The regional and cellular distribution of GABAA receptor subunits in the human amygdala.

GABAergic neurotransmission in the amygdala plays a crucial role in mediating emotional learning, fear, and memory. In this study, expression of five major GABAA receptor subunits (α1, α2, α3, ß2,3, and γ2) was investigated in the normal human amygdala using immunohistochemistry. At the regional level, the amygdala contains a highly heterogeneous distribution of all the subunits investigated. The most intense staining for α1, α2, ß2,3, and γ2 subunits was present in the lateral nucleus (LA), and α3 in the intercalated nuclei (ICM). Six distinct cell populations that express GABAA receptor subunits were identified throughout the amygdala: type 1 aspiny cells in the basolateral nuclear group (BLNG) and superficial cortical-like nuclear region (SCLR) express α1, ß2,3, and γ2; type 2 larger aspiny cells in the paralaminar nucleus (PL) express α1, ß2,3, and γ2; type 3 aspiny cells in the BLNG express α1, ß2,3, and γ2 as well as calcium-binding proteins including parvalbumin (PV), calbindin (CB), and calretinin (CR); type 4 pyramidal cells in the BLNG and SCLR express α2, α3, ß2,3, and γ2 subunits at high levels on proximal specialised spines; type 5 cells in the central nucleus (CE) express α2, α3, and ß2,3; type 6 cells are found closely packed in the intercalated cell masses (ICM) and express α3 and ß2,3. The α1 subunit rarely co-labelled with α2 and α3 in the same cell population, while the α2 and α3 were often expressed within the same type 4 or 5 cell though not at always at the same puncta. The predominant GABAA receptor subunit combinations expressed in the human amygdala are the α1ß2,3γ2 and α2ß2,3γ2. Cells classified as interneuron types (types 1-3) contained GAD and principally expressed α1ß2,3γ2. The major projection neurons of the BLNG (type 4) are non-GABAergic and mainly express α2ß2,3γ2. The α3 subunit was found intracellularly in type 5 cells and decorating the surface of type 6 cells but rarely co-labelled with the subunits investigated. The results reveal a complex and heterogeneous distribution of GABAA receptor subtypes throughout the amygdala as well as on a variety of cell types through which inhibitory processing is carried out to maintain emotional responses, and control anxiety and fear responses in the brain.

Serum vitamin D level correlates with disease activity and health-related quality of life in antineutrophil cytoplasmic antibody-associated vasculitis.

BACKGROUND AND OBJECTIVE: The association between vitamin D levels and disease activity has been established in patients with several autoimmune rheumatic diseases. We aimed to examine the association between vitamin D and disease activity of antineutrophil cytoplasmic antibody-associated vasculitis (AAV). METHODS: Fifty-four AAV patients and 50 age- and sex-matched healthy controls without vitamin D supplements were included. Clinical and laboratory data were evaluated during the assessment of vitamin D levels. Two different forms of vitamin D in the sera-25(OH)D, which is the sum of 25(OH)D2 and 25(OH)D3, and 25(OH)D3, which only includes 25(OH)D in its D3 form-were measured, and the relationship between vitamin D and the obtained data was assessed. Variations in vitamin D levels relative to the season were also evaluated. RESULTS: Patients with AAV demonstrated considerably lower 25(OH)D serum levels than healthy controls (16.0 vs. 20.4 ng/mL, p = 0.016), and the proportion of individuals with vitamin D deficiency was higher in patients with AAV than in healthy controls (68.5% vs. 48.0%, p = 0.035). Both serum 25(OH)D and 25(OH)D3 were positively associated with the 36-item Short-form Health Survey (SF-36) physical component summary and SF-36 mental component summary (MCS) scores. A negative correlation was observed between 25(OH)D and 25(OH)D3 serum levels and Birmingham vasculitis activity score (BVAS), C­reactive protein (CRP), and white blood cell count. Linear regression analysis indicated haemoglobin and 25(OH)D levels to be independently associated with BVAS and CRP and 25(OH)D levels with SF-36 MCS score. No seasonal variations were observed in vitamin D levels. CONCLUSION: The results from this study suggest that vitamin D levels could provide clinically useful information in AAV.

[Differential population screening and efficacy analysis of (125)I implantation after radiofrequency ablation of primary liver cancer].

Objective: To investigate the clinical efficacy radiofrequency ablation (RFA) combined with (125)I radioactive seed implantation in treatment of hepatocellular carcinoma HCC with the tumor diameter 3-5 cm. Methods: One hundred patients with HCC diagnosed clinically or pathologically with Barcelona staging of B or C in Lishui Central Hospital from February 2012 to September 2017 were retrospectively analyzed. Of the included 100 cases, 89 were males and 11 were females with the mean age of 18-80 (57±11) years old.According to the treatment modality, the subjects were divided into control group (RFA, n=67) and combined group (RFA+(125)I, n=33). Patients in control group were only received RFA and cases in combined group received RFA plus sequenced with (125)I implantation therapy. The prognosis of progression free survival (PFS) and overall survival (OS) between the two groups were compared through the Kaplan-Meier curve and Log-rank test. Results: The median follow-up time period was 6-55 months in the last follow-up time point of Dec 30, 2017. The median PFS were 4-55 (23.0±4.7) and 1-53 (12.0±1.6) months for combined and control groups respectively with significant statistical difference (P=0.015). The median OS were 6-55 (42.0±7.9) and 2-55 (38.0±2.8) months for combined and control groups with the trend of improvement in combined group, but without statistical difference (P=0.444). Subgroup analysis further indicated that the PFS was significant improved in patients with residual tumor lesions who received the combined treatment (PFS: 18 vs 9 months, P=0.025). However, there was no statistical difference for PFS between the control and combined treatment groups for cases without residual tumor lesions after RAF treatment(P=0.685). Conclusions: PFS was obviously increased in HCC patients(tumor diameter 3-5 cm) who received(125)I implantation after radiofrequency ablation, especially for cases with residual tumor lesions.

MiR-10b suppresses the growth and metastasis of colorectal cancer cell by targeting FGF13.

OBJECTIVE: MicroRNA (miRNA) plays important roles in the progression of different cancers. In this study, we investigated the precise role of miR-10b in the growth and metastasis of colorectal cancer (CRC) cell. PATIENTS AND METHODS: The levels of miR-10b in CRC cell lines were detected using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay. A series of functional assays, including cell proliferation, colony formation, wound healing and transwell invasion were conducted using miR-10b transfected cells. The expressions of E-cadherin and N-cadherin were assessed by immunofluorescence staining. Xenograft model and lung metastasis model were constructed to investigate the impact of miR-10b on the growth and metastasis of CRC cell in vivo. RESULTS: We revealed that miR-10b was markedly down-regulated in CRC. The up-regulation of miR-10b suppressed the growth, colony formation, migration and invasion of CRC cell. Furthermore, the xenograft model indicated that miR-10b inhibited CRC cell growth and lung metastasis in vivo by targeting fibroblast growth factor 13 (FGF13). In addition, we demonstrated that the overexpression of FGF13 rescued the suppressive effects of miR-10b on CRC cells growth, migration and invasion. Finally, the knockdown of FGF13 was able to mimic the inhibitory effects of miR-10b on the progression of CRC cells. CONCLUSIONS: These results demonstrate that miR-10b plays an important role in growth, migration and invasion of CRC by targeting FGF13.

Selective removal of nicotine from the main stream smoke by using a surface-imprinted polymer monolith as adsorbent.

Using molecularly imprinted polymer as a selective adsorbent for gaseous toxicants is a novel attempt. In present work, a nicotine surface-imprinted monolith (MIM) was used for the selective removal of nicotine from smoke. First, the retention capacity and selectivity for this MIM was tested by using it as the stationary phase in gas chromatography and chromatographic conditions optimized. Then, the gas phase adsorption isotherms of MIM were constructed and the adsorption thermodynamics explored. At last, the applicability for MIM in the removal of nicotine in smoke was explored. Results indicated a stronger retention capacity and a higher selectivity of MIM toward the template vapor, with a capacity factor (87.88) and a selectivity factor (10.15) under the optimized conditions. A higher standard adsorption enthalpy change for this MIM toward the template (ΔHa0 = 65.53 kJ mol-1) than that for the non-imprinted monolith (NIM) column (ΔHa0 = 47.46 kJ mol-1) was observed. The adsorption isotherm for MIM appears the BET type II shape, while that for the NIM was approximately linear. When this MIM was used as the adsorbent, it exhibited a high performance in the selective removal of nicotine from the main stream smoke, with an adsorption percentage of 99.43%.

[Clinical value of (125)I radioactive seed implantation plus transcatheter arterial chemoembolization combined with radiofrequency ablation in treating patient with sub-capsular hepatocellular carcinoma].

Objective: To analyze the clinical efficacy and safety of (125)I radioactive seed implantation in the treatment of sub-capsular hepatocellular carcinoma (sub-HCC) with sequential radiofrequency ablation and transcatheter arterial chemoembolization (TACE). Methods: The clinical data of 76 cases with advanced HCC with sub-capsular nodules including 68 males and 8 females, with an average age of (58±9) years, ranging from 33 to 78 years, enrolled in Lishui Central Hospital from January 2010 to December 2016 were collected.The average maximum diameter of tumor is (5.7±2.3) cm, ranging from 3.1 cm to 12.0 cm.The patients were divided into TACE+ RFA group and (125)I + TACE+ RFA group with 38 cases in each group.The overall survival (OS) and progression free survival(PFS) were calculated.The clinical efficiency and adverse events were evaluated. Results: The disease control rate were 84.2%(32/38) in (125)I + TACE+ RFA group and 63.2% (24/38) in TACE+ RFA group, χ(2)=4.34, P= 0.04.The median PFS were 18 months in (125)I + TACE+ RFA group and 11 months in TACE+ RFA group, χ(2)=4.84, P=0.03.The FPS cumulative rate in (125)I + TACE+ RFA group were higher than that in TACE+ RFA group at 6 months (94.7%±3.6% vs 81.3%±6.4%, Z=24.1>2.58, P=0.00), 1 year (89.2%±5.1% vs 40.7%±8.3%, Z=13.3>2.58, P=0.00) and 2 year (55.9%±8.6% vs 29.6%±8.2%, Z=7.2>2.58, P=0.00). The median OS were 42 months in (125)I + TACE+ RFA group and 30 months in TACE+ RFA group, χ(2)=4.76, P=0.029.The survival cumulative rate in (125)I+ TACE+ RFA group were higher than that in TACE+ RFA group at 1 year (92.1%±4.4% vs 83.8%±6.1%, Z=23.5>2.58, P=0.00), 2 year (75.8%±7.0% vs 59.8%±8.4%, Z=12.43>2.58, P=0.00), 3 year (59.0%±8.2% vs 41.7%±8.9%, Z=8.3>2.58, P=0.00), 5 year (34.2%±8.2% vs 18.2%±8.1%, Z=5.5>2.58, P=0.00). In addition, there was no statistical difference in liver function and complications between TACE+ RFA group and (125)I+ TACE+ RFA group. Conclusion: (125)I radioactive seed implantation plus TACE combined with RFA treatment is an effective and safe treatment for sub-capsular hepatocellular carcinoma.

Prognostic nutritional index is correlated with disease activity in patients with systemic lupus erythematosus.

The prognostic nutritional index (PNI), which is calculated using serum albumin level and total lymphocyte count in the peripheral blood, is regarded as an index that reflects the immunonutritional status of patients. PNI was calculated in 217 systemic lupus erythematosus (SLE) patients according to the following formula: 10 × serum albumin value (g/dL) + 0.005 × peripheral lymphocyte count (/mm3). Pearson's correlation analysis was used to elucidate the correlation between continuous variables. Linear and logistic regression analyses were performed to assess the correlation between laboratory variables and SLE Disease Activity Index-2000 (SLEDAI-2 K) and to differentiate between active and inactive SLE. Ninety-three patients were classified as active SLE (SLEDAI-2 K ≥ 5) and 124 as inactive SLE. Patients with active SLE exhibited lower median PNI than those with inactive SLE (39.0 vs. 49.1, p < 0.001). Multivariable logistic regression analysis revealed PNI as an independent predictor of active SLE. Multivariable linear regression analysis revealed that PNI was significantly correlated with laboratory variables of SLEDAI-2 K, erythrocyte sedimentation rate, C-reactive protein and SLEDAI-2 K. Furthermore, in patients who switched from active to inactive SLE after treatment ( n = 55), PNI increased as disease activity improved ( p < 0.001), which suggests that PNI may be useful for estimating SLE activity.

Renal outcome after kidney-transplantation in Korean patients with lupus nephritis.

We investigated renal outcome of kidney-transplantation in 19 Korean recipients with biopsy-proven lupus nephritis and compared it with 18 Korean age- and gender-matched recipients without lupus nephritis who were diagnosed with end-stage renal disease caused by renal diseases other than lupus nephritis in a single centre. We reviewed histological findings of kidneys and calculated cumulative dose of immunosuppressive agents. We assessed renal flare of systemic lupus erythematosus, recurrence of lupus nephritis and graft failure as prognosis. The mean age of recipients with lupus nephritis was 43.5 years and all patients were female. Six patients had class III, 10 had class IV and three had class V. There were no meaningful differences in demographic data, renal replacement modality, cumulative doses of immunosuppressants and prognosis between recipients with and without lupus nephritis. Eight patients experienced renal flare of systemic lupus erythematosus, but there were no cases of recurrence of lupus nephritis or graft failure in recipients with lupus nephritis. Kidney-recipients with class IV lupus nephritis exhibited a lower cumulative renal flare of systemic lupus erythematosus free survival rate than those with class III lupus nephritis. In conclusion, renal outcome of kidney-transplantation in patients with lupus nephritis is similar to that in those without lupus nephritis, and class IV was associated with renal flare of systemic lupus erythematosus.

Serum Wisteria floribunda agglutinin-positive Mac-2-binding protein can reflect systemic lupus erythematosus activity.

Serum Mac-2-binding protein (M2BP) is elevated in various chronic inflammatory diseases, and evidence suggests that glycosylation of M2BP induces discrete biological effects. However, the role of serum M2BP in systemic lupus erythematosus (SLE) is still unclear. Recently, a Wisteria floribunda agglutinin-positive-M2BP (WFA+-M2BP) immunoassay has shown promise in detecting highly glycosylated M2BP. In this study, by using WFA+-M2BP immunoassay, we measured serum M2BP in 203 SLE patients and evaluated its clinical significance. Eighty patients were classified as having active SLE and 123 patients as having inactive SLE. The median serum M2BP was higher in patients with active SLE than in those with inactive SLE (2.1 vs. 0.9, p < 0.001). In multivariate linear regression analysis, serum M2BP, anti-dsDNA, C3 and erythrocyte sedimentation rate (ESR) were associated with SLEDAI-2K. Serum M2BP also strongly correlated with laboratory variables related to SLEDAI-2K, ESR and C-reactive protein. Furthermore, multivariate logistic regression analysis demonstrated that serum M2BP was useful in predicting active SLE. Finally, following immunosuppressive treatment, elevated serum M2BP significantly decreased along with improvement in disease activity. These findings suggest that serum M2BP might contribute to the inflammatory process in SLE, and measuring serum M2BP might be a useful marker to assess SLE disease activity.

Adhesive internal hernia: multidetector CT findings and clinical relevance.

AIM: To review the clinical and multidetector computed tomography (MDCT) features of adhesive internal hernias (IHs) and to ascertain specific MDCT criteria to assist in the diagnosis of adhesive IHs and the early detection of intestinal strangulation. MATERIALS AND METHODS: Medical records and preoperative abdominal MDCT findings of 34 patients with surgically confirmed abdominal adhesive IHs were analysed retrospectively. RESULTS: The specific MDCT features of adhesive IHs included the following: dislocating and clustering of intestinal segments (100%); stretching and crowding of the mesenteric vessels (100%); presence of hernial orifice (88.2%), peritoneal adhesive bands (76.5%); and the fat notch sign (85.3%). In addition, the significant MDCT features indicative of intestinal strangulation compared with those without intestinal strangulation were bowel wall thickening (p=0.009), intramural haemorrhage (p=0.007), and abnormal bowel wall enhancement (p=0.023). Furthermore, bowel obstruction occurred in 17 (50%) patients, and mesenteric whirl was apparent in 8 (23.5%) patients. CONCLUSION: This article illustrates the specific MDCT criteria of adhesive IHs. Knowledge of MDCT findings in adhesive IHs and their complications is essential for making the correct diagnosis and may help guide early clinical management.

Identification of dipeptidyl peptidase-IV inhibitory peptides from mare whey protein hydrolysates.

Inhibition of dipeptidyl peptidase-IV (DPP-IV) activity is a promising strategy for treatment of type 2 diabetes. In the current study, DPP-IV inhibitory peptides were identified from mare whey protein hydrolysates obtained by papain. The results showed that all the mare whey protein hydrolysates obtained at various hydrolysis durations possessed more potent DPP-IV inhibitory activity compared with intact whey protein. The 4-h hydrolysates showed the greatest DPP-IV inhibitory activity with half-maximal inhibitory concentration of 0.18 mg/mL. The 2 novel peptides from 4-h hydrolysate fractions separated by successive chromatographic steps were characterized by liquid chromatography-electrospray ionization tandem mass spectrometry. The novel peptides Asn-Leu-Glu-Ile-Ile-Leu-Arg and Thr-Gln-Met-Val-Asp-Glu-Glu-Ile-Met-Glu-Lys-Phe-Arg, which corresponded to ß-lactoglobulin 1 f(71-77) and ß-lactoglobulin 1 f(143-155), demonstrated DPP-IV inhibitory activity with half-maximal inhibitory concentrations of 86.34 and 69.84 µM, respectively. The DPP-IV inhibitory activity of the 2 peptides was retained or even improved after simulated gastrointestinal digestion in vitro. Our findings indicate that mare whey protein-derived peptides may possess potential as functional food ingredients in the management of type 2 diabetes.

The Role of MRI in Diagnosing Neurovascular Compression of the Cochlear Nerve Resulting in Typewriter Tinnitus.

BACKGROUND AND PURPOSE: Typewriter tinnitus, a symptom characterized by paroxysmal attacks of staccato sounds, has been thought to be caused by neurovascular compression of the cochlear nerve, but the correlation between radiologic evidence of neurovascular compression of the cochlear nerve and symptom presentation has not been thoroughly investigated. The purpose of this study was to examine whether radiologic evidence of neurovascular compression of the cochlear nerve is pathognomonic in typewriter tinnitus. MATERIALS AND METHODS: Fifteen carbamazepine-responding patients with typewriter tinnitus and 8 control subjects were evaluated with a 3D T2-weighted volume isotropic turbo spin-echo acquisition sequence. Groups 1 (16 symptomatic sides), 2 (14 asymptomatic sides), and 3 (16 control sides) were compared with regard to the anatomic relation between the vascular loop and the internal auditory canal and the presence of neurovascular compression of the cochlear nerve with/without angulation/indentation. RESULTS: The anatomic location of the vascular loop was not significantly different among the 3 groups (all, P > .05). Meanwhile, neurovascular compression of the cochlear nerve on MR imaging was significantly higher in group 1 than in group 3 (P = .032). However, considerable false-positive (no symptoms with neurovascular compression of the cochlear nerve on MR imaging) and false-negative (typewriter tinnitus without demonstrable neurovascular compression of the cochlear nerve) findings were also observed. CONCLUSIONS: Neurovascular compression of the cochlear nerve was more frequently detected on the symptomatic side of patients with typewriter tinnitus compared with the asymptomatic side of these patients or on both sides of control subjects on MR imaging. However, considering false-positive and false-negative findings, meticulous history-taking and the response to the initial carbamazepine trial should be regarded as more reliable diagnostic clues than radiologic evidence of neurovascular compression of the cochlear nerve.

[Association between ultrasound screening frequency and mortality in patients with hepatocellular carcinoma].

Objective: To discuss the association between ultrasound screening frequency and total mortality in patients with HCC before diagnosing HCC, and explore the optimal ultrasound screening frequency for HCC high-risk groups. Methods: Retrospectively collected clinical data of 615 cases of liver cirrhosis who developed to HCC from January 1, 2010 to December 31, 2015. Before diagnosing HCC, all patients were divided into five groups according to ultrasound screening frequency: 0-6, 7-12, 13-24, 25-36 months and not screened within 3 years (never screened). The chance to receive curative therapy, 5-year cumulative mortalities and independent factors of mortality in patients with HCC were analyzed. Results: Chances to receive curative therapy among the 0-6, 7-12, 13-24, 25-36 months and never screened groups were 38.2%, 27.2%, 25.4%, 23.8% and 19.7%, respectively (P<0.05). The 5-year overall mortality rates were 76.4%, 77.7%, 79.3%, 82.5% and 84.6%, respectively. Compared with 0-6 months, the adjusted OR of mortality for the other groups were 1.112, 1.235, 1.305 and 1.451, respectively (all P<0.05). Multivariate analysis showed that ultrasound screening frequency, curative treatment and Child-Pugh (class A/B) were the factors to affect long-term survival in patients with HCC (all P<0.05). Conclusion: For HCC high-risk groups, optimal ultrasound screening frequency is within 6 months, and high-frequency ultrasound screening can increase the chance of receiving curative treatment, reduce total mortality, and improve overall survival.

[Efficacy analysis of radiofrequency ablation combined with 125I seed for multiple nodular hepatocellular carcinoma].

OBJECTIVE: To investigate the clinical efficacy of radiofrequency ablation(RFA)combined with (125)I seed in patients with multiple nodular hepatocellular carcinoma(HCC). METHODS: The clinical data of 47 patients with multiple nodular HCC confirmed clinically or pathologically in the Fifth Affiliated Hospital of Wenzhou Medical University between January 2009 and September 2014 were retrospectively analyzed. All patients were divided into two groups depending on treatment: RFA group(n=29); RFA combined with (125)I seed group(n=18). Survival time was estimated with the Kaplan-Meier analysis. The survival curve was compared by Log-rank test. RESULTS: The clinical data and tumor situation of patients between two groups did not show significant differences. The follow-up time ranged from 2 to 55 months. At the end of the study, the median progression-free survival of two groups were 18 months and 11 months, and the differences were statistically significant(P=0.036). The overall survival rates in RFA combined with (125)I seed group at 1, 2, and 3 years (94.4%, 78.0%, 66.8%, respectively) were higher than those in RFA group (88.5%, 68.6%, 57.1%, respectively). However, the differences between the two groups were not statistically significant(P=0.554). CONCLUSIONS: For multiple nodular HCC, RFA combined with (125)I seed can prolong progression-free survival, have an advantage in local control rate, and is better than single RFA at short-term curative effect.However, its long-term outcomes should be further explored by a large-sample, multi-center and randomized trial.

Gestational weight gain and preterm birth in obese women: a systematic review and meta-analysis.

BACKGROUND: Prepregnant obesity is a global concern and gestational weight gain has been found to influence the risks of preterm birth. OBJECTIVE: To assess the relationship between gestational weight gain and risk for preterm birth in obese women. SEARCH STRATEGY: Four electronic databases were searched from 18 February through to 28 April 2015. SELECTION CRITERIA: Primary research reporting preterm birth as an outcome in obese women and gestational weight gain as a variable that could be compared to the 2009 Institute of Medicine's recommendations. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trials for inclusion. The Newcastle Ottawa Scale was used to assess study bias. MAIN RESULTS: Our search identified six studies meeting the inclusion criteria; five were conducted in the USA and one in Peru. Four studies with a total of 10 171 obese women were meta-analysed. Significant heterogeneity was found between studies in the pooled analysis. Results for indicated preterm birth in obese women with gestational weight gain above the Institute of Medicine's recommendations showed increased risk (adjusted odds ratio 1.54; 95% CI 1.09-2.16). CONCLUSIONS: Available science on this topic is limited to special populations of obese pregnant women. Generalisable research is needed to assess the variation in risk for preterm birth in obese women by differences in gestational weight gain and class of obesity controlling for significant variables in the pathway to preterm birth. This research has the potential to illuminate new science impacting preterm birth and interventions for prevention.

Silencing ECHS1 attenuates the proliferation and induces the autophagy of hepatocellular carcinoma via impairing cell metabolism and activating AMPK.

Hepatocellular carcinoma (HCC) is among the most common cancers in the world with a low survival rate. Our previous study showed Short chain enoyl-CoA hydratase (ECHS1) could bind to HBsAg (HBs) and that ECHS1's localization in mitochondria induced HepG2 cell apoptosis. However, the role of the ECHS1 in energy metabolism and autophagy during hepatocellular carcinoma development remains undefined. We aimed to determine what ECHS1 does to energy metabolism and its effects on HCC progression. We performed CCK-8, EdU assays in hepatocellular carcinoma cell lines (HepG2 and HuH7) with stable ECHS1 knock-down. ATP and NADP+/NADPH levels were measured using an colorimetric assay. Our data demonstrated that ECHS1 silencing inhibited cell proliferation and induced autophagy. ECHS1 knockdown did not increase fatty acid synthesis, but decreased cellular ATP. This resulted in AMP-activated protein kinase (AMPK) activation and induced HCC cell autophagy. Our results showed that silencing ECHS1 to attenuate proliferation and induce autophagy may make it a novel cancer therapy target.

DAMGO depresses inhibitory synaptic transmission via different downstream pathways of µ opioid receptors in ventral tegmental area and periaqueductal gray.

Opioid-induced rewarding and motorstimulant effects are mediated by an increased activity of the ventral tegmental area (VTA) dopamine (DA) neurons. The excitatory mechanism of opioids on VTA-DA neurons has been proposed to be due to the depression of GABAergic synaptic transmission in VTA-DA neurons. However, how opioids depress GABAergic synaptic transmission in VTA-DA neurons remain to be studied. In the present study, we explored the mechanism of the inhibitory effect of [D-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin (DAMGO) on GABAergic synaptic transmission in VTA-DA neurons using multiple approaches and techniques. Our results showed that (1) DAMGO inhibits GABAergic inputs in VTA-DA neurons at presynaptic sites; (2) effect of DAMGO on GABAergic inputs in VTA-DA neurons is inhibited by potassium channel blocker 4-aminopyridine (4-AP) and Gi protein inhibitor N-ethylmaleimide (NEM); (3) phospholipase A2 (PLA2) does not mediate the effect of DAMGO on GABAergic inputs in VTA-DA neurons, but mediates it in the periaqueductal gray (PAG); (4) multiple downstream signaling molecules of µ receptors do not mediate the effect of DAMGO on GABAergic inputs in VTA-DA neurons. These results suggest that DAMGO depresses inhibitory synaptic transmission via µ receptor-Gi protein-Kv channel pathway in VTA-DA neurons, but via µ receptor-PLA2 pathway in PAG neurons.

Dexamethasone therapy for preventing delayed encephalopathy after carbon monoxide poisoning.

We investigated dexamethasone therapy for preventing delayed encephalopathy after carbon monoxide (CO) poisoning. Eighty healthy male rats were exposed to CO and randomly divided into four groups: hyperbaric oxygen treatment (H), treatment (D), combined hyperbaric and dexamethasone treatment (C), and a control (M) group in which the rats inhaled CO to coma in the hyperbaric oxygen chamber, then were removed without further treatment. Twelve rats were put into the hyperbaric oxygen chamber and treated with air for 60 min (N) group. An eight arm maze was used to evaluate cognitive and memory abilities of these mice. Serum myelin basic protein (MBP) levels were evaluated using ELISA, and magnetic resonance imaging was used to observe brain demyelination and morbidity associated with delayed encephalopathy. A sample of the hippocampus from each group was examined by light microscopy. Cognitive and memory functions decreased in the control group M. Three days after CO poisoning, the serum MBP level of each group increased significantly. On Day 10 after CO poisoning, the MBP levels in groups C and D decreased significantly, but returned to normal on Day 18. MBP levels in the M and H groups were elevated at all time points. Brain MRIs showed significant differences among C, D, H and control M groups. Hematoxylin & eosin staining of the hippocampus showed greater damage in the control M and H groups. Early dexamethasone treatment may be useful for preventing delayed encephalopathy after CO poisoning and may reduce serum MBP levels.

Quantitative determination of fluconazole by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) in human plasma and its application to a pharmacokinetic study.

In this study, a simple, rapid and sensitive ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method is described for determination of fluconazole (FLA) in human plasma samples using phenacetin as the internal standard (IS). Sample preparation was accomplished through one-step protein precipitation by methanol, and chromatographic separation was performed on an Acquity BEH C18 column (2.1 mm×50 mm, 1.7 µm) with mobile phase consisted of acetonitrile and water containing 0.1% formic acid (40:60, v/v) at a flow of 0.45 mL/min. Mass spectrometric analysis was performed using a QTrap 5500 mass spectrometer coupled with an electro-spray ionization (ESI) source in the positive ion mode. The MRM transition of m/z 307.2→238.2 was used to quantify for FLA. The linearity of this method was found to be within the concentration range of 10-6 000 ng/mL for FLA in human plasma. Only 1.0 min was needed for an analytical run. The method herein described was superior to previous methods and was successfully applied to the pharmacokinetic study of FLA in healthy Chinese volunteers after oral administration.

Transforming growth factor-ß gene silencing using adenovirus expressing TGF-ß1 or TGF-ß2 shRNA.

Tumor cells secrete a variety of cytokines to outgrow and evade host immune surveillance. In this context, transforming growth factor-ß1 (TGF-ß1) is an extremely interesting cytokine because it has biphasic effects in cancer cells and normal cells. TGF-ß1 acts as a growth inhibitor in normal cells, whereas it promotes tumor growth and progression in tumor cells. Overexpression of TGF-ß1 in tumor cells also provides additional oncogenic activities by circumventing the host immune surveillance. Therefore, this study ultimately aimed to test the hypothesis that suppression of TGF-ß1 in tumor cells by RNA interference can have antitumorigenic effects. However, we demonstrated here that the interrelation between TGF-ß isotypes should be carefully considered for the antitumor effect in addition to the selection of target sequences with highest efficacy. The target sequences were proven to be highly specific and effective for suppressing both TGF-ß1 mRNA and protein expression in cells after infection with an adenovirus expressing TGF-ß1 short hairpin RNA (shRNA). A single base pair change in the shRNA sequence completely abrogated the suppressive effect on TGF-ß1. Surprisingly, the suppression of TGF-ß1 induced TGF-ß3 upregulation, and the suppression of TGF-ß2 induced another unexpected downregulation of both TGF-ß1 and TGF-ß3. Taken together, this information may prove useful when considering the design for a novel cancer immunogene therapy.