Dimethyl Fumarate Protects against Lipopolysaccharide- (LPS-) Induced Sepsis through Inhibition of NF-κB Pathway in Mice.

Sepsis is one of the most severe complications and causes of mortality in the clinic. It remains a great challenge with no effective treatment for clinicians worldwide. Inhibiting the release of proinflammatory cytokines during sepsis is considered as an important strategy for treating sepsis and improving survival. In the present study, we have observed the effect of dimethyl fumarate (DMF) on lipopolysaccharide- (LPS-) induced sepsis and investigated the possible mechanism. By screening a subset of the Johns Hopkins Drug Library, we identified DMF as a novel inhibitor of nitric oxide synthesis in LPS-stimulated RAW264.7 cells, suggesting that DMF could be a potential drug to treat sepsis. To further characterize the effect of DMF on LPS signaling, TNF-α, MCP-1, G-CMF, and IL-6 expression levels were determined by using cytokine array panels. In addition, an endotoxemia model with C57BL/6 mice was used to assess the in vivo efficacy of DMF on sepsis. The survival rate was assessed, and HE staining was performed to investigate histopathological damage to the organs. DMF was found to increase the survival of septic mice by 50% and attenuate organ damage, consistent with the reduction in IL-10, IL-6, and TNF-α (inflammatory cytokines) in serum. In vitro experiments revealed DMF's inhibitory effect on the phosphorylation of p65, IκB, and IKK, suggesting that the primary inhibitory effects of DMF can be attributed, at least in part, to the inhibition of phosphorylation of IκBα, IKK as well as nuclear factor-κB (NF-κB) upon LPS stimulation. The findings demonstrate that DMF dramatically inhibits NO and proinflammatory cytokine production in response to LPS and improves survival in septic mice, raising the possibility that DMF has the potential to be repurposed as a new treatment of sepsis.

STIM1 Regulates Endothelial Calcium Overload and Cytokine Upregulation During Sepsis.

BACKGROUND: Stromal interaction molecule 1 (STIM1)-mediated store-operated Ca2+ entry (SOCE) is now recognized as the main mechanism of the majority of nonexcitable cell calcium influx. Calcium overload is a primary mechanism of endothelial cell injury during systemic inflammatory response and sepsis. Whether STIM1-mediated SOCE plays a role in calcium overload in vascular endothelial cell injury remains unclear. MATERIALS AND METHODS: To explore the role of STIM1-gated SOCE in vascular endothelial cell calcium overload and inflammation, we established a human septic serum or lipopolysaccharide (LPS)-induced human umbilical vein endothelial cell (HUVEC) experimental system and derived ribonucleic acid interference (RNAi)-mediated STIM1, ORAI1 (orai gene [HGNC: 25896 Entrez Gene: 84876] coding protein, ORAI Calcium Release-Activated Calcium Modulator 1), and transient receptor potential channel 1 (TRPC1) (core components of store-operated Ca2+[SOC]) downregulated HUVECs, as well as STIM1 overinduced HUVECs. RESULTS: Our results show that sepsis serum or LPS stimulation increased STIM1 in HUVECs and increased all cytokines except for VEGF and the inflammatory mediators tumor necrosis factor, intercellular cell adhesion molecule-1, and endothelin-1 in a time-dependent manner. RNAi-mediated knockdown of STIM1 significantly inhibited serum or LPS-induced inflammatory cytokine expression, and STIM1 overexpression in HUVECs promoted LPS-mediated induction of these cytokines. Meanwhile, similar to the blocking effect of the specific SOC inhibitors Gd3+ and La3+ on LPS-induced calcium influx, RNAi-mediated depletion of STIM1 or the SOC proteins TRPC1 and ORAI1 could significantly inhibit serum or LPS-induced extracellular calcium influx, as well as the expression of the inflammatory cytokines tumor necrosis factor, intercellular cell adhesion molecule-1, and endothelin-1. Simultaneous downregulation of the SOCE core units TRPC1 and ORAI1 inhibited LPS-induced calcium influx and cytokine expression, which could not be restored by inducing STIM1. Forced expression of nuclear factor-κB (NF-κB) in HUVECs significantly induced STIM1 expression, whereas RNAi-mediated depletion of NF-κB significantly inhibited STIM1 mRNA levels and significantly reduced the thapsigargin-mediated SOCE calcium influx, which was similar to results with the NF-κB inhibitor wogonin. CONCLUSIONS: Septic serum stimulates the expression of STIM1, cytokines, and inflammatory mediators in HUVECs. STIM1-mediated SOCE is required for Ca2+ influx induced by LPS or septic serum and contributes cytokines and inflammatory mediators in septic serum-stimulated HUVECs. In addition, STIM1-mediated SOCE on Ca2+ influx by septic serum or LPS involves NF-κB signaling.

Enlightenment and lesson from the successful treatment of acute superior mesenteric artery embolism.

 Despite recent advances, treatment of acute superior mesenteric artery embolism (SMAE) in emergency is still very difficult. The comparative rarity, the difficulty of early diagnosis, and the extremely high mortality of SMAE give us the sufficient reasons to report our successful experience in curing a patient with acute SMAE. In the present case, the patient was diagnosed early without evidence of intestinal necrosis. The diagnosis of SMAE was verified by computed tomography angiography (CTA). A comprehensive treatment was immediately employed including a combination of intra-arterial thrombolysis and embolectomy operation with emergent laparotomy. We summarize that the successful treatment of superior mesenteric artery embolism depends on early diagnosis and timely reestablishment of arterial flow under the support of general treatments such as prevention of sepsis and control of organic insufficiencies. Through a combined treatment, patient's life could be saved without complications.

Toll-like receptor 4 is involved in bacterial endotoxin-induced endothelial cell injury and SOC-mediated calcium regulation.

Bacterial endotoxins may lead to vascular endothelial cell injury. Our study explored the role of TLR4 (Toll-like receptor 4) and STIM1 (stromal interaction molecule 1) in bacterial endotoxin-induced calcium overload and inflammatory reactions in HUVECs (human umbilical vein endothelial cells). It showed that under LPS (lipopolysaccharide) stimulation, LBP (LPS-binding protein) mRNA levels peaked at 24 h, TLR4 levels at 12 h and NF-κB (nuclear factor κB) levels at 6 h (all P<0.01). LBP levels increased gradually and peaked at 24 h of LPS treatment. TLR4 protein levels increased significantly at 1 h and peaked at 12 h. NF-κB protein levels markedly increased at 1 h and peaked at 6 h. Knockdown of STIM1 alone, TLR4 alone or both STIM1 and TLR4 together, markedly abolished LPS-induced increase in calcium influx into cells (P<0.05, P<0.01 and P<0.01 respectively). LBP-TLR4 and STIM-NF-κB interactions were detected without LPS treatment, enhanced by LPS stimulation, and markedly reduced by knocking down TLR4 and STIM respectively. Both the NF-κB inhibitor, PDTC (pyrrolidine dithiocarbamate) and TLR4 knockdown could block LPS induction of NF-κB, STIM, TNFα (tumour necrosis factor α) and IL-6 (interleukin 6). The data indicate LPS-LBP may activate TLR4 signalling and downstream transcription factor NF-κB, which further can activate STIM1 and eventually lead to calcium influx and injury of HUVECs. Inhibition of TLR4 effectively reverses LPS induction of inflammatory mediator generation and extracellular calcium influx mediated by STIM1.

[Establishment of a high-velocity fragment-induced penetrating liver injury model in landrace pigs].

OBJECTIVE: To establish a stable fragment-induced penetrating liver injury model in landrace pigs and evaluate the characteristics of deep tissue injury. METHODS: According to the different positioning methods of aiming points, twelve healthy adult landrace pigs were divided into group A (the relative height "h" of the aiming point and the highest point of the body surface on the tracing line was set to 5 cm) and group B ("h" was set to 6 cm). Ultrasonography was used to determine the direction of fragment projection, and an experimental ballistic gun was used to project high-velocity fragments to cause injury to animals. The vital signs of the two groups were monitored, and whole blood cell count, blood gas analysis, and liver and renal function were tested. Damages to the liver and adjacent organs, as well as the amount of bleeding and survival time were analyzed. RESULTS: For the overall analysis of the two groups, the liver hit rate of fragment simulating projectiles was 100% (right anterior lobe and right lateral lobe injury), the hit rate of other organs in the abdominal cavity was 25% (3/12), and the incidence of hemothorax or pneumothorax was 8% (1/12). The wounds were mainly characterized by liver lacerations, with total or partial disconnection of the distal liver lobe. There was no significant difference in wound length and bleeding amount between groups A and B [wound length (cm): 9.8±1.7 vs. 11.2±3.8, bleeding amount (g): 597.0±477.1 vs. 1 032.0±390.3, both P > 0.05]. The depth of liver parenchymal laceration in group B with the aiming point closer to the anterior median line was significantly longer than that in group A (cm: 2.8±0.4 vs. 1.9±0.6, P = 0.015). Mean arterial pressure (MAP), pH value, residual arterial blood base (BE), hemoglobin (Hb) and hematocrit (HCT) levels decreased after the fragment-induced injury, and then reached a trough level [MAP (mmHg, 1 mmHg ≈ 0.133 kPa): 87.0±33.6, pH: 7.26±0.15, BE (mmol/L): -6.65±8.48, Hb (g/L): 9.86±1.10, HCT: 0.309±0.029, all P < 0.05] in the first hour. Blood lactate (Lac), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) levels increased over time, and reached a peak level [Lac (mmol/L): 10.21±4.40, LDH (U/L): 1 417.0±223.3, AST (U/L): 234.5 (162.5, 357.5), both P < 0.05] at 1 hour after injury. Pearson's correlation analysis showed that the total amount of bleeding was correlated with the depth of liver parenchyma laceration (r = 0.684,P = 0.014). The Kaplan-Meier survival curve showed that the 3 hours survival rate in group A was higher than that in group B, but the difference was not statistically significant [83.3% (5/6) vs. 33.3% (2/6), P > 0.05]. CONCLUSIONS: The high-velocity fragment-induced penetrating liver injury model established by striking landrace pigs closer to the anterior median line with fragment simulating projectiles is reproducible and the degree of damage is controllable, and the model is applicable to further relevant research of hepatic ballistic trauma.

Novel dopamine-modified oxidized sodium alginate hydrogels promote angiogenesis and accelerate healing of chronic diabetic wounds.

Herein, the dopamine (DA) was grafted with oxidized sodium alginate (OSA) via Schiff base reduction reaction, aiming to fabricate novel DA-grafted OSA (OSA-DA) hydrogels with enhanced biocompatibility and suitable adhesion for clinical applications. The chemical structures of OSA-DA were characterized via UV-Vis, FTIR and 1H NMR spectroscopy analysis. The hydrogel characteristics, biocompatibility, as well as the chronic diabetic wound healing efficacy were investigated. Our results demonstrated that DA was grafted with OSA successfully with highest grafting rate of 7.50%. Besides, OSA-DA hydrogels possessed suitable swelling ratio and appropriate adhesion characteristics. Additionally, OSA-DA exhibited satisfactory cytocompatibility and cell affinity in L-929 cells, and superior biocompatibility in SD rats. Moreover, OSA-DA exerted remarkable promoting effects on migration and tube formation of human umbilical vein endothelial cells (HUVECs). Studies on full-thickness excision chronic diabetic wounds further revealed that OSA-DA hydrogels could accelerate healing via promoting angiogenesis, reducing inflammation response, and stimulating collagen deposition. Overall, our studies would provide basis for SA-based hydrogels as clinical wound dressings.

Acheron regulates vascular endothelial proliferation and angiogenesis together with Id1 during wound healing.

RNA binding protein acheron has proved to be either the mediator of integrin-extracellular matrix interactions or the regulatory factor that participates in vertebrate development, cell differentiation and cell death. We report the role of acheron in vascular endothelial proliferation, angiogenesis and wound healing post-trauma. Co-immunoprecipitation showed that Acheron forms a ternary complex with ß1 integrin and Id1 in human umbilical vein endothelial cells following stimulation with serious trauma serum. Acheron, vascular endothelial growth factor (VEGF), and ß1 integrin mRNA expression was apparently inhibited, and capillary density and wound healing rate also were reduced in Id1-deficient mice trauma model. Acheron together with Id1 significantly induces VEGF, not CD105 level inhibition by serious trauma serum for 24 h. In conclusion, we have demonstrated that acheron may be an effective mediator of promoting endothelial proliferation, angiogenesis and wound healing probably by regulating VEGF together with Id1.

[Effect of early intensive insulin therapy on high mobility group box 1 protein levels and prognosis of patients with severe trauma].

OBJECTIVE: To study the effect of intensive insulin therapy on serum high mobility group box 1 (HMGB1) levels and its relationship with the prognosis in early phase of severe trauma. METHODS: Eighty severe trauma patients [injury severity score (ISS)≥ 16] were divided into groups according to injury to matched anatomical regions. Forty patients of intensive therapy group were given early intensive insulin therapy, while another 40 patients of the conventional treatment group received routine treatment based on clinical experience with insulin treatment. The insulin dose and the blood glucose levels were recorded within 72 hours after treatment. The relationship between HMGB1 levels and prognosis was analyzed by testing serum HMGB1 levels at 24, 36, 48, 60 or 72 hours after treatment, and clinical terminal events such as multiple organ dysfunction syndrome (MODS) and death rate within 1 week were recorded. Results The insulin dose of intensive therapy group was significantly greater than that of conventional treatment group following the blood glucose levels were significantly lower than those of the conventional treatment group after treatment for 72 hours. The levels of HMGB1 (µg/L) lowered after intensive insulin therapy for 36 hours , and were significantly lower than those of conventional treatment group at 36, 48, 60 and 72 hours after intensive treatment (36 hours: 41.3 ± 9.5 vs. 52.7 ± 11.5, 48 hours: 48.6 ± 17.6 vs. 124.1 ± 22.9, 60 hours: 47.7 ± 23.3 vs. 132.9 ± 33.4, 72 hours: 54.3 ± 26.3 vs. 140.6 ± 16.5, P <0.05 or P <0.01). The incidence of MODS and mortality in intensive therapy group was respectively significantly lower than that of the conventional treatment group (20.0% vs. 55.0%, 10.0% vs. 30.0%, both P <0.05). In conventional treatment group the patients with HMGB1 ≥ 132.26 µg/L ( n =22) occurred MODS, and those with HMGB1<132.26 µg/L ( n =18) did not occur MODS. The HMGB1 levels in death patients ( n =12) were ≥ 132.26 µg/L. CONCLUSION: Early intensive insulin treatment could probably reduce MODS and mortality by inhibiting stress hyperglycemia and serum HMGB1 levels effectively. Serum HMGB1 of severe trauma patients can be used for the clinical indicator of prognosis.

LPS-induced vein endothelial cell injury and acute lung injury have Btk and Orai 1 to regulate SOC-mediated calcium influx.

Patients with sepsis and sepsis-related complications have a high mortality. Endothelial cell dysfunction plays a central role in sepsis pathophysiological process. In sepsis patients, endothelial cell apoptosis is associated with intracellular calcium overload. Multiple functions in the apoptotic process have been found to be regulated by calcium signaling. Our previous work had proved that LPS-induced cell injury was associated with store-operated calcium (SOC) entry mediated by stromal interaction molecule-1 (STIM 1) in Human umbilical vein endothelial cells (HUVEC), but the underlying molecular mechanism has not been adequately defined. Here we report that the LPS-induced cell injury is related to the calcium overload in HUVEC. SOC entry mediated by calcium release-activated calcium modulator (Orai) 1 and transient receptor potential canonical (TRPC) 1 was associated with LPS-induced calcium overload and cell apoptosis. Bruton's tyrosine kinase (Btk)/Phospholipase C(PLC) γ/inositol 1,4,5-triphosphate receptor (IP3R) played a major role in regulating calcium overload in LPS-induced HUVEC. Knockdown of Btk markedly inhibited the expressions of Orai 1 and its downstream molecule IP3R but not that of TRPC1 in LPS-induced HUVEC. In mice, knockdown of Btk and Orai 1 inhibited LPS-induced calcium overload, pulmonary vascular endothelial cell (VEC) injury and acute lung injury. These findings demonstrated that Btk acts as a regulator of calcium-dependent signaling, especially in the Orai 1-mediated SOC entry of the LPS-induced VEC.

Ivermectin inhibits cell proliferation and the expression levels of type I collagen, α­SMA and CCN2 in hypertrophic scar fibroblasts.

A hypertrophic scar (HPS) is characterized by abnormal cell proliferation and the overproduction of extracellular matrix. Currently, the treatment options available for this remain unsatisfactory. Innovative treatments are required to attenuate or prevent hypertrophic scarring and the present study searched for a drug capable of becoming a new preventative and therapeutic strategy. Although the underlying mechanisms have not been fully clarified; it is widely accepted that the TGF­ß1/SMAD3 signaling pathway serves an essential role in HPS formation. In the present study, a compound library consisting of clinically used drugs was screened for their inhibitory activity against the SMAD3 protein. The results indicated that ivermectin was able to suppress the phosphorylation of SMAD3. Therefore, the present study further investigated whether ivermectin exhibited antifibrotic effects on HPS fibroblasts. The results demonstrated that ivermectin inhibited the proliferation of HPS fibroblasts and significantly decreased the production of type I collagen, α­smooth muscle actin and cellular communication network factor 2, as determined by analyzing the mRNA and protein expression levels. In conclusion, the results of the present study suggested that ivermectin may be a promising therapeutic agent for HPS.

When there is a will there is a way: The role of proactive personality in combating COVID-19.

The global COVID-19 pandemic has disrupted personal and work lives and created great uncertainty and stress, especially for frontline health care professionals like doctors and nurses who risk personal health while facing increased workloads and new COVID-related tasks. People can passively respond to this disruption, or they can be more active and choose to shape the conditions surrounding their work during the crisis. We designed a multiwave, multisource study examining whether a proactive orientation is a key resource for frontline health care professionals in the COVID-19 pandemic. Drawing from proactive and conservation of resources theories, we studied a sample of 408 doctors and nurses at a COVID-19 hospital in the locked-down area surrounding Wuhan City, China during the first wave of the virus. Our aim is to examine how personal agency contributes to health care professionals' performance and well-being when combating COVID-19. Proactive personality as a dispositional resource was associated with higher levels of perceived strengths use, a job-related motivational resource. This effect was jointly moderated by routine disruption and perceived organizational support. Proactive personality was indirectly associated with performance and two indicators of well-being (resilience and thriving) through perceived strengths use. More frequent physical exposure to the virus magnified the effects of perceived strengths use on an archival indicator of performance during the first wave of the pandemic. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Human calcium/calmodulin-dependent serine protein kinase regulates the expression of p21 via the E2A transcription factor.

CASK (calcium/calmodulin-dependent serine protein kinase) is a kind of scaffolding protein that recruits or organizes other proteins at the plasma membrane to co-ordinate signal transduction pathways within the cytoplasm and nucleus. We have previously found that hCASK (human CASK) binds Id1 (inhibitor of DNA binding 1) through hCASK's GUK (guanylate kinase) domain and inhibits cell growth, probably via interactions with Id1. Overexpression of hCASK resulted in a reduced rate of cell growth, although inhibition of CASK via RNAi (RNA interference) promoted cell proliferation in ECV304 cells. This study revealed that hCASK regulates the protein and mRNA level of p21(wafi/cip1) (referred to throughout as p21), and activated the expression of p21 in a time-dependent manner. Two E-boxes in the proximal region at the TSS (transcription start site) play key roles in regulating hCASK-mediated p21 expression. We suggest that E2A (E12 and E47), a representative of the E proteins that binds the E-box elements, is a participant in the mediation of p21 expression by hCASK. The results of the present study suggest that hCASK regulation of cell growth might involve p21 expression, and that the bHLH (basic helix-loop-helix) transcription factor E2A probably participates in hCASK regulation of p21 expression. From these findings, we propose a novel proliferation signalling pathway mediated by hCASK.

Effects of carboxymethyl chitosan oligosaccharide on regulating immunologic function and inhibiting tumor growth.

Herein, the effects of carboxymethyl chitosan oligosaccharide (CM-COS) on regulating immunologic function and inhibiting hepatocellular tumor growth were evaluated. Results showed that CM-COS caused dramatic viability loss of hepatocellular carcinoma BEL-7402 with non-toxicity towards normal liver L-02 cells. CM-COS repressed tumor growth of hepatoma-22, and elevated the spleen index and thymus index of tumor-bearing mice. Contents of VEGF and MMP-9 were significantly down-regulated by CM-COS. Histological analyses revealed that CM-COS promoted tumor cell necrosis and produced no significant toxicity to spleen tissues. Moreover, expressions of Caspase-3 in tumor tissues and IL-2 in spleen tissues were significantly activated by CM-COS. Additionally, in vitro cell viability, phagocytic capability and NO production of mouse peritoneal macrophages exposed to CM-COS were significantly higher. CM-COS remarkably increased the in vivo phagocytosing capacity of peritoneal macrophages of Kunming mice. Taken together, our findings suggested that CM-COS might be potentially effective and non-toxic candidate as anti-hepatoma agents.

[Bacterial endotoxin-induced endothelial cell injury and calcium overload associated with Toll-like receptor and calcium signal].

OBJECTIVE: To explore the effect of toll-like receptor 4 (TLR4), myeloid differentiation protein-2 (MD2), and stromal interaction molecular 1 (STIM1) for regulating human vascular endothelial calcium overload injury and inflammatory reaction induced by bacterial endotoxin (LPS). METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured in Dulbecco's modification of Eagle's medium (DMEM). (1) The levels of TLR4, MD2 and nuclear factor-κB (NF-κB) were detected by reverse transcriotion-polymerase chain reaction (RT-PCR) before and 0.5, 1, 6, 12, 24 hours after LPS stimulation. (2) Intracellular calcium peak level was detected by confocal following probe fluo-3 AM loading in HUVEC cells induced with LPS and transfected by psiSTIM or psiTLR. (3) MD2, STIM1 or NF-κB protein level was detected by immunoprecipitation (IP) and immuno-blotting in HUVEC cells which were transfected by TLR4 inhibited expression (psiTLR) for 12 hours and followed by LPS stimulation for 6 hours. (4) HUVEC cells were randomly divided into 6 groups: control group, LPS group, PDTC 0.1 mg/L group, PDTC 1 mg/L group, psiTLR 1 h group and psiTLR 12 h group. Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by enzyme linked immunosorbent assay (ELISA) in supernatant. The mRNA levels of STIM1 and NF-κB were detected by RT-PCR. RESULTS: (1) The mRNA levels of TLR4, MD2, and NF-κB gradually increased after LPS induction and peaked at 6 hours (2-ΔΔCt: 23.52±2.88, 17.43±3.43, 18.13±2.99, respectively), which were statistically significant before the stimulation with LPS (2-ΔΔCt: 7.02±2.81, 5.19±3.22, 8.11±1.42, all P < 0.05). (2) Extracellular calcium influx in LPS group was increased significantly higher than control group (nmol/L: 108.13±22.33 vs. 41.57±13.19, P < 0.01). Extracellular calcium influx in psiSTIM+LPS group (nmol/L: 62.61±14.12 vs. 108.13±22.33, P < 0.05) and psiTLR+LPS group (nmol/L: 50.78±8.05 vs. 109.43±20.21, P < 0.01) were both suppressed as compared with LPS group. While extracellular calcium peak level in psiTLR+psiSTIM+LPS group further decreased (nmol/L: 39.31±6.42 vs. 109.43±20.21, P < 0.01). (3) MD2 protein but not STIM1 or NF-κB can be detected in anti-TLR4 precipitates in control (ctrl-) by immunoprecipitation. MD2 protein level increased in anti-TLR4 precipitates in LPS group (ctrl+) and was suppressed in TLR4 inhibiting group (psiTLR). (4) The levels of TNF-α in PDTC 1 mg/L group were significantly lower than those of LPS group (ng/L: 0.60±0.24 vs. 1.77±0.66, P < 0.01). The levels of IL-6 in PDTC 0.1 mg/L, 1 mg/L group and psiTLR 12 h group decreased significantly lower than that of LPS group (ng/L: 232.10±63.54, 134.32±37.23, 284.23±56.14 vs. 510.22±89.23, all P < 0.05). Compared to LPS group, the mRNA levels of NF-κB and STIM1 were obviously inhibited in PDTC 1 mg/L group and psiTLR 12 h group [NF-κB mRNA (2-ΔΔCt): 17.22±2.35, 13.24±3.54 vs. 30.16±2.06; STIM1 mRNA (2-ΔΔCt): 12.57±2.43, 12.21±2.46 vs. 25.12±2.02, all P < 0.05]. CONCLUSIONS: TLR4, MD2, NF-κB signal and SOC calcium channel STIM1 mediate LPS induced-calcium influx and inflammatory mediators level in HUVEC cells. Extracellular calcium overload and inflammatory response by endotoxin induction can be effectively inhibited by down-regulation of TLR4, NF-κB and/or STIM1.

Neutrophil elastase ameliorates matrix metalloproteinase-9 to promote lipopolysaccharide-induced acute lung injury in mice 1.

PURPOSE: To investigate the regulatory roles of neutrophil elastase (NE) and matrix metalloproteinase-9 (MMP-9) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. METHODS: To construct LPS-induced ALI mouse models, wild-type C57BL/6 mice were administered 5.0 mg/kg of LPS through endotracheal, and/or 1.0 mg/kg of ONO-5046, and/or 20.0 mg/kg of chemically modified tetracycline-3 (CMT-3) by gavage. The levels of MMP-9, tissue inhibitor of metalloprotease-1, interleukin (IL)-6 were detected by real time RT-PCR at 6 h, 24 h and 48 h, and tumor necrosis factor (TNF), lung wet-dry weight ratio, white blood cell (WBC) count and polymorphonuclear (PMN) count in bronchoalveolar lavage fluid (BALF) were tested at 48 h after administration. The 5-day survival analysis of the ALI mice was also performed. RESULTS: Both ONO-5046 and CMT-3, regardless of being used individually or combined, significantly reduced the levels of MMP-9, IL-6, and TNF in lung tissue as well as in BALF, and the WBC and PMN count in BALF. Combined treatment with ONO-5046 and CMT-3 remarkably improved the survival rate of ALI mice. CONCLUSION: Neutrophil elastase synergizes with matrix metalloproteinase-9 to promote and regulate the release of inflammatory mediators and the infiltration of inflammatory cells, consequently affecting the survival of lipopolysaccharide-induced acute lung injury mice.

Total ginsenosides synergize with ulinastatin against septic acute lung injury and acute respiratory distress syndrome.

Total ginsenosides synergize with ulinastatin (UTI) against septic acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). We randomly divided 80 cases of severe sepsis-induced ALI and ARDS into a UTI group and a ginsenosides (GS)+UTI group. Continuous electrocardiac monitoring of pulse, respiratory rate, blood pressure, and heart rate; invasive hemodynamic monitoring; ventilator-assisted breathing and circulation support; and anti-infection as well as UTI treatment were given in the UTI group with GS treatment added for 7 consecutive days in the GS+UTI group. The indicators of pulmonary vascular permeability, pulmonary circulation, blood gases, and hemodynamics as well as APACHE II and ALI scores were detected on days 1, 3, and 7. The ALI score in the GS+UTI group was significantly decreased (P < 0.05) compared with that of the UTI group, and the indicators of pulmonary capillary permeability such as pulmonary vascular permeability index, extravascular lung water index, and oxygenation index, in the GS+UTI group improved significantly more than that of the UTI group. The indicators of hemodynamics and pulmonary circulation such as cardiac index, intrathoracic blood volume index, and central venous pressure improved significantly (P < 0.05), and the APACHE II score in the GS+UTI group was lower than that of the UTI group. GS can effectively collaborate with UTI against ALI and/or ARDS.

Liver kinase B1 promoter CpG island methylation is related to lung cancer and smoking.

The aim of this study was to explore the association of CpG islands methylation of liver kinase B1 (LKB1) with primary lung cancer and smoking, providing a theoretical basis for the demethylating drug to treat lung cancer by detecting the LKB1 promoter CpG methylation. mRNA expression of LKB1 were detected by in situ hybridization and methylation status on Hap II locus of the promoter of LKB1 was analyzed by methylation-specific polymerase chain reaction (PCR). 7 of 80 LKB1 positive cases had methylation on CpG islands while 18 of 44 LKB1 negative cases had methylation on CpG islands. The difference was significant between CpG island methylation and LKB1 expression. 8 of 54 cases of early and middle lung cancer were detected LKB1 promoter CpG island methylation while 30 controls were not detected, the difference was significant. 5 of 64 more-than-5-year cases had methylation on CpG islands while 20 of 60 less-than-5-year cases had methylation. The difference was significant between of 5-year survival and CpG island methylation of LKB1. 22 of 74 smoking cases of lung cancer had methylation on CpG islands of LKB1 while only 3 of 50 non-smoking cases had methylation. The difference of smoking and CpG island methylation of LKB1 was significant. LKB1 promoter CpG islands aberrant methylation is closely associated with the occurrence, development and prognosis of lung cancer, especially with smoking history including clinical early diagnosis and prognosis. CpG islands methylation in the promoter of LKB1 is likely important one of the mechanism of smoking-associated lung cancer.

Deoxyribonucleic acid (DNA) methyltransferase contributes to p16 promoter CpG island methylation in lung adenocarcinoma with smoking.

In this study, the relationship between CpG island methylation and smoking and DNA methyltransferase in the occurrence and development of lung adenocarcinoma was explored by detecting p16 promoter methylation status. Protein and mRNA levels of p16 were detected by immunohistochemistry and in situ hybridization assays. p16 gene promoter and exon 1 CpG island locus Hap II sites methylation status was analyzed with the methylation-specific PCR. Only 4 of 40 p16-positive cases were detected to methylate on CpG islands with 10% methylating rate whereas 18 of p16-negative cases were methylated up to 36.73% of methylating rate. The methylating rates of both p16-positive and p16-negative groups were significantly different. 17 of 50 cases with smoking from total 89 lung adenocarcinoma cases were detected to methylate on CpG islands while only 5 of the remaining 39 non-smokers to methylate. The difference of the methylating rates in both smokers and non-smokers was significant to suggest the closely association of CpG island methylation of p16 with smoking. Furthermore, p16 promoter CpG islands were detected to methylate in 15 of 35 cases with higher DNA methyltransferase activity whereas only 7 detected to methylate in the remaining 54 cases with lower DNA methyltransferase activity. p16 promoter CpG island methylation likely made p16 expressing silence thus contributed to the tumorigenesis of lung adenocarcinoma. Smoking is likely to promote p16 CpG island methylation or by its effect of the activity and metabolism of DNA methyltransferase 1 (DNMT) on CpG island methylation status.

The interaction effects of temperature and humidity on emergency room visits for respiratory diseases in Beijing, China.

Few epidemiological studies have been reported as to whether there was any interactive effect between temperature and humidity on respiratory morbidity, especially in Asian countries. The present study used time-series analysis to explore the modification effects of humidity on the association between temperature and emergency room (ER) visits for respiratory, upper respiratory tract infection (URI), pneumonia, and bronchitis in Beijing between 2009 and 2011. Results showed that an obvious joint effect of temperature and humidity was revealed on ER visits for respiratory, URI, pneumonia, and bronchitis. Below temperature threshold, the temperature effect was stronger in low humidity level and presented a trend fall with humidity level increase. The effect estimates per 1 °C increase in temperature in low humidity level were -2.88 % (95 % confidence interval (CI) -3.08, -2.67) for all respiratory, -3.24 % (-3.59, -2.88) for URI, -1.48 % (-1.93, -1.03) for pneumonia, and -3.79 % (-4.37, -3.21) for bronchitis ER visits, respectively. However, above temperature threshold, temperature effect was greater in high humidity level and trending upward with humidity level increasing. In high humidity level, a 1 °C increase in temperature, the effect estimates were 1.84 % (1.55, 2.13) for all respiratory, 1.76 % (1.41, 2.11) for URI, and 7.48 % (4.41, 10.65) for bronchitis ER visits. But, there was no statistically significant for pneumonia. This suggests that the modifying effects of the humidity should be considered when analyzing health impacts of temperature.

VEGF regulates antidepressant effects of lamotrigine.

The anticonvulsant drug lamotrigine has been shown to produce strong antidepressant effects in the treatment of bipolar disorder patients. Our previous studies have demonstrated that brain derived neurotrophic factor (BDNF) signaling plays an important role in regulating its behavioral actions in several rodent models of depression. The current study extends earlier work on BDNF and explores the role of another important neurotrophin vascular endothelial growth factor (VEGF) in regulating the antidepressant actions of lamotrigine. The results showed that chronic administration of 30 mg/kg lamotrigine (14 days) normalized the down-regulated frontal and hippocampal VEGF protein expression as well as the behavioral deficits induced by chronic unpredictable stress. In addition, pharmacological inhibition of VEGF signaling by infusion of SU5416, a selective Flk-1 receptor inhibitor, blocks the antidepressant effects of lamotrigine in all behavioral paradigms. Taken together, this study provides further evidence that VEGF is also an essential regulator for the antidepressant effects of lamotrigine.