Biomechanical study on the treatment of Pauwels type III femoral neck fracture by open reduction and internal fixation with hollow nail internal fixation.

OBJECTIVE: We aimed to explore the therapeutic effect of open reduction and internal fixation with hollow nail internal fixation for Pauwels type Ⅲ femoral neck fracture. PATIENTS AND METHODS: From January 2016 to February 2021, a total of 100 eligible patients with Pauwels type III femoral neck fracture were involved in this study and divided into two groups randomly: the combined remedy group and the closed therapy group, with 50 patients in each group. After that, 50 subjects in the combined remedy group were treated with open reduction and support plate combined with hollow screw internal fixation, and the treatment conditions were observed and recorded. The closed therapy group received routine treatment. RESULTS: Among the 100 patients selected, the operation time of the combined remedy group was significantly lower than that of the closed therapy group, and the intraoperative bleeding was also significantly less. In the closed therapy group, the time of getting out of bed after the operation and the excellent and good rate were better; moreover, the functional score and pain score of three months after the operation were significantly better than that of one month after the operation. The functional score and pain score of one month after the operation were not statistically significant for the combined remedy group or the closed therapy group. CONCLUSIONS: In the treatment of Pauwels type Ⅲ femoral neck fracture with open reduction and internal fixation combined with hollow nail internal fixation, the operation time and intraoperative bleeding volume were significantly decreased, but the postoperative recovery time was enhanced compared to that of total joint replacement. After the operation, the functional score and pain score had a significant relationship with the recovery time, and there was no significant relationship with the type of treatment. Therefore, in clinical treatment, doctors should take appropriate treatment methods for their patients.

Network pharmacological and molecular docking verification of the mechanism of Osteoking in preventing deep vein thrombosis of lower limb.

OBJECTIVE: The aim of this study was to predict the mechanism of Osteoking in preventing deep vein thrombosis (DVT) of the lower limb by network pharmacology and molecular docking. MATERIALS AND METHODS: The relevant active components and targets of Osteoking were collected through the TCMSP database, and the relevant disease targets of DVT were collected through the GeneCards, OMIM, and DisGeNET databases. The intersecting gene targets of Osteoking and DVT were obtained using Venny 2.1.0 software. PPI network construction and core target selection using Cytoscape 3.9.0 software. The Metascape database was used for GO and KEGG enrichment analysis of relevant targets. Finally, the molecular docking of the main active components and key targets was carried out. RESULTS: There are 361 potential targets and 71 core targets of Osteoking in preventing deep vein thrombosis of the lower limb. Signal pathways are involved in various diseases such as cancer, diabetic complications, atherosclerosis, and more. Some of the most common pathways include AGE-RAGE signaling pathway and Calcium signaling pathway. Molecular docking results showed that the main active components of Osteoking had relatively stable binding activities with the key targets. CONCLUSIONS: Osteoking can play a role through multiple targets and multiple signal pathways to prevent the formation of deep venous thrombosis of the lower limb after fracture.

Effect of rehabilitation on the long-term efficacy of botulinum toxin-A for spastic cerebral palsy.

OBJECTIVE: To investigate the long-term effects of botulinum toxin-A (BTX-A) nerve block on relaxation of spasticity in cerebral palsy. PATIENTS AND METHODS: From June 2015 to December 2018, 52 children, aged 20-56 months, with spastic cerebral palsy were treated with BTX-A. The dose of BTX-A was selected based on the weight of the child and the modified Ashworth scale (MAS). The injection dose ranged from 45 IU to 150 IU (average 68.0±31.6 IIU). The muscle tone and motor functions of all children were evaluated before the block. The spasticity was measured using the MAS, and the motor function was measured using the Physician Rating Scale (PRS) and the gross motor function measure (GMFM). After two years, all children were re-evaluated. RESULTS: No significant difference was observed between the trial and control groups in terms of age, weight, MAS, PRS, and GMFM measurements before the block (p>0.05). The PRS and GMFM improved significantly in both groups after two years (p<0.05). The PRS and GMFM in the trial group increased more significantly than those in the control group (p<0.05). CONCLUSIONS: The BTX-A block showed a long-term positive effect. Rehabilitation training after the block could help children to improve their motor functions.

Associations of young onset age and genetic risk of beta cell dysfunction with glycaemic progression in individuals with type 2 diabetes.

AIM: To study the relationship between genetic risk of beta cell dysfunction, young onset age and glycaemic progression in individuals with type 2 diabetes (T2D). MATERIALS AND METHODS: 1385 T2D outpatients were included in cross-sectional sub-study and 730 insulin-naïve outpatients were followed for 3 years in prospective sub-study. Genetic risk score (GRS) was derived from 24 beta cell dysfunction-related single nucleotide polymorphisms, with lower and upper 25 percentiles defined as low and high genetic risk. Glycaemic progression was defined as requirement for sustained insulin therapy. RESULTS: 388 participants in cross-sectional and 128 in prospective sub-study experienced glycaemic progression. Young onset age (T2D diagnosis below 40 year-old) was associated with high risk of glycaemic progression as compared to usual-onset counterparts (adjusted OR 1.64 [95% CI 1.14-2.36], and 2.92 [95% CI 1.76-4.87] in cross-sectional and prospective sub-study, respectively). As compared to those with intermediate risk, a low GRS was associated with lower risk for glycaemic progression (adjusted OR 0.72 [95% CI 0.49-1.06], and 0.51 [95% CI 0.29-0.90]) whereas a high GRS was not significantly associated with glycaemic progression. Notably, the association of young-onset T2D with high risk of glycaemic progression was independent of known clinical risk factors and beta cell dysfunction GRS (P interaction > 0.10). CONCLUSION: Young onset age and low genetic risk of beta cell dysfunction are independently associated with risk of glycaemic progression. Our data do not support that genetic risk modulates the risk of glycaemic progression in individuals with young-onset T2D.

Suberoylanilide hydroxamic acid attenuates epidural fibrosis via inhibiting myofibroblast differentiation and increasing fibroblast apoptosis.

OBJECTIVE: Epidural fibrosis represents a fatal stage of failed back surgery syndrome (FBSS) of known and idiopathic etiology, but no valid therapy is presently available. Previous evidence demonstrated that suberoylanilide hydroxamic acid (SAHA), a histone deacetylases inhibitor, has antifibrotic and anti-inflammatory potential. Current studies have proved that SAHA inhibits myofibroblast differentiation and increases fibroblast apoptosis to attenuate epidural fibrosis. The purpose of this study was to investigate the effect and mechanism of SAHA on repressing epidural fibrosis. PATIENTS AND METHODS: First, the levels of acetylation of histone and α-tubulin in adult human fibroblasts (AHF) and human epidural fibroblasts (HEF) were analyzed following SAHA and transforming growth factor-ß(TGF-ß) treatment. Then, mRNA and protein obtained from human fibroblasts following TGF-ß activation and SAHA treatment in vitro culture were used to test the influence of SAHA on the activation and apoptosis of fibroblasts, so as to further explore the related mechanism of SAHA. Then, a laminectomy model was established in rats to observe the therapeutic effect of SAHA on epidural scar tissue. RESULTS: The present research proved that the increases of HDAC 3 and α-tubulin were observed in AHF and HEF after TGF-ß administration, but SAHA decreased HDAC 3 and α-tubulin expressions. In addition, cell study demonstrated that SAHA inhibited fibroblast activation via decreasing TGF-ß function and accelerated apoptosis by promoting cleaved-caspase-3. In the epidural fibrosis model, it was found that SAHA weakened scar hyperplasia and collagen deposition, and effectively inhibited the process of epidural fibrosis. CONCLUSIONS: These results indicated that SAHA inhibited HDAC 3 expression, decreased TGF-ß effect, and enhanced caspase-3 in fibroblasts, leading reduction of myofibroblast activation and apoptosis elevation. Hence, SAHA ameliorated epidural fibrosis development.

LncRNA DANCR affected cell growth, EMT and angiogenesis by sponging miR-345-5p through modulating Twist1 in cholangiocarcinoma.

OBJECTIVE: LncRNA DANCR has been reported to play an important role in various cancers. Therefore, this study aimed at exploring the function and regulatory mechanism of DANCR in Cholangiocarcinoma (CCA). PATIENTS AND METHODS: qRT-PCR was used to measure the expression of DANCR, miR-345-5p in tissues and cells. Western blot was applied to measure the protein expression of Twist, N-cadherin, Vimentin, E-cadherin, VEGF-A, VEGF-C, PCNA and C-caspase 3. The relationship between DANCR and miR-345-5p was determined by luciferase reporter assay. MTT assay and flow cytometry were used to assess cell proliferation and apoptosis, respectively. Transwell assay was performed to detect cell invasion and migration. RESULTS: We found that the expression of DANCR was significantly induced in CCA tissues and cells. Inhibition of DANCR remarkably suppressed CCA cell proliferation, migration, invasion, EMT and angiogenesis as well as induced cell apoptosis in vitro and in vivo. Luciferase reporter assay determined that DANCR directly targeted miR-345-5p and Twist1 was a target mRNA of miR-345-5p. Otherwise, miR-345-5p down-expression partially reversed the effect induced by the suppression of DANCR in CCA. Moreover, the suppressive effects of high miR-345-5p expression on CCA cells were reversed by improving Twist1 expression. CONCLUSIONS: In this study, we verified that LncRNA DANCR affected cell proliferation, migration, invasion, angiogenesis, epithelial-mesenchymal transition (EMT) and induced apoptosis through modulating miR-345-5p/Twist1 axis in Cholangiocarcinoma.

Topical calcineurin inhibitors in the treatment of oral lichen planus: a systematic review and meta-analysis.

BACKGROUND: TCS (topical corticosteroids) are the first-line drug in the treatment of oral lichen planus (OLP). However, the value of topical calcineurin inhibitors (TCI) including tacrolimus, pimecrolimus and ciclosporin for OLP is still controversial. OBJECTIVES: To compare the efficacy and safety of TCI vs. TCS for OLP. METHODS: The authors searched PubMed, Embase, Cochrane Central Register of Controlled Trials, Web of Science and four Chinese databases from 1950 to May 2018. The randomized controlled trials comparing TCI and TCS for OLP reported at least one of the following outcomes: improvement of clinical signs and/or symptoms, relapse, blood levels of TCI and adverse events. RESULTS: Twenty-one trials involving 965 patients were included in the analysis. For the treatment of OLP (3-8 weeks), TCI including tacrolimus, pimecrolimus and ciclosporin were similar to TCS in efficacy. Tacrolimus-TCS resulted in similar outcomes, with relapse at 3 weeks to 6 months. Blood levels of TCI were usually undetectable. In addition, tacrolimus showed a statistically higher incidence of local adverse events than TCS for short-term treatment. A few systemic adverse events occurred in the tacrolimus and ciclosporin groups, but they were not serious. CONCLUSIONS: The evidence for tacrolimus (n = 12), pimecrolimus (n = 3) and ciclosporin (n = 6) demonstrated that treatment with TCI may be an alternative approach when OLP does not respond to the standard protocols. Tacrolimus 0·1% should be the first drug of choice when selecting TCI for short-term treatment in recalcitrant OLP. Further well-designed trials are warranted to evaluate the long-term efficacy and safety of TCI. What's already known about this topic? The main topical drug for oral lichen planus (OLP) is topical corticosteroids (TCS). Patients with OLP who are not responsive to TCS or are at risk of adverse events from TCS need other alternative drugs. Topical calcineurin inhibitors (TCI), including tacrolimus, pimecrolimus and ciclosporin, have become a hot topic in a variety of mucocutaneous immune-mediated diseases. What does this study add? TCI including tacrolimus, pimecrolimus and ciclosporin were similar to TCS in efficacy for the short-term treatment of OLP. The local adverse events of tacrolimus were higher than with TCS. A few systemic adverse events were reported with TCI, but they were all tolerable and not serious. The limited evidence for pimecrolimus (three trials) and ciclosporin (six trials) requires further studies to evaluate the short-term and long-term efficacy and safety of TCI compared with TCS.

Influence of miR-34a on preeclampsia through the Notch signaling pathway.

OBJECTIVE: The aim of this study was to investigate the influence of micro-ribonucleic acid-34a (miR-34a) on preeclampsia through the Notch signaling pathway. PATIENTS AND METHODS: The expressions of miR-34a, Notch-1, Notch-2, and Notch-3 in the placenta of 39 preeclampsia patients and 42 normal patients were detected by immunohistochemistry and Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The correlations between miR-34a expression with the expressions of Notch-1, Notch-2 and Notch-3 were analyzed, respectively. Besides, placental trophoblasts were isolated from preeclampsia patients and cultured in vitro. The expressions of miR-34a, Notch-1, Notch-2 and Notch-3 in placental trophoblasts were analyzed. Furthermore, the influences of miR-34a on the protein expressions of Notch-1, Notch-2, Notch-3, and hairy and enhancer of split-1 (Hes-1) in the Notch signaling pathway were analyzed by Luciferase reporter gene assay and Western blotting. The role of Notch in trophoblast invasion was investigated through the Notch inhibitors. In addition, its influence on the expression of urokinase-type plasminogen activator (uPA) was studied by miR-34a overexpression. RESULTS: The expressions of miR-34a and Notch-1 were correlated with preeclampsia in the placentas of preeclampsia patients and normal patients to a certain degree. The expression of miR-34a in preeclamptic placenta was significantly higher than that of the normal placenta (p<0.05). However, Notch-1 expression was markedly lower in preeclamptic placenta (p<0.05). No significant differences were found in the expressions of Notch-2 and Notch-3 between the two types of placentas (p>0.05). MiR-34a had a remarkable negative correlation with Notch-1 expression in the Notch family (p<0.001, r=-0.5775). RT-PCR results revealed that the mRNA expression of miR-34a in placental trophoblasts of patients with preeclampsia was notably higher than that of normal people (p<0.01). However, Western blotting demonstrated that the protein expressions of Notch-1, Notch-2 and Notch-3 exhibited the opposite results. Additionally, the protein expression of Notch-1, Notch-2, Notch-3 and Hes-1 in trophoblasts transfected with pre-miR-34a was significantly decreased. The treatment with Notch inhibitors markedly reduced the trophoblast invasion. Furthermore, miR-34a overexpression or intracellular domain of Notch (ICN) overexpression regulated uPA expression. CONCLUSIONS: MiR-34a regulates uPA system through the Notch signal transduction, thereby regulating the invasion of placental trophoblasts in patients with preeclampsia.

A biological study of supernumerary teeth derived dental pulp stem cells based on RNA-seq analysis.

AIM: To identify the basic characteristics and gene expression profiles of supernumerary teeth derived stem cells (SNTSCs) and compare them with those of normal dental pulp stem cells (DPSCs). METHODOLOGY: Flow cytometry was conducted to identify the protein expression of stem cell markers. Cell proliferation, migration and differentiation abilities of both SNTSCs and DPSCs were determined by CCK8, transwell and differentiation assays, respectively. Gene expression profiles were studied by RNA sequencing analyses. After knocking down the expression of certain differential expression genes (DEGs), the function of DEGs was investigated by CCK8 and transwell assays. Statistical differences were determined using a two-tailed t-test and P values below 0.05 were considered significant. RESULTS: Supernumerary teeth derived stem cells were capable of differentiating into adipocyte, chondrocyte and osteoblast lineage cells, and compared to ordinary DPSCs, SNTSCs had a significantly higher cell proliferation rate (P < 0.01) and significantly lower migration rate (P < 0.01). RNA-seq results revealed the differential expression genes (DEGs) between SNTSCs and DPSCs. A principal component analysis (PCA) and cluster analysis revealed that the gene expression patterns of SNTSCs and DPSCs were different from each other. A total of 12 861 genes were differentially expressed at a significant P value (P ≤ 0.01), and 5292 of these increased in SNTSCs and 7569 decreased. Further study on the selected DEGs revealed that FUT11, FAM155A and BRD2 inhibited the cell proliferation rate of SNTSCs, and FUT11 and GLUD1 inhibited the cell migration rate, whilst FAM155A promoted the migration rate. CONCLUSIONS: The biological characteristics and gene expression profile of SNTSCs was revealed. The stem cell properties of SNTSCs were similar to normal DPSCs but they had a high cell proliferation rate and may have greater potential for cell differentiation.

The mechanisms of Ang II-induced hypertensive vascular remodeling under suppression of CD68 in macrophages.

OBJECTIVE: High blood pressure (hypertension) is one of the most common cardiovascular diseases. In recent years, there were more and more studies on the function of inflammation in hypertension. CD68 mainly mediates the activation of cytokine interleukin-17 (IL-17) signaling pathway and participates in inflammatory responses. It has been studied the function of CD68 and IL-17 in hypertension, but it has not been reported whether it affected hypertension and vascular remodeling when macrophage CD68 expression inhibited. In this study, antisense-CD68 mice were used to study the effect and mechanism of angiotensin II-induced hypertensive vascular remodeling under specific suppression of macrophage CD68. MATERIALS AND METHODS: Fifty 8-week-old male antisense-CD681 and C57 mice were divided into control and experimental group (angiotensin II group, 1000 ng•kg-1•min-1). After infusion of angiotensin II for 28 days, hematoxylin-eosin (HE) staining and immunohistochemical staining were used to observe the remodel of vascular. The changes of aortic inflammatory factors were detected by Real-time PCR (RT-PCR) and Western blotting. RESULTS: By specifically inhibiting the expression of macrophage CD68, macrophage infiltration was mitigated in Ang II-induced hypertensive vascular remodeling model mouse, which also down-regulated the expression of vascular tissue inflammatory factor and activation of vascular smooth muscle cell p65. CONCLUSIONS: CD68 regulates the Ang II-induced hypertensive vascular remodeling through mediating macrophage inflammatory factor release.

Melatonin therapy adds extra benefit to varicecelectomy in terms of sperm parameters, hormonal profile and total antioxidant capacity: A placebo-controlled, double-blind trial.

In this study, we aimed at investigating the impact of melatonin supplementation on semen parameters, hormonal profile and total antioxidant capacity after varicocelectomy. Infertile male patients who were diagnosed with varicocele and underwent subinguinal varicocelectomy were included in the study. After performing subinguinal varicocelectomy, the patients were randomised into two groups: 27 receiving melatonin for 3 months and 27 as the placebo-controlled group receiving placebo for 3 months. The pre-operative parameters of semen analyses, hormonal profile and seminal oxidative stress status of both groups were compared with those of post-operative parameters. There were statistically significant improvements in post-operative parameters of semen analyses (sperm concentration, motility and proportions of normally formed spermatozoa), peripheral blood inhibin B and total antioxidant capacity in melatonin group compared with placebo group. In conclusion, melatonin therapy adds extra benefit to varicecelectomy in terms of sperm parameters, peripheral blood inhibin B and total antioxidant capacity; however, further studies including large number of samples are needed to make a proper decision on melatonin supplementation after varicocelectomy.

Methylene blue attenuates renal ischemia-reperfusion injury by negative regulation of NLRP3 signaling pathway.

OBJECTIVE: To investigate the effect of methylene blue (MB) on renal ischemia-reperfusion (IR) injury in mice and its possible relevant mechanisms. MATERIALS AND METHODS: A total of 30 male C57/BL6 mice aged 4 months old were randomly divided into the following three groups: Sham group (n=10), IR group (n=10), and MB group (n=10). Mice in MB group were treated with gavage continuously using methylene blue solution (dosage: 25 mg·kg-1·d-1) until they were 7 months old. Mice in the other two groups were administrated with the same amount of normal saline for gavage. After that, the abdomen of mice in Sham group was opened and closed, and bilateral renal pedicles of mice in IR group and MB group were occluded using a micro-artery clamp for 45 min for modeling. After modeling, the renal tissues and blood samples of mice were taken for detection. The levels of serum creatinine (Scr), blood urea nitrogen (BUN), and malondialdehyde (MDA), and activity of superoxide dismutase (SOD) in mice in each experimental group were detected and statistically analyzed, respectively. The degree of renal tubular necrosis in renal tissues of mice was observed under an optical microscope. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors [interleukin-1ß (IL-1ß), IL-18, IL-10 and transforming growth factor-ß1 (TGF-ß1)] in renal tissues of mice in each experimental group, followed by relevant statistical analyses. The expressions of relevant proteins [NACHT, LRR, and PYD domains-containing protein 3 (NLRP3), nuclear factor-κB (NF-κB), caspase-1, pro-IL-1ß, and IL-1ß] in renal tissues of mice in each experimental group were detected via Western blotting, and the gray value of each band was detected for statistical analysis. RESULTS: It was found in this experimental study that Scr and BUN levels in MB group were significantly lower than those in IR group, and the differences were statistically significant (p<0.05). Compared with that in IR group, the degree of renal tubular necrosis in MB group was significantly alleviated, and the difference was statistically significant (p<0.05). Compared with those in IR group, the levels of inflammatory factors (IL-1ß and IL-18) in MB group were significantly decreased, but the levels of IL-10 and TGF-ß1 in MB group were significantly increased, and the differences were statistically significant (p<0.05). Compared with those in IR group, the activity of SOD in MB group was increased significantly, but the level of MDA was decreased, and the differences were statistically significant (p<0.05). The protein expressions of NLRP3, NF-κB, caspase-1, and pro-IL-1ß in MB group were decreased compared with those in IR group, but the expression of IL-1ß in MB group was increased, and the differences were statistically significant (p<0.05). CONCLUSIONS: We showed that methylene blue can alleviate the apoptosis and inflammatory response induced by renal IR injury in mice, and its relevant mechanism may be related to the fact that methylene blue can negatively regulate NLRP3 signaling pathway.

Sex modulates the association of fibroblast growth factor 21 with end-stage renal disease in Asian people with Type 2 diabetes: a 6.3-year prospective cohort study.

AIM: To study whether plasma fibroblast growth factor 21 independently predicts the risk of end-stage renal disease in Asian people with Type 2 diabetes. METHODS: In this prospective cohort study, 1700 Asian people with Type 2 diabetes were followed for a mean of 6.3 years in a regional hospital in Singapore. Incident end-stage renal disease was identified by linkage with a national renal registry. The association of baseline fibroblast growth factor 21 levels with risk of progression to end-stage renal disease was studied using survival analyses. RESULTS: Participants were aged 60 ± 10 years, with an average diabetes duration of 12 years. Their estimated GFR was 73 ± 28 ml/min/1.73 m2 and 62% had albuminuria at baseline. A total of 179 incident end-stage renal disease cases were identified. Plasma fibroblast growth factor 21 interacted with sex in its association with end-stage renal disease (Pinteraction = 0.003). A 1-sd increment in fibroblast growth factor 21 (natural log-transformed) was associated with a 1.32-fold (95% CI 1.05-1.66, P = 0.02) increased hazard for end-stage renal disease in women, after adjustment for traditional risk factors including estimated GFR and albuminuria. Taking death as a competing risk did not materially change the outcome [sub-distribution hazard ratio 1.35 (95% CI 1.11-1.66, P = 0.003)]. Fibroblast growth factor 21 did not predict end-stage renal disease risk in men after adjustment for baseline estimated GFR and albuminuria [hazard ratio 1.07 (95% CI 0.89-1.28, P = 0.49)]. CONCLUSIONS: Plasma fibroblast growth factor 21 level independently predicted risk of progression to end-stage renal disease in women with Type 2 diabetes. The pathophysiological relationships among FGF21, sex and renal progression warrant further study.

The extracellular matrix protein mindin attenuates colon cancer progression by blocking angiogenesis via Egr-1-mediated regulation.

Mindin, a secreted, highly conserved extracellular matrix (ECM) protein, exerts a broad spectrum of effects on the innate immune system. However, its function in colorectal cancer (CRC) progression is not well established, and its upstream regulation mechanisms remain unclear. Contrary to previous reports, this study used two different enzyme-linked immunosorbent assay (ELISA) kits to show that the serum level of mindin was significantly decreased in CRC patients and that this decreased level is more significantly associated with the early stages of the disease. To explore the regulation of mindin, we used a bioinformatics approach to predict potential transcription factors and determined that early growth response factor (Egr)-1 directly regulates mindin expression at the transcriptional level using dual luciferase, chromatin immunoprecipitation (ChIP) DNA and electrophoretic mobility shift assay (EMSA) methods. Egr-1 regulates mindin mRNA and protein expression in CRC cells, and the protein expression of both Egr-1 and mindin was significantly decreased in tumor lesions of patients compared with adjacent control tissues. Mindin is essential for Egr-1-mediated inhibition of endothelial cell tube formation, and mindin inhibits endotheliocyte proliferation, migration and angiogenic sprouts in vitro. Overexpression of mindin suppressed xenograft tumor growth by blocking angiogenesis instead of directly suppressing CRC cell proliferation. Mechanically, mindin inhibits the hypoxia-induced HIF-1a and VEGFA protein expression in CRC cells and the phosphorylation of VEGFR-2 in endothelial cells. The results suggest that the serum level of mindin can be used as a novel biomarker for early detection of CRC and that the Egr-1/mindin axis is a potential therapeutic target for the inhibition of angiogenesis in CRC development.

Apocynin ameliorates pressure overload-induced cardiac remodeling by inhibiting oxidative stress and apoptosis.

Oxidative stress plays an important role in pressure overload-induced cardiac remodeling. The purpose of this study was to determine whether apocynin, a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, attenuates pressure overload-induced cardiac remodeling in rats. After abdominal aorta constriction, the surviving rats were randomly divided into four groups: sham group, abdominal aorta constriction group, apocynin group, captopril group. Left ventricular pathological changes were studied using Masson's trichrome staining. Metalloproteinase-2 (MMP-2) levels in the left ventricle were analyzed by western blot and gelatin zymography. Oxidative stress and apoptotic index were also examined in cardiomyocytes using dihydroethidium and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), respectively. Our results showed that abdominal aorta constriction significantly caused excess collagen deposition and cardiac insult. Treatment with apocynin significantly inhibited deposition of collagen and reduced the level of MMP-2. Furthermore, apocynin also decreased the NADPH oxidase activity, reactive oxygen species production and cardiomyocyte apoptotic index. Interestingly, apocynin only inhibited NADPH oxidase activity without affecting its expression or the level of angiotensin II in the left ventricle. In conclusion, apocynin reduced collagen deposition, oxidative stress, and inhibited apoptosis, ultimately ameliorating cardiac remodeling by mechanisms that are independent of the renin-angiotensin system.

Serological biomarker screening and host factor analysis elucidating immune response heterogeneity in active pulmonary tuberculosis

While mortality and morbidity from pulmonary tuberculosis (PTB) have improved, diagnosis of this infectious disease remains suboptimal without a point-of-care test. Antibody/ antigen-based serodiagnostics is the most amenable for point-of-care translation but hampered by a lack of validated biomarkers and a heterogeneous patient antibody response. Using a case-control design, we assessed serodiagnostic potential of immunoglobulins G, A, and dimeric IgA responses against 18 antigenic preparations, followed by antibody-subclass responses against antigen 60 (A60), and four markers of host innate immunity by enzymelinked immunoassay using sera samples (n=110) collected from April to October 2007 in VietNam from human immunodeficiency-negative patients with provisional diagnosis of PTB. We further analyzed host variables to investigate factors driving biomarker heterogeneity observed in patients. Among active pulmonary tuberculosis patients, low correlation was observed between anti-A60 antibody-classes, and between anti-A60 immunoglobulin G subclasses, but anti-A60 immunoglobulin A subclasses were significantly correlated. The best diagnostic combination of anti-A60 immunoglobulin G/A and a C-reactive protein “ruleout” remains insufficient at 82%/92% sensitivity/specificity (95%CI: 72-92%/82-98%). Heterogeneity of anti-A60 immunoglobulins G2, G3, M, as well as C-reactive protein and serum amyloid A levels observed in this study population appeared to be significantly associated with history of previous tuberculosis, hemoptysis, age, vaccination, night sweats, smoking, chest pain, fever, alcohol, and solid culture count. Further research on tuberculosis serological biomarkers may require consideration of host factors and new approaches using multiple biomarkers.

Anxiety-like behavior and dysregulation of miR-34a in triple transgenic mice of Alzheimer's disease.

OBJECTIVE: MicroRNAs (miRNAs) play an important role in the development of the brain and also implicated in the pathogenesis of neurological diseases such as Alzheimer's disease (AD). Recent studies implied that dysregulation of miRNAs is involved in neuropsychiatric disorders such as anxiety disorder in AD. MATERIALS AND METHODS: In this study, behavioral experiments such as open field test, elevated plus maze test and light-dark box test were performed to evaluate anxiety-like behaviors in a triple transgenic mouse model of AD (3xTg-AD mice), and Q-PCR was used to measure the change of miR-34a expression. RESULTS: Behavioral tests revealed anxiety-like behaviors in 3xTg-AD mice. Q-PCR assay showed significantly elevated expression of miR-34a in the hippocampus of 3xTg-AD mice compared with the age- and gender-matched wild-type mice. Western-blot analysis showed that the expression of metabotropic glutamate receptor 7 (GRM7) but not fibroblast growth factor-2 (FGF2), two anxiety disorder-related target genes of miR-34a, was significantly decreased in hippocampus of 3xTg-AD mice compared with the wild-type mice. CONCLUSIONS: We concluded that anxiety-like behavior occurred in 3xTg-AD mice with an involvement of miR-34a/GRM7.

Protective effects of estrogen combined with sevoflurane in an experimental model of cerebral infarction and focal cerebral ischemia-reperfusion injury.

OBJECTIVE: Potential additive effects of estrogens and sevoflurane against cerebral infarction after transient or permanent middle cerebral artery occlusion (MCAO) have not been addressed. We evaluated these using a rat model of MCAO. MATERIALS AND METHODS: 60 adult female Sprague-Dawley rats were used in the experiments after ovariectomy. Animals were divided into placebo/no MCAO, placebo + MCAO, and estrogen + MCAO groups. Each group was further subdivided into subgroups exposed to sevoflurane or oxygen. Animals in the placebo group received intraperitoneal injections of saline, whereas the estrogen group animals received intraperitoneal injections of estradiol (1 mg per day). MCAO was performed 1 week after the ovariectomy. Sevoflurane and oxygen subgroups breathed either sevoflurane or oxygen for 30 min during the surgery. Outcomes were the levels of serum estradiol (E2), interleukin (IL)-6, and beta-amyloid protein (ß-AP) (all by ELISA), neurological deficit scores (24 hours, 7 and 28 days after the operation), spatial learning and memory (both by the Morris water maze test on days 7 and 28). RESULTS: MCAO significantly up-regulated serum levels of IL-6 and ß-AP (p < 0.05 for both comparisons). The animals that received the combined treatment with estrogen and sevoflurane showed less extensive up-regulation of these markers (p < 0.05 vs. placebo-treated animals). Furthermore, MCAO induced severe neurological dysfunction and disorders of spatial learning and memory. All these were attenuated by the combined treatment. CONCLUSIONS: We demonstrate neuroprotective effects of pre-conditioning with estrogen and post-conditioning sevoflurane in experimental animal undergoing MCAO.

Hypoxic markers in non-small cell lung cancer (NSCLC) - A review.

Surgery, chemotherapy and radiotherapy have been the main pillars of cancer treatment. Some of the recent improvements in survival of other cancers can be attributed to novel treatment therapies. Such therapies mostly target specific molecules involved in cancer progression and metastasis. Development and clinical introduction of targeted therapies involve identification of new and potentially important molecules in cancer progression. The next important step is to evaluate its prognostic value. Prognostication by molecular markers is also important as this may identify subgroups of patients in need for additional treatment or not, which was not possible with the traditional clinic-pathological prognosticators. Hypoxic markers have been widely explored in the recent past for their prognostic efficacy in non-small cell lung cancer. The present review article will enlighten importance of hypoxic markers with special reference to non-small cell lung carcinoma.

Loss of Fas apoptosis inhibitory molecule leads to spontaneous obesity and hepatosteatosis.

Altered hepatic lipogenesis is associated with metabolic diseases such as obesity and hepatosteatosis. Insulin resistance and compensatory hyperinsulinaemia are key drivers of these metabolic imbalances. Fas apoptosis inhibitory molecule (FAIM), a ubiquitously expressed antiapoptotic protein, functions as a mediator of Akt signalling. Since Akt acts at a nodal point in insulin signalling, we hypothesize that FAIM may be involved in energy metabolism. In the current study, C57BL/6 wild-type (WT) and FAIM-knockout (FAIM-KO) male mice were fed with normal chow diet and body weight changes were monitored. Energy expenditure, substrate utilization and physical activities were analysed using a metabolic cage. Liver, pancreas and adipose tissue were subjected to histological examination. Serum glucose and insulin levels and lipid profiles were determined by biochemical assays. Changes in components of the insulin signalling pathway in FAIM-KO mice were examined by immunoblots. We found that FAIM-KO mice developed spontaneous non-hyperphagic obesity accompanied by hepatosteatosis, adipocyte hypertrophy, dyslipidaemia, hyperglycaemia and hyperinsulinaemia. In FAIM-KO liver, lipogenesis was elevated as indicated by increased fatty acid synthesis and SREBP-1 and SREBP-2 activation. Notably, protein expression of insulin receptor beta was markedly reduced in insulin target organs of FAIM-KO mice. Akt phosphorylation was also lower in FAIM-KO liver and adipose tissue as compared with WT controls. In addition, phosphorylation of insulin receptor substrate-1 and Akt2 in response to insulin treatment in isolated FAIM-KO hepatocytes was also markedly attenuated. Altogether, our data indicate that FAIM is a novel regulator of insulin signalling and plays an essential role in energy homoeostasis. These findings may shed light on the pathogenesis of obesity and hepatosteatosis.