Influência de um método de treinamento físico militar sobre marcadores indiretos de dano muscular / Influence of a military physical training method on indirect markers of muscle damage

O Exército Brasileiro (EB) desenvolveu recentemente o método de treinamento físico militar Cross Operacional (CO), caracterizado por exercícios combinados aeróbios e resistidos de moderada/alta intensidade. O caráter de moderada/alta intensidade do CO pode levar ao comprometimento das fibras musculares, aumentando o risco de lesão e exacerbação de uma resposta inflamatória aguda. Nesse contexto, a análise de marcadores indiretos de dano muscular pode ser utilizada para avaliação do nível de intensidade do CO e recuperação pós-exercício. O objetivo desse estudo foi observar o efeito agudo do CO sobre os marcadores indiretos de dano muscular em militares do EB. Vinte e quatro militares voluntários, do sexo masculino, com idade média de 20,8 ± 1,8 anos participaram desse estudo. As quatro sessões correspondentes aos níveis do CO foram executadas conforme delineamento cruzado, com washout de sete dias, e as amostras sanguíneas foram coletadas no repouso, imediatamente após, 24 e 48 horas após as sessões de CO. Os marcadores dosados nas amostras foram creatinoquinase (CK), lactato desidrogenase (LDH), aspartato aminotransferase (AST), mioglobina (Mb), lactato (Lac), proteína C reativa (PCR) e haptoglobina (Hp). Em todos os níveis do CO, a CK teve um aumento significativo após 24 horas e o Lac, a LDH e a Mb no momento imediatamente após o CO. Já a AST teve aumento significativo nos níveis 2, 3 e 4, com pico de concentração após 24 horas. Os níveis séricos de PCR aumentaram apenas no momento 24 horas do nível 4 e a Hp após 48 horas do nível 3 do CO. O presente estudo demonstrou que o CO foi capaz de alterar os marcadores indiretos de dano muscular. Todavia, após 48 horas de repouso, os marcadores indiretos de dano muscular apresentaram redução, indicando um processo de recuperação. Apesar das elevações nos marcadores musculares indicadores de dano, a ausência de alterações conclusivas nos níveis séricos das proteínas inflamatórias de fase aguda sugere que o CO pode não ter uma duração ou intensidade suficiente para induzir uma resposta inflamatória sistêmica substancial

The Brazilian Army (EB) recently developed a military physical training method called Cross Operational (CO), which contains combined aerobic and resistance exercises at moderate / high intensity. The moderate / high intensity character of CO can cause impairment of muscle fibers, increasing risk of injury and exacerbation of an acute inflammatory response. In this context, analysis of indirect markers of muscle damage can be used to assess the level of intensity of CO and post-exercise recovery. The aim of this study was to observe the acute effect of CO on indirect markers of muscle damage in military personnel of EB. Twenty-four volunteers, military, male, 20,8 ± 1,8 years old participated in this study. The four sessions corresponding to the CO levels were performed according to a crossover design, with a seven-day washout and the blood samples were collected at rest, after 24 hours and 48 hours after CO sessions. The markers measured in the samples were creatinekinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), myoglobin (Mb), lactate (Lac), C-reactive protein (PCR) and haptoglobin (Hp). At all levels of CO, CK had a significant increase after 24 hours and Lac, LDH and Mb increased at the moment immediately after CO. AST had a significant increase in levels 2, 3 and 4 and a peak of concentration after 24 hours. The PCR levels increased only at the 24-hour moment after level 4 and Hp after 48 hours of level 3 of CO. This study demonstrates that CO was able to change the indirect markers of muscle damage. However, after 48 hours of rest, indirect markers of muscle damage were reduced, indicating a recovery process. Despite the elevations in muscle markers indicating damage, the absence of conclusive changes in the levels of the acute phase inflammatory proteins suggests that CO may not have sufficient duration or intensity to induce a substantial systemic inflammatory response

Effects of two nanotopographies of ultraviolet-treated titanium implant surface on macrophage behaviour and inflammatory cytokines secretion / 中华口腔医学杂志

Objective@#To investigate the effects of two nanotopographies of ultraviolet (UV)-treated titanium surface on macrophage biological behaviour and inflammatory cytokines secretion, and to provide basis for clinical application of UV-treatment in dental implant modification.@*Methods@#Titanium disks were allocated into two groups. Samples in one group were acid-etched in hydrofluoric acid (Acid Ti group), and those in the other group were acid-etched and anodized (Anodization group) to form two nanotopographies respectively. The surface morphology was evaluated by field-emission scanning electron microscopy (FE-SEM). The samples were stored in the dark for 8 weeks. Thirteen samples from each group were exposed to UV-irradiation for 48 h (Acid Ti+UV group and Anodization+UV group), UV-untreated samples from Acid Ti and Anodization groups served as control. Hydrophilicity of samples was measured using contact angle measuring device. After 4, 24 and 72 h of incubation, macrophage cell adhesion and proliferation were conducted using cell counting kit-8. Cytokine/chemokine secretions [tumor necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1) and macrophage inflammatory protein-1α (MIP-1α)] were measured from cell culture supernatants at 24 and 72 h using magnetic luminex assay. Cell morphology was examined using FE-SEM after 2 h of incubation.@*Results@#Micropitted/nanopillar and micropitted/nanotubular topographies were observed in Acid Ti group and Anodization group respectively. Contact angles in Acid Ti+UV and Anodization+UV groups (20.2°±2.8° and 0.0°±0.0°) were significantly smaller than those in the Acid Ti and Anodization groups (P<0.05). Cell adhesion and proliferation in all groups at 4 and 24 h showed no difference (P>0.05). Cell proliferation in Acid Ti+UV and Anodization+UV groups at 72 h were (0.92±0.13) and (1.10±0.08) respectively, which were significantly higher than those in Acid Ti and Anodization groups. TNF-α concentration in Acid Ti+UV and Anodization+UV groups at 72 h were (1.03±0.11) and (0.87±0.10) ng/L, MCP-1 were (301.7±50.3) and (240.8±18.7) ng/L, MIP-1α were (224.9±30.6) and (233.9±14.9) ng/L respectively, which were significantly lower than those in Acid Ti and Anodization groups (P<0.05).@*Conclusions@#UV treatment can increase hydrophilicity of two titanium surface topographies, especially of Anodization+UV group. UV-treated titanium surfaces can promote macrophage proliferation and reduce the inflammatory response in vitro.

Relação entre adiposidade, perfil energético, proteínas inflamatórias e lesões osteoarticulares em equinos jovens sobre diferentes sistemas de criação / Relationship between adiposity, energy profile, inflammatory proteins and osteoarticular lesions in young horses on different rearing systems

Os atuais sistemas de criação de equinos estão associados às dietas ricas em carboidratos que resultam em sobre peso e acúmulo de gordura em animais ainda muito jovens. Nesses animais com sobre peso submetidos exercício físico intenso há aumento na incidência de osteoartrite juvenil e outras afecções inflamatórias. O objetivo deste estudo foi associar a adiposidade corporal e a forma de criação de potros com o perfil energético sanguíneo, as concentrações sanguíneas de proteínas inflamatórias e lesões osteoarticulares na região társica comparando animais criados em regimes intensivo ou extensivo. Foram avaliados 40 potros com 18 meses de idade da raça crioula, 23 fêmeas e 17 machos, sendo 20 animais criados exclusivamente em sistema extensivo e 20 animais criados em sistema intensivo. Foram efetuadas coletas de sangue para avaliação bioquímica e eletroforese proteica. Foram efetuadas através de ultrassonografia a mensuração da gordura na crista do pescoço, região retroperitoneal e na base da cauda. Em 17 animais do grupo intensivo e nove animais do grupo extensivo foi efetuado o estudo radiográfico da região do tarso esquerdo. Foi observado maiores níveis de colesterol total e LDL, glicemia, Amilóide A sérica (SAA), transferrina, haptoglobina, ceruloplasmina, glicoproteína ácida e uma proteína de 23Kda de peso molecular (não identificada) nos animais do grupo intensivo com relação aos do grupo extensivo. O grupo intensivo também apresentou maior depósito de gordura na região da crista do pescoço, região retroperitoneal e base da cauda. Em 100% dos animais do grupo intensivo foram observados lesões compatíveis com osteoartrite juvenil, enquanto que em apenas 23% dos animais do grupo extensivo apresentaram tais alterações. No teste exato de Fisher foi observado que os animais do grupo intensivo apresentaram 105% mais chance de desenvolver osteoartrite que os animais do grupo extensivo. Ainda, no teste de Pearson foi observada correlação positiva entre a gordura na crista do pescoço com o grau de comprometimento articular dos potros. A gordura na crista do pescoço apresentou correlação positiva com as alterações osteoarticulares, com os níveis séricos de colesterol LDL, níveis séricos de glicose, níveis de glicoproteína ácida, haptoglobina, transferrina e SAA. A SAA apresentou correlação com a espessura de gordura retroperitoneal. Conclui-se que o depósito de gordura na crista do pescoço apresenta correlação com as alterações no perfil energético, inflamatório e no comprometimento osteoarticular dos animais avaliados. Os níveis de glicose, colesterol LDL, glicoproteína ácida, haptoglobina, ceruloplasmina, transferrina e SAA estiveram correlacionados ao depósito de gordura na crista do pescoço. Potros em sistema intensivo apresentam, em relação aos criados em sistema extensivo, 105 % mais chance de apresentarem lesões articulares degenerativas crônicas compatíveis com osteoartrite juvenil.(AU)

High carbohydrate diets are increasingly used in horse rearing systems. This can result in weight gain and fat accumulation in young horses. There is a growing incidence of juvenile osteoarthritis and other inflammatory conditions in overweight young horses that undergo intense physical exercise. The aim of this study was to associate corporal adiposity with energy profile, serum concentration of acute phase proteins and presence of osteo-articular lesion in the tarsal region of young horses raised in two different rearing systems: intensive system and extensive system. We evaluated 40 young horses 18 months old, 23 of them were females and 17 were males. Twenty horses were raised in the extensive rearing system and twenty horses were raised in intensive rearing system. Blood samples were collected for biochemical analysis and protein electrophoresis. Fat deposition on the crest of the neck, peritoneum and tailhead was measured by ultrasonography. Radiographic examination of the left tarsus was performed in 17 horses of the farm rearing system and in nine horses of the extensive rearing system. We observed higher levels of total cholesterol, LDL, glucose, serum amyloid A (SAA), transferrin, haptoglobin, acid glycoprotein and unidentified protein 23Kda in horses of the intensive system. These horses also showed higher fat deposition on the crest of the neck, peritoneum and tailhead than horses raised on extensive system. All horses on the intensive system group that underwent radiographic examination had lesions compatible with juvenile osteoarthritis while only 23% of the animals of the extensive system group showed such changes. With Fisher's exact test we observed that horses of the intensive rearing system are 105% more likely to develop osteoarthritis than horses of the extensive rearing system. With the Pearson correlation test we found a positive correlation between fat deposition on the crest of the neck and degree of articular injury. Fat deposition on the crest of the neck also showed a positive correlation with serum levels of LDL, glucose, acid glycoprotein, haptoglobin, transferrin and SAA. The SAA correlated with the thickness of retroperitoneal fat. There was a positive correlation between retroperitoneal fat deposition and presence of osteoarticular abnormalities. In conclusion, fat deposition on the crest of the neck has a correlation with energetic profile changes, cute phase proteins changes and with articular injuries. Levels of glucose, LDL cholesterol, acid glycoprotein, haptoglobin, ceruloplasmin, transferrin and SAA have a correlation with fat deposition on the crest of the neck. In addition, young horses of the intensive rearing system are 105% more likely to have chronic degenerative joint lesions compatible with juvenile osteoarthritis than horses of the extensive rearing system.(AU)

Study on detection biomarkers of Alzheimer's disease based on plasma immune and inflammatory proteins / 上海交通大学学报（医学版）

Objective·To explore plasma immune and inflammatory proteins that could serve as potential screening markers for Alzheimer's disease (AD).Methods·Healthy controls (n=19) and AD patients (n=19) were enrolled.Plasma samples were collected and 70 kinds of immune and inflammatory proteins were detected.The immune and inflammatory proteins associated with AD were screened by Mann-Whitney U test and partial correlation analysis.Discriminant analysis was used to develop multi-protein combined algorithm to distinguish plasma samples of AD patients from those of healthy controls.Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficacy for the multi-protein combined algorithm.Results·Among the 70 proteins analyzed,23 were significantly higher in AD patients (P＜0.05),among which 19 were strongly correlated with AD (P＜0.05).These 19 proteins were analyzed with Wilks' lambda stepwise analysis to develop discriminant algorithm for detecting plasma samples of AD.Finally,the discriminant algorithm established by 11 plasma immune and inflammatory proteins (EGF,GRO,MDC,MCP-1,MCP-2,MCP-4,TARC,SCF,TRAIL,CTACK,GCP-2) was found to have an optimal diagnostic efficacy (AUC=0.994).The optimal cutoff value of the algorithm was-0.609.When the optimal cutoff value was obtained,the sensitivity of the equation could reach 100％ and the specificity could reach 94.7％.Conclusion·The discriminant equation composed of the above 11 plasma immune and inflammatory proteins has the potential to assist AD screening.

Bone marrow mesenchymal stem cells regulate nuclear factor kappaB expression in alveolar macrophages of acute lung injury rats with sepsis / 中国组织工程研究

BACKGROUND:Bone marrow mesenchymal stem cels have a therapeutic effect on acute lung injury, but the mechanism is unclear. If the mechanism is understood, the majority of patients with acute lung injury can obtain a benefit. OBJECTIVE:To explore the possible mechanism underlying bone marrow mesenchymal stem cels in the treatment of acute lung injury with sepsis in rats. METHODS: (1) Thirty-six adult Wistar rats were randomly divided into three groups, sham operation group (sham group), sepsis group and bone marrow mesenchymal stem cels group (cel treatment group). In the sepsis and cel treatment groups, animal models of sepsis with acute lung injury were established by cecal ligation and puncture, while in the sham group, the cecum was not ligated and punctured. Then, 1 mL normal saline was injected via the femoral vein in the sepsis and sham groups, and 1 mL bone marrow mesenchymal stem cel suspension (1×109/L) was injected into the cel treatment group. After 6 hours, interleukin 10 and macrophage inflammatory protein-2 levels in serum were measured in the three groups. Lung tissues were taken for pathological observation using hematoxylin-eosin staining. (2) Rat alveolar macrophages were obtained by bronchoalveolar lavage, seeded into 24-wel culture plates, and divided into three groups: control group (group A), sepsis model group (group B) and intervention group of bone marrow mesenchymal stem cels (group C). Normal saline, septic plasma, and co-intervention of septic plasma and mesenchymal stem cels were used in the groups A, B, C, respectively. Then, cels in the three groups were cultured in a 5% CO2 incubator at 37℃ for 1 hour. After that, alveolar macrophages were taken to detect whether nuclear factor-κB (P65) protein entered into the nucleus using laser scanning confocal microscopy. RESULTS AND CONCLUSION: (1) The results of animal experiments showed that compared with the sham group, the macrophage inflammatory protein-2 levels in the sepsis group and cel treatment group were significantly increased (P 0.05); inflammatory cel infiltration, interstitial pulmonary edema and pulmonary hemorrhage existed in the sepsis and cel treatment groups, but these symptoms were significantly reduced in the cel treatment group compared with the sepsis group. (2) Results from cel experiments showed that compared with the group A, in group B and group C, the number of nuclear factor-κB (P65) proteins into the nucleus was significantly higher (P < 0.05), but it was lower in the group C than the group B (P < 0.05). These findings indicate that bone marrow mesenchymal stem cels in acute lung injury with sepsis can regulate nuclear factor-κB (P65) protein of alveolar macrophages into the nucleus, reduce expression of macrophage inflammatory protein-2, and thereby play a protective role in the lungvia reducing neutrophil infiltration. Temporarily, this study cannot explain whether bone marrow mesenchymal stem cels have an effect on interleukin 10.

Evaluation of rat plasma proteins after bothropic venom inoculation and treatment with Mesenquimal stem cells

The aim of this work was to evaluate the mesenchymal stem cells treatment of rats with myonecrosis caused by Rhinocerophis alternatus venom through acute phase proteins (APP) profile. The animals were distributed into three experimental groups (G1, G2 and G3). G1 and G2 were inoculated with 120 μg of R. alternatus venom diluted in 200 µL of ultra-pure water in gastrocnemic muscle, while G3 received 200 µL of ultra-pure water. Three days after, G1 was treated with 5 X 10(6) MSC diluted in PBS and G2 and G3 only with PBS. Each three days after the treatments (3rd, 6th, 9th, 12th 15th days), blood of five animals in each group was collected in order to evaluate the APP. A decrease (P<0.05) in α2-globulin fraction was observed in G1 on the 6th day. In G1 and G2, a raise (P<0.05) was observed in β globulin, a common occurrence in the late phases of inflammatory process, although no significant difference was observed between them. Concerning gamma globulins levels, on the 6th day after the treatments, in G1 and G2 groups, increase in the levels was observed. These data showed that the MSC treatment after bothropic envenomation in the rats caused alteration in APP.

Peptidoglycan modulation of chemokine production in normal human epidermal kerafinocytes and the role of Toll-like receptor 2 in this process / 中华皮肤科杂志

Objective To investigate the effect ofpeptidoglycan from Staphylococcus aureus on the release of several chemokines including intedeukin 8 (IL-8), regulated upon activation, normal T cell expressed and secreted (RANTES), macrophage-derived chemokine (MDC) by normal human epidermal keratinocytes (KCs) and the role of Toll-like receptor 2 (TLR2) in this process. Methods KCs were derived from the foreskin of a healthy boy and propagated. After 2 - 4 passages, KCs were collected and treated with various concentrations (3, 10, 30 and 100 mg/L) of peptidoglycan for 24 hours or with peptidoglycan of 100 mg/L for varying durations (3, 6, 12, 36 hours). A fi'action of KCs were pretreated with functional grade purified anti-TLR2 monoclonal antibody before the treatment with peptidoglycan of 100 mg/L. After additional 12-hour culture following the treatment, enzyme linked immunosorbent assay was used to detect the level of IL-8, RANTES and MDC in culture supernatants of KCs. Results KCs spontaneously released IL-8 and RANTES. Peptidoglycan increased the production of IL-8 but decreased that of RANTES by KCs. The levels of IL-8 were 209.96 ± 10.31 ng/L, 250.28 ± 9.52 ng/L, 285.11 ± 10.28 ng/L, 359.40 ± 6.93 ng/L in KCs treated with peptidoglycan of 3, 10, 30, 100 mg/L, respectively, compared to 135.41 ± 14.37 ng/L in untreated KCs (all P < 0.05). On the contrary, a significant decrement was seen in the secretion of RANTES by KCs treated with peptidoglycan of 10, 30, 100 mg/L compared with untreated KCs (110.72 ± 8.51 ng/L, 90.50 ±2.45 ng/L, 49.89 ± 13.74 ng/L vs 149.94 ± 18.71 ng/L, all P < 0.05). The monoclonal antibody to TLR-2 could markedly suppress the promotion of IL-8 production by peptidoglycan, but had no obvious influence on the inhibition of RANTES production by peptidoglycan. MDC could not be detected in the culture super-natants of KCs with or without peptidoglycan stimulation. Conclusion Peptidoglycan could inhibit RANTES secretion but induce IL-8 production by KCs likely via TLR2.

Expression of MCP-1 and MIP-2 mRNA in Vagina of Murine Model of Vulvovaginal Candidiasis / 中华皮肤科杂志

Objective To investigate the kinetic expression level of chemokines (MCP-1 and MIP-2) in vagina of a murine model of vulvovaginal candidiasis (VVC).Methods The estrogen-treated murine model of VVC was set up.Vaginal specimens were obtained in different duration after inoculation of C.albicans intravaginally.Semi-quantitative RT-PCR was applied to determine MCP-1 and MIP-2 mRNA levels in these tissues.Results Compared with the control mice treated with olive oil,persistent growth of C.albicans was found from the 2nd day to 21st day after inoculation in estrogen-treated mice.Significantly higher levels of MCP-1 mRNA were observed in vaginal tissues in infected estrogen-treated mice than those in other 2 groups,infected but non estrogen-treated mice and estrogen-treated but uninfected mice.The high level of MCP-1 mRNA maintained from the 4th day to 21st day in infected estrogen-treated mice.It was also found that levels of MIP-2 mRNA were significantly higher in the vagina in the 2nd day in 3 groups of mice than those in naive mice,however,no significant difference was shown among 3 groups throughout the study period.Conclusion High level of MCP-1,rather than MIP-2,may be associated with susceptibility to VVC.

Protective effects of NF-kappa B inhibitor on ischemia/reperfusion injury in rat liver transplantation / 中国普通外科杂志

Objective To study the effect of nuclear factor-?B(NF?B) on cytokins expression and neutrophil accumulation in murine liver transplantation. Methods Donor and recipient Wistar rats were pretreated by introperitoneal injection of proline dithiocarbamates(ProDTC, 15 mg/kg) 15 min before transplantation. Results Compared with NS control rats the level of NF?B p65 significantly decreased suppressing the transcription of TNF-?,MIP-2 and ICAM-1 and the release of serum TNF-? and MIP-2 as well as the activity of myeloperoxidase(MPO) in rats receiving ProDTC(all P

Surfactant protein A regulates the expression of MIP-2 and inhibits NF-?B binding activity in tubular epithelial cells / 中华肾脏病杂志

Objective To investigate the effect of surfactant protein A (SP-A) on the production of MIP-2 and binding activity of NF-?B in rat tubular epithelial cells, and evaluate its possible role in renal inflammation. Methods Confluent cultures of NRK-52E cells (a renal tubular epithelial cell line of rat origin) were pretreated with various concentrations of SP-A(0 to 80 ?g/ml) and stimulated by lipopolysaccharide (LPS) (10 ?g/ml) with 2% serum. MIP-2 expression was measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). The effect of SP-A on NF-?B binding activity was assessed by electrophoretic mobility shift assay (EMSA). Results MIP-2 mRNA and protein was expressed and up-regulated in NRK-52E cells stimulated by LPS. The expression of MIP-2 was down-regulated by SP-A. NF-?B binding activity was inhibited by SP-A in a concentration-dependent manner. Conclusion SP-A binding activity and down-regulates the expression of MIP-2 in renal tubular epithelial cells, which may play an important role in the modulation of renal tissue inflammation.

The Expression of Interleukin-17,Interferon-?,and Macrophage Inflammatory Protein-3?mRNA in Patients with Psoriasis Vulgaris / 中华皮肤科杂志

Objective To investigate the expression levels of of interleukin-17(IL-17),interferon-gamma(IFN-?),and macrophage inflammatory protein-3alpha(MIP-3?)mRNA in skin lesions of pa-tients with psoriasis vulgaris.Methods The skin biopsies were collected from skin lesions in31cases of psoriasis vulgaris and16normal controls.The expressions of IL-17,IFN-?,and MIP-3?mRNA were semi-quantitatively analyzed with reverse transcriptase-polymerase chain reaction(RT-PCR)in psoriatic lesions and normal skin tissues.Results Expressions of IL-17,IFN-?,and MIP-3?were presented in all speci-mens of both psoriatic lesions and normal controls.The expression levels were1.142?0.059for IL-17mRNA,1.114?0.056for IFN-?mRNA,1.140?0.052for MIP-3?mRNA,in skin lesions,respectively,which were greatly higher than those in normal controls(0.879?0.034,0.905?0.026and0.868?0.031,respectively)(all P

Effects of recombinant viral chemokine vMIP-II on cellular immunity stimulated by LPS / 中国病理生理杂志

AIM: To investigate the produce of intracellular cytokine following short-term in vitro stimulation with vMIP and LPS, and discuss the effect of vMIP to cellular immunity. METHODS: The methods of Cross-linking of radioactivity, ELISA and four-colors flow cytometer were used to test the level of the secretion of chemokine IL-12 and intracellular cytokine IFN-? and IL-4. RESULTS: After treated the PBMCs with vMIP-II, the levels of secretion of IL-12, IFN-? and IL-4 were reduced in the present of LPS by competitively combining chemokine receptor; vMIP promoted CD4+T cell to secrete IL-12, IFN-? and IL-4. CONCLUSION: vMIP-II can protect systemic response of immunity and reduce extremely inflammation by down-regulating proinflammation.