[Exosomes derived from miR-133a-3p engineered mesenchymal stem cells promote myocardial repair in rats after acute myocardial infarction].

Objective: To investigate the effects of exosome derived from miR-133a-3p engineered human umbilical cord blood mesenchymal stem cells (ucMSC) on myocardial repair after acute myocardial infarction (AMI) in rats. Methods: UcMSC was amplified and cultured in vitro. Lentiviral carrying miR-133a-3p and negative control vectors were transfected into ucMSC. Exosomes secreted by the transfected ucMSC were named miR-133a-3p-Exo and miR-NC-Exo, respectively. The AMI model of rats was established by ligation of the left anterior descending coronary artery. MiR-133a-3p-Exo or miR-NC-Exo were then injected into the border zone of the infarct area. Cardiac function was assessed by echocardiography after twenty-eight days of intervention, and Masson staining was used to evaluate the area of myocardial fibrosis post-AMI. The myocardial apoptosis after infarction was evaluated by TUNEL staining and the angiogenesis after infarction was evaluated by immunofluorescence staining in the current study. Results: Compared with the miR-NC-Exo group, the left ventricular ejection fraction in the miR-133a-3p-Exo group was significantly increased ((47.4%±9.8%) vs. (64.2%±8.9%), P<0.05). While the myocardial fibrosis area ((31.2%±7.3%) vs. (18.0%±1.5%), P<0.01) and the percentage of apoptotic cardiomyocytes ((25.6%±3.6%) vs. (15.1%±4.4%), P<0.05) was significantly reduced in the miR-133a-Exo group. Besides, the expression of CD31 and α-smooth muscle actin (α-SMA) were also increased significantly in the miR-133a-3p-Exo group compared to the miR-NC-Exo group (CD31: (2.9±0.9) vs. (13.9±2.0), P<0.000 1, α-SMA: (3.5±0.9) vs. (11.0±1.6), P<0.000 1). Conclusion: Exosome derived from miR-133a-3p engineered ucMSC effectively inhibited myocardial apoptosis and promoted angiogenesis, thus improving the cardiac function after myocardial infarction in rats.

[Comparison of three dimensional CT venography and ascending phlebography based on propensity score matching in the evaluation of recurrent varicose veins].

Objective: To compare the efficacy of lower extremity three dimensional CT venography (CTV) and lower extremity ascending phlebography in evaluating recurrent varicose veins. Methods: A retrospective analysis was conducted on clinical data from 235 patients with unilateral recurrent varicose veins who were treated at the Department of Vascular Surgery,Beijing Shijitan Hospital,Capital Medical University, between January 2015 and December 2020.There were 112 males and 123 females, with an age of (62.5±11.4)years (range:24 to 75 years).Patients were stratified into two groups based on preoperative imaging examination:the CTV group (utilizing lower extremity venous ultrasound+lower extremity CTV) and the control group (employing lower extremity venous ultrasound+lower extremity ascending phlebography).The two groups were matched in a 1∶1 ratio using propensity score matching, resulting in 43 cases per group.Comparative analyses between the groups at the one-year postoperative follow-up were performed using independent sample t tests, Wilcoxon rank-sum tests, χ2 tests, and linear regression analysis. Results: One year post-surgery,the CTV group exhibited a lower venous clinical severity score (VCSS) compared to the control group(M(IQR),3.0(4.3) vs.4.0(5.8),Z=-2.038,P=0.040).Additionally, the chronic venous insufficiency patients' quality of life questionnaire (CIVIQ-20) scores were significantly higher in the CTV group than in the control group (89.0(8.0) vs.82.5(17.0), Z=-2.627, P=0.010).Patients in the CTV group also experienced a shorter ulcer healing time compared to the control group (4.0(4.0) weeks vs.12.0(7.0) weeks, Z=-3.217,P<0.01).Both groups showed no clinically symptomatic recurrent varicose veins or ulcers.However, they exhibited ultrasound-detectable varicose vein recurrence, with no statistically significant difference (χ2=0.453,P=0.500).The number of diseased vessels requiring management based on ultrasound supplemented by CTV was 16, while the number supplemented by ascending phlebography was 7,with a statistically significant difference (χ2=4.800,P=0.030).Linear regression analysis demonstrated that clinical-etiology-anatomy-pathology clinical grading and the preoperative imaging examination method exerted independent influences on VCSS and CIVIQ-20 during the one-year postoperative assessment. Conclusions: CTV-assisted ultrasound enables a direct and comprehensive evaluation and localization of diseased veins in patients with recurrent varicose veins.The utilization of lower extremity vein ultrasound combined with CTV-guided management of lower extremity vessels in minimally invasive treatment significantly improves patient prognosis, surpassing the assessment provided by ascending phlebography.

Effects of ciprofol for the induction of general anesthesia in patients scheduled for elective surgery compared to propofol: a phase 3, multicenter, randomized, double-blind, comparative study.

OBJECTIVE: Ciprofol is a newly developed intravenous sedative-hypnotic drug. The objective of the study was to prove whether ciprofol was non-inferior to propofol for the successful induction of general anesthesia. The ideal post-induction sedation level was assessed by comparing patients' clinical symptoms and their hemodynamic effects in responding to noxious stimuli, mostly tracheal intubation and bispectral index (BIS) alterations following ciprofol/propofol administration. PATIENTS AND METHODS: In this multi-center, randomized, double-blind phase 3 trial, selective surgery patients were randomly assigned in a 1:1 ratio to either ciprofol 0.4 mg/kg (n = 88) or propofol 2.0 mg/kg (n = 88) groups. The primary endpoint was the percentage of patients with successful anesthesia inductions. Secondary endpoints included the times to successful induction of general anesthesia and loss of the eyelash reflex, changes in BIS, as well as safety indicators. RESULTS: The anesthesia induction success rates for both ciprofol 0.4 mg/kg and propofol 2 mg/kg groups were 100.0%, with a 95% CI lower success limit of -4.18% difference between the two groups, indicating that ciprofol was non-inferior to propofol. For secondary outcomes, the average time to successful anesthesia and loss of the eyelash reflex were 0.91 min and 0.80 min for ciprofol and 0.80 min and 0.71 min for propofol, respectively. The pattern of BIS changes with ciprofol was similar to propofol and stable during the anesthesia maintenance period. Safety was comparable with 88.6% TEAEs in the ciprofol group compared to 95.5% in the propofol group. The incidence of injection pain was significantly lower in the ciprofol group compared to the propofol group (6.8% vs. 20.5%, p < 0.05). In addition, the patients treated with ciprofol had a lesser increase in blood pressure and heart rate, and fewer cases with BIS > 60 within 15 min of intravenous administration, which indicated that ciprofol may provide a better ideal sedation level during the post-induction period under an equivalent dosing regimen to propofol. CONCLUSIONS: Ciprofol for patients undergoing selective surgery is a new option for the induction of general anesthesia.

[Long-term efficacy of radiofrequency closure in the treatment of great saphenous vein varicose].

Objective: To examine the long-term efficacy of radiofrequency closure in the treatment of great saphenous vein varicose. Methods: The clinic data of 185 patients with varicose veins of lower limbs treated with radiofrequency closure admitted at Department of Vascular Surgery, Beijing Shijitan Hospital, Capital Medical University from July 2016 to January 2017 was analyzed retrospectively. A total of 203 limbs were treated by radiofrequency closure. The long-term efficacy of radiofrequency closure was evaluated by analyzing the closure rate, clinical-etiology- anatomy-pathophysiology (CEAP) grading, venous clinical severity score (VCSS), chronic venous insufficiency questionnaire (CIVIQ) score, and complications, using repeated measures analysis of variance. Results: All procedures were successful. The closure rate was 98.0% (199/203) at one year and two years postoperative, which was still maintained at 97.5% (198/203) at 3 years of follow-up. Postoperative CEAP grading was significantly downgraded compared with that before the operation. Totally 88.4% (76/86) of C5 to C6 grade patients downgraded to C2 to C4 grade at 6 months, and 95.3% (82/86) downgraded to C0 to C2 garde at 3 years postoperative. VCSS and CIVIQ score in both groups significantly improved at all follow-up time points compared to preoperative scores (VCSS: F=1 064.7, P=0.003; CIVIQ score: F=2 984.3, P=0.001). The most common complication was subcutaneous blood stasis (10.8%), most of which disappeared within 1 month after the surgery. Other complications included pigmentation and thrombophlebitis (5.9% and 3.9%, respectively). Conclusion: The long-term efficacy of radiofrequency closure of the great saphenous vein is satisfactory.

Neuroprotective effect of DUSP14 overexpression against isoflurane-induced inflammatory response, pyroptosis and cognitive impairment in aged rats through inhibiting the NLRP3 inflammasome.

OBJECTIVE: Postoperative cognitive dysfunction (POCD) is a common complication after general anesthesia in the elderly people. Dual-specificity phosphatase 14 (DUSP14, also known as MKP6) has been implicated in the pathogenesis of various inflammatory diseases. However, the exact role and mechanism of DUSP14 in POCD remains unclear. MATERIALS AND METHODS: An isoflurane exposure induced POCD aged rat model was successfully constructed. The pathological changes of hippocampal tissues of aged rats were detected by Nissl staining. Evaluation of learning and memory abilities in aged rats was measured using Morris water maze task test. The DUSP14 level was detected by immunohistochemistry (IHC) assay, quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot, respectively. Levels of brain injury markers [S-100ß and neuron specific enolase (NSE)] and inflammatory cytokines [interleukin (IL)-1ß (tumor necrosis factor (TNF)-α and IL-6] were detected using Enzyme Linked Immunosorbent Assay (ELISA) or qRT-PCR. The apoptosis of hippocampal nerve cells was assessed by Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Western blot assay was used to measure the expression of proteins related to apoptosis, pyroptosis and NOD-like receptor family pyrin domain-containing 3 (NLRP3)-Caspase-1 pathway. RESULTS: Isoflurane exposure led to brain injury, inflammatory response, cognitive dysfunction in aged rats and decreased the expression of DUSP14. Overexpression of DUSP14 could inhibit apoptosis, inflammation, pyroptosis, brain tissue damage, and improve cognitive dysfunction of aged rats after isoflurane anesthesia. Further mechanism studies revealed that DUSP14 may play a neuroprotective effect on POCD by regulating NLRP3 inflammasome-mediated pyroptosis. CONCLUSIONS: DUSP14 may effectively protect against isoflurane-induced neuro-inflammation, brain damage and cognitive dysfunction, indicating that DUSP14 may be a potential predictor and therapeutic target for POCD.

Carbachol alleviates myocardial injury in septic rats through PI3K/AKT signaling pathway.

OBJECTIVE: To explore the effect of carbachol on myocardial injury in septic rats, and to further study its influence on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway. MATERIALS AND METHODS: A total of 48 healthy male Sprague-Dawley rats were randomly divided into sham group (n=16), model group (n=16), and carbachol group (n=16). The rat model of sepsis was established via cecal ligation and puncture. Carbachol was intraperitoneally injected (10 µg/kg) immediately after operation in carbachol group, and no cecal ligation was performed in sham group. At 48 h after operation, the survival rate of rats in each group was recorded, the activity of plasma creatine kinase-MB (CK-MB) was detected, and the cardiac function in each group was determined. Moreover, the heart was isolated, and the myocardial tissues were taken to detect the apoptosis level using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) apoptosis kit. The content of inflammatory factors in myocardial tissues was determined using enzyme-linked immunosorbent assay (ELISA) kits, and the expression levels of apoptosis-related proteins and the PI3K/AKT signaling pathway-related proteins were detected via Western blotting. RESULTS: Carbachol could significantly raise the survival rate of septic rats (p<0.01), remarkably decrease the activity of CK-MB (p<0.01), markedly reduce the left ventricular internal diameter at end-systole (LVIDs), and markedly increase the left ventricular ejection fraction (LVEF, %) and left ventricular fractional shortening (LVFS, %). Besides, carbachol could evidently lower the apoptosis level of myocardial cells of septic rats (p<0.01), reduce the content of inflammatory factors including tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and IL-6 (p<0.01), notably decrease the expression of Caspase-3 in myocardial tissues (p<0.01), remarkably increase the expression of Bcl-2/Bax (p<0.01), and distinctly inhibit the expressions of phosphorylated (p-)PI3K, p-AKT, Nod-like receptor protein 3 (NLRP3), and Caspase-1 (p<0.01). CONCLUSIONS: Carbachol can reduce the release of inflammatory factors in myocardial cells, the expression of apoptotic proteins and the apoptosis of myocardial cells, and improve the cardiac function and survival rate of septic rats by inhibiting the PI3K/AKT signaling pathway.

Effect of lncRNA GAS5 on rats with acute myocardial infarction through regulating miR-21.

OBJECTIVE: The aim of this study was to investigate the effect of long non-coding ribonucleic acid (lncRNA) growth arrest specific 5 (GAS5) on acute myocardial infarction (AMI) model rats and to explore its regulatory mechanism. MATERIALS AND METHODS: The rat model of AMI was established by subcutaneous injection of isoproterenol (ISO). 30 Sprague Dawley (SD) rats were randomly divided into three groups, including Control group, Model group, and lncRNA GAS5 inhibitor [small interfering ribonucleic acid (siRNA) GAS5] group. Hematoxylin and eosin (H&E) staining was used to detect the pathological damage of myocardial tissues in rats of each group. Myocardial cell apoptosis in each group determined via terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The levels of matrix metalloproteinase (MMP)-2 and MMP-9 in the serum of rats in each group were examined by enzyme-linked immunosorbent assay (ELISA). Reverse transcription-polymerase chain reaction (RT-PCR) was adopted to measure the expression level of miR-21 in rat myocardial tissues. RESULTS: Compared with Control group, rats in Model group had significantly poor cardiac function, serious pathological damage of myocardial tissues, as well as increased apoptosis rate of myocardial cells. Meanwhile, the levels of MMP-2 and MMP-9 were significantly elevated in serum of Model group, while miR-21 level was down-regulated. In comparison with Model group, rats in siRNA GAS5 group exhibited significantly improved cardiac function, alleviated pathological damage to myocardial tissues, as well as decreased apoptosis rate of myocardial cells. Furthermore, the levels of MMP-2 and MMP-9 decreased significantly in serum of siRNA GAS5 group, whereas the expression level of miR-21 in myocardial tissues was down-regulated. CONCLUSIONS: SiRNA GAS5 can enhance the cardiac function of AMI model rats, relieve pathological damage, reduce myocardial cell apoptosis, and inhibit the occurrence of myocardial fibrosis. The possible underlying mechanism may be associated with up-regulation of miR-21.

LncRNA KCNQ1OT1 is overexpressed in non-small cell lung cancer and its expression level is related to clinicopathology.

OBJECTIVE: The aim of this study was to explore the expression of long non-coding RNA (lncRNA) KCNQ1OT1 in non-small cell lung cancer (NSCLC), and to elucidate its clinical significance. PATIENTS AND METHODS: The quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of lncRNA KCNQ1OT1 in NSCLC tissues and para-cancer tissues (5 cm or above away from the tumor). The relation between lncRNA KCNQ1OT1 expression and the clinical-pathological data was analyzed by the multivariate logistic regression analysis. Furthermore, the survival analysis was performed by the Kaplan-Meier method. RESULTS: The expression of lncRNA KCNQ1OT1 increased significantly in NSCLC tissues than that of in para-cancer tissues. According to the median expression of lncRNA KCNQ1OT1, NSCLC patients were divided into two groups, including high expression group and low expression group. Meanwhile, the lncRNA KCNQ1OT1 expression was correlated with tumor size, tumor node metastasis (TNM) staging, and lymph node metastasis of NSCLC patients. Both univariate analysis and multivariate analysis indicated that the high expression of lncRNA KCNQ1OT1 was closely related to TNM staging and lymph node metastasis. In addition, the Kaplan-Meier analysis showed that the overall survival and progression-free survival time of patients with higher lncRNA KCNQ1OT1 expression were significantly worse than those with lower lncRNA KCNQ1OT1 expression. CONCLUSIONS: LncRNA KCNQ1OT1 might contribute to the development of NSCLC.

CircRNA_100876 promote proliferation and metastasis of breast cancer cells through adsorbing microRNA-361-3p in a sponge form.

OBJECTIVE: This study was designed to investigate the expression level of circRNA_100876 in breast cancer (BC) tissues or cells, and to further explore whether it can promote cell metastasis and proliferative capacity via targeting microRNA- 361-3 p. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine the expression of circRNA_100876 in 50 pairs of BC tissue specimens and corresponding adjacent ones, and the correlation between circRNA_100876 expression and prognosis of patients with BC was analyzed. Meanwhile, qRT-PCR was further performed to verify circRNA_100876 level in BC cell lines. In addition, circRNA_100876 knockdown model was constructed using lentivirus and transfected in BC cells. Subsequently, the impact of circRNA_100876 on BC cell function was analyzed using Cell Counting Kit-8 (CCK-8), transwell and clone formation assays. The interplay between circRNA_100876 and microRNA- 361-3 p was verified using the Luciferase reporter gene assay and cell reverse experiment. RESULTS: QRT-PCR results showed that circRNA_100876 level in BC tissues was conspicuously higher than that in the adjacent tissues, and the patients with distant metastasis had higher expression than those without. Moreover, patients with a high expression of circRNA_100876 had a relatively lower overall survival rate. Compared with the NC group, the cell proliferation and invasion ability of circRNA_100876 knockdown group was conspicuously decreased. QRT-PCR revealed that microRNA-361-3p and circRNA_100876 showed a negative correlation in the expression level of genes in BC tissues. In addition, the results of the Luciferase reporter gene assay confirmed that circRNA_100876 can be targeted by microRNA-361-3p through their binding site. CONCLUSIONS: High expression of circRNA_100876 is conspicuously positively relevant to poor prognosis of BC patients. Additionally, circRNA_100876 is able to promote BC metastasis as well as proliferative capacity by modulating microRNA-361-3p expression.

Apatinib resensitizes cisplatin-resistant non-small cell lung carcinoma A549 cell through reversing multidrug resistance and suppressing ERK signaling pathway.

OBJECTIVE: To observe the reversal effect of apatinib on the resistance to cisplatin (DDP) of A549/cisplatin (A549/DDP) cells and its relevant mechanism. MATERIALS AND METHODS: A549/DDP cells were treated with the control method, apatinib alone, DDP alone and DDP combined with apatinib. The cell proliferation was detected by the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the cell clone formation assay. The cell apoptosis was detected by Hoechst 33258 staining and annexin V and propidium iodide (PI) double labeling. The changes in apoptotic proteins, multidrug resistance protein 1 (MDR1) and extracellular signal-regulated kinase (ERK) signaling pathway proteins in each group after treatment were detected by Western blotting. RESULTS: MTT assay results showed that compared with A549 cells, A549/DDP cells had obvious resistance to DDP. MTT assay and cell clone formation assay revealed that the tumor inhibition rate of the sub-lethal dose of apatinib (10 µM) combined with DDP was higher than that of DDP alone. The apoptosis detection results indicated that the proportion of apoptotic cells in the apatinib (10 µM) combined with DDP group was significantly increased. Western blotting results revealed that compared with that in parental A549 cells, the expression level of MDR1 in A549/DDP cells was significantly increased, and the ERK signaling pathway was activated. In the apatinib combined with DDP group, the levels of cleaved caspase-3, cleaved caspase-9 and B-cell lymphoma-2 (Bcl-2)-associated X (BAX) proteins were significantly upregulated, while the level of Bcl-2 proteins was downregulated. Apatinib could inhibit the expression of MDR1 and the activity of the ERK signaling pathway in a dose-dependent manner. CONCLUSIONS: Apatinib can restore the sensitivity of A549/DDP cells to DDP by down-regulating the expression level of MDR1 and inhibiting the activity of the ERK signaling pathway.

[Validation of the Caprini risk assessment model for venous thromboembolism in Chinese hospitalized patients in a general hospital].

Objective: To assess the validity of Caprini risk assessment model in prediction of venous thromboembolism in Chinese hospitalized patients in a general hospital. Methods: Medical record review was performed in Beijing Shijitan Hosital for all eligible hospitalized patients who underwent screening for venous thromboembolism between January and December 2015. The Caprini score of patients with or without venous thromboemboilism and incidence of venous thromboembolism in patients with various Caprini risk levels, surgery and medical patients was compared. Results: A total of 6 966 inpatients were enrolled. Three hundred and ninety-six patients developed venous thromboembolism. The Caprini median score of patients with venous thromboemboilism was 5 (3-7), which higher than 3(2-5) of patients without venous thromboembolism(Z=-13.68, P<0.01). Incidence of venous thromboembolism of patients in low, moderate, high, highest risk level was 1.0%, 1.8%, 5.7%, 10.6%, respectively. There was no statistically significant difference of incidence between low and moderate risk patients (OR=1.88, 95%CI: 0.89-3.99, P>0.05), but significant difference between moderate and high risk (OR=3.23, 95%CI: 2.06-5.06, P<0.01), high and highest risk patients (OR=1.97, 95%CI: 1.59-2.45, P<0.01). There was no incidence difference of venous thromboembolism between surgery and medical patients in the same Caprini level of low (χ(2)=3.58 , P>0.05), moderate(χ(2)=2.89, P>0.05), high(χ(2)=0.46, P>0.05), highest risk(χ(2)=1.61, P>0.05). Conclusion: Caprini risk assessment model can effectively predict the occurence of venous thromboembolism in Chinese hospitalized patients with high risk of VTE(Caprini score >2)in a general hospital.

Cardiovascular disease in rheumatoid arthritis: medications and risk factors in China.

This study aims to assess the risk factors of cardiovascular disease (CVD) and to determine the association of traditional and biologic disease-modifying anti-rheumatic drugs (DMARDs) with risk for CVD in Chinese rheumatoid arthritis (RA) patients. A cross-sectional cohort of 2013 RA patients from 21 hospitals around China was established. Medical history of CVD was documented. The patients' social background, clinical manifestations, comorbidities, and medications were also collected. Of the 2013 patients, 256 had CVD with an incidence of 12.7%. Compared with non-CVD controls, RA patients with CVD had a significantly advanced age, long-standing median disease duration, more often male and more deformity joints. Patients with CVD also had higher rates of smoking, rheumatoid nodules, interstitial lung disease, and anemia. The prevalence of comorbidities, including hypothyroidism, diabetes mellitus (DM), hypertension, and hyperlipidemia, was also significant higher in the CVD group. In contrast, patients treated with methotrexate, hydroxychloroquine (HCQ), and TNF blockers had lower incidence of CVD. The multivariate analysis showed that the use of HCQ was a protective factor of CVD, while hypertension, hyperlipidemia, and interstitial lung disease were independent risk factors of CVD. Our study shows that the independent risk factors of CVD include hypertension, hyperlipidemia, and interstitial lung disease. HCQ reduces the risk of CVD in patients with RA.

Chronic alcohol administration affects purine nucleotide catabolism in vivo.

AIMS: To investigate the relationship between chronic alcohol administration and purine nucleotide metabolism in vivo. MAIN METHODS: Rat models of alcohol dependence and withdrawal were used. The concentrations of uric acid (UAC), urea nitrogen (UREA), creatinine (CREA), and beta-2-microglobulin (ß2-M) and creatinine clearance rate (CCR) in plasma were measured. The PLC method was used to detect the absolute content of purine nucleotides in different tissues. Enzymatic activities of adenosine deaminase (ADA), xanthine oxidase (XO), ribose 5-phosphate pyrophosphokinase (RPPPK), glutamine phosphoribosylpyrophosphate amidotransferase (GPRPPAT), hypoxanthine-guanine phosphate ribose transferase (HGPRT), and adenine phosphoribosyltransferase (APRT) in the tissues were analyzed. Real-time PCR was used to determine the relative level of ADA and XO. KEY FINDINGS: The renal function of rats with alcohol dependence was normal. Further, the content of purine nucleotides (GMP, AMP, GTP, and ATP) in tissues of the rats was decreased, which indicated that the increased uric acid should be derived from the decomposition of nucleotides in vivo. The activity of XO and ADA increased, and their mRNA expression was enhanced in the alcohol dependence group, but there was no significant difference in the activity of RPPPK and GPRPPAT in the liver, small intestine, and muscle; furthermore, no significant difference in the activity of HGPRT and APRT was observed in the brain. SIGNIFICANCE: These results indicate that chronic alcohol administration might enhance the catabolism of purine nucleotides in tissues by inducing gene expression of ADA and XO, leading to elevation of plasma uric acid levels.

Integrated analysis of gene expression profile and genetic variations associated with ovarian cancer.

OBJECTIVE: Ovarian cancer is the most lethal gynecologic cancer worldwide, since most patients are diagnosed at an advanced stage. To improve the early diagnosis and treatment of ovarian cancer, we performed a integrated analysis of transcription profile and genetic variations to study on the molecular pathogenesis in ovarian cancer. METHODS: mRNA expression profiles of ovarian cancer and normal controls downloaded from ArrayExpress database were applied to identify differentially expressed genes (DEGs). The chromosomal distributions of these DEGs were established using DAVID. Then, DNASeq data from the Cancer Genome Atlas (TCGA) were extracted to analyze gene mutational information including the number of mutations (mut), the number of mutational genes (mutG) and chromosomal distributions of mutations. Statistical method was offered to carrying on correlation analysis of gene mutations and differential expression. RESULTS: A total of 1732 DEGs were identified, and the chromosomal distributions of 97 genes were unknown. These DEGs were most significantly distributed on chromosome 4 with p value = 1.34E-7. Chromosome 1 enriched the most DEGs (11.56%). Statistical algorithm showed that DEGs presented significantly positive correlation with mut (p = 0.000009) and mutG (p = 0.00001). In 48.7% DEGs, gene mutations were found. CONCLUSIONS: We conducted scientific analysis on integration of DEGs in expression profiles and genetic mutations in ovarian cancer, displayed the correlation of differential expression and genetic variations. The result indicated that expression profiles were significantly correlated to genotype.

Downregulation of spinal endomorphin-2 correlates with mechanical allodynia in a rat model of tibia cancer.

The endogenous tetrapeptide endomorphin-2 (EM2) participates in pain modulation by binding to pre- and/or post-synaptic µ opioid receptor (MOR). In the present study, pathological expression and antinociceptive effects of EM2 at the spinal level were investigated in a rat model of bone cancer pain. The model was established by introducing Walker 256 mammary gland carcinoma cells into the tibia medullary cavity. Immunohistochemical staining for EM2 showed a markedly reduced EM2-immunoreactivity in the ipsilateral spinal dorsal horn on days 6, 12 and 18 post Walker 256 inoculation (p < 0.05). Intrathecal injection (i.t.) of EM2 significantly attenuated cancer-induced mechanical allodynia (p < 0.05) which could be blocked by ß-funaltrexamine (ß-FNA), the µ receptor antagonist (p < 0.05). Furthermore, topical application of EM2 dose-dependently inhibited the electrically evoked C-fiber responses and postdischarge of wide dynamic range (WDR) neurons within the spinal cord (p < 0.05), and pretreatment with ß-FNA abolished the hyperactivity of these neurons. Compared with the antinociception of morphine which took effect from 40 min to 100 min post application, the analgesic action of EM2 was characterized by quick onset and short-lived efficacy (p < 0.05), being most potent at 10 min and lasting about 20 min. These findings indicate that the down-regulated spinal EM2 is an important contributor to the neuropathological process of bone cancer pain and enhancing activation of EM2/µ receptor signaling might provide a therapeutic alternative to optimizing the treatment of cancer-induced bone pain.

Expression of Trk receptors in otolith-related neurons in the vestibular nucleus of rats.

The expression of the three Trk receptors (TrkA, TrkB, and TrkC) in otolith-related neurons within the vestibular nuclei of adult Sprague-Dawley rats was examined immunohistochemically. Conscious animals were subjected to sinusoidal linear acceleration along either the anterior-posterior (AP) or interaural (IA) axis on the horizontal plane. Neuronal activation was defined by Fos expression in cell nuclei. Control animals, viz labyrinthectomized rats subjected to stimulation and normal rats that remained stationary, showed only a few sporadically scattered Fos-labeled neurons. Among experimental rats, the number of Fos-labeled neurons and their distribution pattern in each vestibular subnucleus in animals stimulated along the antero-posterior axis were similar to those along the interaural axis. No apparent topography was observed among neurons activated along these two directions. Only about one-third of the Trk-immunoreactive neurons in the vestibular nucleus expressed Fos. Double-labeled Fos/TrkA, Fos/TrkB and Fos/TrkC neurons constituted 85-98% of the total number of Fos-labeled neurons in vestibular nuclear complex and its subgroups x and y. Our findings suggest that Trk receptors and their cognate neurotrophins in central otolith neurons may contribute to the modulation of gravity-related spatial information during horizontal head movements.

Morphine alleviates early stages of apoptosis in cultured CEM x174 cells infected with simian immunodeficiency virus.

The reduction in apoptosis caused by short-term exposure of CEM x174 cells infected with SIVmac239 to morphine was investigated. Eeffects of morphine on the viability of normal and infected CEM x174 cells were determined by MTS assay. Apoptosis induced by SIVmac239 and the effects of morphine were analyzed by flow cytometry. cAMP levels, PKA activity, and the resulting histone H3 phosphorylation levels were measured. The results show a pronounced decrease in numbers of infected SIVmac239 cells compared to controls. Morphine elevated cell viability in the infected groups. Annexin V binding assays showed that 1 microM l(-1) morphine increased the percentage of viable cells and decreased apoptotic cells. Morphine also downregulated cAMP and PKA activity in both groups, but more markedly in the infected group. Histone H3 phosphorylation was elevated after virus infection and decreased in the presence of morphine. The results indicate that the cAMP-PKA signal transduction cascade is involved in morphine regulation of early SIVmac239-induced apoptosis.

Neurotrophin receptor immunostaining in the vestibular nuclei of rats.

The distribution of high-affinity neurotrophin receptors in cells of the vestibular nuclear complex and its subnuclei of adult rats was examined. We noted a high density of tyrosine kinase (Trk) A- and B- and a lower density of TrkC-immunostained cells. In particular, long, intensely labelled immunostained-TrkB fibres formed networks in the neuropil. Both TrkA- and TrkB-immunostained cells were widely distributed in the lateral, medial and spinal vestibular nuclei, and were less frequently seen in the superior vestibular nucleus, x and y subnuclei. However, immunostaining for TrkC was weak in many cells within the vestibular nuclei. The widespread and abundant neuronal distribution of Trk receptors predicts that their associated neurotrophins exert significant effects on individual cells within the vestibular nuclei.

Bacterial lipopolysaccharide activates nuclear factor-kappaB through interleukin-1 signaling mediators in cultured human dermal endothelial cells and mononuclear phagocytes.

Bacterial lipopolysaccharide (LPS)-mediated immune responses, including activation of monocytes, macrophages, and endothelial cells, play an important role in the pathogenesis of Gram-negative bacteria-induced sepsis syndrome. Activation of NF-kappaB is thought to be required for cytokine release from LPS-responsive cells, a critical step for endotoxic effects. Here we investigated the role and involvement of interleukin-1 (IL-1) and tumor necrosis factor (TNF-alpha) signal transducer molecules in LPS signaling in human dermal microvessel endothelial cells (HDMEC) and THP-1 monocytic cells. LPS stimulation of HDMEC and THP-1 cells initiated an IL-1 receptor-like NF-kappaB signaling cascade. In transient cotransfection experiments, dominant negative mutants of the IL-1 signaling pathway, including MyD88, IRAK, IRAK2, and TRAF6 inhibited both IL-1- and LPS-induced NF-kappaB-luciferase activity. LPS-induced NF-kappaB activation was not inhibited by a dominant negative mutant of TRAF2 that is involved in TNF signaling. LPS-induced activation of NF-kappaB-responsive reporter gene was not inhibited by IL-1 receptor antagonist. TLR2 and TLR4 were expressed on the cell surface of HDMEC and THP-1 cells. These findings suggest that a signal transduction molecule in the LPS receptor complex may belong to the IL-1 receptor/toll-like receptor (TLR) super family, and the LPS signaling cascade uses an analogous molecular framework for signaling as IL-1 in mononuclear phagocytes and endothelial cells.