PM2.5-induced inflammation and myocardial cell injury in rats.

OBJECTIVE: Numerous studies show association of particular matter (PM) in air pollution with cardiovascular dysfunction, and increased morbidity and mortality. The main mechanisms of this adverse effect involve increasing oxidative stress, inflammatory responses, and genotoxicity. Several recent studies investigated the ability of PM2.5 to cause myocardial injury in animal models using various methods, such as intratracheal instillation, intraperitoneal injection or tail vein injection. The purpose of this study is to explore the PM2.5-induced myocardial inflammatory reaction in rats through the new technology of multi-functional aerosol concentration and enrichment system. MATERIALS AND METHODS: Thirty Wistar rats were divided into two groups, 15 in each group. In the exposure group, PM2.5 multi-functional aerosol concentration and enrichment system was used for PM2.5 online oral and nasal exposure (5 times a week, 4 hours exposure, for the duration of 3 months). Histopathological examination of the left ventricular myocardial tissue of both groups was done using hematoxylin and eosin (H&E) staining. Ultrastructural changes of the heart specimens were assessed using electron microscopy. The levels of CRP and ICAM-1 were detected by immunohistochemistry. RESULTS: Compared with the control group, myocardial tissue of the exposure group exhibited edema, widened myocardial space and infiltration of inflammatory cells. There was nuclear pyknosis, mitochondrial membrane and spinal fusion, rough endoplasmic reticulum expansion, degranulation and cell swelling in the exposed group. The area of CRP positive staining in the exposed group was 3.7-fold higher than that in the control group (p < 0.05), and the ICAM-1 positive staining area of the exposed group was 12-fold higher than that of the control group (p < 0.05). CONCLUSIONS: Prolonged exposure to PM2.5 inhalation promotes significant upregulation of ICAM-1 and CRP expression in myocardial tissues, ultrastructural alterations in myocardial cells, and influx of inflammatory cells.

MicroRNAs target on cartilage extracellular matrix degradation of knee osteoarthritis.

OBJECTIVE: Knee osteoarthritis (KOA) is currently indicated to be characterized by destruction of articular cartilage. The destruction can be described as an imbalance between synthesis and degradation of extracellular matrix (ECM) components. It is accompanied with changes of pro-inflammatory cytokines and degradation enzymes dominated by matrix metalloproteinase (MMP) and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS). Recent studies have revealed that microRNAs (miRNAs) are associated with synthesis and degradation of extracellular matrix (ECM). They play an important role in articular cartilage homeostasis of knee osteoarthritis (KOA). The related mechanisms include mediating the relevant enzymes and pro-inflammatory cytokines. The aim of this study is to reveal the potential microRNAs (miRNAs) and their corresponding upstream or downstream targeting on cartilage extracellular matrix (ECM) degradation in knee osteoarthritis (KOA). MATERIALS AND METHODS: 7 databases were extensively searched with a theme of MicroRNAs (miRNAs) and knee osteoarthritis (KOA). The articles were searched regardless of publication status and language. The databases include PubMed, Cochrane Library, EMbase, China Biology Medicine (CBM), China National Knowledge Infrastructre (CNKI), WanFang Data and Chinese Scientific Journal Database (VIP). RESULTS: This article reviews the microRNAs (miR-140, miR-146a, miR-25, miR-543, miR-19, miR-125b, miR-92a, miR-27b, miR-448, miR-558, miR-155) and their corresponding upstream or downstream in mediating cartilage extracellular matrix (ECM) degradation. CONCLUSIONS: MicroRNAs (miRNAs) have been involved in the pathogenesis of KOA. They can directly regulate cartilage homeostasis by targeting on ECM degradation via corresponding upstream/downstream.

Inhibition of microRNA-802-5p inhibits myocardial apoptosis after myocardial infarction via Sonic Hedgehog signaling pathway by targeting PTCH1.

OBJECTIVE: The incidence of acute myocardial infarction (AMI) has increased significantly in recent years, seriously threatening human life and health. This paper focused on the role of microRNA-802-5p (miR-802-5p) in myocardial infarction (MI) and its underlying mechanisms. MATERIALS AND METHODS: Quantitative Real-Time Polymerase Chain Reaction (RT-PCR) was employed to detect miR-802-5p expression. Western blot was performed to detect protein expression. Flow cytometry and terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining were performed to observe myocardial apoptosis. Hematoxylin-eosin (HE) staining was used to observe the morphology of myocardial tissue. The cardiac function of rats was detected using echocardiography. RESULTS: The expression of miR-802-5p was increased in hypoxic-treated H9c2 cells and infarcted myocardium in MI rats. Hypoxia treatment reduced the viability of cardiomyocytes and increased the level of lactate dehydrogenase (LDH) in the cell supernatant. Hypoxia treatment increased Bax expression in myocardial cells while Bcl-2 expression decreased, and the number of apoptotic cells increased. MiR-802-5p silencing reversed these effects. Moreover, miR-802-5p silencing reduced myocardial damage in MI rats, and significantly improved cardiac function. Through the Luciferase activity assay, we proved that miR-802-5p could directly target PTCH1. The knockdown of PTCH1 reversed the protective effect of miR-802-5p silencing on hypoxic myocardium. CONCLUSIONS: MiR-802-5p expression was increased in hypoxia-treated H9c2 cells and infarcted myocardium in MI rats. MiR-802-5p silencing could inhibit apoptosis after MI via activating Sonic Hedgehog signaling pathway by targeting PTCH1, thereby reducing myocardial injury and improving cardiac function of MI rats.

Association of major adverse cardiovascular events and cardiac troponin-I levels following percutaneous coronary intervention: a three-year follow-up study at a single center.

OBJECTIVE: Major adverse cardiovascular events occurrences of patients with different cardiac troponin-I (cTnI) levels following percutaneous coronary intervention (PCI) remained controversial. The prognostic relevance and risk factors of PCI-related myocardial infarction (MI) were not very clear as well. PATIENTS AND METHODS: Our study included 249 coronary artery disease patients without preoperative cTnI elevation who successfully accepted PCI from 2013 to 2014. A three-year follow-up was conducted for each patient. The patients were divided into PCI-related MI group and non-PCI-related MI group. Risk factors of PCI-related MI were first explored. The occurrence of MACE was recorded. The prognostic relevance between PCI-related MI (PMI) group and non-PCI-related MI group, as well as different postoperative cTnI levels, were compared. RESULTS: Low-density lipoprotein cholesterol (LDL-C), age, Gensini Score, total stent length, and intra-operative complication were found positively correlated with PCI-related MI occurrence, while hemoglobin and prior PCI history were negatively correlated. After 3-year follow-up, the Kaplan-Meier survival curve showed MACE occurrence was significantly increased in PCI-related MI group. Comparing to patients with normal postoperative cTnI, MACE occurrence was increased in patients with a 10×upper limit of normal (ULN)≤cTnI<70×ULN and cTnI≥70×ULN, while there was no difference in patients with 1×ULN≤cTnI<5×ULN and 5×ULN≤cTnI<10×ULN. Cox proportional hazard regression analysis revealed PMI, NT-proBNP, and left ventricular ejection function (LVEF)<50% were positively correlated with MACE occurrence, while maximum inflation pressure and apoA-I were negatively correlated. CONCLUSIONS: Prognosis of PCI-related MI was poor, as well as in patients with postoperative cTnI≥10×ULN. Among the risk factors of PMI, LDL-C, age, Gensini Score, total stent length, and intra-operative complication were positively correlated with PCI-related MI occurrence, while hemoglobin and prior PCI history were negatively correlated.

MicroRNA-150 alleviates acute myocardial infarction through regulating cardiac fibroblasts in ventricular remodeling.

OBJECTIVE: The aim of this study was to investigate the effect of microRNA-150 on the regulation of myocardial fibrosis and ventricular remodeling in rats with acute myocardial infarction (AMI). MATERIALS AND METHODS: The AMI rats model was established by the ligation of the left anterior descending coronary artery (LAD) in vivo. After AMI procedures, the rats were injected with microRNA-150 lentivirus overexpression or negative control, respectively. Cardiac function of rats was evaluated by echocardiography. Hematoxylin and eosin (HE) staining and Masson trichrome were performed to evaluate myocardial fibrosis in each rat. Meanwhile, cardiomyocyte apoptosis was detected by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) method. The expression levels of microRNA-150, col1α1, col1α2, col3 and α smooth muscle actin (α-SMA) in the border zone of rat infarct myocardium were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot, respectively. RESULTS: MicroRNA-150 expression in the border zone of infarct myocardium decreased significantly at day 28 after AMI (p<0.05). Overexpressing microRNA-150 significantly improved cardiac function, decreased collagen volume fraction (CVF) and attenuated cardiomyocyte apoptosis in rats. Furthermore, the expression levels of col1ɑ1, col1ɑ2, col3 and α-SMA in the border zone of infarct myocardium were remarkably down-regulated in rats overexpressing microRNA-150 compared with those of controls (p<0.001). CONCLUSIONS: MicroRNA-150 expression in the border zone of rat infarct myocardium decreased at day 28 after AMI. In addition, the upregulation of microRNA-150 in myocardial tissue could inhibit myocardial fibrosis and improve ventricular remodeling at post-AMI.

LncRNA PEG10 aggravates cardiac hypertrophy through regulating HOXA9.

OBJECTIVE: To uncover the role of long non-coding RNA (lncRNA) PEG10 in the progression of cardiac hypertrophy by regulating HOXA9. MATERIALS AND METHODS: In vivo cardiac hypertrophy model was established by performing transverse aortic constriction model (TAC) procedures in mice. Relative levels of PEG10, ANP and BNP in mice undergoing TAC procedures or sham operations were determined. In vitro cardiac hypertrophy model was established by phenylephrine (PE) treatment in primary cardiomyocytes. Relative levels of PEG10, ANP and BNP in cardiomyocytes were determined as well. Regulatory effects of HOXA9 on surface area of cardiomyocytes and relative levels of ANP and BNP were assessed. Finally, potential influences of PEG10/HOXA9 regulatory loop on cell surface area and relative levels of ANP and BNP were explored. RESULTS: Compared with mice in sham group, those in TAC group presented higher levels of PEG10, ANP and BNP. PE treatment markedly upregulated PEG10, ANP and BNP in primary cardiomyocytes, which were downregulated by transfection of si-PEG10. Besides, surface area of cardiomyocytes was enlarged by PE treatment, which was reduced after silence of PEG10. Silence of HOXA9 presented a similar effect as that of PEG10 in cardiomyocytes. Transfection of si-HOXA9 reversed the expanded cell surface area, and upregulated ANP and BNP in cardiomyocytes overexpressing PEG10. CONCLUSIONS: PEG10 is upregulated in hypertrophic cardiomyocytes. PEG10 aggravates cardiac hypertrophy by positively regulating HOXA9.

Effects of human umbilical cord mesenchymal stem cells-derived exosomes on fracture healing in rats through the Wnt signaling pathway.

OBJECTIVE: To investigate the role of human umbilical cord mesenchymal stem cell (hucMSC)-derived exosomes in the Wnt signaling pathway and their effects on fracture healing in rats. MATERIALS AND METHODS: A total of 24 healthy male Sprague-Dawley (SD) rats were randomly divided into 3 groups, of which the experimental groups were injected with Phosphate-Buffered Saline (PBS) and hUCMSC-derived exosomes, respectively, at the fracture site, and a blank control group was set. At 2 and 3 w after treatment, respectively, the healing condition at the fracture site in the rats was detected by micro-computed tomography (CT). The protein expressions of ß-catenin and Wnt3a of the Wnt signaling pathway in the bone tissue were measured via Western blotting (WB) assay. Quantitative Real Time-fluorescence Polymerase Chain Reaction (qRT-PCR) was performed to determine the expressions of osteogenic marker genes [collagen type I (COL-1), osteopontin (OPN) and runt-related transcription factor 2 (RUNX2)]. RESULTS: The results of the micro-CT scan showed that the rats treated with exosomes had better apposition of the fracture site, and the appearance of cortical bone was continuous. The fracture sites in the blank control group and PBS injection group were not healed, and the appearance of cortical bone was discontinuous, with significant fracture lines. According to the WB results, the protein expression levels of ß-catenin and Wnt3a in exosome treatment group were significantly higher than those in the blank control group and PBS injection group (p<0.01). The qRT-PCR results indicated that the expression levels of COL-1, OPN and RUNX2 in exosome treatment group were increased evidently compared with those in the other two groups (p<0.01). CONCLUSIONS: HucMSC-derived exosomes are probably involved in the repair of fracture in rats through the Wnt signaling pathway.

Glucose consumption and alpha-glucosidase inhibitory activities of aqueous root extract of Helicteres angustifolia.

OBJECTIVE: The root of Helicteres angustifolia L. (Sterculiaceae) has been used as tea to treat diabetics effectively by local people in Laos. However, no scientific evidence is available for this ethnomedicinal usage. This study was undertaken to explore the hypoglycemic effect of aqueous extract from Helicteres angustifolia root. PATIENTS AND METHODS: The effect of aqueous extract from Helicteres angustifolia root on glucose consumption in C2C12 myotubes was investigated at a dose of 25, 50 and 100 µg/mL, respectively. The alpha-glucosidase inhibitory activity of the extract was evaluated using rat intestinal maltase and sucrose. Moreover, oral sucrose tolerance test (OSTT) in normal and streptozotocin induced diabetic rats was performed. Finally, their cytotoxicity in C2C12 cells and acute oral toxicity in rats was analyzed. RESULTS: Aqueous root extract of Helicteres angustifolia significantly enhanced glucose consumption in C2C12 myotubes. The extract also significantly inhibited rat intestinal maltase (IC50 = 1.44 ± 0.24 mg/mL) and sucrase activity (IC50 = 0.54 ± 0.12 mg/mL), respectively. The OSTT results showed that the extract significantly suppressed the increase of blood glucose levels in normal and diabetic rats. The extract was also proven to have low acute toxicity (LD50 > 5 g/kg) and low cytotoxicity in C2C12 cells (IC50 > 0.4 mg/mL). CONCLUSIONS: The findings from this study indicate that aqueous root extract of Helicteres angustifolia possesses significant alpha-glucosidase inhibitory activity and moderate enhanced glucose consumption activity, while with low cytotoxic and acute toxicity.

DLC-1 is an independent prognostic marker and potential therapeutic target in hepatocellular cancer.

BACKGROUND: The 5-year survival rate of patients with hepatocellular cancer (HCC) was very low because of invasion and metastasis in the early stage. Biomarkers might help predict early occurrence of invasion and metastasis. Accumulating evidence has shown that deleted in liver cancer-1 (DLC1) may be considered as a metastasis suppressor gene in numerous solid and hematological cancers. However, its prognostic role and mechanisms that regulate and coordinate these activities remain poorly understood. METHODS: With the method of immunohistochemistry, the expression of DLC-1 as well as Rho A, ROCK2, moesin had been characterized in 80 HCC tissues and adjacent noncancerous tissues. The correlation between their expression and their relationships with clinicopathological characteristics of HCC were also investigated. In addition, the prognostic value of DLC1 expression within the tumor tissues was assessed by Cox regression and Kaplan-Meier analysis. RESULTS: DLC1 expression was significantly lower in HCC tissues than in adjacent noncancerous tissues, and DLC-1 expression was found to be negatively correlated with tumor differentiation, TNM stage and lymph node metastasis. Furthermore, DLC-1 expression was found to inversely correlate with Rho A, ROCK2 and moesin which were all highly expressed in HCC tissues. Kaplan-Meier analysis showed that significantly longer 5-year survival rate was seen in HCC patients with higher DLC1 expression, compared to those with lower expression of DLC1. Multivariate Cox proportional hazard analyses revealed that DLC1 was an independent factor affecting the overall survival probability. CONCLUSION: DLC1 could be served as a tumor suppressor gene in the progression especially in the invasion and metastasis of HCC. DLC1 perhaps played its role by regulating the expression of Rho A, ROCK2 and moesin. Evaluation of the expression of DLC-1 might be a good prognostic marker for patients with HCC.

Investigation of modified platelet-rich plasma (mPRP) in promoting the proliferation and differentiation of dental pulp stem cells from deciduous teeth

Stem cells from human exfoliated deciduous teeth (SHEDs) have great potential to treat various dental-related diseases in regenerative medicine. They are usually maintained with 10% fetal bovine serum (FBS) in vitro. Modified platelet-rich plasma (mPRP) would be a safe alternative to 10% FBS during SHEDs culture. Therefore, our study aimed to compare the proliferation and differentiation of SHEDs cultured in mPRP and FBS medium to explore an optimal concentration of mPRP for SHEDs maintenance. Platelets were harvested by automatic blood cell analyzer and activated by repeated liquid nitrogen freezing and thawing. The platelet-related cytokines were examined and analyzed by ELISA. SHEDs were extracted and cultured with different concentrations of mPRP or 10% FBS medium. Alkaline phosphatase (ALP) activity was measured. Mineralization factors, RUNX2 and OCN, were measured by real-time PCR. SHEDs were characterized with mesenchymal stem cells (MSCs) markers including vimentin, CD44, and CD105. mPRP at different concentrations (2, 5, 10, and 20%) enhanced the growth of SHEDs. Moreover, mPRP significantly stimulated ALP activity and promoted expression of RUNX2 and OCN compared with 10% FBS. mPRP could efficiently facilitate proliferation and differentiation of SHEDs, and 2% mPRP would be an optimal substitute for 10% FBS during SHEDs expansion and differentiation in clinical scale manufacturing.

PHF2 histone demethylase acts as a tumor suppressor in association with p53 in cancer.

Plant homeodomain finger 2 (PHF2) has a role in epigenetic regulation of gene expression by demethylating H3K9-Me2. Several genome-wide studies have demonstrated that the chromosomal region including the PHF2 gene is often deleted in some cancers including colorectal cancer, and this finding encouraged us to investigate the tumor suppressive role of PHF2. As p53 is a critical tumor suppressor in colon cancer, we tested the possibility that PHF2 is an epigenetic regulator of p53. PHF2 was associated with p53, and thereby, promoted p53-driven gene expression in cancer cells under genotoxic stress. PHF2 converted the chromatin that is favorable for transcription by demethylating the repressive H3K9-Me2 mark. In an HCT116 xenograft model, PHF2 was found to be required for the anticancer effects of oxaliplatin and doxorubicin. In PHF2-deficient xenografts, p53 expression was profoundly induced by both drugs, but its downstream product p21 was not, suggesting that p53 cannot be activated in the absence of PHF2. To find clinical evidence about the role of PHF2, we analyzed the expressions of PHF2, p53 and p21 in human colon cancer tissues and adjacent normal tissues from patients. PHF2 was downregulated in cancer tissues and PHF2 correlated with p21 in cancers expressing functional p53. Colon and stomach cancer tissue arrays showed a positive correlation between PHF2 and p21 expressions. Informatics analyses using the Oncomine database also supported our notion that PHF2 is downregulated in colon and stomach cancers. On the basis of these findings, we propose that PHF2 acts as a tumor suppressor in association with p53 in cancer development and ensures p53-mediated cell death in response to chemotherapy.

Prognostic value of clinical and immunological markers in acute phase of SFTS virus infection.

SFTS virus (SFTSV) is a novel bunyavirus that causes severe fever with thrombocytopenia syndrome (SFTS), an emerging infectious disease that occurred in China in recent years, with an average case fatality rate of 10-12%. Intervention in the early clinical stage is the most effective measure to reduce the mortality rate of disease. To elucidate the natural course of and immune mechanisms associated with the pathogenesis of SFTSV, 59 laboratory-confirmed SFTS patients in the acute phase, who were hospitalized between October 2010 and September 2011, were enrolled in this study, and the patients sera were dynamically collected and tested for SFTSV viral RNA load, 34 cytokines or chemokines and other related laboratory parameters. All clinical diagnostic factors in the acute phase of SFTS were evaluated and assessed. The study showed that the severity of the disease in 11 (18.6%) patients was associated with abdominal pain (p 0.007; OR = 21.95; 95% CI, 2.32-208.11) and gingival bleeding (p 0.001; OR=122.11; 95% CI, 6.41-2328). The IP-10, TNF-α, IL-6, IL-10, granzyme B and HSP70 levels were higher over the 7-8 days in severe cases, accompanied by altered AST, CK and LDH levels. HSP70 (p 0.012; OR=8.29; 95% CI, 1.58-43.40) was independently correlated with the severity of the early acute phase of SFTSV infection. The severity of SFTS can be predicted based on the presence of symptoms such as abdominal pain and gingival bleeding and on the level of HSP70 in the acute phase of the disease.

Using multiple cytokines to predict hepatocellular carcinoma recurrence in two patient cohorts.

BACKGROUND: Cytokines are tightly linked to the carcinogenesis, development and prognosis of hepatocellular carcinoma (HCC). We determined the prognostic value of 39 circulating cytokines in HCC patients after radical resection and then developed a novel cytokine-based prognostic classifier (CBPC) for the prediction of patient prognosis. METHODS: A total of 179 patients were divided into two cohorts based on the date of radical resection. Thirty-nine cytokines were simultaneously analysed in patient serum samples using multiplex bead-based Luminex technology. Support vector machine-based methods and Cox proportional hazard models were used to develop a CBPC from the training cohort, which was then validated in the validation cohort. RESULTS: Among seven cytokines significantly correlating with the disease-free survival (DFS) in the training cohort, six of them were validated to be significant prognostic factors to predict DFS and overall survival (OS) in the validation cohort, namely fibroblast growth factor 2 (FGF-2), growth-regulated oncogene (GRO), interleukin 8 (IL-8), interferon gamma-induced protein 10 (IP-10), vascular endothelial growth factor (VEGF), and interferon alpha-2 (IFN-α2). By integrating six cytokines and three clinical characteristics, we developed a CBPC to predict the recurrence and 3-year OS of HCC patients (sensitivity, 0.648; specificity, 0.918). In the validation cohort, the CBPC were confirmed to have significant predictive power for predicting tumour recurrence and OS (sensitivity, 0.585; specificity, 0.857). Interestingly, IFN-α2 was the only cytokine being independent prognostic factor in both patient cohorts. CONCLUSION: Our study verifies the presence of specific cytokine-phenotype associations with patient prognosis in HCC. The CBPC developed include multiple circulating cytokines and may serve as a novel screening approach for identifying HCC patients with a high risk of post-resection recurrence and shorter OS. These individuals may also be suitable for cytokine-targeted therapies.

Postprandial total and HMW adiponectin following a high-fat meal in lean, obese and diabetic men.

BACKGROUND/OBJECTIVES: Recent work suggests that macronutrients are pro-inflammatory and promote oxidative stress. Reports of postprandial regulation of total adiponectin have been mixed, and there is limited information regarding postprandial changes in high molecular weight (HMW) adiponectin. The aim of this study was to assess the effect of a standardised high-fat meal on metabolic variables, adiponectin (total and HMW), and markers of inflammation and oxidative stress in: (i) lean, (ii) obese non-diabetic and (iii) men with type 2 diabetes mellitus (T2DM). SUBJECTS/METHODS: Male subjects: lean (n=10), obese (n=10) and T2DM (n=10) were studied for 6 h following both a high-fat meal and water control. Metabolic variables (glucose, insulin, triglycerides), inflammatory markers (interleukin-6 (IL6), tumour necrosis factor (TNF)α, high-sensitivity C-reactive protein (hsCRP), nuclear factor (NF)κB expression in peripheral blood mononuclear cells (p65)), indicators of oxidative stress (oxidised low density lipoprotein (oxLDL), protein carbonyl) and adiponectin (total and HMW) were measured. RESULTS: No significant changes in TNFα, p65, oxLDL or protein carbonyl concentrations were observed. Overall, postprandial IL6 decreased in subjects with T2DM but increased in lean subjects, whereas hsCRP decreased in the lean cohort and increased in obese subjects. There was no overall postprandial change in total or HMW adiponectin in any group. Total adiponectin concentrations changed over time following the water control, and the response was significantly different in lean subjects compared with subjects with T2DM (P=0.04). CONCLUSIONS: No consistent significant postprandial inflammation, oxidative stress or regulation of adiponectin was observed in this study. Findings from the water control suggest differential basal regulation of total adiponectin in T2DM compared with lean controls.

CREG: a possible candidate for both prevention and treatment of proliferative vascular disease.

Cellular repressor of E1A-stimulated genes (CREG), a novel cellular protein, was discovered in 1998. Accumulating evidence, mainly from our laboratory, has suggested that CREG plays critical roles in reducing neointimal hyperplasia, maintaining vascular homeostasis, and promoting endothelial restoration. The study of CREG has the potential to offer new insights into both prevention and treatment of proliferative vascular disease, and will help us understand the processes of vascular repair after injury. It will also contribute to the development of new therapeutic strategies and devices, such as anti-in-stent restenosis stents. The present review summarizes our research on the molecular identity of CREG, and reviews the biological activities of CREG in regulating cell differentiation, proliferation, migration, and apoptosis of vascular smooth muscle cells and endothelial cells.

The mode of reproductive-derived Spink (serine protease inhibitor Kazal-type) action in the modulation of mammalian sperm activity.

The reproductive-derived serine protease inhibitor Kazal-type (Spink) has been identified in seminal plasma, and Spink-spermatozoa binding has been illustrated in many mammalian species including human. We used mice as experimental animal to study the mode of Spink action in the modulation of mammalian sperm activity. A Spink3-binding zone was cytochemically stained on the sperm head at apical hook separated from intact acrosome, whether the cells were capacitated or not. The Spink3-spermatozoa binding neither changed the population of cells in the uncapacitated, capacitated and acrosome-reacted status nor affected the capacitation-related protein phosphorylation and cell motility enhancement. Despite that, the Spink-spermatozoa interaction resulted in decreasing the intracellular calcium concentration ([Ca(2+)](i)) of the cell head and suppressing both the acrosome reaction induced by Ca(+2) ionophore A23187 and the cell fertility. Furthermore, Spink3 seen on the head of spermatozoa in the uterine cavity after coitus could be removed by the trypsin-like activity in the uterine fluid of oestrous females, and free Spink3 in the uterine cavity suppressed the protease activity. We integrated our data to shed light on the molecular mechanism of how Spink and its inhibiting protease are interplayed to modulate the activity of mammalian spermatozoa during their transit in the reproductive tract.

Significant volume reduction and shape abnormalities of the basal ganglia in cases of chronic liver cirrhosis.

BACKGROUND AND PURPOSE: Chronic liver disease frequently includes cognitive and movement disorders, suggesting an alteration of the striatum. With the exception of hyperintensities evident on T1-weighted images indicative of Mn deposition, radiographic findings of the BG are nonspecific. Volumetric and morphometric analysis of DGM is limited. Whether DGM undergoes degeneration and whether this change is associated with pallidal hyperintensity and cognitive performance are currently unknown in patients with cirrhosis. MATERIALS AND METHODS: The DGM volumes of 28 patients with chronic cirrhosis and 28 control patients were compared. Using 3D high-resolution MR images, the volume and shape of each structure were automatically analyzed by the FSL. Correlations between the DGM volume and other clinical variables, including the pallidal signal intensity, were assessed by multiple regression analysis. RESULTS: Patients with Child B and Child C liver disease had significantly smaller bilateral putaminal volumes than control patients, and patients with Child C also demonstrated smaller left caudate nucleus and left amygdala volumes than control patients. Pallidal hyperintensity correlated with smaller striatum volume, which was linearly related to worse cognitive performance. The nonuniform distributed shape abnormalities in the striatum further support the ascending spiral interconnecting theory of the striatum. CONCLUSIONS: These findings strongly suggest lower DGM volume develops according to the severity of the liver cirrhosis. The Mn deposition might contribute the striatum deficit. These findings support the value of additional psychomotor research associated with liver cirrhosis.

Effect of trychostatin A treatment on gene expression in cloned mouse embryos.

Histone deacetylation occurs upon the transfer of somatic nuclei into enucleated oocytes, but its role in reprogramming somatic chromatin to the totipotent state is unknown. To investigate the importance of histone deacetylation in reprogramming, we constructed embryos by electrofusing breast cancer cells with enucleated mouse oocytes. The reconstructed embryos were then cultured before and/or after activation for 6h in the presence of trychostatin A (TSA), a potent inhibitor of histone deacetylase. Total RNA was isolated from these TSA-treated and untreated embryos and real-time reverse transcription PCR was conducted to monitor transcription of ErbB2, Muc1, eIF-4C, MuERV-L, and c-mos genes. The nuclear-cytoplasmic interaction inhibited typical expression of ErbB2 and Muc1 in the somatic cells. Moreover, the inhibition of histone deacetylation prior to activation did not increase the levels of eIF-4C, MuERV-L, and c-mos expression in the nuclear transfer (NT) embryos (P>0.05), whereas additional treatment with 100nM TSA beyond the activation point improved expression of these genes (P<0.05). Trychostatin A treatment also improved the development rates of NT embryos at the 2-cell, 4-cell, and blastocyst stages (78.6% vs. 90.2%, 45.2% vs. 68.9%, and 16.7% vs. 30.3%, respectively, P<0.05). We hypothesized that the reprogramming of gene expression in NT embryos is independent of somatic histone deacetylation, and that hyperacetylation may have a positive effect on NT embryo development.

Autopsy report of four cases who died from Streptococcus suis infection, with a review of the literature.

There was an outbreak of human Streptococcus suis (S. suis) infection in southwest China in 2005. The total number of documented patients was 204 and 38 of them died. Four cases were autopsied and are reported in this paper. The autopsies showed that multiple organs were involved, with a prominent injury of the lung, kidney, and intestine. The essential pathologic changes were multiple microthrombi (hyaline thrombi) formation in the capillaries of various organs and tissues, accompanied by congestion and hemorrhage. The pathogen of the disease was S. suis, serotype 2, which was confirmed by means of germ culture of the heart, blood, and tissue samples at the Chinese Center for Disease Control and Prevention (China CDC). The autopsy diagnosis of all four cases was septicemia with disseminated intravascular coagulation (DIC). The cause of death was toxic shock with multi-organ dysfunction. Combined with the epidemic features, clinical manifestations, etiological, and autopsy findings, it accorded with streptococcal toxic shock syndrome (STSS).

The role of pro-inflammatory factors in mediating the effects on the fetus of prenatal undernutrition: implications for schizophrenia.

Exposure to prenatal undernutrition or malnutrition increases the risk of schizophrenia, although little is known about the mechanism. Pro-inflammatory factors are critical in brain development, and are believed to play an important role in neurodevelopmental disorders associated with prenatal exposure to infection, including schizophrenia. However it is not known whether pro-inflammatory factors also mediate the effects on the fetus of prenatal malnutrition or undernutrition. In this study, we established a new prenatal undernourished rat model induced by maternal exposure to a diet restricted to 50% of the low (6%) protein diet (RLP50). We observed the disappearance of maternal nest-building behavior in the RLP50 dams, increased levels of TNFA and IL6 in the placentas (P<0.001; P=0.879, respectively) and fetal livers (P<0.001; P<0.05, respectively), and a decrease in the fetal brains (P<0.05; P<0.01, respectively). Our results are similar to previous studies of maternal infection, which implies that a common pathway mediated by pro-inflammatory factors may contribute to the brain development, consequently increasing the risk of schizophrenia and other psychiatric diseases programmed by varied maternal adversities. We also provide a new prenatal undernourished model for researching prenatal problems, which differs from previous malnourished model in terms of the maternal behavior of dams and of observed pro-inflammatory factor levels in fetal tissues.