A Rare 2D Framework Cu(atz)2 with Ferrimagnetic and High Proton Conductivity.

Materials combining proton conductivity and magnetism have attracted great attention in recent years due to their intriguing application in sensors and fuel cells. Herein a two-dimensional metal-organic framework, [Cu(atz)2 (H2 O)2 ]⋅H2 O (1) (Hatz=5-aminotetrazole), has been obtained in a green synthesis method. The single-crystal structure revealed that the atz- ligands as linkers coordinate with copper ions to sql networks, between which water molecules are immobilized through hydrogen bonds. The resulting complex 1 exhibits a high proton conductivity of 1.11×10-4 and 6.19×10-4  S cm-1 at room temperature and 333 K, respectively, under 98% RH with an activation energy of 0.56 eV. Upon dehydration, the proton conductivity of 1_dg drops by an order of magnitude. Furthermore, the magnetic behavior changes from long-range ferrimagnetic ordering of 1 to canted antiferromagnetic behaviour of 1_dg.

In vivo tracking on longer retention of transplanted myocardin gene-modified adipose-derived stem cells to improve erectile dysfunction in diabetic rats.

BACKGROUND: Stem cell therapy has revealed a promising future for treating erectile dysfunction (ED), but the fate and curative mechanism of intracavernosal transplanted stem cells are under further exploration. This study aimed to demonstrate the effects of myocardin gene modification on improving erectile function and prolonging the retention of implanted adipose-derived stem cells (ASCs) using in vivo small animal imaging. METHODS: ASCs were isolated, cultured, and identified by flow cytometry and osteogenic and adipogenic induction. The effects of gene modification on cell proliferation, apoptosis, and contraction were determined by CCK-8, EdU, flow cytometry, and collagen gel lattice contraction assays as well as confocal microscopy. A total of 20 normal and 60 diabetes mellitus ED to (DMED) Sprague-Dawley rats were recruited to the 7 day and 21 day groups. Each group contained subgroups of 10 rats each: the negative control (NC), DMED + ASCs plus Ad-Luc-Myocardin, DMED + ASCs plus Ad-Luc, and DMED + phosphate buffer solution (PBS) groups. Erectile function was evaluated with the intracavernosal pressure/mean arterial pressure (△ICP/MAP) ratio. In vivo small animal imaging and an EdU cell tracking strategy were introduced to detect the transplanted ASCs, and IHC and WB were performed to assess smooth muscle cell protein levels. RESULTS: The ASCs expressed high CD29 and CD90 and scant CD45, while the multi-induction potential was verified by oil red O and alizarin red staining. Gene transfection of myocardin had no significant influence on ASC apoptosis but inhibited cell proliferation and promoted cell contraction. Myocardin combined with ASCs enhanced the therapeutic potential of ASCs for improving the △ICP/MAP ratio as well as α-SMA and calponin expression. In vivo imaging confirmed that ASCs resided within the cavernous body in 21 days, while only a few red EdU dots were detected. CONCLUSIONS: Myocardin induced ASC differentiation towards smooth muscle-like cells and enhanced the therapeutic potential of ASCs for ameliorating ED in STZ-induced diabetic rats. Notably, in vivo small animal tracking was an effective strategy for monitoring the implanted stem cells, and this strategy might have advantages over traditional EdU assays.

Effects of small interfering RNA targeting TLR4 on expressions of adipocytokines in obstructive sleep apnea hyponea syndrome with hypertension in a rat model.

We explored the effects of RNA interference-mediated silencing of TLR4 gene on expressions of adipocytokines in obstructive sleep apnea hyponea syndrome (OSAS) with hypertension in a rat model. Systolic blood pressure of caudal artery and physiological changes were observed when establishing rat models of OSAS with hypertension. Mature rat adipocytes were induced from separated and cultured primary rat adipocytes. To transfect rat mature adipocytes, TLR4 siRNA group and negative control (NC) siRNA group were established. Expressions of TLR4 mRNA of adipocytes were examined after silenced by siRNA by quantitative real-time polymerase chain reaction (qRT-PCR). By enzyme-linked immunosorbent assay (ELISA), expressions of inflammatory cytokines, and adipocytokines of adipocytes were detected. Blood pressure in rat caudal artery was higher in the intermittent hypoxia group than that of the blank control group by 29.87 mmHg, and cardiocytes in the former group showed physiological changes, which indicated successful establishment of rat models of OSAS with hypertension. Red particles could be seen in mature rat adipocytes when stained with Oil Red O. Transfection of TLR4 mRNA was significantly suppressed in the TLR4 siRNA group, which didn't happen in the untransfected control group. Rats in the TLR4 siRNA group had significantly reduced expressions of such inflammatory cytokines as interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) and such adipocytokines as visfatin, adiponectin (ADN), and leptin than those in the untransfected control group. RNA interference-mediated silencing of TLR4 gene could regulate occurrence and development of OSAS with hypertension in rats by downregulating expressions of adipocytokines.

Downregulated SOCS1 expression activates the JAK1/STAT1 pathway and promotes polarization of macrophages into M1 type.

Macrophage polarization is flexible, and involves in different signaling pathways and various transcription factors. Suppressor of cytokine signaling (SOCS) is an important inhibitor of cytokine signaling pathways and also a key physiological regulator for natural and acquired immunity systems. Following transfection of SOCS1 short hairpin (sh)RNA into mouse macrophage cells, reverse transcription­quantitative polymerase chain reaction demonstrated that the mRNA levels of Janus kinase (JAK)1 and signal transducer and activator of transcription (STAT)1 increased significantly. In addition, western blotting indicated that JAK1, STAT1 and p­STAT1 expression was significantly enhanced. Fludarabine can inhibit phosphorylation of STAT1 and SOCS1 expression. When fludarabine was added and SOCS1 shRNA was transfected, the inhibition of fludarabine was weakened, and p­STAT1 expression was upregulated. Flow cytometry detection indicated that, following the downregulation of SOCS1 expression, M1­type cells significantly increased, but the proportion of M2­type cells did not change significantly. Fludarabine can reduce the effect of SOCS1 shRNA on promoting M1­type cell polarization, and macrophages can polarize into both M1 and M2 phenotypes. Further ELISA results presented that, when downregulating SOCS1 expression, interleukin (IL)­4 and IL­10 expression was both downregulated, and tumor necrosis factor (TNF)­α and interferon (IFN)­Î³ expression was significantly upregulated. When adding fludarabine or injecting with the traditional Chinese medicine Xuebijing, IL­4 and IL­10 expression was both significantly upregulated, and TNF­α and IFN­Î³ expression was significantly downregulated. When adding fludarabine and downregulating SOCS1, IL­4, IL­10, TNF­α and IFN­Î³ expression presented no significant changes. The above results indicated that, when SOCS1 expression is downregulated, it will activate the JAK1/STAT1 pathway, and thereby promote the polarization of macrophages into M1 type. The findings are of great importance for understanding occurrence, development and treatment of various immune­related diseases.

Soluble Egg Antigen Activates M2 Macrophages via the STAT6 and PI3K Pathways, and Schistosoma Japonicum Alternatively Activates Macrophage Polarization to Improve the Survival Rate of Septic Mice.

Sepsis is one of the most challenging health problems worldwide. Our previous study showed that chronic schistosoma japonica (SJ) infection might increase serum anti-inflammatory factors to play a protective role, thus improving the survival rate of septic mice. Further research revealed that SJ infection promoted J774A.1 macrophage differentiation into M2 macrophages; suppressed LPS-induced activation of M1 macrophages; up-regulated CD163, IL-10, and TGF-ß1 expression; inhibited TNF-α and iNOS expression; and blocked the effect of LPS-promoted TNF-α and iNOS expression. Furthermore, adoptive transfer of ex vivo programed M2 macrophages significantly increased the survival rate of septic mice. In vitro studies suggested that soluble egg antigen (SEA) from SJ played the same role as worm infection but had no impact on M1 macrophages. SEA reduced LPS-induced TNF-α and iNOS expression, decreased the inhibitory effect of LPS on IL-10 and TGF-ß1 expression, increased STAT6 phosphorylation, and up-regulated PI3K and Akt expression but inhibited SOCS1 expression. When PI3K inhibitors were added, SEA-induced expression of CD163, IL-10, and arg1 might be reduced. Therefore, worm infection has a protective effect in septic mice in which SEA may play a key role via the STAT6 and PI3K pathways. This finding may provide a favorable solution for the treatment of sepsis, especially early cases. J. Cell. Biochem. 118: 4230-4239, 2017. © 2017 Wiley Periodicals, Inc.

Recombinant Trichinella spiralis 53-kDa protein activates M2 macrophages and attenuates the LPS-induced damage of endotoxemia.

Sepsis is a serious clinical condition of excessive systemic immune response to microbial infection. The pro-inflammatory stage of sepsis is generally launched by innate cells such as macrophages. They release inflammatory cytokines, activate other immune cells and cause severe tissue/organ damage. In this study, we have revealed that recombinant Trichinella spiralis (TS) excretory-secretory protein (rTsP53) exhibited anti-inflammatory properties and rescued mice from LPS-induced endotoxemia, which is a common model for sepsis study, potentially through the induction of M2 macrophages. rTsP53 treatment significantly decreased inflammatory cytokines (IL-6, IFN-γ and TNF-α) and increased IL-4, IL-10, IL-13 and TGF-ß secretion, both in circulation and in tissues. rTsP53 also induced the activation and infiltration of F4/80(+)CD163(+) macrophages to inflammatory tissues, increased M2 macrophage-related Arg1 and Fizz1 expression, and decreased M1 macrophage-related iNOS expression. PCR array showed that rTsP53 activated several genes that involve the survival of macrophages and also anti-inflammatory genes such as SOCS3. Together, our results show that rTsP53 activates M2 macrophages, which has strong anti-inflammatory potential to prevent LPS-induced lethal sepsis.

[Phenotypic modulation of bladder smooth muscle in diabetic rats].

OBJECTIVE: To investigate whether phenotypic modulation of bladder smooth muscle occurs in diabetic rats. METHODS: Thirty-two male SD rats were randomly assigned into diabetic group and control group. Diabetic rat models were established by a single intraperitoneal injection of streptozotocin (60 mg/kg). Nine weeks later, the bladder tissues of the rats were examined for structural changes using HE and Masson's trichrome staining , and the expressions of myocardin, α-SMA, and SMMHC in bladder smooth muscles were detected with RT-PCR and Western blotting. RESULTS: Compared with the control group, the diabetic rats showed obvious polydipsia and polyuria with significantly increased collagenous fibers and lowered expressions of myocardin, α-SMA, and SMMHC in the bladder tissue (P<0.05). CONCLUSION: s In rats at 9 weeks after diabetic model establishment, phenotypic transition of the bladder smooth muscles occurs to cause bladder contractile dysfunction, which may play an important role in the pathology of diabetic bladder dysfunction.

Micrometam C Protects against Oxidative Stress in Inflammation Models in Zebrafish and RAW264.7 Macrophages.

Micrometam C is a core of novel marine compound isolated from the mangrove associates Micromelum falcatum. In this study, we investigated the protective effects of micrometam C in inflammation models in the transgenic zebrafish line Tg (corola: eGFP) and RAW264.7 macrophages. We found that micrometam C significantly suppressed the migration of immune cells in tail-cutting-induced inflammation in transgenic zebrafish and reduced lipopolysaccharide (LPS)-induced reactive oxygen species (ROS) in both zebrafish and macrophages. In addition, micrometam C also restored LPS-induced reduction of endogenous antioxidants, such as catalase (CAT), glutathione (GSH) and superoxide dismutase (SOD). The protective effects of micrometam C were in parallel to its inhibition of NADPH oxidase and nuclear factor-kappa-binding (NF-κB) activity. Thus, the present results demonstrate that micrometam C protects against LPS-induced inflammation possibly through its antioxidant property.

Association between the PARP1 Val762Ala polymorphism and cancer risk: evidence from 43 studies.

BACKGROUND: Poly (ADP-ribose) polymerase-1 (PARP-1) plays critical roles in the detection and repair of damaged DNA, as well as cell proliferation and death. Numerous studies have examined the associations between PARP1 Val762Ala (rs1136410 T>C) polymorphism and cancer susceptibility; nevertheless, the findings from different research groups remain controversial. METHODS: We searched literatures from MEDLINE, EMBASE and CBM pertaining to such associations, and then calculated pooled odds ratio (OR) and 95% confidence interval (CI) by using random-effects model. The false-positive report probability (FPRP) analysis was used to confirm the validity of significant findings. Moreover, potential effects of rs1136410 variants on PARP1 mRNA expression were analyzed for three ethnicities by combining data from HapMap (genotype) and SNPexp (mRNA expression). RESULTS: The final meta-analysis incorporated 43 studies, consisting of 17,351 cases and 22,401 controls. Overall, our results did not suggest significant associations between Ala variant (Ala/Ala or Ala/Val genotype) and cancer risk. However, further stratification analysis showed significantly increased risk for gastric cancer (Ala/Ala vs. Val/Val: OR = 1.56, 95% CI = 1.01-2.42, Ala/Val vs. Val/Val: OR = 1.34, 95% CI = 1.14-1.58, dominant model: OR = 1.41, 95% CI = 1.21-1.65 and Ala vs. Val: OR = 1.29, 95% CI = 1.07-1.55). On the contrary, decreased risk for brain tumor (Ala/Val vs. Val/Val: OR = 0.77, 95% CI = 0.68-0.87, dominant model: OR = 0.77, 95% CI = 0.68-0.87 and Ala vs. Val: OR = 0.82, 95% CI = 0.74-0.91). Additionally, we found that the Ala carriers had a significantly increased risk in all models for Asians. Our mRNA expression data provided further biological evidence to consolidate this finding. CONCLUSIONS: Despite some limitations, this meta-analysis found evidence for an association between the PAPR1 Val762Ala and cancer susceptibility within gastric cancer, brain tumor and Asian subgroups.

Dexamethasone down-regulates the expression of microRNA-155 in the livers of septic mice.

To investigate the expression of microRNA-155 (miRNA-155) in the livers of mice with lipopolysaccharide (LPS)-induced sepsis and to determine the role of dexamethasone (DXM) in the regulation of miRNA-155 expression, we pretreated mice with or without DXM prior to LPS exposure. Our study demonstrated that the expression of miRNA-155 and inflammatory factors increased in the liver tissues of mice with LPS-induced sepsis and that DXM down-regulated their expression in a dose-dependent manner. Moreover, DXM alone inhibited the expression of miRNA-155 to below the baseline level, but did not impact the expression of inflammatory factors, suggesting that the down-regulation of miRNA-155 by DXM may partially, but not completely, depend on the suppression of pro-inflammatory cytokines by DXM. Our data indicate that the overexpression of miRNA-155 in the livers of mice with LPS-induced sepsis may play an important role in the pathological processes of sepsis and that the down-regulation of miRNA-155 by DXM may be a novel mechanism regulating inflammation and immunity.

[The research of establishing discriminant function for patients with angina pectoris by stepwise analysis based on serum inflammatory factors].

OBJECTIVE: To improve cost-efficiency, discriminant functions in stepwise method was founded for the differential diagnosis of angina pectoris by detecting the serum level of high-sensitivity C-reactive protein (hs-CRP), macrophage migration inhibitory factor (MIF), interleukin-4 (IL-4) and interleukin-10 (IL-10) in patients with stable angina pectoris (SAP) and unstable angina pectoris (UAP). METHODS: Thirty-nine SAP patients and 47 UAP patients were enrolled into the study, while 39 healthy volunteers were enrolled into the controlled group forming the entire set of training samples. The serum levels of hs-CRP, MIF, IL-4 and IL-10 were measured by enzyme linked immunosorbent assay (ELISA). Data was analyzed by software to define discriminant functions in the ways of "entering" and "stepwise". Both functions were evaluated by the results of validation. RESULTS: By the way of "enter independent together", the following discriminant functions were defined based on the data of training samples' age, hs-CRP, MIF, IL-4, IL-10: healthy control group =-129.858 + 2.869×age -2.451×hs-CRP + 1.393×MIF + 6.001×IL-4 + 4.848×IL-10; SAP group=-161.037 + 2.896×age-2.022×hs-CRP + 1.662×MIF + 6.703×IL-4 + 6.287×IL-10; UAP group=-199.087 + 2.468×age-1.440×hs-CRP + 3.404×MIF-13.875×IL-4 + 7.752×IL-10. Retrospective validation showed 4.8% of total miss-grouping, while cross-validation showed 5.6% of total miss-grouping. By the way of "stepwise", the above data was screened by software and training samples' age, MIF and IL-10 were suggested to define the following functions: healthy control group = - 125.218 + 2.659 × age + 0.599×MIF + 5.040 × IL-10; SAP group=-157.864 + 2.721×age + 1.008×MIF + 6.468×IL-10; UAP group=- 197.327 + 2.360×age + 2.932×MIF + 7.640×IL-10. Both retrospective and cross validation showed 6.4% of total miss-grouping. Both sets of discriminant functions had the same efficiency (100%) for differential diagnosis of SAP and UAP. CONCLUSION: The discriminant functions based on samples' age, MIF and IL-10, which were screened and suggested by stepwise method, may contribute to the differential diagnosis of atypical SAP and UAP, and therefore demonstrate better cost-efficiency.

[Expression and effects of microRNA-155 in the livers of septic mice].

OBJECTIVE: To evaluate the effects of microRNA-155 (miR-155) on liver injury in mice with sepsis. METHODS: One hundred and twenty BALB/c mice were randomly divided into two groups of equal number according to random number table. Sepsis was induced by intraperitoneal injection of lipopolysaccharide (LPS,20 mg/kg). The mice were sacrificed at the time-points of 0, 2, 6, 12, 24, 48 hours. Blood and liver tissue were collected, and the levels of tumor necrosis factor- α (TNF- α ), interleukin (IL-6, IL-10) in serum and liver homogenate and alanine transaminase (ALT) in serum were determined by enzyme linked immunosorbent assay(ELISA). The injury of liver tissue was evaluated by histopathology. The expression of miR-155 in liver tissue was assessed by fluorescent quantitation reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The levels of TNF- α , IL-6 and IL-10 in serum and liver homogenate of septic mice increased with passage of time, and then the levels of TNF-α and IL-6 lowered after reaching the peak value, but remained higher than that of control group.TNF-α (ng/L) reached the peak value at 2 hours post-LPS-injection (serum: 1538.46 ± 102.12 vs. 64.52 ± 18.44,liver homogenate: 255.26 ± 41.23 vs. 60.21 + 13.55, both P<0.05). The level of IL-6 (µg/L) reached the peak value at 6 hours post-LPS-injection (serum: 875.33 ± 102.37 vs. 153.72 ± 20.67, liver homogenate: 9.22 + 0.82 vs. 3.35 ±0.36, both P<0.05), and that of IL-10 (ng/L) reached the peak value at 48 hours post-LPS-injection (serum: 520.13 ± 88.52 vs. 23.43 3.01, liver homogenate: 260.12 + 50.38 vs. 16.37 ± 3.71, both P<0.05). There were significant differences in above indexes between septic and control group (all P<0.05). The serum level of ALT (U/L) rose with passage of time, reaching the peak value at 48 hours post-LPS-injection (603.26 + 70.21 vs. 45.84 + 5.64, P<0.05). The values showed significant differences between septic and control group (P<0.05). A large number of leucocytic infiltration was found in liver. Hepatic tissue showed architectural distortion. Hepatocyte vacuolation and nodular necrosis were obvious at 12 hours post-LPS-injection. Relative expression of miR-155 was found to be increased at 2 hours post-LPS-injection, reaching its peak value at 12 hours post-LPS-injection [(72.96 ± 9.34)-fold of control group, P<0.05]. CONCLUSION: The increase in miR-155 expression might play an important role in the mechanism of liver injury during sepsis.

[The protective effect of chronic schistosoma japonica infestation against sepsis in mice].

OBJECTIVE: To preliminarily study the protective effect of chronic schistosoma japonica (SJ) infestation against sepsis in mice and its mechanism. METHODS: BALB/c male mice were used, and the experiment was divided into three parts. Experiment 1: chronic SJ infestation model was reproduced by SJ cercaria inoculation through abdominal skin for 8 weeks. Twenty mice were randomly grouped into normal group (n=10) and SJ group (n=10). The levels of interleukins (IL-4, IL-10),tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in serum were detected by enzyme linked immunosorbent assay (ELISA). Real-time polymerase chain reaction (PCR) was employed to detect the levels of IL-10 mRNA and TNF-αmRNA in abdominal macrophages. This experiment was meant to evaluate immune state in mice with chronic SJ infestation. Experiment 2: lipopolysaccharide (LPS) was intraperitoneally injected to reproduce sepsis model. Thirty mice were randomly grouped into LPS group (n=15) and SJ-LPS group (n=15). The levels of cytokines were determined by ELISA at 0, 24, 48 and 72 hours after LPS injection. This experiment was meant to detect the effect of chronic SJ infestation in mice during the septic process. Experiment 3: two types of sepsis model were reproduced by cecal ligation and puncture (CLP) and LPS injection, respectively. The survival rate of mice with chronic SJ infestation in 72 hours in either type of sepsis was evaluated. RESULTS: Experiment 1: compared with normal group [IL-4 (56.32±8.66) ng/L, IL-10 (48.17±7.23) ng/L], chronic SJ infestation showed an increase in serum IL-4 [(151.35±12.24) ng/L] and IL-10 [(133.22±11.09) ng/L, both P<0.05]. Chronic SJ infestation also resulted in an increase in IL-10 mRNA expression (SJ group 4.46±1.82, normal group 1.52±0.60) and inhibited TNF-α mRNA expression (SJ group 1.61±0.93, normal group 2.32±1.03) in abdominal macrophages (both P<0.05), indicating that macrophages could be differentiated into alternative activated macrophages. Experiments 2 and 3 showed that the levels of serum IL-4 and IL-10 were increased at 0 hour after LPS injection, and then gradually decreased in SJ-LPS group, but the levels were still higher than those in LPS group at 72 hours [IL-4 (ng/L): 92.2±7.6 vs. 41.5±4.5; IL-10 (ng/L): 92.1±7.8 vs. 35.6±4.0, both P<0.05]; the levels of TNF-α and IFN-γ were increased at 24 hours, and then decreased in SJ-LPS group, and the levels were lower than those in LPS group at 72 hours [TNF-α (ng/L): 82.9±5.6 vs. 91.5±5.2; IFN-γ (ng/L): 44.1±4.8 vs. 52.6±4.0, both P<0.05]. Therefore, chronic SJ infestation could improve the survival rate of mice with sepsis induced by CLP or LPS (CLP: 80% vs. 20%, LPS: 70% vs. 30%, both P<0.05). CONCLUSION: Chronic SJ infestation could elevate anti-inflammatory factors in septic mice, thus ameliorating the survival rate, so it has protective effect on mice with sepsis.

[Polymorphism and structural analysis of suppressor of cytokine signaling-1 of rat].

OBJECTIVE: To identify the suppressor of cytokine signaling-1 (SOCS-1) of rat from the amplified gene with the help of bioinformatics to predict the deduced protein's structure and function in order to lay the foundation for further theoretical study. METHODS: The full-length rat SOCS-1 gene was amplified and identified from the GeneBank Nucleotide database, and the corresponding structure and function of its deduced protein was predicted by the bioinformatics analyzing tools online and the complicated bioinformatics software package Vector NTI suite 8.0, meanwhile the molecular cladogram was reconstructed. RESULTS: Two sequences were obtained by polymerase chain reaction (PCR) amplification of different SOCS-1 gene. The gene was comprised of 639 base pairs in the length, deduced 212 amino acids (aa), contained a SOCS box (aa172-aa208), a SH2 domain (aa80-aa155) and a nuclear localization sequence (aa160-aa174). The primary structure contained two linear B cell epitopes, all of them were on the surface of the protein and far away from the spatial structure. CONCLUSION: SOCS-1 gene has a polymorphism, its conservative SH2 region and SOCS box are related to its inhibitory effect on the signal transduction pathway. The nucleic localization sequence may affect other nuclear transduction factors. The B cell linear epitopes may be a candidate of immunodiagnosis with promising prospects.

[The significance of the changes in expressions of heat shock protein 70 and heme oxygenase-1 in the myocardium in patients died suddenly due to acute myocardial infarction].

OBJECTIVE: To investigate the changes and its clinical significance of the expressions of the mRNA and protein of heat shock protein 70 (HSP70) and heme oxygenase-1 (HO-1) genes in the myocardium of acute myocardial infarction (AMI) areas in patients who died suddenly due to AMI. METHODS: Specimens of myocardial tissue at infarct site was obtained during autopsy from 18 patients who died suddenly due to AMI, and they were enlisted as study group, and that of myocardial tissue from 17 normal hearts of patients died rapidly after accidents were as control group. The levels of mRNA expression of HSP70 and HO-1 genes were measured in all the specimens by reverse transcription-polymerase chain reaction (RT-PCR) using cDNA samples, and the levels and locations of HSP70 and HO-1 protein expressions in myocardial cells of all specimens were examined by immunohistochemistry (IHC), and the pathological changes in the myocardial tissue were observed. RESULTS: The expressions of HSP70 mRNA (0.841+/-0.058), HO-1 mRNA (0.918+/-0.161) and HSP70 protein (3 556.68+/-574.19), HO-1 protein (4 336.68+/-865.30) in the myocardium of the AMI areas in the study group were significantly higher than those in control group (the expressions of HSP70 mRNA: 0.105+/-0.034, HO-1 mRNA: 0.086+/-0.053, HSP70 protein: 289.21+/-68.51, HO-1 protein: 1 556.78+/-506.26, all P<0.01), and a weak expression of HSP70 and HO-1 protein was found in the cardiocytes of the AMI area in study group. In the control group, HSP70 protein expression was negative in the cardiocytes, and there was a weak expression of HO-1 protein in the cardiocytes. There was a significant positive correlation between the expressions of mRNA and protein of HSP70 and HO-1 genes at the cardiocytes of patients died suddenly of AMI in study group (r(1)=0.865, r(2)=0.816, both P<0.01). CONCLUSION: HSP70 and HO-1 probably were both involved in the pathological and physiological processes of AMI, while HO-1 may express in cardiocytes, but it is more abundant in the cardiocytes in the AMI area.

Expressions of SOCS-1 and SOCS-3 in the myocardium of patients with sudden cardiac death.

BACKGROUND: As the regulators of cytokines, suppressors of cytokine signaling (SOCS) play an important role in the inflammation reaction. Some studies found that SOCS-1 and SOCS-3 were involved in the pathogenesis of some inflammatory diseases such as rheumatoid arthritis, inflammatory bowel disease. But the expressions of SOCS in coronary heart disease have not yet been reported. This study aimed to investigate the expression and clinical significance of SOCS-1 and SOCS-3 in the myocardium of patients with sudden cardiac death (SCD). METHODS: Myocardial autopsy specimens were collected from 24 patients at the Forensic Medicine Department of Sun Yat-Sen University, Guangzhou, China between 2005 and 2006. Of them, 9 patients had autopsy findings consistent with coronary atherosclerosis (non-myocardial infarction) leading to SCD (non-MI group), 7 died of acute myocardial infaction (MI group), and 8 died from traffic accidents and trauma (control group). The expressions of SOCS-1 mRNA and SOCS-3 mRNA in the myocardium of the non-MI, MI and control groups were detected using RT-PCR. The levels of SOCS-1 and SOCS-3 proteins were detected using immunohistochemistry. Statistical analyses were performed using SPSS version 13.0 software and the data were analyzed by ANOVA. RESULTS: The expressions of SOCS-1 mRNA and SOCS-3 mRNA in the non-MI and MI groups were significantly higher than those in the control group[(0.788±0.101), (0.741±0.111) vs. (0.436±0.044), (P<0.01); (0.841±0.092), (0.776±0.070) vs. (0.454±0.076), (P<0.01)] respectively. The antibody-positive cells of SOCS-1 protein in the myocardium of the non-MI and MI groups were significantly higher than those in the myocardium of the control group[(320.00±48.48), (347.14±70.88) vs. (42.50±10.35), (P<0.01)] respectively. The antibody-positive cells of SOCS-3 protein in the myocardium of the non-MI and MI groups were significantly higher than those in the myocardium of the control group[(381.11±59.25) vs. (40.00±10.69), (P<0.01)] and[(332.86±111.91) vs. (40.00±10.69), (P=0.001)]. CONCLUSION: The expressions of SOCS-1 and SOCS-3 in the myocardium of patients with SCD from coronary heart disease are significantly increased and contribute to the pathogenesis of SCD.

[Study of the effect of p53 and Bcl-2 protein expression in myocardial apoptosis in septic rats].

OBJECTIVE: To explore the significance of p53 and Bcl-2 gene in myocardial apoptosis in septic rats. METHODS: Adult male Sprague-Dawley (SD) rats were randomly divided into normal control group (n=6), sham operation group (n=6) and cecal ligation and puncture (CLP) group (n=24). The sepsis model was reproduced by CLP. Rats in CLP group were divided into four subgroups according to the time points of 3, 9, 12, and 24 hours after operation, with 6 rats in each subgroup, and their hearts were examined according to the experimental protocol. Myocardial apoptosis was detected with electron microscopy and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) technique. Bcl-2 and p53 protein expression was measured by immunohistochemistry method. RESULTS: Myocardial apoptosis index (AI) in septic rats were increased, higher than that in normal control group [(1.500+/- 0.141)%] and sham operation group [(1.567+/-0.258)%, all P<0.05]. The myocardial AI peaked at 12 hours [(55.633+/-2.073)%], and lowered at 24 hours [(33.683+/-2.070)å]. The expression level of p53 protein in CLP group was lowest at 3 hours [(13.817+/-0.964)%], peaked at 12 hours [(80.567+/-5.055)%], and higher than that in normal control group [(0.617+/-0.232)%] and sham operation group [(0.600+/-0.297)%, all P<0.05 ]. The trend was parallel with that of the results of TUNEL. However, the expression level of Bcl-2 protein peaked at 3 hours [(31.650+/-1.799)%], and was lowest at 12 hours [(0.650+/-0.308)%], and lower than that in normal control group [(47.017+/-0.691)%] and sham operation group [(46.817+/-0.567)%, all P<0.05]. The trend was opposite with that of the results of TUNEL. CONCLUSION: Myocardial apoptosis may be a mechanism of the pathogenesis of myocardial injury in sepsis. The change in the modulating genes p53 and Bcl-2 of apoptosis may serve as indicators of myocardial injury. p53 and Bcl-2 gene may be used as an interventional means in sepsis to change its outcome.

[The expression and clinical implication of advanced oxidized protein products in patients with multiple organ dysfunction syndrome].

OBJECTIVE: To investigate the expression and clinical implication of advanced oxidized protein products (AOPP) in patients with multiple organ dysfunction syndrome (MODS). METHODS: Serum concentrations of C-reactive protein (CRP) and AOPP were determined in 180 patients with systemic inflammatory response syndrome (SIRS) or MODS (90 patients, respectively). The acute physiology and chronic health evaluation III (APACHE III) scoring system was applied to assess severity of patients' condition. The contents of serum CRP and AOPP in MODS group, SIRS group and normal control group, and also in survivor and dead patients in MODS group were determined and compared. The correlation between CRP and AOPP levels and the correlation between AOPP levels and severity of MODS were also observed. Ninety healthy volunteers who matched with study subjects in age and gender comprised the normal control group. RESULTS: The CRP [(22.22+/-4.32) mg/L] and AOPP [(130.66+/-18.08) micromol/L] levels in patients with MODS were significantly higher than those in normal control group [(2.38+/-0.89) mg/L and (33.20+/-5.32) micromol/L, respectively] and SIRS group [(5.32+/-1.22) mg/L and (48.58+/-6.03) micromol/L, respectively, all P < 0.05], and were positively correlated with APACHE III scores [(98.66+/-20.87) scores] of the patient (r1 = 0.469, r2 = 0.528, both P < 0.01). However, there was no significant difference between SIRS group and normal control group. The CRP and AOPP levels were found to be significantly higher in the patients who eventually died (47 cases) as compared to those in the patients who survived (43 cases, both P < 0.05). Positive correlations were noted between AOPP and CRP level (r = 0.448, P < 0.01). The serum concentrations of CRP and AOPP levels were elevated with the increase of the number of failed organs in MODS patients(all P < 0.05). CONCLUSION: The data show that AOPP might participate in the process of pathogenesis of MODS. The serum AOPP level may be taken as a diagnostic and prognostic indicator for MODS.

[Study on the relationship between the expression of SOCS-1/3 in the peripheral blood mononuclear cells in patients with multiple organ dysfunction syndrome and their prognosis].

OBJECTIVE: To observe the mRNA and protein expression of suppressor of cytokines signaling protein 1/3 (SOCS-1/3) in the peripheral blood mononuclear cells (PBMCs) in patients with multiple organ dysfunction syndrome (MODS), and to explore the relationship between the SOCS expression and the patients' prognosis. METHODS: Peripheral blood specimens of 32 patients with MODS and 24 healthy volunteers (controls) were collected and the PBMCs were isolated by centrifugation and Ficoll-Hypaque sedimentation. The mRNA of SOCS-1/3 was determined by reverse transcription-polymerase chain reaction (RT-PCR). And the protein content of SOCS-1/3 was determined by Western blotting. The relationship between the SOCS expression and the patients' prognosis was analyzed. RESULTS: There was no significant difference in the SOCS-1 mRNA expression between control group and MODS group (0.526+/-0.044 vs. 0.466+/-0.047, P>0.05). The SOCS-1 expression of MODS group was significantly higher than that of control group (0.814+/-0.045 vs. 0.479+/-0.021, P<0.05). In the MODS group, the SOCS-1 mRNA expression and protein content of dead patients were significantly lower than those of survived patients (mRNA 0.487+/-0.032 vs. 0.532+/-0.028, protein 0.787+/-0. 029 vs. 0.838+/-0.040, both P<0.05). There was significant negative correlation between the SOCS-1 mRNA expression and the MODS score (r1=-0.731,P<0.01). There was also significant negative correlation between the SOCS-1 content and the MODS score (r2=-0.526,P<0.01). The SOCS-3 mRNA expression of MODS group was higher than that of control group (0.993+/-0.415 vs. 0.461+/-0.046, P<0.05). The SOCS-3 content in the PBMCs of MODS groups was significant higher than that of control group (0.458+/-0.033 vs. 0.403+/-0. 024, P<0.05). In the MODS group, the SOCS-3 mRNA expression and protein content of dead patients were significant higher than those of survived patients (mRNA 1. 245+/-0.408 vs. 0.805+/-0.326, protein 0.486+/-0.021 vs. 0.425+/-0.016, both P<0.05). There was positive correlation between the SOCS-3 mRNA expression and the MODS score but the correlation was not significant (r=0. 468, P>0.05). And there was significant positive correlation between the SOCS-3 content and the MODS score (r=0.783, P<0.01). CONCLUSION: SOCS-1 may protect the tissue from further damage while SOCS-3 may cause tissue damage indirectly. The detection of SOCS-1/3 may help to predict the prognosis of MODS.