RESUMO
Objective: To investigate the safety and efficacy of retroperitoneal laparoscopic selective renal artery branch occlusion with nephron sparing surgery in patients with renal carcinoma of stage ≥ T1b. Methods: From July 2016 to September 2020, 35 patients with renal cancer ≥T1b underwent retroperitoneoscopic nephron sparing surgery in the First Affiliated Hospital of Shenzhen University. The surgical methods were retroperitoneoscopic nephron sparing surgery with total renal artery occlusion (group A) or selective renal artery branch occlusion (group B). Operation time, heat ischemia time, blood transfusion rate, positive margin rate, intraoperative blood loss, postoperative complications and length of hospital stay were compared between the two groups, and the total glomerular filtration rate (GFR) and the single-nephron glomerular filtration rate (sGFR) of the offected kidneys were compared between the two groups before, 3 months after and 12 months after surgery. Results: Among the 35 patients, 19 were male and 16 were female, aged (55.7±8.4) years and the body mass index is (24.6±3.1) kg/m2. The tumor diameter was (54.7±10.3) mm. The difference was statistically significant of operative time between group A and B [(103.5±14.3) vs (123.2±14.1) min,P=0.003]. There were no significant differences in thermal ischemia time, blood transfusion rate, positive margin, intraoperative blood loss, incidence of postoperative complications and length of hospital stay between the two groups (all P>0.05). The decrease of renal sGFR in the group A was significantly higher than group B at 3 months and 12 months after surgery [(23.1±3.6) vs (29.1±7.1) ml/min;(25.9±4.7) vs (30.7±7.2),both P<0.05]. Conclusion: Retroperitoneal laparoscopic selective renal artery branch occlusion and neon-sparing surgery for patients with ≥ T1b stage renal carcinoma is a safe and effective surgical method, which can well protect the renal function of patients in the early postoperative stage without increasing intraoperative blood loss and postoperative complications.
Assuntos
Carcinoma de Células Renais , Neoplasias Renais , Laparoscopia , Carcinoma de Células Renais/cirurgia , Feminino , Humanos , Neoplasias Renais/cirurgia , Masculino , Nefrectomia , Néfrons , Artéria Renal , Estudos Retrospectivos , Resultado do TratamentoRESUMO
OBJECTIVE: To compare the effects on wound bed of deep burn following eschar excision with different wound management in rabbits. METHODS: Eighteen full-thickness burns models of Japanese white rabbits were established. They were randomly divided into 3 groups of traditional dressing, biological dressing and negative pressure wound therapy (NPWT) (n=6 each), according to the random number table. Eschar excision was performed three days later. The wound bed was observed and wound tissue was harvested for counting the quantity of bacteria, tissue dry wet ratio, measuring the level of tumor necrosis factor alpha (TNF-α), interleukin (IL)-1ß and IL-6, the amount of collagen fibers and the microvessel density instantly and again seven days later. Statistical analyses were performed. RESULTS: The NPWT group was better than other groups by observing the wound bed. The quantity of bacteria of traditional dressing group, biological dressing group and NPWT group at the time point of seven days after escharectomy turned out to be (9.4±1.5)×10(4,) (8.1±2.7)×10(4,) (3.9±0.7)×10(4) cfu/g, the NPWT group was significantly lower than traditional dressing group and biological dressing group (both P<0.05), and all lower than that at the time point of the day when escharectomy was performed (576.9±169.5)×10(4,) (589.9±99.6)×10(4,) (583.0±160.4)×10(4) cfu/g ( all P<0.05). There were no statistically significant differences among three groups at two time points in tissue dry wet ratio (all P>0.05). The IL-6 of biological dressing group was higher than that of traditional dressing group at the time point of seven days after the eschar excision was performed[(94±10) vs (76±8) ng/L, P<0.05]. The amount of collagen fibers of three group at the time point of seven days after escharectomy turned out to be (60±9), (55±12), (77±17). The NPWT group was significantly higher than traditional dressing group and biological dressing group (P<0.05), and all higher than that at the time point of the day when escharectomy was performed[(39±6), (39±11), (38±6)](all P<0.05). The microvessel density of three groups at the time point of seven days after escharectomy turned out to be (42±6), (53±4), (82±10). The NPWT group was higher than that of the other two groups, and biological dressing group was higher than that of traditional dressing group (all P<0.05). The biological dressing group and NPWT group were both higher than that of the day when the eschar excision was performed (36±5) and (36±5) (P<0.05). CONCLUSIONS: NPWT is the optimal selection for wound to inhibit the growth of bacteria, promote the accumulation of collagen and tissue vascularization. But these managements have similar effects on reducing tissue edema and inflammatory reaction.
Assuntos
Queimaduras , Cicatrização , Animais , Inflamação , Tratamento de Ferimentos com Pressão Negativa , CoelhosRESUMO
Acetaminophen (APAP)-induced liver injury remains the main cause of acute liver failure in the United States. Our previous work demonstrated that LPS binding protein (LBP) knockout mice are protected from APAP-induced hepatotoxicity. LBP is known to bind avidly to LPS, facilitating cellular activation. In this study, we sought to specifically inhibit the interaction between LBP and LPS to define the role of this interaction in APAP-induced liver injury. The peptide LBPK95A was able to inhibit LBP-mediated LPS activation of RAW 267.4 cells in a dose-dependent manner in vitro. In vivo, C57Bl/6 mice were treated with either LBPK95A or vehicle control concurrently with the administration of APAP (350 mg/kg). Mice treated with LBPK95A had significantly lower serum aspartate aminotransferase and alanine aminotransferase levels. Morphometric analysis of the liver tissue showed significantly less liver injury in mice treated with LBPK95A. To assess whether the LBPK95A altered glutathione depletion and APAP metabolism, we measured total glutathione levels in the liver after APAP. We found no difference in the glutathione levels and APAP-adduct formation between LBPK95A vs. vehicle control both at baseline and after APAP. In conclusion, our results support the hypothesis that LBP-induced liver injury after APAP is due to its ability to mediate activation by endogenous LPS. Our results suggest that blocking LBP-LPS interactions is a potential therapeutic avenue for the treatment of APAP-induced liver injury.
Assuntos
Acetaminofen/toxicidade , Antídotos/farmacologia , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Peptídeos/farmacologia , Sequência de Aminoácidos , Animais , Antídotos/química , Linhagem Celular , Citocinas/genética , Citocinas/metabolismo , Glutationa/metabolismo , Lipopolissacarídeos/metabolismo , Fígado/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Peptídeos/químicaRESUMO
OBJECTIVE: To explore the effect of LncRNA MEG3 in the subarachnoid hemorrhage (SAH) and its underlying mechanism. PATIENTS AND METHODS: The expressions of lncRNA MEG3 in SAH patients and animal model were detected by quantitative real-time PCR (qRT-PCR). After LncRNA MEG3 was overexpressed in neurons by lentivirus, viability and apoptosis abilities were detected by cell counting kit-8 (CCK-8) assay, flow cytometry, and TUNEL assay, respectively. The apoptosis-related genes and Pi3k/Akt pathway-related proteins were further detected by a Western blot. RESULTS: The expressions of lncRNA MEG3 in SAH patients were remarkably higher than normal controls, which were positively correlated with SAH severity. After lncRNA MEG3 overexpression, neuronal cell activity was decreased and cell apoptosis was increased. Moreover, the expressions of Bax, p53, and cleaved Caspase-3 were increased, whereas the expression of Bcl-2 and Pi3k/Akt pathway-related proteins were decreased after lncRNA MEG3 overexpression. CONCLUSIONS: LncRNA MEG3 is up-regulated in SAH, which may promote SAH-induced neuronal cell injury via inhibition of the Pi3k/Akt pathway.
Assuntos
Neurônios/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Longo não Codificante/biossíntese , Hemorragia Subaracnóidea/metabolismo , Idoso , Animais , Células Cultivadas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neurônios/patologia , Inibidores de Fosfoinositídeo-3 Quinase , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Ratos , Transdução de Sinais/efeitos dos fármacos , Hemorragia Subaracnóidea/patologiaRESUMO
Objective: To explore the influence of different inner dressings in negative-pressure wound therapy (NPWT) on escharectomy wound of full-thickness burn rabbits. Methods: Eighteen Japanese white rabbits were inflicted with full-thickness burn on unilateral back. They were divided into polymer dressing group (PD), biological dressing group (BD), and silver biological dressing group (SBD), according to the random number table, with 6 rabbits in each group. On 3 days post burn, the wounds were performed with escharectomy, and then wounds of rabbits in group PD were covered with polyurethane foam. Wounds of rabbits in group BD were covered with porcine acellular dermal matrix (ADM) and wounds of rabbits in group SBD were covered with silver porcine ADM. Then continuous NPWT was performed on rabbits of the three groups for 7 days. Immediately after surgery and on post surgery day (PSD) 7, general observation of wound was conducted and tissue around the wound was harvested for determination of dry to wet weight ratio. The content of bacteria was counted and the content of tumor necrosis factor α (TNF-α), interleukin-1ß (IL-1ß), and IL-6 in wound was determined by enzyme-linked immunosorbent assay. Fibroblasts in wound were counted after Masson staining and number of microvessels was counted after CD31 antibody immunohistochemical staining. Data were processed with analysis of variance for repeated measurement, LSD-t test, paired samples t test, and Bonferroni correction. Results: (1) Immediately after surgery, there was no granulation tissue in basal wound of rabbits in the three groups, with rich blood supply and obvious edema. On PSD 7, much granulation tissue was found in basal wound of rabbits in the three groups, with no or mild edema and no obvious redness and swelling in wound edge. (2) There were no significant differences in dry to wet weight ratios of tissue around the wound among and within the three groups immediately after surgery and on PSD 7 (with F values respectively 0.70 and 0.09, t values from 0.17 to 0.52, P values above 0.05). (3) Immediately after surgery, the content of bacteria in wounds of rabbits in groups PD, BD, and SBD was respectively (603.0±146.0) ×10(4,) (573.0±63.0) ×10(4,) and (590.0±100.0)×10(4) colony-forming unit (CFU)/g, with no significant difference among them (F=0.13, P>0.05). On PSD 7, the content of bacteria in wounds of rabbits in groups PD, BD, and SBD were respectively (5.4±0.8) ×10(4,) (4.6±0.9) ×10(4,) and (3.5±0.9)×10(4) CFU/g. Among them, the content of bacteria in wounds of rabbits in group SBD was lower than that in groups PD and BD, respectively (with t values respectively 3.78 and 2.29, P<0.05 or P<0.01). The content of bacteria in wounds of rabbits in the three groups on PSD 7 was decreased compared with that immediately after surgery (with t values from 10.05 to 21.81, P values below 0.01). (4) There was no significant difference in content of TNF-α, IL-1ß, and IL-6 in wounds of rabbits in the three groups immediately after surgery and on PSD 7 (with F values from 0.10 to 1.89, P values above 0.05). The content of TNF-α in wounds of rabbits in the three groups on PSD 7 was significantly higher than that immediately after surgery (with t values from 2.93 to 5.01, P<0.05 or P<0.01). (5) There was no significant difference in amount of fibroblasts in wounds of rabbits in the three groups immediately after surgery and on PSD 7 (with F values respectively 0.01 and 0.81, P values above 0.05). The amount of fibroblasts in wounds of rabbits in the three groups on PSD 7 was larger than that immediately after surgery (with t values from 4.78 to 11.58, P values below 0.01). (6) There was no significant difference in number of microvessels in wounds of rabbits in the three groups immediately after surgery and on PSD 7 (with F values respectively 2.42 and 2.49, P values above 0.05). The number of microvessels in wounds of rabbits in the three groups on PSD 7 was larger than that immediately after surgery (with t values from 7.17 to 11.14, P values below 0.01). Conclusions: SBD is better at inhibiting the growth of bacteria. PD, BD, and SBD have almost the same effects on reducing tissue edema and inflammatory reaction, and on promoting the accumulation of collagen fibers and tissue vascularization.
Assuntos
Derme Acelular , Bandagens , Queimaduras/terapia , Tratamento de Ferimentos com Pressão Negativa , Cicatrização , Animais , Curativos Biológicos , Ensaio de Imunoadsorção Enzimática , Tecido de Granulação , Inflamação , Interleucina-1beta , Interleucina-6 , Coelhos , Lesões dos Tecidos Moles , Resultado do Tratamento , Fator de Necrose Tumoral alfaRESUMO
BACKGROUND: Sarcopenia is defined as the loss of skeletal muscle mass and function associated with aging. Muscle mass can be reliably and accurately quantified using clinical CT scans but reference measurements are lacking, particularly in healthy US populations. METHODS: Two-phase CT scans from healthy kidney donors (age 18-40) at the University of Michigan between 1999-2010 were utilized. Muscle mass was quantified using two thoracic and two lumbar muscle cross-sectional area (CSA) measures. Indexed measurements were computed as area divided by height-squared. Paired analyses of non-contrast and contrast phases and different Hounsfield Unit (HU) ranges for muscle were conducted to determine their effect on CSA muscle measures. We report the means, standard deviations, and 2SD sarcopenia cutoffs from this population. RESULTS: Healthy population CSA (cm2) cutoffs for N=604 males/females respectively were: 34.7/20.9 (T12 Dorsal Muscle), 91.5/55.9 (T12 Skeletal Muscle), 141.7/91.2 (L3 Skeletal Muscle), 23.5/14.3 (L4 Total Psoas Area), and 23.4/14.3 (L4 Psoas Muscle Area). Height-indexed CSA (cm2/m2) cutoffs for males/females respectively were: 10.9/7.8 (T12 Dorsal Muscle), 28.7/20.6 (T12 Skeletal Muscle), 44.6/34.0 (L3 Skeletal Muscle), 7.5/5.2 (L4 Total Psoas Area), and 7.4/5.2 (L4 Psoas Muscle Area). We confirmed that a mask of -29 to 150 HU is optimal and shows no significant difference between contrast-enhanced and non-contrast CT scan CSA measurements. CONCLUSIONS: We quantified reference values for lumbar and thoracic muscle CSA measures in a healthy US population. We defined the effect of IV contrast and different HU ranges for muscle. Combined, these results facilitate the extraction of clinically valuable data from the large numbers of existing scans performed for medical indications.
Assuntos
Músculo Esquelético/patologia , Músculos Psoas , Sarcopenia , Tomografia Computadorizada por Raios X/normas , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valores de Referência , Estudos Retrospectivos , Sarcopenia/complicações , Sarcopenia/patologiaRESUMO
Objective: To explore the effects of different fluid resuscitation program on renal function in swine during shock stage of severe burn. Methods: Twenty-four Guangxi Bama miniature swine were inflicted with 40% total body surface area on the back, and then they were divided into four groups according to the random number table, with 6 swine in each group. At post injury hour (PIH) 2, swine in succinylated gelatin group (S), hydroxyethyl starch group (H), and allogeneic plasma group (A) were respectively resuscitated with succinylated gelatin, hydroxyethyl starch 130/0.4, and plasma according to burn shock " domestic general" resuscitation formula, and swine in Parkland group (P) were resuscitated with lactated Ringer's solution according to Parkland formula. Hemodynamic indexes including heart rate, blood pressure, urine volume, pulmonary capillary wedge pressure, and central venous pressure before injury, at the first and second PIH 24 were recorded. The volume of resuscitation fluid was calculated at the first and second PIH 24. Blood and urine samples were collected before injury and at PIH 4, 8, 24, and 48, and then serum creatinine and urea nitrogen were detected by automatic biochemical analyzer, urine microalbumin and urine creatinine were detected by automated urine analyzer and the ratio of which was calculated. The renal tissue of swine in each group was obtained at PIH 48, and the pathologic changes were observed by optical microscopy and electron microscope. Data were processed with analysis of variance of repeated measurement, one-way analysis of variance, and LSD test. Results: (1) The hemodynamic indexes of swine in each group were similar before injury and at the first and second PIH 24 (with P values above 0.05). Compared with those before injury, except that the heart rate of swine in group A had no significant change at the first PIH 24 (P>0.05), the heart rate of swine in each group was significantly increased at the first and second PIH 24 (with P values below 0.01); except that the systolic blood pressure of swine in group P was significantly increased at the first and second PIH 24 (P<0.05 or P<0.01), there were no significant changes of systolic blood pressure and diastolic blood pressure of swine in each group at the first and second PIH 24 (with P values above 0.05); except that urine volume of swine in groups S and A was significantly decreased at the first PIH 24 (P<0.05 or P<0.01), there were no significant change of urine volume of swine in each group at the first and second PIH 24 (with P values above 0.05); pulmonary capillary wedge pressure and central venous pressure of swine in each group were significantly increased at the first and second PIH 24 (P<0.05 or P<0.01). (2) Compared with that in group A, the volume of resuscitation fluid of swine in groups S and H had no significant change in the first and second PIH 24 (with P values above 0.05), while it was significantly increased in group P in the first PIH 24 and significantly decreased in the second PIH 24 (with P values below 0.05). (3) Compared with those in group A, except that serum creatinine of swine in group H was significantly increased at PIH 24 and significantly increased in group P at PIH 4, 8, 24, and 48, urea nitrogen of swine in group P was significantly decreased at PIH 4 and 8 and significantly increased at PIH 48, the ratio of urine microalbumin to urine creatinine of swine in group P was significantly increased at PIH 8, 24, and 48 (P<0.05 or P<0.01), serum creatinine, urea nitrogen, and the ratio of urine microalbumin to urine creatinine of swine in each group had no significant change at each time point (with P values above 0.05). Serum creatinine of swine in group P was (125±16) µmol/L at PIH 24, which was significantly higher than that before injury [(75±13) µmol/L, P<0.05]. Urea nitrogen of swine in group S was (2.90±1.17) µmol/L at PIH 48, which was significantly lower than that before injury [(4.60±0.47) µmol/L, P<0.05]; urea nitrogen of swine in group H was (4.82±0.82) µmol/L at PIH 4, which was significantly higher than that before injury [(3.80±0.73) µmol/L, P<0.05]; urea nitrogen values of swine in group A were respectively (4.80±0.33), (4.92±0.35), and (2.60±0.27) µmol/L at PIH 4, 8, and 48, while those at PIH 4, 8 were significantly higher and at PIH 48 was significantly lower than the value before injury [(3.93±0.32) µmol/L, with P values below 0.01]. The ratios of urine microalbumin to urine creatinine of swine in group P were respectively (106.7±16.4) and (171.6±36.9) mg/mmol at PIH 24 and 48, which were significantly higher than the ratio before injury [(59.0±3.0) mg/mmol, with P values below 0.01]. The serum creatinine, urea nitrogen, and the ratio of urine microalbumin to urine creatinine of swine in each group at the other time points were similar to those before injury (with P values above 0.05). (4) The renal tissue of swine in the four groups had no obvious pathological change. Conclusions: According to the renal function results, fluid resuscitation with electrolyte and colloids are better than with lactated Ringer's solution in swine during shock stage of burn injury, while natural colloids and succinylated gelatin have similar effects, and both are superior to hydroxyethyl starch 130/0.4.
Assuntos
Queimaduras/complicações , Hidratação , Choque , Animais , Pressão Sanguínea , Superfície Corporal , Coloides , Derivados de Hidroxietil Amido , Soluções Isotônicas , Ressuscitação , Lactato de Ringer , SuínosRESUMO
Lipopolysaccharide binding protein (LBP) is a plasma protein that plays an important intermediary role in host-endotoxin (LPS) interactions. LBP binds with high affinity to the lipid A portion of LPS and then interacts with the monocytic differentiation antigen CD14 to markedly up-regulate TNF-alpha production by mononuclear phagocytes. In the presence of LBP, 100-fold less LPS is required to trigger this cytokine response. LBP has been implicated in the interaction of LPS with both CD14+ (monocytes, macrophages, neutrophils) and CD14-cells (endothelial and epithelial cells) to promote such varying responses as secretion of cytokines and nitric oxide (NO), expression of adhesion molecules, production of tissue factor, and activation of neutrophils. Non-CD 14-bearing cells, such as endothelial cells, can also respond to LPS through this pathway via the soluble form of CD14, an interaction that is similarly enhanced by LBP. Recently, it has been proposed that LBP cannot only potentiate host responses to LPS but can also facilitate the neutralization of LPS under certain conditions. LBP has been described as acting as a lipoprotein transfer protein facilitating the transfer of LPS both to the target receptor (CD14) and lipoprotein (HDL).
Assuntos
Proteínas de Fase Aguda/farmacologia , Proteínas de Transporte/farmacologia , Lipopolissacarídeos/farmacologia , Ativação de Macrófagos/efeitos dos fármacos , Glicoproteínas de Membrana , Ativação de Neutrófilo/efeitos dos fármacos , Animais , HumanosRESUMO
Chronic rejection remains a major cause of late graft dysfunction. Although much research has focused on acute rejection, little is known about the mechanisms of chronic rejection. Our group has recently reported evidence of significant intestinal smooth muscle hypertrophy and hyperplasia associated with abnormal contractile and electrical activities in a rat model of chronic intestinal rejection. The changes in the smooth muscle layer are associated with a significant inflammatory infiltrate. In order to further delineate the immune mechanisms of chronic rejection, we sought to clarify the nature of this infiltrate. Orthotopic small bowel transplantation was performed using an allogeneic (ACI-Lewis) rat combination. The rats only received immunosuppression for the first 28 days posttransplantation (cyclosporine 15 mg/kg daily from postoperative day 0 to 6 and every other day from postoperative day 7 to 28). This led to chronic rejection of the graft by day 90, at which time the rats were sacrificed. Analysis by immunohistochemistry revealed NK and CD5+ leukocytes infiltrating the muscular layer. Examination of cytokine production by radiolabeled polymerase chain reaction showed high levels of steady state interferon-gamma mRNA in full thickness intestinal segments and within the isolated muscularis of chronically rejecting intestinal allografts as compared to syngeneic and control grafts. Interferon-gamma mRNA was localized to both the muscularis and mucosa. Interestingly, positively hybridized cells within the muscularis tended to preferentially localize to the myenteric and submucosal plexuses suggesting potential role for this cytokine in chronic intestinal ejection.
Assuntos
Rejeição de Enxerto/metabolismo , Interferon gama/biossíntese , Intestino Delgado/transplante , Animais , Sequência de Bases , Divisão Celular/fisiologia , Células Cultivadas , Doença Crônica , DNA Complementar/genética , Modelos Animais de Doenças , Imuno-Histoquímica , Hibridização In Situ , Interferon gama/genética , Interferon gama/farmacologia , Intestino Delgado/imunologia , Intestino Delgado/metabolismo , Jejuno/metabolismo , Masculino , Dados de Sequência Molecular , Músculo Liso/efeitos dos fármacos , Músculo Liso/fisiologia , Reação em Cadeia da Polimerase/métodos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos ACI , Ratos Endogâmicos Lew , Transcrição GênicaRESUMO
Skin is an especially attractive target for genetic manipulation because it is readily accessible and easily monitored for both the presence and the expression of inserted genes. This study was designed to assess the feasibility of particle mediated gene transfer to burned skin and to compare the transfection efficiency, anatomic distribution, and duration of transgene expression achievable in normal versus burned skin. Two days following scald injury of varying depths in 60 degrees C water (10 s: superficial partial; 20 s: deep partial; 40 s: full thickness) reporter gene (beta-galactosidase) constructs were delivered using a gene gun at various helium pressures (200-600 psi) to normal and burned skin. A time course study was performed to examine the kinetics of transgene expression. Animals received a superficial partial thickness burn and were sacrificed 12 h, 1, 3, 5, 7, 14, or 21 days after gene transfer. India Ink injection and immunohistochemistry were used to assess the depth of the scald injury. Transfection efficiency was measured in skin homogenates 24 h after gene transfer by morphometric and chemoluminescent assays. We found that the extent of tissue damage was directly related to the duration of heat source exposure. Reporter gene activity was significantly higher in superficial partial thickness burns compared to normal controls and gradually declined with increasing tissue injury. No activity was seen in the full thickness burn group. Beta-galactosidase activity reached a maximum level 12 h after gene transfer in both normal and superficial partial thickness burned skin with no levels seen after 5 days post-transfection. These findings indicate that particle-mediated gene transfer in thermally injured skin is feasible and may provide a means of introducing biologic agents into injured tissue capable of enhancing bacterial clearance and improving wound healing.
Assuntos
Biolística , Queimaduras/terapia , Terapia Genética , Animais , Queimaduras/patologia , DNA Recombinante/administração & dosagem , Estudos de Viabilidade , Expressão Gênica , Genes Reporter , Vetores Genéticos/administração & dosagem , Ouro , Óperon Lac , Medições Luminescentes , Masculino , Microesferas , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes de Fusão/análise , Organismos Livres de Patógenos Específicos , Transfecção , Transgenes , beta-Galactosidase/análiseRESUMO
BACKGROUND: Many of the physiologic derangements resulting in septic shock are caused by inflammatory mediators such as nitric oxide (NO) and cytokines produced in response to bacterial endotoxin or, more specifically, lipopolysaccharide. The recent development of a novel class of lipopolysaccharide antagonists offers the opportunity to block this response selectively. In this article we investigated the ability of one of these antagonists, B464 (Eisai), to block lipopolysaccharide-induced release of macrophage NO and cytokines. METHODS: The mouse macrophage cell line RAW264.7 was grown in vitro and exposed to (1) media control, (2) B464 alone, (3) lipopolysaccharide alone, or (4) lipopolysaccharide plus graded concentrations of B464. Supernatants were assayed for nitrite plus nitrate, the stable end products of NO, as well as tumor necrosis factor-alpha and interleukin-6. Total cellular RNA was examined for inducible NO synthase and interleukin-6 mRNA. RESULTS: Lipopolysaccharide-stimulated increases in NO, tumor necrosis factor, and interleukin-6 production were blocked by B464. Reduction of NO was also seen at the level of inducible NO synthase mRNA. Induction of interleukin-6 mRNA was also suppressed. CONCLUSION: B464 is a novel potent specific antagonist of lipopolysaccharide-induced macrophage NO and cytokine production.
Assuntos
Interleucina-6/biossíntese , Lipídeo A/análogos & derivados , Lipopolissacarídeos/antagonistas & inibidores , Óxido Nítrico/biossíntese , Fator de Necrose Tumoral alfa/biossíntese , Aminoácido Oxirredutases/genética , Animais , Sequência de Carboidratos , Linhagem Celular , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Interleucina-6/genética , Lipídeo A/análise , Macrófagos/metabolismo , Camundongos , Dados de Sequência Molecular , Óxido Nítrico Sintase , RNA Mensageiro/análiseRESUMO
Lipopolysaccharide binding protein (LBP) is a serum glycoprotein that complexes with lipopolysaccharide (LPS) to facilitate macrophage response to endotoxin. To determine the conditions that stimulate LBP production in vivo, we measured the induction of LBP in models of inflammation produced by LPS, Corynebacterium parvum, and turpentine injection. Plasma aspartate aminotransferase and alanine aminotransferase concentrations and hepatocyte fibrinogen synthesis were elevated in all models. Northern blot analysis revealed 17-, 14-, and 20-fold upregulation of hepatocyte LBP mRNA following treatment with LPS, C parvum, and turpentine, respectively. Peritoneal macrophage interleukin 6 and tumor necrosis factor production following endotoxin stimulation was augmented by cultured hepatocyte supernatants, suggesting increased LBP synthesis in these groups. The results show that LBP mRNA is induced during hepatic inflammation and suggest that LBP is an acute-phase protein important in regulating the in vivo response to endotoxin.
Assuntos
Proteínas de Fase Aguda , Bacteriemia/imunologia , Proteínas de Transporte/biossíntese , Hepatopatias/imunologia , Macrófagos/efeitos dos fármacos , Glicoproteínas de Membrana , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Bacteriemia/sangue , Bioensaio , Northern Blotting , Proteínas de Transporte/imunologia , Modelos Animais de Doenças , Endotoxinas/imunologia , Estudos de Avaliação como Assunto , Fibrinogênio/análise , Inflamação , Interleucina-6/biossíntese , Interleucina-6/química , Interleucina-6/imunologia , Hepatopatias/sangue , Hepatopatias/patologia , Macrófagos/química , Macrófagos/imunologia , Masculino , Peritônio/citologia , Sondas RNA , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/química , Fator de Necrose Tumoral alfa/imunologiaRESUMO
In response to a burn injury, skin can have an inflammatory response characterized by the production of inflammatory cytokines, recruitment of immune cells, containment of invading organisms, and clearance of noxious substances from the wound. Lipopolysaccharide-binding protein (LBP) is a molecule that is capable of coordinating all 4 functions; we previously found evidence that suggested that LBP is produced within surgical wounds. Because of the central role of LBP in the response to bacterial infection, as well as in the high rate of infection after burn injuries, we sought to determine whether a thermal injury could affect wound LBP production and thereby affect host responses against bacterial infection. Rats were given either a burn or a sham burn and were killed 24, 48, and 72 hours after the injuries. Wound specimens were assayed for bacterial counts and for the presence of LBP, messenger (m)RNA, and interleukin (IL)-1beta mRNA. Wound LBP mRNA was significantly upregulated at 24 hours in the group with burn injuries (P < .05; burn vs sham burn); this was followed by decreases at 48 and 72 hours. Immunohistochemistry showed LBP protein in the epidermis of animals with burns. Bacterial counts increased in the group with burn injuries (P < .05; burn vs sham burn) and continued to rise for 72 hours. IL-1beta mRNA levels were elevated at all time points in the group with burn injuries (P < .05). These results suggest an inverse correlation between burn wound LBP expression and bacterial wound counts. This failure to maintain local LBP production after severe thermal injury despite localized inflammation shown by high IL-1beta levels may predispose local wounds to bacterial invasion.
Assuntos
Proteínas de Fase Aguda/biossíntese , Queimaduras/metabolismo , Proteínas de Transporte/biossíntese , Interleucina-1/biossíntese , Glicoproteínas de Membrana , Pele/metabolismo , Animais , Infecções Bacterianas/imunologia , Proteínas de Transporte/genética , Contagem de Colônia Microbiana , Imuno-Histoquímica , Interleucina-1/genética , Masculino , RNA Mensageiro/análise , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Pele/lesões , Fatores de Tempo , Regulação para Cima , Infecção dos Ferimentos/imunologiaAssuntos
Doenças Ósseas/cirurgia , Fêmur/cirurgia , Prótese de Quadril , Próteses e Implantes , Adulto , Feminino , Humanos , MasculinoAssuntos
Escoliose/cirurgia , Adolescente , Adulto , Feminino , Humanos , Masculino , Dispositivos de Fixação Ortopédica , TraçãoRESUMO
The binding energy spectra and momentum distributions of all valence orbitals of propene were studied by electron momentum spectroscopy (EMS) as well as Hartree-Fock and density functional theoretical calculations. The experiment was carried out at impact energies of 1200 eV and 600 eV on the state-of-the-art EMS spectrometer developed at Tsinghua University recently. The experimental momentum profiles of the valence orbitals were obtained and compared with the various theoretical calculations. Moreover, the experiment with a new analysis method presents a strong support for the correct ordering of the orbital 8a' and 1a'', i.e., 9a' < 8a' < 1a'' < 7a'.
RESUMO
The electron binding energy spectra and momentum profiles of the valence orbitals of difluoromethane, also known as HFC32 (HFC-hydrofluorocarbon) (CH(2)F(2)), have been studied by using a high resolution (e,2e) electron momentum spectrometer, at an impact energy of 1200 eV plus the binding energy, and by using symmetric noncoplanar kinematics. The experimental momentum profiles of the outer valence orbitals and 4a(1) inner valence orbital are compared with the theoretical momentum distributions calculated using Hartree-Fock and density functional theory (DFT) methods with various basis sets. In general, the shapes of the experimental momentum distributions are well described by both the Hartree-Fock and DFT calculations when large and diffuse basis sets are used. However, the result also shows that it is hard to choose the different calculations for some orbitals, including the methods and the size of the basis sets employed. The pole strength of the ionization peak from the 4a(1) inner valence orbital is estimated.
RESUMO
We report here the direct measurements of electron momentum distributions for ethylene using the (e,2e) reaction at different impact energies from 400 to 2400 eV. The "turn up" effects in the (e,2e) cross sections of the 1b(3g) orbital compared with the plane-wave impulse approximation calculations were observed at low and high momentum regions, and such discrepancies become smaller with the increase of the impact electron energies. It is suggested that the observed discrepancies are due to the distorted-wave effects in molecules, while appropriate theoretical calculations using distorted waves in molecules could not be achieved until now.