful for developing effective countermeasures to Stx-associated HUS, it is important to have clinically relevant animal models of this disease. Multiple studies over the last few decades have shown that mice injected with purified Stxs develop some of the pathophysiological features seen in HUS patients infected with the Stx-producing bacteria. These features are also efficiently recapitulated in a non-human primate model (baboons). In addition, rats, calves, chicks, piglets, and rabbits have been used as models to study symptoms of HUS that are characteristic of each animal. These models have been very useful for testing hypotheses about how Stx induces HUS and its neurological sequelae. In this review, we describe in detail the current knowledge about the most well-studied in vivo models of Stx-induced HUS; namely, those in mice, piglets, non-human primates, and rabbits. The aim of this review is to show how each human clinical outcome-mimicking animal model can serve as an experimental tool to promote our understanding of Stx-induced pathogenesis.
l. Backgrounds of Studies Cerebrovascular accident. one of the three major causes of death among audults with cardiopathy and malignancy, has been on the increase in korea while it is on the decrease in European countries and Japan. Types of stroke undergo changes caused by prolongment of life expectancy. and social and economic variety. More patients of ischemic stroke show a tendency to increase now than those of hemorrhagic stroke in the past. Many clinical studies on medical cerebrovascular and oriental stroke of paralysis have been published. but few clinical studies on therapeutics of integrated oriental and western medicine are to be found. So I have made an attempt to study clinical observations and therapeutic responses of ischemic stroke under integrated oriental and western medicine. 2.Methods. Among the patients admitted into the clinic of Joong-Poong, Woo-Suk University Hospital from May 1. 1993 until April 30. 1994 those 56 patients who were diagnosed as ischemic stroke on Computed Tomography(CT) and showed no dubious symptom after examination of coagulation and bleeding time were classified into the following six steps and treated: l)diagnosis 2)emergency treatment 3)basic treatment 4)treatment of risk and provoking factors. and preceeding disease 5)complications and conservative therapy 6)rehabilitation. For a period of basic treatment both herb medication and urokinase therapy were applied at the same time. Intravenous injection has been given at a unit of 300.000 dosage a day as urokinase therapy during basic treatment. If they showed any dubious symptom in glucose tolerance test. fructose 500ml and urokinase 300.000 dosage were mixed and injected. In case of no symptom 5% DW 500ml was mixed with urokinase 300.000 unit. and injected at a speed of 15gtt per minute. 3. Results and Conclusions 1) The level of ambulation has been improved from 42.9% when admitted to 73.2% when discharged in the degree of recovery. The level of severe function disorder has been remarkably decreased from 55.4% when admitted to 19.6% when discharged. 2) The treatment effect on the basis of therapeutic response of clinical and subjective symptom shows as follows: 7.1% Excellent. 35.7% Good. 37.5% Effective. 10.7% Stationary. and 8.9% Aggravated. The total recovery above effective shows 80.3%. Judging from the above results I think it proper to develop the model of better preventing and treating ischemic stroke through effective therapeutic and clinical studies of integrated oriental and western medicine.
Park, Jae-Heung;Jang, Kyung-Jun;Kim, Cheol-Hong;Kim, Jung-Hee;Kim, Young-Kyun;Yoon, Hyun-Min
Journal of Pharmacopuncture
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v.18
no.1
/
pp.72-78
/
2015
Objectives: The stomach is a sensitive digestive organ that is susceptible to exogenous pathogens from the diet. In response to such pathogens, the stomach induces oxidative stress, which might be related to the development of both gastric organic disorders such as gastritis, gastric ulcers, and gastric cancer, and functional disorders such as functional dyspepsia. This study was accomplished to investigate the effect of Ganoderma lucidum pharmacopuncture (GLP) on chronic gastric ulcers in rats. Methods: The rats were divided into 4 groups of 8 animals each: the normal, the control, the normal saline (NP) and the GLP groups. In this study, the modified ethanol gastritis model was used. The rats were administrated 56% ethanol orally every other day. The dose of ethanol was 8 g/kg body weight. The normal group received the same amount of normal saline instead of ethanol. The NP and the GLP groups were treated with injection of saline and GLP respectively. The control group received no treatment. Two local acupoints CV12 (中脘) and ST36 (足三里) were used. All laboratory rats underwent treatment for 15 days. On last day, the rats were sacrificed and their stomachs were immediately excised. Results: Ulcers of the gastric mucosa appeared as elongated bands of hemorrhagic lesions parallel to the long axis of the stomach. In the NP and GLP groups, the injuries to the gastric mucosal injuries were not as severe as they were in the control group. Wound healings of the chronic gastric ulcers was promoted by using GLP and significant alterations of the indices in the gastric mucosa were observed. Such protection was demonstrated by gross appearance, histology and immunehistochemistry staining for Bcl-2-associated X (BAX), B-cell lymphoma 2 (Bcl-2) and Transforming growth factor-beta 1 (TGF-${\beta}1$). Conclusion: These results suggest that GLP at CV12 and ST36 can provide significant protection to the gastric mucosa against an ethanol induced chronic gastric ulcer.
Kim Kwang-Taik;Chung Bong-Kyu;Lee Sung-Ho;Cho Jong-Ho;Son Ho-Sung;Fang Young-Ho;Sun Kyung;Park Sung-Min
Journal of Chest Surgery
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v.39
no.7
s.264
/
pp.520-526
/
2006
Background: The clinical application of cryosurgery the management of lung cancer is limited because the response of lung at low temperature is not well understood. The purpose of this study is to investigate the response of the pulmonary tissue at extreme low temperature. Material and Method: After general anesthesia the lungs of twelve Mongrel dogs were exposed through the fifth intercostal space. Cryosurgical probe (Galil Medical, Israel) with diameter 2.4 mm were placed into the lung 20 mm deep and four thermosensors (T1-4) were inserted at 5 mm intervals from the cryoprobe. The animals were divided into group A (n=8) and group B (n=4). In group A the temperature of the cryoprobe was decreased to $-120^{\circ}C$ and maintained for 20 minutes. After 5 minutes of thawing this freezing cycle was repeated. In group B same freezing temperature was maintained for 40 minutes continuously without thawing. The lungs were removed for microscopic examination on f day after the cryosurgery. In four dogs of the group A the lung was removed 7 days after the cryosurgery to examine the delayed changes of the cryoinjured tissue, Result: In group A the temperatures of T1 and T2 were decreased to the $4.1{\pm}11^{\circ}C\;and\;31{\pm}5^{\circ}C$ respectively in first freezing cycle. During the second freezing period the temperatures of the thermosensors were decreased lower than the temperature during the first freezing time: $T1\;-56.4{\pm}9.7^{\circ}C,\;T2\;-18.4{\pm}14.2^{\circ}C,\;T3\;18.5{\pm}9.4^{\circ}C\;and\;T4\;35.9{\pm}2.9^{\circ}C$. Comparing the temperature-distance graph of the first cycle to that of the second cycle revealed the changes of temperature-distance relationship from curve to linear. In group B the temperatures of thermosensors were decreased and maintained throughout the 40 minutes of freezing. On light microscopy, hemorrhagic infarctions of diameter $18.6{\pm}6.4mm$ were found in group A. The infarction size was $14{\pm}3mm$ in group B. No viable cell was found within the infarction area. Conclusion: The conductivity of the lung is changed during the thawing period resulting further decrease in temperature of the lung tissue during the second freezing cycle and expanding the area of cell destruction.
Adult respiratory distress syndrome (ARDS) is of particular interest because of its severity of the associated lung injury and its high mortality. However, the pathophysiologies of ARDS in infant and childhood groups are still not well clarified inspite of many previous investigations. To investigate the time course of pathophysiology of ARDS in infant and childhood groups, this study was designed with experimental endotoxin-induced ARDS model using young rabbits (8 week-old). Material and Method: Rabbits were divided into the control group (n=8) and the endotoxin-treated group (n=32). The endotoxin group was subdivided into 4 groups by the sampling times as 3, 6, 12 and 24 hr-groups (G- $E_{3,6,12,24,}$ each n=8). The experimental ARDS was made by a bolus injection of endotoxin (Escherichia coli serotype 055 : B5, 0.50 mg/kg) via rabbit ear vein. For evaluation of the hematologic and inflammatory markers, and superoxide dismutase (SOD) concentrations, the blood samples were taken from the heart. The bronchoalveolar lavage fluid (BALF) were obtained for analysis of the leukocytes and protein concentration. With biopsy of the lung, histopathologic changes of the lung were also evaluated. Result: In the endotoxin groups, significant leukopenia (owing to pancytopenia) occurred in 3 and 6-hr groups, which was followed by significant leukocytosis (owing to neutrophilia) in the 12 and 24-hr groups (p<0.05). Serum levels of tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and interleukin-1 $\beta$ (IL-1 $\beta$) in the endotoxin groups were higher than those of control group (p<0.05). Serum levels of superoxide dismutase (SOD) of G- $E_{3}$ and G- $E_{6}$ were higher than those of control group, whereas those of G- $E_{12}$ were lower than those of control groups (p<0.05). Total leukocyte counts and protein con-centrations in BALF were significantly elevated in the endotoxin groups compared to the control group (p < 0.05). The hemorrhagic pattern of BALF showed occurred in the endotoxin groups. The endotoxin groups (in G- $E_{6}$) had severe infiltration of inflammatory cells (lymphocyte and monocyte) in the pulmonary interstitium and parenchyma, migrations of neutrophil and eosinophil into alveolar spaces and interstitial widening, which are the evidences of acute lung injury. In the endotoxin groups, there were significant positive correlations between the BALF findings and the immunologic markers (TNF-$\alpha$, IL-1$\beta$, SOD) (p<0.05). Conclusion: Severe acute lung injury occurred in all the endotoxin-treated rabbits. The pathophysiologic findings were so progressive until 6-hr by time dependant pattern, and then recovered slowly, Variable hematologic, immuno-logic, and pathologic factors were well correlated in the development and progression of endoxin-induced lung injury. The pathophysiologic responses were sensitive and rapid in young rabbit Young rabbit seemed to be a useful experimental animal model for infant and childhood groups.roups.
The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.
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