Neuroprotection of interleukin-6 against NMDA-induced neurotoxicity is mediated by JAK/STAT3, MAPK/ERK, and PI3K/AKT signaling pathways.

We have previously shown that interleukin-6 (IL-6) has neuroprotective effect against N-methyl-D-aspartate (NMDA)-induced excitotoxicity. The current study aimed to reveal signal transduction pathways involved in the IL-6 neuroprotection. Cerebellar granule neurons (CGNs) from postnatal 8-day infant rats were exposed to IL-6 (120 ng/ml) for 8 days and stimulated with NMDA (100 µM) for 15 or 30 min. Dynamic intracellular Ca(2+) fluorescence intensity, cytosolic Ca(2+)-dependent phospholipase A2 (cPLA2) expression, and apoptosis and necrosis in cultured CGNs were measured by laser scanning confocal microscope, real-time PCR and Western blot, and annexin V-FITC/propidium iodide staining, respectively. NMDA stimulation of neurons evoked an intracellular Ca(2+) overload, an upregulated expression of cPLA2, and an increase in cell death. Chronic IL-6 exposure prevented the NMDA-evoked neuronal Ca(2+) overload, cPLA2 expression upregulation, and apoptosis and necrosis. Anti-gp130 monoclonal antibody (mAb), a blocker of gp130 that is a 130-kDa signal-transducing ß-subunit of IL-6 receptor complex, blocked these effects of IL-6 preventing NMDA neurotoxicity. AG490, PD98059, or LY294002, inhibitors specific for the intracellular signals, JAK, MAPK, and PI3K, respectively, partially blocked these IL-6 neuroprotective effects. Phosphorylation levels of STAT3, ERK1/2, and AKT, the downstream proteins for these enzymes of JAK, MAPK, and PI3K, respectively, were elevated by IL-6 pretreatment. The enhanced activation of STAT3, ERK1/2, and AKT by IL-6 was abolished by AG490, PD98059, and LY294002, respectively. Anti-gp130 mAb attenuated the activation of all the three detected signaling molecules. The present findings suggest that IL-6 neuroprotection is jointly mediated by the cellular signal transduction pathways, gp130-JAK-STAT3, gp130-MAPK-ERK, and gp130-PI3K-AKT.

Lymphocyte-derived catecholamines induce a shift of Th1/Th2 balance toward Th2 polarization.

AIMS: Our previous work has shown that lymphocytes synthesize and secrete catecholamines (CAs), which regulate lymphocyte proliferation and apoptosis. In the present study, we explored the effect of the lymphocyte-derived CAs on differentiation and function of T helper (Th) cells. METHODS: Lymphocytes were separated from the mesenteric lymph nodes of mice and stimulated by concanavalin A (Con A). These cells were treated with alpha-methyl-p- tyrosine (α-MT), an inhibitor of tyrosine hydroxylase (TH) that is a rate-limiting enzyme for synthesis of CAs, and pargyline, an inhibitor of monoamine oxidase that degrades CAs. RESULTS: Treatment of Con A-stimulated lymphocytes with α-MT (10(-6) M) reduced CAs both in the cultured lymphocytes and in the culture supernatants. Simultaneously, α-MT upregulated expression of mRNAs and proteins of T-box expressed in T cells (T-bet) and interferon-γ (IFN-γ) but downregulated expression of mRNAs and proteins of GATA binding protein 3 (GATA-3) and interleukin-4 (IL-4) in Con A-activated lymphocytes. In contrast, pargyline (10(-6) M) increased intracellular and supernatant CA contents in Con A-activated lymphocytes. Meanwhile, the treatment with pargyline downregulated expression of T-bet and IFN-γ but upregulated expression of GATA-3 and IL-4 in these lymphocytes. CONCLUSION: CAs synthesized and secreted by lymphocytes regulate differentiation and function of Th cells, with an effect facilitating the shift of Th1/Th2 balance toward Th2 polarization.

Effectiveness and Safety of Palbociclib Plus Endocrine Therapy in Patients with Advanced Breast Cancer: A Multi-Center Study in China.

BACKGROUND: Palbociclib has been approved for marketing in China. However, its effectiveness, safety, and latent variables in the Chinese population require further investigation. METHODS: Information was retrieved from 397 patients with metastatic breast cancer (mBC) who received at least two cycles of palbociclib plus endocrine therapy (PAL plus ET) at eight clinical sites in China. The patients' demographic characteristics, treatment patterns, and adverse events (AEs) were analyzed. RESULTS: The objective response rate (ORR) and clinical benefit rate (CBR) for PAL plus ET were 28.97% and 66.25%, respectively. The median PFS was 14.2 months in the whole population. In addition to protein Ki-67 status and sensitivity to ETs, no liver metastases, fewer metastatic sites, an earlier line of therapy, and treatment combined with AI instead of FUL were also considered as independent prognostic factors for PAL treatment. Administration of PAL was generally well tolerated in patients with hormone-receptor-positive and human-epidermal-growth-factor-receptor-2-negative (HR+/HER2-) advanced breast cancer (ABC). The therapy was safe in the elderly population, which is consistent with the outcomes of the whole population and previous reports. CONCLUSIONS: In this most widely distributed study in China to date, palbociclib combined with ET proved its effectiveness for HR+/HER2- ABC treatment, and adverse events were manageable. Here, we identified some independent prognosis factors, but the mechanism by which these factors influence effectiveness requires further verification.

The hypothalamus mediates the effect of cerebellar fastigial nuclear glutamatergic neurons on humoral immunity.

OBJECTIVES: We explored effect of glutamatergic neurons in the fastigial nucleus (FN), one of three cerebellar nuclei, on humoral immunity and revealed that this effect was mediated by the hypothalamus via FN-hypothalamic glutamatergic transmission. METHODS: Rats were immunized with bovine serum albumin (BSA). On the third day after the immunization, 6-diazo-5-oxo-L-norleucine (DON), an inhibitor of glutaminase for glutamate synthesis, was microinjected in bilateral FN and D,L-threo-ß-hydroxyaspartic acid (THA), an inhibitor of glutamate transporters on plasma membrane, was microinjected in both sides of lateral hypothalamic area (LHA). Glutamate content in the hypothalamus was examined by high-performance liquid chromatography (HPLC). Flow cytometry and enzyme-linked immunosorbent assay (ELISA) were used to measure B lymphocyte percentage in mononuclear cells of peripheral blood and levels of anti-BSA IgM and IgG antibodies in the serum, respectively. RESULTS: DON injection in bilateral FN reduced B lymphocyte percentage and anti-BSA IgM and IgG levels, and simultaneously decreased glutamate content in the hypothalamus. Combined treatment with DON in the FN and with THA in the LHA elevated B cell number and anti-BSA IgM and IgG levels and increased hypothalamic glutamate content compared with DON treatment alone. However, combined treatment with DON in the FN and with THA in the ventrolateral thalamic nuclei (VL) did not significantly alter DON-dependent changes in B cell number and antibody levels, although the co-treatment altered DON-dependent glutamate content in the thalamus. CONCLUSION: Cerebellar FN glutamatergic neurons participate in modulation of humoral immunity and this effect is mediated by the hypothalamus via FN-hypothalamic glutamatergic transmission.

Estimation Strategy Selection Is Modulated by Snapshot Emotional Priming, but Not Math Anxiety.

The present study explored the role of snapshot emotional priming and math anxiety in estimation strategy selection. Participants were asked to complete a two-digit multiplication estimation task (e.g., 34 × 67) under explicit (Experiment 1) and implicit (Experiment 2) snapshot emotional priming conditions by freely choosing to use DU (down-up, e.g., doing 30 × 70 = 2100 for 34 × 67) or UD (up-down, e.g., doing 40 × 60 = 2400 for 34 × 67) strategies to arrive as close as possible to the correct answer. In Experiment 1, individuals' estimation performance was positively influenced by explicit happy priming (shorter RT (reaction time)), while not affected by explicit fear priming. In Experiment 2, individuals' estimation ACC (accuracy) when using the UD strategy was negatively affected by both implicit happy and fear priming, but their RT when using DU and UD strategies was positively impacted by implicit happy priming. In both experiments, the correlations between math anxiety and estimation performance (ACC, RT, and strategy selection adaptivity) was not significant. The present study suggests that fear priming was not always detrimental to individuals' estimation performance, and happy priming did not always universally improve individuals' estimation performance. Additionally, estimation strategy selection was not influenced by math anxiety.

[Expression of CAR in myocardial of viral myocarditis and dilated cardiomyopathy].

OBJECTIVE: In order to improve accuracy and reliability of forensic diagnosis of sudden cardiac death, pathogenesis and relationship between the viral myocarditis (VMC) and dilated cardiomyopathy (DCM) were investigated. METHODS: Improved immunohistochemical technique was used to detect the expression of the CAR in myocardium samples, including 22 deceased with VMC, 20 deceased with DCM and 16 control deceased. RESULTS: The brown staining on the cell membrane of myocardium showed positive result. There was a prominent CAR expression in VMC group and DCM group, which were statistically significant difference compared with control group (P < 0.05). CONCLUSION: The CAR expression showed significantly higher in VMC and DCM groups. The viral infection can result in myocardial necrosis and impaired cardiac functions. These abnormalities can trigger a cascade of events that contributed to the progress of VMC to DCM.

[High mobility group box 1 contributes to the endoplasmic reticulum stress of liver in rats with trauma].

OBJECTIVE: To investigate the role of high mobility group box 1 (HMGB1) in hepatic endoplasmic reticulum stress (ERS) in rats with trauma. METHODS: Sixty SPF Sprague-Dawley (SD) rats were randomly divided into groups (n = 6). The rat model of liver injury following traumatic stress was established by continuous compressing the bilateral hind-limbs of rats for 3 hours and then intermittent compressing and decompressing for 30 minutes respectively three times with standard weight of 15 kg. The experiment 1 was divided into two groups: control group and 6, 18, 30 hours after crush. The experiment 2 was divided into control group, crush model group (18 hours after crush), HMGB1 inhibitor sodium butyrate (SB) or ethyl pyruvate (EP) groups, and SB or EP treatment groups (500 mg/kg SB solution or 40 mg/kg EP solution was injected intraperitoneally after 3 hours crush). The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were measured with automatic biochemistry analyzer. Histopathological severity of liver injury was assessed by hematoxylin and eosin (HE) staining. The expressions of HMGB1 and ERS-related proteins were detected with Western Blot. The expression and translocation of HMGB1 in liver tissue were evaluated by immuno-histochemical technique. RESULTS: (1) Compared with the control group, the pathological changes of liver injury, the levels of AST and ALT in serum and protein expression of HMGB1 as well as ERS-related proteins such as glucose regulated protein 78 (GRP78), caspase-12, and inositol-requiring enzyme 1α (IRE1α) in liver tissue were significantly increased after traumatic stress, and reached the peak at 18 hours. The expression of C/EBP-homologous protein (CHOP) was increased in a time-dependent manner and peaked at 30 hours after crush. Immunohistochemistry showed that HMGB1 expression increased at 6 hours after crush, some HMGB1 shifted from nucleus to cytoplasm, and the expression was more obvious at 18 hours. (2) Compared with crush model group, the expressions of HMGB1 and ERS-related proteins were significantly decreased following the administration of HMGB1 inhibitors SB or EP (HMGB1/ß-actin: 0.703±0.213, 0.512±0.075 vs. 1.041±0.186; GRP78/ß-actin: 0.614±0.052, 0.450±0.115 vs. 0.847±0.120; caspase-12/ß-actin: 0.636±0.066, 0.812±0.142 vs. 1.086±0.130; CHOP/ß-actin: 0.314±0.046, 0.621±0.123 vs. 0.996±0.764; IRE1α/ß-actin: 0.473±0.033, 0.519±0.094 vs. 0.742±0.054, all P < 0.05), the levels of serum AST and ALT were significantly decreased [AST (U/L): 1 030.50±427.73, 1 414.50±347.86 vs. 2 122.20±322.76; ALT (U/L): 285.75±11.30, 368.50±80.58 vs. 473.80±33.54, all P < 0.01], the degree of acute liver injury was reduced. Only SB or EP could not affect the parameters mentioned above. CONCLUSIONS: HMGB1-ERS pathway was involved in mediating traumatic stress-induced acute liver injury in rats.

Circulating soluble form of LR11, a regulator of smooth muscle cell migration, is a novel marker for intima-media thickness of carotid arteries in type 2 diabetes.

BACKGROUND: Smooth muscle cell (SMC) migration from the media to the intima, a process affecting plaque stability in advanced-stage atherosclerosis, is under the control of LR11. To delineate the clinical significance of the circulating soluble form of LR11 (sLR11) in patients with type 2 diabetes (T2D), we analyzed the correlation of sLR11 levels with intima-media thickness (IMT) of carotid arteries. METHODS: Plasma sLR11 levels were measured in 165 patients with T2D (mean age 56.2±10.4 y, 58.2% males, and BMI 24.6±3.6) by ELISA. Averaged IMT levels of common carotid arteries were determined by ultrasonography. RESULTS: Circulating sLR11 levels were 9.8±3.5ng/ml, and correlated positively with the classical atherosclerosis risk factors age, sex, systolic blood pressure, low-density lipoprotein-cholesterol (LDL-C), fasting plasma-glucose (FPG), and glycosylated hemoglobin. Multivariate linear regression analysis indicated that only FPG was associated with sLR11; sLR11 correlated positively with IMT, together with age and FPG, but less with LDL-C. Among the serum risk factors for IMT, multivariate linear regression analysis uncovered that sLR11 was independently associated with IMT. Subsequent logistic analysis revealed that FPG correlated best with IMT values at a cut-off of 0.80mm and sLR11 at a cut-off of 0.90mm, respectively, while LDL-C showed lower discriminatory power at any IMT cut-off values. CONCLUSION: Increased sLR11 concentrations are highly associated with increased IMT as well as with FPG in middle-aged, non-obese patients with T2D. Circulating sLR11 may be a novel marker representing the pathophysiology of intimal SMCs in patients with T2D.

[Effect of matrine on regulating renal tubulointerstitial expression of MMP-3, TIMP-1 and FN].

OBJECTIVE: To explore the effect and mechanism of matrine on regulating renal tubulointerstitium expression of MMP-3, TIMP-1 and FN. METHODS: Seventy male SD rats were randomly allocated to five groups: normal group, shame UUO group, UUO group, UUO group treated with fusinopril (F group), UUO group treated with large dose of matrine (E group) and UUO group treated with small dose of matrine (D group). The expression levels of MMP-3, TIMP-1 and FN of the rats were determined with immunohistochemistry at 7, 14 days of the experiment. RESULTS: The expression levels of MMP-3 of the rats in the UUO group and treatment groups decreased significantly compared with the normal group and shame UUO group (P<0.05). The treatment groups had higher expression of MMP-3 than that of the UUO group (P< 0.05). The expression levels of TIMP-1 and FN of the rats in the normal group and shame UUO group were amongst the lowest. The treatment groups had lower expression of TIMP-1 and FN than that of the UUO group (P<0.05). No significant difference of expression of MMP-3, FN and TIMP-1 were found between E group and F group (P>0.05). The expression level of MMP-3 was negatively correlated with those of TIMP-1 and FN in the UUO group (P < 0.01). CONCLUSION: Matrine decreases the expression of TIMP-1 and FN in the tubulointerstitium and increases the expression of MMP-3, which would delay the progression of renal tubulointerstitial fibrosis.