Overexpression of SMS in the tumor microenvironment is associated with immunosuppression in hepatocellular carcinoma.

Disorders of polyamine metabolism may contribute to the development of hepatocellular carcinoma (HCC), but the precise mechanism remains unknown. This study reports that spermine synthase (SMS), an enzyme involved in polyamine biosynthesis, is overexpressed in HCC and not associated with hepatitis virus infection in HCC patients. The results of analyzing the clinical data of HCC patients showed that SMS level as a categorical dependent variable was related to clinicopathological features of poor prognosis. Furthermore, the Kaplan-Meier survival analysis and ROC curve indicated that increased SMS level is associated with poor survival rate in HCC and may be a potential biomarker to discriminate HCC tissues. However, SMS overexpression limited the therapeutic effect of immune checkpoint blockade (ICB), which seemed to be related to the immunosuppressive effect of the HCC immune microenvironment formed by higher mRNA transcript levels of immune checkpoints and higher infiltration levels of immunosuppressive cells. In samples with high and low SMS expression, functional enrichment analysis of the differentially expressed genes (DEGs) showed that SMS may be linked to the occurrence and development of HCC by affecting a variety of immune-related pathways, such as Intestinal immune network for IgA production, Fc gamma R-mediated phagocytosis, Antigen processing and presentation, Th1 and Th2 cell differentiation. Subsequently, analysis of the co-expression network of SMS in the liver hepatocellular carcinoma (LIHC) cohort revealed that SMS has a broad impact on multiple important immune- and metabolic-related processes in HCC. In summary, SMS is a promising biomarker to differentiate the prognosis, immune characteristics, and holds promise as a potential target for ICB therapy to improve HCC.

Ganoderic Acid A Promotes Amyloid-ß Clearance (In Vitro) and Ameliorates Cognitive Deficiency in Alzheimer's Disease (Mouse Model) through Autophagy Induced by Activating Axl.

Alzheimer's disease (AD) is thought to be caused by amyloid-ß (Aß) accumulation in the central nervous system due to deficient clearance. The aim of the present study was to investigate the effect of ganoderic acid A (GAA) on Aß clearance in microglia and its anti-AD activity. Aß degradation in BV2 microglial cells was determined using an intracellular Aß clearance assay. GAA stimulated autophagosome formation via the Axl receptor tyrosine kinase (Axl)/RAC/CDC42-activated kinase 1 (Pak1) pathway was determined by Western blot analyses, and fluorescence-labeled Aß42 was localized in lysosomes in confocal laser microscopy images. The in vivo anti-AD activity of GAA was evaluated by object recognition and Morris water maze (MWM) tests in an AD mouse model following intracerebroventricular injection of aggregated Aß42. The autophagy level in the hippocampus was assayed by immunohistochemical assessment against microtubule-associated proteins 1A/1B light-chain 3B (LC3B). Intracellular Aß42 levels were significantly reduced by GAA treatment in microglial cells. Additionally, GAA activated autophagy according to increased LC3B-II levels, with this increased autophagy stimulated by upregulating Axl and Pak1 phosphorylation. The effect of eliminating Aß by GAA through autophagy was reversed by R428, an Axl inhibitor, or IPA-3, a Pak1 inhibitor. Consistent with the cell-based assay, GAA ameliorated cognitive deficiency and reduced Aß42 levels in an AD mouse model. Furthermore, LC3B expression in the hippocampus was up-regulated by GAA treatment, with these GAA-specific effects abolished by R428. GAA promoted Aß clearance by enhancing autophagy via the Axl/Pak1 signaling pathway in microglial cells and ameliorated cognitive deficiency in an AD mouse model.

Administration of isoliquiritigenin prevents nonalcoholic fatty liver disease through a novel IQGAP2-CREB-SIRT1 axis.

Isoliquiritigenin (ISO) is a flavonoid extracted from the root of licorice, which serves various biological and pharmacological functions including antiinflammatory, antioxidation, liver protection, and heart protection. However, the mechanism of its action remains elusive and the direct target proteins of ISO have not been identified so far. Through cell-based screening, we identified ISO as a potent lipid-lowering compound. ISO treatment successfully ameliorated fatty acid-induced cellular lipid accumulation and improved nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) by increasing PPARα-dependent lipid oxidation and decreasing SREBPs-dependent lipid synthesis. Both these signaling required the activation of SIRT1. Knockdown of SIRT1 resulted in the reversal of ISO beneficiary effects suggesting that the lipid-lowering activity of ISO was regulated by SIRT1 expression. To identify the direct target of ISO, limited proteolysis combined with mass spectrometry (LiP-SMap) strategy was applied and IQGAP2 was identified as the direct target for ISO in regulating lipid homeostasis. In the presence of ISO, both mRNA and protein levels of SIRT1 were increased; however, this effect was abolished by blocking IQGAP2 expression using siRNA. To explore how IQGAP2 regulated the expression level of SIRT1, proteome profiler human phospho-kinase array kit was used to reveal possible phosphorylated kinases and signaling nodes that ISO affected. We found that through phosphorylation of CREB, ISO transduced signals from IQGAP2 to upregulate SIRT1 expression. Thus, we not only demonstrated the molecular basis of ISO in regulating lipid metabolism but also exhibited for the first time a novel IQGAP2-CREB-SIRT1 axis in treating NAFLD/NASH.

Novel hepatoprotective role of Leonurine hydrochloride against experimental non-alcoholic steatohepatitis mediated via AMPK/SREBP1 signaling pathway.

BACKGROUND AND AIMS: Non-alcoholic steatohepatitis (NASH) is the hepatic manifestation of metabolic syndrome and is characterized by steatosis, inflammation, and fibrosis. We aim to characterize the hepatoprotective effects of Leonurine hydrochloride (LH) and the possible pathway in a cell and rodent model of diet-induced steatohepatitis (NASH). METHODS: For in vitro studies, Palmitic acid (PA) and free fatty acid (FFA) induced HepG2 and HL7702 steatosis cell models were used. For in vivo studies, NASH was induced by feeding mice MCD diet. These mice received either placebo or LH at three different doses (50、100、200 mg/kg/day) for 6 weeks. Histological staining's, and commercially available kits for ALT and AST and hepatic contents of TG, TC, MDA, SOD, and GSH were used to assess NASH. Furthermore, relative liver protein and gene expression levels were determined by Western Blot and qPCR, respectively. RESULTS: After establishing NASH models, LH treatment improved lipid accumulation, hepatic contents of TG, TC, and expression levels of ALT and AST in dose-dependent manner. Also, LH improved MDA, SOD, and GSH expression levels. The results of RT-PCR and Western blotting showed that LH upregulated the expression of AMPK phosphorylation and downregulated SREBP-1c and its target genes expression level. CONCLUSIONS: Our data reveal the promising role of Leonurine hydrochloride in the prevention and treatment of NASH, in vitro and in vivo. This effect may be partially mediated by the AMPK/SREBP1 pathway. These findings provide a novel therapeutic target for the clinical treatment of NASH.

Administration of methyl palmitate prevents non-alcoholic steatohepatitis (NASH) by induction of PPAR-α.

BACKGROUND AND AIMS: The lack of valid therapeutic approach that can ameliorate the manifestations of NASH is a barrier to therapeutic development. Therefore, we investigate the novel role of Methyl Palmitate (MP) in preventing NASH and the possible mechanism involved. METHODS: 50 Male C57BL/6 J mice were randomly divided into 5 groups (n = 10). The control group was fed control diet; model group was fed MCD diet; MP 1 group was fed MCD diet supplemented with MP (75 mg/kg/day); MP 2 group was fed MCD plus MP diet (150 mg/kg/day); and MP 3 group was fed MCD plus MP diet (300 mg/kg/day). Histological staining's, and commercially available kits for serum ALT and AST and hepatic contents of TG, TC, MDA, SOD, and GSH were used to assess NASH. Furthermore, relative liver protein and gene expression levels were determined by Western Blot and qPCR, respectively. RESULTS: Mice fed MCD diet developed NASH, which was markedly improved by MP in a dose-dependent manner. MP treatment improved hepatic content of TG, TC, MDA, SOD and GSH and serum levels of ALT and AST. In vivo studies showed that MP treatment activated PPARα expression, that in turns, promoted ß-oxidation protein and gene expressions, suppressed TNFα, MCP1, TGFß1 and Colla1 protein and gene expression levels, contributing to the prevention of NASH. CONCLUSIONS: Our results indicated that MP could successfully prevent NASH. This effect of MP was mediated through induction of PPARα pathway. This study provides a novel therapeutic target that plays pivotal role in the prevention of NASH.

Study of cochlear hydrops analysis masking procedure in patients with Meniere's disease and otologically normal adults.

BACKGROUND: The cochlear hydrops analysis masking procedure (CHAMP) is a new diagnostic technique for Meniere's disease (MD). But its value has not been well proven. This study aimed to evaluate the diagnostic value of CHAMP for MD. METHODS: CHAMP test was taken in three populations using the Auditory Evoked Potential system delivered by Bio-logic Systems Corporation: (1) otologically normal subjects; (2) patients clinically diagnosed with definite MD; (3) patients clinically diagnosed with probable and possible MD. RESULTS: According to the comparison between the normal and definite MD group, if the abnormal criterion of CHAMP was defined as latency delay less than 0.3 ms, then the corresponding sensitivity was only 52%. However, if the abnormal criterion was defined as latency delay between 0.6 and 3.8 ms, then a sensitivity of 93% and a specificity of 100% can be achieved. The complex amplitude ratio showed a significant overlap between normal and definite MD group. If the abnormal criterion was defined as a complex amplitude ratio less than 0.95, the corresponding specificity was only 50%. However, if the abnormal criterion was defined as less than 0.80, the corresponding sensitivity was 60%, and the specificity was 97%. If the abnormal criterion of CHAMP was defined as latency delay less than 0.6 ms or the complex amplitude ratio less than 0.80, CHAMP result can be obtained in all subjects with good sensitivity and specificity. CONCLUSIONS: CHAMP can differentiate patients with Meniere's disease from otologically normal subjects with high sensitivity and specificity. The recommended criterion of abnormal CHAMP was a latency delay less than 0.6 ms or a complex amplitude ratio less than 0.80.

[Prevalence of blindness and moderate and severe visual impairment among adults aged 50 years or above in the Shunyi District of Beijing: the China Nine-Province Survey].

UNLABELLED: OBJECTIVE To assess the prevalence of blindness and moderate and severe visual impairment among adults aged 50 years or above in the Shunyi district of Beijing,China. METHODS: It was a population-based cross-section study. Geographically defined cluster sampling was used in 5840 randomly selected individuals aged > or = 50 years in 28 clusters in the Shunyi district from April to August 2006. The survey was preceded by a pilot study where operational methods were refined and quality assurance evaluation was carried out. All participants were enumerated through village registers followed door-to-door visits. Eligible individuals were invited to receive visual acuity measurement and eye examination. Statistical analyses were performed using Stata/SE Statistical Software, release 9. 0. Chi-square test was used to investigate the association of age, gender and education with presenting and best corrected visual acuity. RESULTS: Five thousands eight hundreds and forty individuals were recruited and 5118 persons were examined,the response rate was 87.64%. Based on the criteria of World Health Organization visual impairment classification in 1973,57 persons were diagnosed as blindness, 160 persons were diagnosed as moderate and severe visual impairment defined as best corrected visual acuity, the prevalence of blindness and moderate and severe visual impairment were 1.11% and 3.12% respectively. Sixty-five persons were diagnosed as blindness, 428 persons were diagnosed as moderate and severe visual impairment defined as presenting visual acuity, the prevalence of blindness and moderate and severe visual impairment were 1.27% and 8. 36% respectively. The prevalence of blindness and moderate and severe visual impairment was higher in aged (trend Chi2 = 784.23, P = 0.000), female (Chi2 = 28.35, P = 0.000), and illiterate persons (trend Chi2 = 397.13, P = 0.000). Cataract was still the first leading cause of blindness and visual impairment, the retinal diseases was the second. Un-corrected refractive error also was the main cause of visual impairment. CONCLUSIONS: The prevalence of blindness and moderate and severe visual impairment has been decreasing among older adults aged 50 years or above in the Shunyi district since 1996. The first leading cause of blindness and visual impairment is still cataract.

[Prevalence and surgery status of cataract among adults aged 50 years or above in the Shunyi district of Beijing: the China Nine-Province Survey].

OBJECTIVE: To assess the prevalence of cataract, the outcomes of cataract surgery, the rate of cataract surgical coverage, and the rate of cataract blindness social burden. METHODS: Cluster sampling was used in 5118 randomly selected individuals aged > or = 50 years in the Shunyi district of Beijing from April to August 2006. All individuals were received visual acuity and eye examination, including the evaluation of lens status and cataract surgical status by slit lamp biomicroscopy. Data bank was established by Epi-info Software. Statistical analyses were performed using Stata/SE Statistical Software, release 9.0 (Stata Corp, College Station,TX). Multiple logistic regression analysis was employed to analyzed the prevalence of cataract,cataract surgical coverage rate,cataract blindness social burden rate and outcomes of surgery among different ages, sex and educational background. RESULTS: In 5118 individuals, 797 cases of cataract were found, the prevalence of cataract was 15.57% among adults aged 50 or above. The prevalence of cataract was higher in the aged, female, illiterate persons (P < 0.01). In 189 eyes with cataract surgery, 65.08% and 23.28% of eyes were performed by the modern extra-capsular surgery and Phacoemulsification respectively. The rate of intraocular lens implantation was 89.42%. Post-operative presenting and best corrected visual acuity more than 0.7 was 30.16% and 61.38% of operated eyes respectively. The main causes of the post-operated eyes with worse visual acuity were post-capsular opacity and refractive error. The cataract surgical coverage rate was 62.22%, and the cataract blindness social burden rate was 2.64%. The cataract surgical rate was lower and cataract blindness social burden rate was higher in the aged persons (P < 0.01). CONCLUSIONS: Cataract is the most common eye disease that may lead into blindness and severe visual impairment among older adults aged equal or more than 50 years. The cataract surgical coverage rate is high in the Shunyi district, but still is lower in the aged persons. The visual outcomes of the surgery should be further improved in the future.

[Study of characteristic of the cochlear hydrops analysis masking procedure in normal adults].

OBJECTIVE: To study the characteristic of the cochlear hydrops analysis masking procedure (CHAMP) in normal adults, and to evaluate the diagnostic values of its parameters for membranous labyrinth hydrops. METHODS: Twenty otologically normal adults were recruited (male:female = 10:10), and their auditory brainstem responses (ABR) were obtained to six stimulus conditions using Bio-logic auditory evoked potential system: clicks presented alone (unmasked condition) and clicks presented with ipsilateral pink noise high-pass filtered at 8, 4, 2, 1, and 0.5 kHz respectively. RESULTS: The wave V latency of ABR to the high-pass masking pink noise clicks were longer than ABR to clicks alone. The latency delays of wave V for clicks presented with ipsilateral pink noise high-pass filtered at 8, 4, 2, 1, and 0.5 kHz compared to clicks alone were (0.30 ± 0.18), (0.97 ± 0.43), (1.65 ± 0.64), (3.21 ± 0.56), (4.66 ± 0.37) ms respectively. The complex amplitude ratio between ABR to click + 0.5 kHz high-pass noise and click alone was 0.95 ± 0.11. CONCLUSIONS: CHAMP is a promising diagnostic method for membranous labyrinth hydrops, and the latency delay of wave V might be used as the normal criterion. The specificity of the complex amplitude ratio need further evaluation in clinical work.

[Comparison of tone burst evoked auditory brainstem responses with different filter settings for referral infants after hearing screening].

OBJECTIVE: Auditory brainstem responses (ABR) evoked by tone burst is an important method of hearing assessment in referral infants after hearing screening. The present study was to compare the thresholds of tone burst ABR with filter settings of 30 - 1500 Hz and 30 - 3000 Hz at each frequency, figure out the characteristics of ABR thresholds with the two filter settings and the effect of the waveform judgement, so as to select a more optimal frequency specific ABR test parameter. METHODS: Thresholds with filter settings of 30 - 1500 Hz and 30 - 3000 Hz in children aged 2 - 33 months were recorded by click, tone burst ABR. A total of 18 patients (8 male/10 female), 22 ears were included. RESULTS: The thresholds of tone burst ABR with filter settings of 30 - 3000 Hz were higher than that with filter settings of 30 - 1500 Hz. Significant difference was detected for that at 0.5 kHz and 2.0 kHz (t values were 2.238 and 2.217, P < 0.05), no significant difference between the two filter settings was detected at the rest frequencies tone evoked ABR thresholds. The waveform of ABR with filter settings of 30 - 1500 Hz was smoother than that with filter settings of 30 - 3000 Hz at the same stimulus intensity. Response curve of the latter appeared jagged small interfering wave. CONCLUSIONS: The filter setting of 30 - 1500 Hz may be a more optimal parameter of frequency specific ABR to improve the accuracy of frequency specificity ABR for infants' hearing assessment.

Potential roles of Cdk5/p35 and tau protein in hippocampal mossy fiber sprouting in the PTZ kindling model.

BACKGROUND: The most well-documented synaptic reorganization associated with temporal lobe epilepsy is mossy fiber sprouting (MFS), which is believed to play a critical role in epileptogenesis. However, the molecular mechanisms underlying this phenomenon remain unclear. Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/threonine kinase which is found to be crucial in axon growth and synaptic plasticity. We hypothesized that Cdk5 contributed to MFS via phosphorylating its substrate tau protein, which was known to facilitate microtubule stabilization and axonal elongation. METHODS: 240 male SD rats were randomly divided into the control group and PTZ group. The epileptic models were established by intraperitoneal PTZ injection, while the control rats were injected with an equal dose of saline. At different time points, Cdk5/p35 mRNA and protein, total tau protein and its phosphorylation at Ser202 (p-tau) and Cdk5 activity were analyzed in different regions of hippocampus by in situ hybridization, immunohistochemistry, Western blot, immuno-precipitation and liquid scintillation counter. Hippocampus was also evaluated for MFS with Timm stain. RESULTS: Prominent MFS was observed in area CA3 rather than the inner molecular layer in PTZ treated rats and the degree of MFS progressed with the development of behavioral kindled seizures. The expression of Cdk5/p35 mRNA and protein, tau protein and its phosphorylation at Ser202 significantly increased from 3 days to 4 weeks in the PTZ group, which was in accordance with the progression of MFS in area CA3. CONCLUSIONS: Cdk5/p35 and its substrate tau protein may be involved in MFS. Understanding the molecular mechanisms underlying MFS may lead to therapeutic interventions that limit epileptogenesis.

Mossy fiber sprouting, hippocampal damage and spontaneous recurrent seizures in pentylenetetrazole kindling rat model.

AIM: The aim of this study was to determine the correlations among hippocampal damage, spontaneous recurrent seizures (SRS), and mossy fiber sprouting (MFS) using pentylenetetrazole (PTZ) kindling model. METHODS: Chronic epileptic model was established by administration of PTZ. Behaviour and EEG seizure activity were recorded. Rats' hippocampus were analyzed with haematoxylin and eosin (H&E) stain for histological lesions and evaluated for MFS with Timm stain. RESULTS: Prominent MFS was observed in area CA3 rather than the inner molecular layer in PTZ treated rats and the degree of MFS progressed with the development of behavioral kindled seizures. MFS preceded the occurrence of spontaneous seizures. No obvious neuronal necrosis and loss were observed in different regions of the hippocampus during kindling progression. CONCLUSION: MFS is not the outcome of SRS. Severe hippocampal damage is not required in the development of MFS and SRS.

[Analysis of abnormality of the middle ear in infant and young children].

OBJECTIVE: To discuss the diagnosis of middle ear abnormality in infants and young children. METHODS: To analyze retrospectively the data of audiology (including ABR, tympanometry) and CT scanning in 31 infants and young children who presented middle ear abnormality. RESULTS: Wave I latencies of ABR were delayed in 38 of 62 ears and not delayed in 15 ears, but CT scanning showed high density in 6 ears of these 15 ears. Wave I could not be elicited in 9 ears. Tympanometries were tested in 16 cases and were abnormal in 17 ears. CT scanning was carried out in 15 cases who's ABR and tympanometries showed abnormal. High signal intensity was present in mastoids and middle ear cavities in both ears of 12 cases and unilateral ear of 3 cases. Wave I latency of ABR was delayed and High signal intensity was present in mastoids and middle ear cavities in CT scanning of 13 ears. Wave I latency of ABR was normal, but high signal intensity was present in mastoids and middle ear cavities in CT scanning of 4 ears, there was no any ear which Wave I latency was delayed but CT scanning was normal. And disaccord among ABR, Tympanometry and CT scanning were showed. A typical case was reported. CONCLUSIONS: The most abnormality of the middle ear could be found used the tympanometry and I latency of ABR in infant and young children, but still there were some abnormality of the middle ear could not be showed. Some quandaries were existed and more sensitivity tests were needed in the diagnosis of abnormality in middle ears of infant and young children.

9-(2-Thien-yl)-9H-carbazole.

In the title compound, C(16)H(11)NS, the dihedral angles between the fused ring system and the pendant thienyl ring are 86.37 (5) and 57.14 (5)°.

[The use of muscle autograft denatured by microwave for repair of gaps in removal of facial neuromas].

OBJECTIVE: To study the clinical value of muscle autograft denatured by microwave for repair of gaps in removal of facial neuromas. METHODS: Two cases of patients with facial nerve Schwann cell neuromas were reported. The operations for removal of facial neuromas were completed, and the gaps of the nerves were repaired with muscle autograft denatured by microwave of 250 W for 120 sec. RESULTS: The patients were followed up for two years, and the recovery of facial function on the affected sides were satisfactory. CONCLUSION: Muscle autograft denatured by microwave technique is convenient, highly efficient for repairing facial nerve gap after removal of facial neuroma.