[Voxel-based morphometry (VBM) MRI analysis of gray matter in patients with occupational noise-induced hearing loss].

Objective: To investigate the changes of brain gray matter volume in patients with occupational noise-induced hearing loss by voxel based morphometry (VBM) . Methods: 16 age-and education-matched healthy controls and 42 patients with occupational noise induced hearing loss, including 27 in mild group and 15 in severe group, received MRI 3D-FSPGR sequence T1WI sagittal scan, and then underwent VBM of brain gray matter volume data analysis. Results: The brain gray matter volume of the left occipitotemporal lateral gyrus, the anterior cingulate gyrus, the bilateral angular gyrus, the precuneus and the near midline area of cerebellum differed between experimental group and control group (P<0.01) . Conclusion: The volume of gray matter in specific brain areas of patients with occupational noise-induced hearing loss was changed, and the effect of noise on brain structure was revealed from the perspective of imaging.

Increased human epididymis protein 4 in benign gynecological diseases complicated with chronic renal insufficiency patients.

We examined the serum concentration of human epididymis protein (HE4) in patients with benign gynecological diseases complicated with chronic renal deficiency and its significance in the differential diagnosis of benign and malignant gynecological diseases. Serum HE4 and cancer antigen 125 concentrations were detected by chemiluminescence. Clinically or pathologically confirmed gynecological diseases were grouped and retrospectively analyzed, including 50 cases of gynecological benign diseases, 35 cases of non-mucinous epithelial ovarian carcinoma, 36 cases of endometrial adenocarcinoma, 15 cases of gynecological benign diseases patients complicated with chronic renal deficiency, 15 cases of gynecological diseases without chronic renal deficiency, and 30 normal controls. Serum HE4 values in the ovarian cancer group, endometrial cancer group, gynecological benign diseases with chronic renal deficiency group, and chronic renal deficiency group were significantly increased compared with the benign gynecological diseases and normal control groups, showing a significant difference (P < 0.001). A comparison of 4 groups with high HE4 showed that the HE4 level in the 2 groups with renal deficiency were higher than those in the ovarian cancer and endometrial cancer groups, but the difference was not significant (P > 0.05); there was no significant difference between 2 groups with renal deficiency (P > 0.05). Serum concentration of HE4 was high in patients with chronic renal deficiency, which should be distinguished during differential diagnosis of gynecological benign and malignant tumors in patients with chronic renal deficiency to avoid misdiagnosis.

Effect of hypertonic sodium chloride hydroxyethyl starch 40 on ET, TXB2, 6-keto-PGF1α, and ANP of preeclampsia in caesarean section.

OBJECTIVE: Preeclampsia is a unique disease of pregnancy. Delivery via caesarean section is the most important way of terminating the pregnancy and treating preeclampsia. Perioperative fluid therapy is performed to maintain the circulatory volume and reduce tissue edema. This study evaluated the effects of hypertonic sodium chloride hydroxyethyl starch 40 (HSH40) as perioperative fluid therapy for preeclampsia patients. MATERIALS AND METHODS: Forty preeclamptic women were randomly divided into two groups: the Ringer's solution group and the HSH40 group. Their ECG, HR, MAP, and SPO2 were monitored. Their MVP and HR were recorded at five, eight, and ten minutes after anesthesia induction and at the end of the caesarean section. The corresponding volume of infusion, blood loss, and urine output during the operation were also recorded. Venous samples were collected before HSH40 infusion and 30 min after infusion to measure the plasma concentrations of ET, TXB2, 6-keto-PGF1α, and ANP via a radioimmunoassay. RESULTS: HSH40 infusion significantly decreased the plasma ET levels (p < 0.01), significantly changed the plasma ANP and TXB2 levels (p < 0.05), and significantly increased the plasma 6-keto-PGF1α levels (p < 0.01) in the experimental group compared with those before infusion. The plasma levels of ET, ANP, TXB2, and 6-keto-PGF1α did not significantly change in the control group. Compared with T1, MAP decreased significantly at T2, T3, T4, and T5 within groups (p < 0.05) and between the two groups. MAP significantly changed at T2, T3, T4, and T5 (p < 0.05). HR did not significant change at T1, T2, T3, T4, and T5 within or between groups. Volume of infusion and urine volume significantly differed between groups (p < 0.05). CONCLUSION: Low-dose HSH40 lowers the plasma levels of vasoconstrictor substances (ET and TXB2) and increases the levels of vasodilator substances (6-keto-PGF1α and ANP) during preeclampsia. It effectively maintains and stabilizes the circulating blood volume, increasing renal blood flow, which improves renal function and increases urine output.

Association between metabolic syndrome and vascular endothelium dysfunction in children and adolescents.

We aimed at investigating the association between metabolic syndrome (MS) and vascular endothelial cell dysfunction (ECD) in children and adolescents. Sixty children (30 obese children and 30 children with MS) were included in this retrospective analysis. Thirty healthy subjects were randomly selected as the control group. A series of indices/biomarkers known to be related to MS/ECD were determined using ELISA. Correlations between the variables measured were analyzed. Compared with the control group, PAI-1, vWF, VE-cad, TM, and VEGF were significantly increased in the MS group (P < 0.05). Adolescents in the obese group had significantly increased levels of serum PAI-1, VE-cad, TM, and VEGF as compared with the control group (P < 0.05). Further, vWF in the obese and control groups did not differ significantly (P = 0.556). Our results suggest that ECD is correlated with MS in children and adolescents. Pathophysiological changes of the vascular endothelium may exist in obese children who have yet to develope MS. PAI-1, vWF, VE-cad, TM, and VEGF could be used as biomarkers for predicting ECD. ECD that develops in patients with MS may be associated with obesity, elevated blood lipid, elevated blood glucose, and higher blood pressure.

Tissue distribution, expression, and antimicrobial activity of Anas platyrhynchos avian ß-defensin 6.

In this study, a novel avian ß-defensin (AvBD) was isolated from the Chaohu duck. The complete nucleotide sequence of the gene contained a 204-bp open reading frame that encoded 67 amino acids (aa), including a signal peptide of 20 aa, a propiece of 5 aa, and a mature peptide of 42 aa. The homology, characterization, and comparison of this gene with AvBD from other avian species confirmed that it was duck AvBD6. Also, the preproprotein of AvBD6 from chicken, goose, and duck was highly conserved with 100% aa homology. The AvBD6 mRNA was widely expressed in the investigated tissues of healthy 5-mo-old ducks, with the exception of the skin, kidney, and bursa. The AvBD6 mRNA was highly expressed in the spleen, lung, stomach, tongue, and egg yolk, successively; moderately expressed in the bone marrow and liver; and expressed to lower degrees in the trachea, heart, intestine, muscle, esophagus, and testis. We produced recombinant AvBD6 by expressing the gene in Escherichia coli. The yield of soluble glutathione S-transferees (GST)-AvBD6 in the inclusion bodies increased significantly as the incubation temperature was decreased from 37 to 30°C. As expected, the GST-AvBD6 exhibited strong bactericidal properties [minimum inhibitory concentration (MIC) = 0.5 to 2 µM] against the Aeromonas veronii biovar sobria strain BJCP-5, E. coli, and Enterococcus faecalis [ATCC 29212); it possessed medium bactericidal properties (MIC = 4 to 8 µM) against Staphylococcus aureus, Salmonella enteritidis, Pseudomonas aeruginosa (ATCC 27853), and Bacillus subtilis (CMCC 63501;; and it possessed low bactericidal properties (MIC = 16 µM) against Candida albicans (ATCC 10231). Additionally, the antimicrobial activity of GST-AvBD6 proved to be the same as that of synthetic AvBD6. The A. veronii biovar sobria strain BJCP-5 cells that were treated with GST-AvBD6 showed lysis and shrinkage under scanning electron microscopy. The mechanisms of AvBD6-mediated killing of A. veronii biovar sobria involved both cell lysis and nonlysis. Our results indicate that AvBD6 plays an important role in the innate immunity of the Chaohu duck.

Pure docosahexaenoic acid can improve depression behaviors and affect HPA axis in mice.

OBJECTIVE: Aim of this study was to identify whether docosahexaenoic acid (DHA) has a vital antidepressant by hypothalamic-pituitary-adrenal axis (HPA axis) or not. METHODS: Mice were divided into 2 groups: control group and DHA dietary group. DHA dietary group was treated with DHA dietary everyday for consecutive 50 days. The forced swimming test and tail suspension test were conducted. Hypothalamic and erythrocyte fatty acids and monoamine neurotransmitters levels in hypothalamus were assayed; corticosterone, adrenocorticotropic hormone and corticotropin-releasing factor in serum, hypothalamus and pituitary were assayed, respectively. RESULTS: (1) In the forced swimming test, DHA dietary significantly decreased immobility time, whereas swimming time and climbing time were increased. In tail suspension test, DHA dietary significantly shortened immobility time. (2) DHA dietary increased the ration of n-3/n-6 (polyunsaturated fatty acids) and DHA level in hypothalamic and erythrocyte fatty acids; (3) DHA dietary significantly increased 5-hydroxytryptamine, 5-hydroxyindoleacetic acid and dopamine levels in hypothalamus; (4) DHA dietary significantly decreased serum corticosterone level by 20.23% and serum adrenocorticotropic hormone level by 25.13%; significantly increased serum corticotropin releasing factor level by 21.92%. Besides, DHA dietary decreased arginine vasopressin level by 20.11% in hypothalamus, by 23.76% in pituitary tissues, respectively; (5) DHA dietary decreased corticotropin-releasing factor levels by 30.83% in hypothalamus, by 29.75% in pituitary tissues, respectively. In hypothalamus, DHA dietary decreased significantly adrenocorticotropic hormone level by 19.14%, but insignificantly decreased adrenocorticotropic hormone level in pituitary. CONCLUSIONS: DHA shows an antidepressant property. Moreover, DHA has multiple effects on depression including the monoamine neurotransmitter systems, red blood cell membranes and HPA axis.

Exogenous sodium hydrosulfide can attenuate naloxone-precipitated withdrawal syndromes and affect cAMP signaling pathway in heroin-dependent rat's nucleus accumbens.

BACKGROUND: H2S is a novel type of endogenous neural regulatory factor and gaseous mediator. Exogenous H2S can increase heroin-induced learning and memory damage in rat and alleviates heroin-induced rat hippocampus damage through antioxidant and anti-apoptosis effects. OBJECTIVE: Aim of this study was to identify whether hydrogen sulfide (H2S) protects heroin withdrawal rat is related with adenylate cyclase (AC)-cAMP-protein kinase A (PKA)-cAMP response element-binding protein (CREB) signaling pathway in heroin-dependent rat's nucleus accumbens or not. METHODS: Male Spragne-Dawley rats were randomly divided into Saline + Saline group, Saline + sodium hydrosulfide (NaHS) group, Saline + Heroin group, NaHS + Heroin group according to the principle of increasing heroin dosage day by day, with the establishment of heroin-naloxone-induced withdrawal symptoms determined at day 10. Then the levels of H2S and cAMP and AC and PKA activities were assayed, and the level of phosphorylated CREB (p-CREB), the levels of phosphorylated N-methyl-D-aspartate receptor 1 subunit (p-NR1), phosphorylated N-methyl-D-aspartate receptor 2a subunit (p-NR2A) and phosphorylated N-methyl-D-aspartate receptor 2b subunit (p-NR2B) were assayed in nucleus accumbens. RESULTS: Exogenous H2S can alleviate heroin withdrawal symptoms by increasing the level of H2S level in nucleus accumbens. Exogenous H2S can decrease the high activities of AC, PKA and the high levels of cAMP, p-CREB caused by heroin. Furthermore, exogenous H2S can decrease the high level of p-NR1 and can increase the low levels of p-NR2A and p-NR2B caused by heroin. It is surprising that exogenous H2S treatment alone was able to raise the activities of AC and PKA as well as the levels of cAMP, p-CREB, p-NR1, p-NR2A and p-NR2B. CONCLUSIONS: Exogenous H2S decreases naloxone-precipitated withdrawal signs, maybe through decreasing AC/cAMP/PKA/CREB/NMDR signaling pathway in heroin-dependent rats' nucleus accumbens.

Oral oligofructose challenge reduces expression of glucose transport-1 and 5'-adenosine monophosphate-activated protein kinase in lamellar wall of Holstein heifer claw.

The laminar tissue of bovine laminitis may undergo energy failure. The expression of glucose transport protein-1 (GLUT-1) and 5'-adenosine monophosphate-activated protein kinase (AMPK) affects the energy metabolism of digital laminar tissue. This study aimed to determine the expression of glucose uptake and AMPK in laminar wall corium of Holstein heifer claw by oral administration of oligofructose. A total of twelve clinically healthy Holstein heifers were selected and divided into two groups, including control (CON, n = 6) and experimental (OF, n = 6) groups. The heifers of OF group were given 17 g/kg BW oligofructose dissolved in water (20 mL/kg BW) and the heifers of CON group were given water only (20 mL/kg BW). The laminar tissues were collected after euthanasia. The amount of protein and transcript expression of AMPK and GLUT-1 were determined by western blot and quantitative reverse transcription-polymerase chain reaction (qRT-PCR), respectively. Expressions of phosphoenolpyruvate carboxy-kinase (PEPCK), receptor-c coactivator1-α (PGC-1α) and peroxisome proliferator-activated receptor-γ (PPAR-γ) were determined by qRT-PCR. The heifers of OF group showed no significant change in the expression and concentration of AMPK. The phosphor-(Thr172) AMPK and GLUT-1 were significantly decreased, while the gene contents of PPAR-γ and PGC-1α were significantly increased. The activation of AMPK and GLUT-1 in digital laminar tissues of heifers was inhibited, which may contribute to digital laminar tissue damage.

Meta-analysis of epidemiological studies of association of P53 codon 72 polymorphism with bladder cancer.

Although there have been many studies investigating a possible association between p53 codon 72 polymorphism and risk of bladder cancer, the results have been inconsistent. We conducted a meta-analysis of six epidemiological studies, which included 597 bladder cancer cases and 731 controls. Patients with bladder cancer had a significantly lower frequency of Pro/Arg [odds ratio (OR) = 0.80, 95% confidence interval (CI) = 0.64-0.99], when compared to controls. Stratifying for race, we found that among Caucasians, patients with bladder cancer had a significantly higher frequency of Arg/Arg (OR = 1.64, 95%CI = 1.18-2.28) and a lower frequency of Pro/Arg (OR = 0.62, 95%CI = 0.44-0.86), compared to controls. Stratifying various studies by the stage of bladder cancer, we found that invasive bladder cancers had a significantly lower frequency of Arg/Arg (OR = 0.58, 95%CI = 0.36-0.93) and a higher frequency of Pro/Arg (OR = 0.62, 95%CI = 0.44-0.86) than did non-invasive bladder cancers. No significant association was found between this genotype and human papilloma virus. Based on our meta-analysis, we suggest that p53 codon 72 polymorphism is associated with bladder cancer and that genotypic distribution of this polymorphism varies with the stage of bladder cancer.

[Clinical application of the gasless unilateral axillary approach in endoscopic thyroid surgery].

Objective: To investigate the efficacy, safety and advantages of gasless unilateral axillary approach (GUAA) in endoscopic thyroid surgery. Methods: A total of 334 patients who underwent the GUAA endoscopic thyroid surgery (GUAA group) or conventional open thyroid surgery (OS group) in the Department of Head and Neck Surgery of Zhejiang Cancer Hospital from January 2017 to June 2018 were retrospectively analyzed. There were 45 males and 289 females, aged from 12 to 72 years old, of whom 139 patients were assigned to GUAA group and 195 patients to OS group. Pathological results included papillary thyroid carcinoma (282 cases), nodular goiter (41 cases) and thyroid adenoma (11 cases). Surgical exploration development curve of GUAA group was drawn and was divided into two parts: the technical exploration stage and the technical stable stage. Surgical efficiency, incidences of complications, and incision satisfaction were compared between GUAA group in technical stable stage and OS group. SPSS 25.0 software was adopted for statistical analysis. Results: The mean age in GUAA group was younger than that in OS group, with a significant difference [(35.3±9.5) years vs. (48.1±10.6) years, t=11.31, P<0.01]. The cases in the endoscope group were divided into technical exploration stage for 51 cases and technical stable stage for 88 cases according to the exploration and development curve. In unilateral radical thyroidectomy and unilateral thyroid lobectomy, the mean operation time [(90.6±18.6) min and (93.5±22.0) min] and postoperative drainage volumes [(121.5±87.6) ml and (155.5±69.1) ml] of GUAA group in the stable stage were more than those of OS group [(61.6±15.6) min and (46.5±8.4) min] and [(93.2±42.3) ml and (78.9±48.7) ml]. The difference was statistically significant (t=12.28, 7.23, 3.35 and 3.05 respectively, all P<0.05), but there were no significant differences in surgical bleeding volumes between two groups [(12.7±6.8) ml vs. (13.5±7.7) ml, t=0.74, P>0.05 and (16.3±14.1) ml vs. (11.9±5.1) ml, t=1.05, P>0.05]. Compared with OS group, GUAA group had the lower incidence of anterior cervical discomfort during swallowing (2.3% vs. 29.2%, P<0.01) and the higher incision satisfaction score (1.1±0.5 vs. 2.8±0.7, t=21.12, P<0.01), however, GUAA group had the higher incidence of supraclavicular (or infraclavicular) numbness after surgery (5.7% vs. 0, P<0.01). And there was no significant difference in the incidences of temporary recurrent laryngeal nerve injury, bleeding, hematoma, infection, lymphatic leakage or chylous leakage after surgery between two groups (P>0.05). Conclusion: GUAA endoscopic thyroid surgery is a safe method with high cosmetic satisfaction.

Influence of miR-34a on myocardial apoptosis in rats with acute myocardial infarction through the ERK1/2 pathway.

OBJECTIVE: To study the influence of micro-ribonucleic acid (miR)-34a on myocardial apoptosis in rats with acute myocardial infarction (AMI) through the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway. MATERIALS AND METHODS: A total of 24 Sprague-Dawley (SD) rats were randomly divided into sham group (n=12) and model group (n=12). The heart was exposed in the sham group, while the AMI model was established in the model group. After sampling, the morphology of myocardial tissues was observed via hematoxylin-eosin (HE) staining, the expressions of B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) were detected via immunohistochemistry, and the protein expression levels of ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2) were detected via Western blotting. Moreover, the expression of miR-34a was detected via quantitative Polymerase Chain Reaction (qPCR), the apoptosis was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), and the myocardial injury indexes were detected using a fully-automatic biochemical analyzer. RESULTS: The morphology of myocardial tissues was normal with a complete structure in the sham group, while there was damage to myocardial tissues in different degrees in the model group. The immunohistochemical results revealed that the Bax expression was increased and the Bcl-2 expression was decreased in the model group compared with those in the sham group (p<0.05). The results of Western blotting showed that the protein expression levels of both ERK1/2 and p-ERK1/2 were significantly increased in the model group compared with those in the sham group (p<0.05). The qPCR results manifested that the expression of miR-34a in the model group markedly declined compared with that in the sham group (p<0.05). Besides, the TUNEL detection showed that the apoptosis rate in the model group was remarkably increased compared with that in the sham group (p<0.05), and the content of cardiac troponin T and creatine kinase isoenzyme in the model group was significantly higher than that in the sham group ((p<0.05). CONCLUSIONS: MiR-34a affects the apoptosis in AMI by regulating the ERK1/2 signaling pathway.

Effects of platelet-rich plasma on angiogenesis and osteogenesis-associated factors in rabbits with avascular necrosis of the femoral head.

OBJECTIVE: Platelet-rich plasma (PRP) contains various growth factors and cytokines that can enhance the recovery of the damaged tissues. The present study aimed to examine the effects of PRP on the recovery of avascular necrosis of the femoral head (ANFH), and to provide novel insights into the clinical treatment of this disease. MATERIALS AND METHODS: A total of 24 New Zealand white rabbits were randomly divided into the normal control group, ANFH model and PRP-treated groups (n =1 2 each). Blood samples were extracted from the auricular vein at 4, 8 and 12 weeks after establishing the model to determine the hemorheological indexes, as well as the content of serum osteocalcin bone Gla-protein (BGP) and vascular endothelial growth factor (VEGF). In addition, femoral head tissue was collected, with part of it used for hematoxylin and eosin (HE) staining to observe the histological changes. The remaining was used to detect the mRNA expression levels of alkaline phosphatase (AKP), basic fibroblast growth factor (bFGF), transforming growth factor-ß1 (TGF-ß1), bone morphogenetic protein 2 (BMP-2) and platelet-derived growth factor B (PDGF-B) by reverse transcription-polymerase chain reaction. RESULTS: Compared with the model group, PRP treatment significantly improved the hemorheological indexes, as well as significantly increased the contents of BGP and VEGF. In the PRP group, the expression levels of TGF-ß1, bFGF, BMP-2 and PDGF-B were significantly upregulated, while AKP expression was downregulated compared with the model group. Furthermore, PRP evidently improved the histological structure of the ANFH tissue. CONCLUSIONS: PRP was able to improve the hemorheological indexes following femoral neck fracture, repair the local blood vessels, and promote the expression of osteoblast-associated and angiogenesis-associated factors, which suggested a high efficiency in repairing ANFH.

The application of PRP combined with TCP in repairing avascular necrosis of the femoral head after femoral neck fracture in rabbit.

OBJECTIVE: In view of the high occurrence of avascular necrosis of the femoral head (ANFH) after femoral neck fracture and the difficulties in the treatment, our work aimed to explore the effects of platelet-rich plasma (PRP) combined with tri-calcium phosphate (TCP) on the repair of ANFH after femoral neck fracture and to provide reference for clinical treatment. MATERIALS AND METHODS: Thirty New Zealand white rabbits were randomly divided into control group, TCP group, and PRP+TCP group. The rabbit ANFH model was established and femoral head tissues were collected. HE staining was used for histological observation. Image analysis and statistical analysis were used to calculate the New Bone Area fraction (NBA %). The levels of bone morphogenetic protein (BMP)-7, transforming growth factor (TGF)-ß1, basic fibroblast growth factor (bFGF), interleukin (IL)-6 and tumor necrosis factor (TNF)-a in serum were detected by Enzyme-Linked ImmunoSorbent Assay (ELISA). RESULTS: The new bone area of TCP group was significantly lower than that of PRP+TCP group (p<0.05). Compared with the control group, the levels of BMP-7, TGF-ß1 and bFGF were significantly increased in both TCP and PRP+TCP groups (p<0.05), and the increase in PRP+TCP group was higher than that in TCP group. TCP and PRP+TCP can both significantly reduce the content of IL-6 and TNF-a (p<0.05); however, higher decrease was found in PRP+TCP group compared with the TCP group at 8 weeks after injection. CONCLUSIONS: PRP combined with TCP, which can promote new bone formation and inhibit inflammatory response, showed higher efficiency in repairing ANFH than internal fixation alone.

Characterization of a selective and potent antagonist of human P2X(7) receptors, AZ11645373.

BACKGROUND AND PURPOSE: The ATP-gated P2X(7) receptor has been shown to play a role in several inflammatory processes, making it an attractive target for anti-inflammatory drug discovery. We have recently identified a novel set of cyclic imide compounds that inhibited P2X(7) receptor-mediated dye uptake in human macrophage THP-1 cells. In this study the actions and selectivity of one of these compounds, AZ11645373, were characterized. EXPERIMENTAL APPROACH: We measured membrane currents, calcium influx, and YOPRO-1 uptake from HEK cells expressing individual P2X receptors, and YOPRO1 uptake and interleukin-1beta release from THP-1 cells in response to ATP and the ATP analogue benzoylbenzoyl ATP (BzATP). KEY RESULTS: AZ11645373 up to 10 microM, had no agonist or antagonist actions on membrane currents due to P2X receptor activation at human P2X(1), rat P2X(2), human P2X(3), rat P2X(2/3), human P2X(4), or human P2X(5) receptors expressed in HEK cells. AZ11645373 inhibited human P2X(7) receptor responses in HEK cells in a non-surmountable manner with K (B) values ranging from 5 - 20 nM, with mean values not significantly different between assays. K (B) values were not altered by removing extracellular calcium and magnesium. ATP-evoked IL-1beta release from lipopolysaccharide-activated THP-1 cells was inhibited by AZ11645373, IC(50) = 90 nM. AZ11645373 was > 500-fold less effective at inhibiting rat P2X(7) receptor-mediated currents with less than 50% inhibition occurring at 10 microM. CONCLUSIONS AND IMPLICATIONS: AZ11645373 is a highly selective and potent antagonist at human but not rat P2X(7) receptors and will have much practical value in studies of human cells.

Effects of propofol on lipopolysaccharide-induced expression and release of HMGB1 in macrophages.

This study aimed to determine the effects of different concentrations of propofol (2,6-diisopropylphenol) on lipopolysaccharide (LPS)-induced expression and release of high-mobility group box 1 protein (HMGB1) in mouse macrophages. Mouse macrophage cell line RAW264.7 cells were randomly divided into 5 treatment groups. Expression levels of HMGB1 mRNA were detected using RT-PCR, and cell culture supernatant HMGB1 protein levels were detected using enzyme-linked immunosorbent assay (ELISA). Translocation of HMGB1 from the nucleus to the cytoplasm in macrophages was observed by Western blotting and activity of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in the nucleus was detected using ELISA. HMGB1 mRNA expression levels increased significantly in the cell culture supernatant and in cells after 24 h of stimulating RAW264.7 cells with LPS (500 ng/mL). However, HMGB1 mRNA expression levels in the P2 and P3 groups, which received 500 ng/mL LPS with 25 or 50 µmol/mL propofol, respectively, were significantly lower than those in the group receiving LPS stimulation (P<0.05). After stimulation by LPS, HMGB1 protein levels were reduced significantly in the nucleus but were increased in the cytoplasm (P<0.05). Simultaneously, the activity of NF-κB was enhanced significantly (P<0.05). After propofol intervention, HMGB1 translocation from the nucleus to the cytoplasm and NF-κB activity were inhibited significantly (each P<0.05). Thus, propofol can inhibit the LPS-induced expression and release of HMGB1 by inhibiting HMGB1 translocation and NF-κB activity in RAW264.7 cells, suggesting propofol may be protective in patients with sepsis.

Potent antitumoral efficacy of a novel replicative adenovirus CNHK300 targeting telomerase-positive cancer cells.

PURPOSE: Telomerase reverse transcriptase (hTERT) is the key determinant of telomerase activity and plays a crucial role in cellular immortalization and oncogenesis. It will be a promising target for cancer gene therapy. We constructed a novel replicative adenovirus CNHK300 in which hTERT promoter with three extra E-boxes downstream of the promoter was introduced and used to regulate adenoviral E1a gene, and studied its properties of selective replication in cancer cells and antitumoral activity. METHODS: Luciferase assay was used to detect hTERT promoter activity. The selective replication of CNHK300 in cancer cells was investigated by E1a Western blot and green fluorescent protein (GFP) reporter gene assay. The antitumoral activity of CNHK300 and its toxicity were measured on animal models. RESULTS: Luciferase assay showed that introducing extra E-boxes downstream of hTERT promoter is beneficial to decreasing the promoter activity in normal cells without affecting its strong activity in cancer cells. Experiments in vitro and in vivo demonstrated that CNHK300 can selectively target to hTERT-positive cancer cells and replicate in them, resulting in oncolytic or antitumoral effect. CNHK300 is superior to ONYX-015 in terms of selective replication and oncolytic or antitumoral effect. The toxicity assay showed no signs of toxicity to liver cells even at the higher dosage of CNHK300 in vivo. CONCLUSION: The hTERT promoter-controlled, replication-competent adenovirus CNHK300 is a promising system for targeted cancer gene therapy.

Cholecystokinin antagonist lorglumide reverses chronic haloperidol-induced effects on dopamine neurons.

Intravenous administration of the cholecystokinin (CCK) antagonist lorglumide (LORG) reversed chronic haloperidol (CHAL)-induced depolarization inactivation (DI) of dopamine (DA) cells in both the A9 and A10 areas. Moreover, microinjection of LORG, but not naloxone, directly into the medial nucleus accumbens (mNAc) dose-dependently reversed CHAL-induced effect. LORG injected into other brain regions was without effect. These results suggest that CCK receptors in the mNAc form an important link for maintaining CHAL-induced DI of DA cells and that CCK is involved in the therapeutic action of antipsychotic drugs.

The effect of intraventricular administration of the 5-HT3 receptor agonist 2-methylserotonin on the release of dopamine in the nucleus accumbens: an in vivo chronocoulometric study.

In the present study we have examined the effects of the serotonin3 (5-HT3) agonist 2-methylserotonin (2-Me-5HT) on the dopamine (DA) release in the nucleus accumbens (NAc) of rats using in vivo chronocoulometric recording. The intraventricular (i.c.v.) administration of 2-Me-5HT dose-dependently increased the DA release in the NAc. This effect was blocked by the selective 5-HT3 antagonist BRL-43694 (granisetron), but not by the 5-HT1/5-HT2 antagonist metergoline. The i.c.v. injection of 8-hydroxydipropylaminotetraline (8-OHDPAT, a selective 5-HT1a agonist) or (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI, a 5-HT2/5-HT1c agonist) failed to alter the DA release in the NAc. The increase in the DA release produced by 2-Me-5HT was abolished in animals that had received acute bilateral injections of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle. Our results suggest that the 2-Me-5HT-induced DA release in the NAc is mediated by 5-HT3 receptors. In addition, 2-Me-5HT induced effect is dependent upon the impulse flow of DA cells.

Chronic BRL 43694, a selective 5-HT3 receptor antagonist, fails to alter the number of spontaneously active midbrain dopamine neurons.

In this study, we examined the effect of chronic administration of the selective 5-HT3 receptor antagonist BRL 43694 on the number of spontaneously active A9 (substantia nigra pars compacta) and A10 (ventral tegmental area) dopamine (DA) cells using the technique of extracellular single unit recording. Overall, chronic BRL 43694 (21 days, 5 mg/kg per day or 2.5 mg/kg q.i.d) failed to alter the number of spontaneously active DA cells in either area compared to saline-injected controls. Furthermore, the acute i.v. administration of BRL 43694 did not change A10 DA cell baseline firing rate. These data suggest that if BRL 43694 possesses antipsychotic activity, it is not the result of its action on midbrain DA neurons.

Chronic treatment with high doses of haloperidol fails to decrease the time course for the development of depolarization inactivation of midbrain dopamine neurons.

Using extracellular single unit recording techniques, we investigated the effects produced by chronic treatment with high doses of haloperidol (CHAL, 5 mg/kg/day, s.c.) on midbrain dopamine (DA) neuronal activity. This regimen of HAL treatment produced a time-dependent reduction in the number of spontaneously active DA neurons. Additionally, this dose regimen induced an irregular firing pattern in many of the remaining active DA neurons in both the ventral tegmental area (A10) and substantia nigra pars compacta (A9) regions. These effects were comparable to those obtained previously in rats treated chronically with lower doses of HAL (0.5 mg/kg/day, s.c.). However, there was a greater decrease in the number of spontaneously active DA cells detected in rats treated with high doses of HAL for three weeks compared to those receiving the low doses. On the other hand, higher doses of apomorphine (200 micrograms/kg, i.v.) were required to reverse both the reduction of DA activity and irregular discharge pattern in rats treated chronically with high doses of HAL. In conclusion, the results of the present study substantiate the view that CHAL-induced depolarization inactivation (DI) of DA neurons is a time-dependent process and chronic treatment with high doses of HAL did not shorten the time course required for the development of DI on the majority of midbrain DA neurons.