Berberine Inhibits HIV Protease Inhibitor-Induced Inflammatory Response by Modulating ER Stress Signaling Pathways in Murine Macrophages

Background

HIV protease inhibitor (PI)-induced inflammatory response plays an important role in HIV PI-associated dyslipidemia and cardiovascular complications. This study examined the effect of berberine, a traditional herb medicine, on HIV PI-induced inflammatory response and further investigated the underlying cellular/molecular mechanisms in macrophages.

Methodology and Principal Findings

Cultured mouse J774A.1 macrophages and primary mouse macrophages were used in this study. The expression of TNF-α and IL-6 were detected by real-time RT-PCR and ELISA. Activations of ER stress and ERK signaling pathways were determined by Western blot analysis. Immunofluorescent staining was used to determine the intracellular localization of RNA binding protein HuR. RNA-pull down assay was used to determine the association of HuR with endogenous TNF-α and IL-6. Berberine significantly inhibited HIV PI-induced TNF-α and IL-6 expression by modulating ER stress signaling pathways and subsequent ERK activation, in turn preventing the accumulation of the RNA binding protein HuR in cytosol and inhibiting the binding of HuR to the 3′-UTRs of TNF-α and IL-6 in macrophages.

Conclusions and Significance

Inhibition of ER stress represents a key mechanism by which berberine prevents HIV PI-induced inflammatory response. Our findings provide a new insight into the molecular mechanisms of berberine and show the potential application of berberine as a complimentary therapeutic agent for HIV infection.     

Identification of a novel mutation of the gene for gap junction protein α3 (GJA3) in a Chinese family with congenital cataract

Cataract, defined as any opacity of the crystallin lens, can be divided into early onset (congenital or infantile) and age-related. It is the leading cause of visual disability in children, and mutations in many genes have currently been linked with this disorder. In the present study, we identified a genetic defect in a Chinese family with congenital cataract. Genomic DNA was extracted from the venous blood of the family and 100 normal controls. To screen for the disease-causing mutation, we sequenced eight candidate genes, and to predict the functional consequences of the mutation, a structural model of the protein was developed using the Protein Data Bank and PyMOL 1.1r1. We found a novel variant (c.163 A > G transition) in the gene for gap junction protein α3, or the connexin46 gene. This mutation resulted in the substitution of a highly conserved asparagine at codon 55 by aspartic acid (p.N55D). There were no nucleotide polymorphisms in the other candidate genes sequenced.     

Three novel homozygous mutations in the GNPTG gene that cause mucolipidosis type III gamma

Background

Mucolipidosis type III gamma (MLIII gamma) is an autosomal recessive disease caused by a mutation in the GNPTG gene, which encodes the γ subunit of the N-acetylglucosamine-1-phosphotransferase (GlcNAc-1-phosphotransferase). This protein plays a key role in the transport of lysosomal hydrolases to the lysosome.

Methods

Three Chinese children with typical skeletal abnormalities of MLIII were identified, who were from unrelated consanguineous families. After obtaining informed consent, genomic DNA was isolated from the patients and their parents. Direct sequencing of the GNPTG and GNPTAB genes was performed using standard PCR reactions.

Results

The three probands showed clinical features typical of MLIII gamma, such as joint stiffness and vertebral scoliosis without coarsened facial features. Mutation analysis of the GNPTG gene showed that three novel mutations were identified, two in exon seven [c.425G>A (p.Cys142Val)] and [c.515dupC (p.His172Profs27X)], and one in exon eight [c.609+1G>C]. Their parents were determined to be heterozygous carriers when compared to the reference sequence in GenBank on NCBI.

Conclusions

Mutation of the GNPTG gene is the cause of MLIII gamma in our patients. Our findings expand the mutation spectrum of the GNPTG gene and extend the knowledge of the phenotype–genotype correlation of the disease.     

外源CO及NO对干旱胁迫下水稻糊粉层细胞死亡的影响

采用荧光显微技术结合药理学方法,以水稻(Oryza sativa L.)种子及其糊粉层为实验材料,研究外源CO、NO对干旱胁迫下水稻种子萌发过程中糊粉层细胞DNA降解及死亡的影响。结果表明:(1)干旱胁迫促进糊粉层细胞的死亡,且近胚端糊粉层细胞的死亡进程早于远胚端的细胞。(2)外源CO及NO供体处理能缓解干旱胁迫下水稻糊粉层细胞DNA的降解,延迟细胞死亡进程;CO专一性抑制剂及NO清除剂能逆转CO及NO的效应,缩短细胞死亡进程。(3)外源CO及NO供体促进干旱胁迫下水稻种子的萌发,CO专一性抑制剂及NO清除剂能抑制干旱胁迫下水稻种子的萌发。(4)CO合成酶抑制剂并不能抑制外源NO对干旱胁迫伤害的缓解效应,即CO能通过NO介导调节干旱胁迫下水稻种子糊粉层细胞的死亡及种子萌发。     

Flavonoid Apigenin Inhibits Lipopolysaccharide-Induced Inflammatory Response through Multiple Mechanisms in Macrophages

Background

Apigenin is a non-toxic natural flavonoid that is abundantly present in common fruits and vegetables. It has been reported that apigenin has various beneficial health effects such as anti-inflammation and chemoprevention. Multiple studies have shown that inflammation is an important risk factor for atherosclerosis, diabetes, sepsis, various liver diseases, and other metabolic diseases. Although it has been long realized that apigenin has anti-inflammatory activities, the underlying functional mechanisms are still not fully understood.

Methodology and Principal Findings

In the present study, we examined the effect of apigenin on LPS-induced inflammatory response and further elucidated the potential underlying mechanisms in human THP-1-induced macrophages and mouse J774A.1 macrophages. By using the PrimePCR array, we were able to identify the major target genes regulated by apigenin in LPS-mediated immune response. The results indicated that apigenin significantly inhibited LPS-induced production of pro-inflammatory cytokines, such as IL-6, IL-1β, and TNF-α through modulating multiple intracellular signaling pathways in macrophages. Apigenin inhibited LPS-induced IL-1β production by inhibiting caspase-1 activation through the disruption of the NLRP3 inflammasome assembly. Apigenin also prevented LPS-induced IL-6 and IL-1β production by reducing the mRNA stability via inhibiting ERK1/2 activation. In addition, apigenin significantly inhibited TNF-α and IL-1β-induced activation of NF-κB.

Conclusion and Significance

Apigenin Inhibits LPS-induced Inflammatory Response through multiple mechanisms in macrophages. These results provided important scientific evidences for the potential application of apigenin as a therapeutic agent for inflammatory diseases.     

Contourlet Textual Features: Improving the Diagnosis of Solitary Pulmonary Nodules in Two Dimensional CT Images

Objective

To determine the value of contourlet textural features obtained from solitary pulmonary nodules in two dimensional CT images used in diagnoses of lung cancer.

Materials and Methods

A total of 6,299 CT images were acquired from 336 patients, with 1,454 benign pulmonary nodule images from 84 patients (50 male, 34 female) and 4,845 malignant from 252 patients (150 male, 102 female). Further to this, nineteen patient information categories, which included seven demographic parameters and twelve morphological features, were also collected. A contourlet was used to extract fourteen types of textural features. These were then used to establish three support vector machine models. One comprised a database constructed of nineteen collected patient information categories, another included contourlet textural features and the third one contained both sets of information. Ten-fold cross-validation was used to evaluate the diagnosis results for the three databases, with sensitivity, specificity, accuracy, the area under the curve (AUC), precision, Youden index, and F-measure were used as the assessment criteria. In addition, the synthetic minority over-sampling technique (SMOTE) was used to preprocess the unbalanced data.

Results

Using a database containing textural features and patient information, sensitivity, specificity, accuracy, AUC, precision, Youden index, and F-measure were: 0.95, 0.71, 0.89, 0.89, 0.92, 0.66, and 0.93 respectively. These results were higher than results derived using the database without textural features (0.82, 0.47, 0.74, 0.67, 0.84, 0.29, and 0.83 respectively) as well as the database comprising only textural features (0.81, 0.64, 0.67, 0.72, 0.88, 0.44, and 0.85 respectively). Using the SMOTE as a pre-processing procedure, new balanced database generated, including observations of 5,816 benign ROIs and 5,815 malignant ROIs, and accuracy was 0.93.

Conclusion

Our results indicate that the combined contourlet textural features of solitary pulmonary nodules in CT images with patient profile information could potentially improve the diagnosis of lung cancer.     

吡哆胺对糖尿病大鼠视皮质AGE-RAGE系统的影响

目的观察吡哆胺对糖尿病大鼠视皮质高级糖基化终末产物(AGE)及其受体(RAGE)表达的影响,探讨吡哆胺对视皮质的保护作用。方法健康SD大鼠随机分为正常对照组(NC组)、糖尿病未治疗组(DM组)、糖尿病吡哆胺治疗组(PM组)和氨基胍治疗对照组(AG组)各20只,用链脲佐菌素(STZ)建立糖尿病模型,PM组和AG组分别于造模成功后第二天开始予吡哆胺和氨基胍灌胃。各组于治疗4w和12w后取材,用酶联免疫吸附(ELISA)法定量检测大鼠视皮质中AGEs含量,荧光免疫组化及图像分析半定量检测各组视皮质RAGE的表达。结果糖尿病治疗组和未治疗组血糖无显著性差异。4w时各组AGEs含量无明显差异,12w时PM组视皮质中AGEs含量与AG组、NC组比较差异无统计学意义,与DM组相比显著降低,差异具有统计学意义(P0.05)。PM组视皮质中RAGE表达比DM组显著减少,差异具有统计学意义(P0.05),但高于NC组(P0.05)。结论糖尿病大鼠12w后视皮质中AGEs含量和RAGE的表达高于正常对照组,吡哆胺类似氨基胍可减少AGEs的堆积,还能抑制RAGE的表达,减轻AGEs-RAGE通路作用导致的组织损伤,对视皮质具有一定的保护作用。     

Basic FGF regulates interstitial collagenase gene expression in human smooth muscle cells

Basic fibroblast growth factor (bFGF) is a mitogenic factor that is implicated in smooth muscle cell growth in atherosclerosis and vascular restenosis. In this study, we examined the effect of bFGF on the expression of the interstitial collagenase gene in human vascular smooth muscle cells. Results from Northern transfer analysis showed that bFGF increased collagenase mRNA levels greater than threefold as early as 24 h. Collagenase pre-mRNA levels were elevated approximately threefold by bFGF, according to RT-PCR analysis. Transient transfections of the smooth muscle cells with a 4.4-kb human collagenase promoter-CAT reporter gene, however, failed to show upregulation of the promoter activity by bFGF. Interestingly, transfections with deleted fragments containing promoter sequences from -1047 to -2271 resulted in modest stimulation of the collagenase-CAT promoter activity by bFGF. bFGF did not alter the stability of the collagenase mRNA, as demonstrated by degradation studies. The enhanced collagenase mRNA levels elicited by bFGF were reflected in increased amounts of collagenase protein that were detected by Western blot analysis. In summary, bFGF upregulates the interstitial collagenase expression, resulting in turnover of the extracellular matrix, an event that could facilitate smooth muscle cell migration and proliferation during the early stages of atherosclerosis and restenosis. J. Cell. Biochem. 65:32–41. © 1997 Wiley-Liss, Inc.     

Human infection with a further evolved avian H9N2 influenza A virus in Sichuan,China

正Dear Editor,Avian influenza A (H9N2) virus plays a crucial role in interspecies transmission between animals and humans due to its wide host range, adaptation in mammals and the diversified gene reassortments. Since 1990s, two lineages of H9N2 virus, A/Duck/HK/Y280/97 (Y280-like)and A/Quail/HK/G1/97 (G1) have established in domestic poultry in China and are now widespread in the whole