Identification of gene modules and hub genes associated with Colletotrichum siamense infection in mango using weighted gene co-expression network analysis.

Colletotrichum siamense is a hemibiotrophic ascomycetous fungus responsible for mango anthracnose. The key genes involved in C. siamense infection remained largely unknown. In this study, we conducted weighted gene co-expression network analysis (WGCNA) of RNA-seq data to mine key genes involved in Colletotrichum siamense-mango interactions. Gene modules of Turquoise and Salmon, containing 1039 and 139 respectively, were associated with C. siamense infection, which were conducted for further analysis. GO enrichment analysis revealed that protein synthesis, organonitrogen compound biosynthetic and metabolic process, and endoplasmic reticulum-related genes were associated with C. siamense infection. A total of 568 proteins had homologs in the PHI database, 370 of which were related to virulence. The hub genes in each module were identified, which were annotated as O-methyltransferase (Salmon) and Clock-controlled protein 6 (Turquoise). A total of 24 proteins exhibited characteristics of SCRPs. By using transient expression in Nicotiana benthamiana, the SCRPs of XM_036637681.1 could inhibit programmed cell death (PCD) that induced by BAX (BCL-2-associated X protein), suggesting that it may play important roles in C. siamense infection. A mango-C. siamense co-expression network was constructed, and the mango gene of XM_044632979.1 (auxin-induced protein 15A-like) was positively associated with 5 SCRPs. These findings help to deepen the current understanding of necrotrophic stage in C. siamense infection.

MeDIP-seq reveals the features of mitochondrial genomic methylation in immature testis of Chinese mitten crab Eriocheir sinensis.

In this study, the methylation of mitochondrial genome in the immature testis of Chinese mitten crab Eriocheir sinensis of the Yangtze River system was determined for the first time using MeDIP-seq. Our methylated DNA fragments covered more than 99% of the mitochondrial genome in E. sinensis loaded from GenBank. There were 8 mutated bases and 42 SNPs in the crab mitochondrial genome. The methylation presented in all genes as well as in an A + T region, but less in intergenic regions in the mitochondrial genome. However, the level of methylation of most genes coding proteins and the A + T region were high. But, the majority of genes encoding tRNAs were hypomethylated, and both the rRNA genes also showed methylation of low or median frequency. Especially, the level of methylation of the intergenic regions is the lowest. Those features indicated that the methylation of DNA may play an important role in gene expressing regulation in the mitochondrial genome of immature testis in E. sinensis.

H3K9ac involved in the decondensation of spermatozoal nuclei during spermatogenesis in Chinese mitten crab Eriocheir sinensis.

As a well-known crustacean model species, the Chinese mitten crab Eriocheir sinensis presents spermatozoa with decondensed DNA. Our aim was to analyze structural distribution of the histone H3 and its acetylated lysine 9 (H3K9ac) during spermatogenesis for the mechanistic understanding of the nuclear decondensation of the spermatozoa in E. sinensis. Using specific antibodies, we followed the structural distribution and acetylated lysine 9 of the histone H3 during spermatogenesis, especially spermiogenesis, of E. sinensis. Various spermary samples at different developmental stages were used for histological immunofluorescence and ultrastructural immunocytochemistry. Our results demonstrate a wide distribution of the histone H3 and H3K9ac during spermatogenesis, including spermatogonia, spermatocytes, spermatids, and immature and mature spermatozoa except for absence of H3K9ac in the secondary spermatocytes. Especially during the initial stage of nuclear decondensation, histone H3 lysine 9 was acetylated and then an amount of H3K9ac was removed from within to outside of the nuclei of late spermatids. The portion of remaining H3K9ac was gradually transferred from the nuclei during the stages of spermatozoa maturation. Our findings suggest both the acetylation of histone H3 lysine 9 and the remain of H3K9ac to contribute to the nuclear decondensation in spermatozoa of E. sinensis.

Spautin-1 Ameliorates Acute Pancreatitis via Inhibiting Impaired Autophagy and Alleviating Calcium Overload.

Acute pancreatitis is characterized by zymogen pre-activation. Severe inflammation caused by zymogen activation can eventually lead to multiple organ dysfunctions, which contributes to the high mortality rate of severe acute pancreatitis. However, there is no specific treatment available for acute pancreatitis therapy. Here, we show that spautin-1, which effectively inhibits autophagy flux, ameliorated the pathogenesis of acute pancreatitis induced by cerulein or L-Arginine. CaMKII phosphorylation due to cytosolic calcium oeverload was revealed in this paper. It was also demonstrated that autophagic protein aggregates degradation blockade accompanying with impaired autophagy correlated positively to intra acinar cells digestive aymogen activation sitimulated by cerulein or L-Arginine. The role of spautin-1 in ameliorating acute pancreatitis was shown here to be associated with impaired autophagy inhibition and Ca2+ overload alleviation. We provided a promising therapy for acute pancreatitis here through targeting both impaired autophagy and increased cytosolic calcium.

[Solanum lyratum Thunberg alkaloid induces human lung adenoearcinoma A549 cells apoptosis by inhibiting NF-kappaB signaling pathway].

OBJECTIVE: To investigate the effects of solanum lyratum Thunberg alkaloid (STA) on induction of apoptosis and the expression of NF-kappaB signaling pathway related genes in A549 cells. METHODS: A549 cells was treated with STA in vitro. The proliferation inhibitory effect was evaluated by MTT assay. Induction of cell apoptotic rate was determined by flow cytometry method (FCM) after Annexin V-FITC/PI double staining. The expression of NF-kappaB/p65 in nuclei, Survivin, IkappaBalpha and p-1kaapaBalpha in cytosol were detected by western blot. RESULTS: STA exhibited strong proliferation inhibitory effect in a dose-and -time-dependent manner against A549 cells. After treated with STA for 24 h, the apoptotic rate was increased significantly. The expression of IkappaBalpha protein was increased markedly,while those of NF-kappaB/p65, Survivin and p-IkappaBalpha proteins were decreased markedly. CONCLUSION: STA can induce apoptosis of lung adenocarcinoma A549 cells, its mechanisms may be related to inhibition of NF-kappaB signaling pathway.

[Apoptosis-inducing effect of Scutellaria barbata extract on human lung cancer SPC-A-1 cells and the expression of apoptosis associated genes].

OBJECTIVE: To explore the apoptosis-inducing effect of Scutellaria barbata extract (SBE) and the expression of apoptosis associated genes survivin and caspase-3 on human lung cancer SPC-A-1 cells. METHODS: Lung cancer SPC-A-1 cells were treated with 2.5, 5 and 10 mg/L for 48 h,and the cells were treated with 2.5 mg/L DDP as positive control. The inhibitory rat was evaluated by MTT assay. Apoptotic rate was determined by TUNEL method. The expression of survivin and caspase-3 mRNA were detected by semi-quantitive RT-PCR. RESULTS: Compared with control group, the inhibitory rate was increased obviously (P < 0.001), the apoptotic rate was increased markedly (P < 0.01), the expression of caspase-3 mRNA was increased significantly (P < 0.05 or P < 0.01), while survivin mRNA was decreased markedly (P < 0.05 or P < 0.01) in SBE groups. CONCLUSION: SBE can induce apoptosis of SPCA-1 cells. The molecular mechanism may be related to up-regulating expression of caspase-3 and down-regulating expression of survivin genes.

[The influence of Solanum lyratum Thunb extract on apoptosis and the expression of fas/fasL genes in Hela cells].

OBJECTIVE: To explore the effects of Solanum lyratum Thunb (SL) extract on the apoptosis and the expression of fas and fasL genes in Hela cells. METHODS: The proliferation inhibitory rate was evaluated by MTF assay. Induction of cell apoptosis rate was determined by flow cytometry. The expression of Fas protein was detected by two-step immunhistochemical staining. The expression of fas and fasL mRNA was detected by semi-quantitive RT-PCR. RESULTS: SL extract displayed strong proliferation inhibitory effect in a dose-and-time-dependent manner against Hela cell. The rate of apoptosis was increased obviously. The expression of fas mRNA and protein was increased significantly, and fasL mRNA was decreased markedly. CONCLUSION: SL can induce apoptosis by up-regulating expression of fas and fasL genes, and inhibit the development of Hela cells.

Study of the protective effect of L-arginine on relative ischemic/reperfusion injury in myocardium of rat / 中国应用生理学杂志

<p><b>AIM</b>To explore the protective effect of L-arginine on isolated rat heart with ischemia/reperfusion injury.</p><p><b>METHODS</b>24 wistar rats were randomly divided into 3 groups (each 8): control group, ischemia group, L-arginine group. The myocardiac relatively ischemia/reperfusion models in vitro were set up by using weak current stimulating isolated rat hearts. During the pre-ischemia, post-ischemia 15 min and post-ischemia 30 min, the coronary fluid was collected for testing contents of MDA and activities of both CK and LDH. Cardiac functional indexes were recorded through Pclab. At the time of 5 min, 10 min, 20 min, 30 min after ischemia, the recovery of PRP, + DP/dt(max) and - DP/dt(max) were calculated.</p><p><b>RESULTS</b>(1) During the reperfusion, L-arginine group achieved better recovery of cardiac function than that of the ischemia group. (2) MDA content, CK and LDH activities both in the coronary fluid and in the myocardium of L-arginine group were lower than those of the ischemia group, while SOD activities in the myocardium of L-arginine group were higher than that of the ischemia group.</p><p><b>CONCLUSION</b>To some extent, L-arginine could protect the myocardium from ischemia/reperfusion injury.</p>

Detection of RNA of SARS coronavirus in hospital sewage / 中华预防医学杂志

<p><b>OBJECTIVE</b>In order to explore the existence of SARS coronavirus (Co-V) and/or its RNA in sewage of hospitals administered SARS patients.</p><p><b>METHODS</b>A novel electropositive filter was used to concentrate the SARS-CoV from the sewage of two hospitals administered SARS patients in Beijing, including twelve 2,500 ml sewage samples from the hospitals before disinfection, and ten 25,000 ml samples after disinfection; as well as cell culture, RT-PCR and sequencing of gene to detect and identify the viruses from sewage.</p><p><b>RESULTS</b>There was no live SARS-CoV detected in the sewage in this study. The nucleic acid of SARS-CoV had been found in the 12 sewage samples before disinfection from both hospitals by semi-nested PCR. After disinfection, SARS-CoV RNA could only be detected from the samples from the 309th Hospital, and the others were negative.</p><p><b>CONCLUSION</b>It provides evidence that there is no live SARS-Cov in the sewage from hospitals with SARS patients though SARS-CoV RNA can be detected.</p>

A study on detecting and identifying enteric pathogens with PCR / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To develop a rapid and definite diagnostic test of bacterial enteritis caused by pathogenic enterobacteria, the most frequent etiologic agent of infectious enteritis in the world.</p><p><b>METHODS</b>A set of conventional PCR assays were applied to detect and identify salmonella, shigella, and E. coli O157:H7 directly from pure culture and fecal samples. The general primers of pathogenic enterobacteria were located on the uidA gene, which were found not only in E. coli nuclear acid, but also in shigella and salmonella genes. Shigella primer was from ipaH gene whose coded invasive plasmid relative antigen existed both in plasmid and in genome. The primers of salmonella were designed from the 16SrRNA sequence. The primer of E. coli O157:H7 was taken from eaeA gene. Five random primers were selected for RAPD. The detection system included common PCR, semi-nested PCR and RAPD.</p><p><b>RESULTS</b>This method was more sensitive, specific and efficient and its processing was rapid and simple. For example, the method could be used to specifically detect and identify salmonella, shigella, and E. coli O157:H7, and its sensitivity ranged from 3 to 50 CFU, and its detection time was 4 hours.</p><p><b>CONCLUSION</b>This PCR method, therefore, can serve as a routine and practical protocol for detecting and identifying pathogenic microorganisms from clinical samples.</p>

Study on Escherichia coli Growth Stimulated by Yeast Extract and Echlonia kurome Okum Water Extract / 微生物学通报

The means of OD value measurement and plate counting were used to choose the bacterial growth stimulants. Among the 120 substances sorted to 13 kinds (the trace elements, REE, carbohydrate, amino acids and amino acids derivatives, vitamins, nucleosides, plant hormones, animal hormones, plant and animal extracts, Echlonia Kurome Okum water extract and yeast extract), Echlonia Kurome Okum water extract and yeast extract were found to stimulate the growth of Escherichia coli significantly.