Regulation network of serum cytokines induced by tuberculosis-specific antigens reveals biomarkers for tuberculosis diagnosis.

In this study, we identified potential serum biomarkers for the diagnosis of active tuberculosis (TB) and screening for latent TB infections (LTBIs). Peripheral blood samples from 40 healthy individuals, 40 patients with TB, and 40 LTBI individuals were stimulated with the TB-specific antigens ESAT-6 and CFP-10. Human inflammatory cytokine arrays were used to detect the expression of inflammatory cytokines. Cytokines with significant changes were screened to construct a cytokine regulation network. The levels of the cytokines CCL1 (I-309), CXCL9 (MIG), IL-10, IL-6, CSF2, CSF3, IL-8, IL-1α, IL-7, TGF-ß1, CCL2, IL-2, IL-13, and TNFα were significantly upregulated in the active TB group. The levels of CCL3, IL-1ß, CCL8, IFNγ, and CXCL10 were significantly increased in the TB groups compared to those in the healthy control group. sTNF RII was upregulated in the LTBI group. CCL4 and MIP1d were significantly increased in all groups.The upregulated cytokines were mainly found in the IFNγ and IL-1α regulatory networks. Importantly, we found that CXCL10 (IP-10), CCL3, CCL8, and IL-1ß may be more suitable than IFNγ for active or latent TB infection screening. Furthermore, we found that levels of CCL1 (I-309), CXCL9 (MIG), IL-10, IL-6, CSF2, CSF3, IL-8, IL-1α, IL-7, TGF-ß1, CCL2, IL-2, and IL-13 after TB antigen stimulation may help distinguish between active and latent TB.

MiR-808 inhibits cardiomyocyte apoptosis and expressions of caspase-3 and caspase-9 in rats with myocardial infarction by regulating TGF-ß1 signaling pathway.

OBJECTIVE: To investigate the effects of micro ribonucleic acid (miR)-808 on cardiomyocyte apoptosis and expressions of caspase-3 and caspase-9 in rats with myocardial infarction (MI) by regulating the transforming growth factor-ß1 (TGF-ß1) signaling pathway. MATERIALS AND METHODS: A total of 24 specific pathogen-free female Sprague-Dawley rats were enrolled and randomly divided into normal group, model group, and miR-808 group, 8 rats in each group. In the model group and miR-808 group, MI model was prepared by ligation of the left anterior descending coronary artery in the rats. The miR-808 group was transfected with miR-808 lentivirus after the model was established. After one week of intervention, the expression of TGF-ß1 was detected by reverse transcription-polymerase chain reaction (RT-PCR). The cardiac function of rats was determined by echocardiography. The myocardium of rats was observed by Masson staining. The cardiomyocyte apoptosis of rats was examined by TdT-mediated dUTP-biotin nick end labeling (TUNEL) method. The expression levels of caspase-3 and caspase-9 were detected by Western blotting. RESULTS: The expression of TGF-ß1 mRNA was higher in the model group than that in the normal group (p<0.05), but compared with that in the model group, it was lower in the miR-808 group. The myocardial function and cardiomyocyte survival rate in the miR-808 group was better and higher than those in the model group (p<0.05). The expression levels of caspase-3 and caspase-9 in the miR-808 group were lower than those in the model group (p<0.05). CONCLUSIONS: MiR-808 can inhibit cardiomyocyte apoptosis in rats with MI by down-regulating TGF-ß1 expression and inhibiting the expressions of caspase-3 and caspase-9.

Downregulated miRNA-26a-5p induces the apoptosis of endothelial cells in coronary heart disease by inhibiting PI3K/AKT pathway.

OBJECTIVE: Multiple microRNAs (miRNAs) are abnormally expressed in endothelial cells during the occurrence of coronary artery disease (CAD). Previous researches have demonstrated that miRNA-26a-5p participates in regulating the proliferation of vascular smooth muscle cells and angiogenesis. The aim of this study was to clarify the role of miRNA-26a-5p in regulating cellular performances of endothelial cells in the progression of CAD. PATIENTS AND METHODS: In vivo CAD model was successfully established by feeding high-fat diet in 8-week-old female ApoE/LDLR-/- mice. CAD mice were administered with miRNA-26a-5p NC or miRNA-26a-5p inhibitor, respectively. Meanwhile, coronary endothelial cells were isolated from CAD mice and normal controls. Relative levels of miRNA-26a-5p, the gene of phosphate and tension homology deleted on chromosome ten (PTEN) and vascular endothelial growth factor (VEGF) in CAD patients and coronary endothelial cells isolated from CAD mice were examined. The regulatory effect of miRNA-26a-5p on atherosclerosis-related genes in primary endothelial cells and HUVECs were detected as well. Moreover, the viability and apoptosis of primary endothelial cells with miRNA-26a-5p knockdown were assessed by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. Dual-luciferase reporter gene assay was conducted to identify the relationship between miRNA-26a-5p and PTEN. Furthermore, the regulatory role of miRNA-26a-5p in phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway was examined in endothelial cells. RESULTS: MiRNA-26a-5p and VEGF were significantly downregulated in CAD patients and primary endothelial cells isolated from CAD mice. However, PTEN was significantly upregulated. CAD mice administrated with miRNA-26a-5p inhibitor exhibited remarkably upregulated ET-1, TxA2, and ANG II, as well as downregulated eNOS and PGI2. Conversely, transfection of miRNA-26a-5p mimics in HUVECs obtained the opposite trends. PTEN was identified as the direct target gene of miRNA-26a-5p. Moreover, significantly reduced viability and enhanced apoptotic rate were observed in endothelial cells isolated from CAD mice administrated with miRNA-26a-5p inhibitor. In addition, the protein level of p-AKT in endothelial cells with miRNA-26a-5p knockdown was significantly down-regulated. CONCLUSIONS: MiRNA-26a-5p influences the proliferative and apoptotic abilities of endothelial cells isolated from CAD mice by targeting PTEN to activate PI3K/AKT pathway.

CIRBP protects H9C2 cells against myocardial ischemia through inhibition of NF-κB pathway.

Myocardial ischemia is a major cause of death and remains a disease with extremely deficient clinical therapies and a major problem worldwide. Cold inducible RNA-binding protein (CIRBP) is reported to be involved in multiple pathological processes, including myocardial ischemia. However, the molecular mechanisms of myocardial ischemia remain elusive. Here, we first overexpressed CIRBP by transfection of pc-CIRBP (pcDNA3.1 containing coding sequenced for CIRBP) and silenced CIRBP by transfection of small interfering RNA targeting CIRBP (siCIRBP). pcDNA3.1 and the negative control of siCIRBP (siNC) were transfected into H9C2 cells to act as controls. We then constructed a cell model of myocardial ischemia through culturing cells in serum-free medium with hypoxia in H9C2 cells. Subsequently, AlamarBlue assay, flow cytometry and western blot analysis were used, respectively, to assess cell viability, reactive oxygen species (ROS) level and apoptosis, and expression levels of IκBα, p65 and Bcl-3. We demonstrated that CIRBP overexpression promoted cell proliferation (P<0.001), inhibited cell apoptosis (P<0.05), reduced ROS level (P<0.001), down-regulated phosphorylated levels of IκBα and p65 (P<0.01 or P<0.001), and up-regulated expression of Bcl-3 (P<0.001) in H9C2 cells with myocardial ischemia. The influence of CIRBP knockdown yielded opposite results. Our study revealed that CIRBP could protect H9C2 cells against myocardial ischemia through inhibition of NF-κB pathway.

Non-serotonergic dorsal and median raphe projection onto parvalbumin- and calbindin-containing neurons in hippocampus and septum.

The median raphe nucleus is involved in controlling and maintaining hippocampal activity through its projection to inhibitory neurons in medial septum and hippocampus. It has been shown that anterogradely axonal-traced fibers originating in the median raphe nucleus project onto calbindin-containing neurons in hippocampus and parvalbumin-containing neurons in medial septum. Parallel immunohistochemistry studies showing serotonin fibers contacting calbindin- and parvalbumin-positive neurons have led to the assumption that raphe fibers projecting on these types of neurons are mainly serotonergic. However, in both dorsal and median raphe nucleus there is a large amount of non-serotonergic neurons which also are projecting neurons, indicating that a part of the raphe fibers projecting to hippocampus and septum may be non-serotonergic. Our aim was to determine whether there is a non-serotonergic projection from the raphe nucleus onto calbindin- and parvalbumin-containing neurons in hippocampus and septum. Biotin dextran amine was used as the anterograde neuronal tracer and injected into either dorsal or median raphe nucleus. By use of triple immunofluorescence-labeling we analyzed the serotonergic content of the biotin dextran amine-labeled fibers contacting parvalbumin- and calbindin-positive neurons. Surprisingly, we found a significant non-serotonergic projection from both dorsal and median raphe nuclei onto calbindin- and parvalbumin-containing interneurons in septum and hippocampus, with a preference in hippocampus for projecting onto calbindin-positive neurons. These results indicate that the raphe nuclei may exert their control on hippocampal and septal activity not only through a serotonergic projection, but also through a significant non-serotonergic pathway.

Application of biotin-avidin system, determination of circulating immune complexes, and evaluation of antibody response in different hydatidosis patients.

The avidin-biotin-peroxidase complex enzyme-linked immunosorbent assay (ABC-ELISA) and standard ELISA were used for the detection of Echinococcus granulosus antibody in sera of 101 patients operated on for hydatid disease, 40 patients with miscellaneous nonhydatid diseases, and 61 normal subjects. Sensitivity and specificity of the two procedures were comparable and the geometric mean antibody titer detected with ABC-ELISA was higher than with standard ELISA. The ABC-ELISA is a sensitive, specific, simple, and convenient method for diagnosing hydatidosis.

Thoracic hydatid cysts: a report of 842 cases treated over a thirty-year period.

From 1957 to 1985, 842 patients were diagnosed as having thoracic hydatid cysts; 810 cysts were intrathoracic, 29 occurred on the "liver roof," 2 were cardiac, and 1 was on the chest wall. A total of 1,010 surgical procedures were performed in 807 patients (35 refused operation). There was a total operative mortality of 0.6% (5 deaths). Procedures became more conservative as experience was gained, and 79% of the procedures were endocystectomies. Intact endocystectomy (Barrett's technique) without preliminary aspiration was the approach of choice. Careful protection of the operating field, suturing of all the bronchial openings, and capitonnage were the keys to successful treatment. One hundred six patients with intact endocystectomies done before July, 1975, were followed for 3 to 20 years. Ruptures occurred during cyst manipulation in 35 patients (33%). Recurrence after operation was seen in 2 patients (1.9%). There were no deaths among the patients undergoing intact endocystectomy. In comparison, we followed 136 patients who underwent aspiration endocystectomy and the recurrence rate was 3.7% (5 patients).

[Effects of bromocriptine microinjection into VTA on the DOPAC level in nucleus accumbens].

The experiment was performed in sodium pentobarbital anesthetized male SD rats. In vivo differential pulse voltammetry (DPV) was used to monitor a metabolite of dopamine-3,4-dihydroxyphenylacetic acid (DOPAC) level in the ipsilateral nucleus accumbens following microinjection of bromocriptine (2.5 micrograms/0.5 microliters, n = 5) into left ventral tegmental area (VTA). The results indicated that after microinjection the amplitude of DOPAC peak increased gradually and reached to 128% and 143% compared to the control at the 50th and 80th min respectively (P < 0.01). The duration of the effect was about two hours. One hundred and eighty min after injection, the amplitude of DOPAC decreased to 111% of control (P > 0.05). SCH23390 (2 nmol/0.5 microliters, n = 4), a selective D1 antagonist was given into left VTA 40 min prior to microinjection of bromocriptine. In this group, the above-mentioned effect of bromocriptine was blocked, the amplitude of DOPAC peak showed no increase. The results suggest that the microinjection of bromocriptine into VTA induces an increase of activity of mesolimbic dopaminergic neurons probably mediated by D1 receptor.

[Regulatory effect of ventral tegmental area on sleep-wakefulness in rats].

Experiments were performed on 31 male SD rats. The results were as follows: (1) The time of waking was increased during the second and third hours after microinjection of 3.3 and 6.6 nmol bromocriptine into bilateral VTA (P < 0.01), but the 1.33 nmol group was without significant effect. (2) The time of waking was decreased during the second and third hours after microinjection of 2 and 4 nmol SCH23390 into VTA (P < 0.01 and P < 0.05 respectively), while 3.4 nmol sulpiride group was without effect. (3) One week after administration of kainic acid (0.3 microgram), the rats presented less time of waking in the day (P < 0.05), while sleep-wakefulness cycle at night showed no change. The results suggest that bromocriptine could increase wakefulness by activating VTA DA neurons via D1 receptor and that the DA neurons might exert a tonic effect for maintaining the wakefulness at the daytime.