Genome-Wide and Expression Pattern Analysis of the HIT4 Gene Family Uncovers the Involvement of GHHIT4_4 in Response to Verticillium Wilt in Gossypium hirsutum.

Chromatin remodelers are essential for regulating plant growth, development, and responses to environmental stresses. HIT4 (HEAT-INTOLERANT 4) is a novel stress-induced chromatin remodeling factor that has been less studied in abiotic stress and stress resistance, particularly in cotton. In this study, we conducted a comprehensive analysis of the members of the HIT4 gene family in Gossypium hirsutum using bioinformatics methods, including phylogenetic relationships, gene organization, transcription profiles, phylogenetic connections, selection pressure, and stress response. A total of 18 HIT4 genes were identified in four cotton species, with six HIT4 gene members in upland cotton. Based on the evolutionary relationships shown in the phylogenetic tree, the 18 HIT4 protein sequences were classified into four distinct subgroups. Furthermore, we conducted chromosome mapping to determine the genomic locations of these genes and visually represented the structural characteristics of HIT4 in G. hirsutum. In addition, we predicted the regulatory elements in HIT4 in G. hirsutum and conducted an analysis of repetitive sequences and gene collinearity among HIT4 in four cotton species. Moreover, we calculated the Ka/Ks ratio for homologous genes to assess the selection pressure acting on HIT4. Using RNA-seq, we explored the expression patterns of HIT4 genes in G. hirsutum and Gossypium barbadense. Through weighted gene co-expression network analysis (WGCNA), we found that GHHIT4_4 belonged to the MEblue module, which was mainly enriched in pathways such as DNA replication, phagosome, pentose and glucuronate interconversions, steroid biosynthesis, and starch and sucrose metabolism. This module may regulate the mechanism of upland cotton resistance to Verticillium wilt through DNA replication, phagosome, and various metabolic pathways. In addition, we performed heterologous overexpression of GH_D11G0591 (GHHIT4_4) in tobacco, and the results showed a significant reduction in disease index compared to the wild type, with higher expression levels of disease resistance genes in the transgenic tobacco. After conducting a VIGS (virus-induced gene silencing) experiment in cotton, the results indicated that silencing GHHIT4_4 had a significant impact, the resistance to Verticillium wilt weakened, and the internode length of the plants significantly decreased by 30.7% while the number of true leaves increased by 41.5%. qRT-PCR analysis indicated that GHHIT4_4 mainly enhanced cotton resistance to Verticillium wilt by indirectly regulating the PAL, 4CL, and CHI genes. The subcellular localization results revealed that GHHIT4_4 was predominantly distributed in the mitochondria and nucleus. This study offers preliminary evidence for the involvement of the GHHIT4_4 in cotton resistance to Verticillium wilt and lays the foundation for further research on the disease resistance mechanism of this gene in cotton.

Metagenomic analysis of the soil microbial composition and salt tolerance mechanism in Yuncheng Salt Lake, Shanxi Province.

The soil in Yuncheng Salt Lake has serious salinization and the biogeographic environment affects the composition and distribution of special halophilic and salt-tolerant microbial communities in this area. Therefore, this study collected soils at distances of 15, 30, and 45 m from the Salt Lake and used non-saline soil (60 m) as a control to explore the microbial composition and salt tolerance mechanisms using metagenomics technology. The results showed that the dominant species and abundance of salt-tolerant microorganisms changed gradually with distance from Salt Lake. The salt-tolerant microorganisms can increase the expression of the Na+/H+ antiporter by upregulating the Na+/H+ antiporter subunit mnhA-G to respond to salt stress, simultaneously upregulating the genes in the betaine/proline transport system to promote the conversion of choline into betaine, while also upregulating the trehalose/maltose transport system encode genes to promote the synthesis of trehalose to resist a high salt environment.

Comparative transcriptome analysis of melon (Cucumis melo L.) reveals candidate genes and pathways involved in powdery mildew resistance.

Powdery mildew is a major disease in melon, primarily caused by Podosphaera xanthii (Px). Some melon varieties were resistant to powdery mildew, while others were susceptible. However, the candidate genes associated with resistance and the mechanism of resistance/susceptibility to powdery mildew in melon remain unclear. In this study, disease-resistant melon cultivar TG-1 and disease-susceptible melon cultivar TG-5 were selected for comparative transcriptome analysis. The results suggested that the numbers of differentially expressed genes (DEGs) in TG-5 was always more than that in TG-1 at each of the four time points after Px infection, indicating that their responses to Px infection may be different and that the active response of TG-5 to Px infection may be earlier than that of TG-1. Transcription factors (TFs) analysis among the DEGs revealed that the bHLH, ERF, and MYB families in TG-1 may play a vital role in the interaction between melon and powdery mildew pathogens. GO enrichment analysis of these DEGs in TG-5 showed that the SBP, HSF, and ERF gene families may play important roles in the early stage of melon development after Px infection. Finally, we speculated on the regulatory pathways of melon powdery mildew and found PTI and ABA signaling genes may be associated with the response to Px infection in melon.

Integrative transcriptomic and gene co-expression network analysis of host responses upon Verticillium dahliae infection in Gossypium hirsutum.

Worldwide, Verticillium wilt is among the major harmful diseases in cotton production, causing substantial reduction in yields. While this disease has been extensively researched at the molecular level of the pathogen, the molecular basis of V. dahliae host response association is yet to be thoroughly investigated. In this study, RNA-seq analysis was carried out on V. dahliae infected two Gossypium hirsutum L. cultivars, Xinluzao-36 (susceptible) and Zhongzhimian-2 (disease resistant) for 0 h, 24 h, 72 h and 120 h time intervals. Statistical analysis revealed that V. dahliae infection elicited differentially expressed gene responses in the two cotton varieties, but more intensely in the susceptible cultivar than in the resistant cultivars. Data analysis revealed 4241 differentially expressed genes (DEGs) in the LT variety across the three treatment timepoints whereas 7657 in differentially expressed genes (DEGs) in the Vd592 variety across the three treatment timepoints. Six genes were randomly selected for qPCR validation of the RNA-Seq data. Numerous genes encompassed in disease resistance and defense mechanisms were identified. Further, RNA-Seq dataset was utilized in construction of the weighted gene co-expression network and 11 hub genes were identified, that encode for different proteins associated with lignin and immune response, Auxin response factor, cell wall and vascular development, microtubule, Ascorbate transporter, Serine/threonine kinase and Immunity and drought were identified. This significant research will aid in advancing crucial knowledge on virus-host interactions and identify key genes intricate in G. hirsutum L. resistance to V. dahliae infection.

Synergistic effect of resveratrol and HSV-TK/GCV therapy on murine hepatoma cells.

Despite its low transfer efficiency, suicide gene therapy with HSV-TK is known for its bystander killing effect. The connexin-based gap junction is believed to mediate the bystander effect. Recently, we found that resveratrol, a polyphenol compound, increased the expression of Cx26 and Cx43, which are connexins and important constituents of gap junctions, in murine hepatoma cells. Hypothetically, the resveratrol-induced upregulation of gap junctions may improve the bystander effect that HSV-TK/GCV has on hepatoma cells. Our present investigation revealed that resveratrol could enhance intercellular communication at the gap junctions in CBRH7919 hepatoma cells and thereby enhance the bystander killing effect of GCV on CBRH7919TK cells. However, inhibition of gap junction using its long-term inhibitor alpha-glycyrrhetinic acid had a negative influence on the bystander effect of gene therapy with HSV-TK/GCV. In addition, combined resveratrol and GCV treatment in tumor-bearing mice with CBRH7919TK and CBRH7919WT cells at a ratio of 2:3 resulted in a significant decrease in the volume and weight of the tumor in comparison to GCV or only resveratrol. The present results demonstrate that resveratrol can enhance the bystander effect exerted by the HSV-TK/GCV system by enhancing connexin-mediated gap junctional communication.

[Impact of hepatitis B virus infection on liver function after hematopoietic stem cell transplantation].

To analyze the impact of hepatitis B virus (HBV) infection on liver function of patients after hematopoietic stem cell transplantation (HSCT), the transplantation outcome of 48 patients infected with HBV prior to transplantation among 185 patients received HSCT was investigated retrospectively. The results showed that during a follow-up for 6 months after HSCT, the alanine aminotransferase (ALT) peak average values of the patients with HBsAg(+), HBsAb(+) and control groups were (281.6 ± 414.6), (95.4 ± 79.9) and (65.1 ± 44.2) U/L, respectively. The incidences of abnormal liver function of the patients with HBsAg(+), HBsAb(+) and control groups were 61.54%, 40.00% and 30.23% respectively. There were no significant differences between any two groups (P > 0.05). The lethality of those patients at late period after transplantation was not related to HBV infection. The hepatocirrhosis and hepatocarcinoma caused by HBV infection have not become major problems in long-term survivors. It is concluded that in HBsAg(+) patients received HSCT, the damage of liver function is more severe than control group, possibly increasing the development of abnormal liver function. The measures against the liver function damage should be taken. The prophylactic administration of ganciclovir for virus may be effective to prevent the activation of HBV.

Oral epidemic diseases of exposure personnel in long-term low dose radiation / 中国医学科学杂志(英文版)

Oral epidemic diseases of exposure personnel in long-term low-dose radiation yet have rarely been studied. Referred to WHO oral health survey method and symptom grading standard, data of 341 exposure persons in long-term low-dose radiation including α particle, β particle, and γ rays, etc., were collected from one camp in China in 2011 with cluster sampling and analyzed? with Foxpro 6.0 and SPSS 16.0 software. The exposure persons worked in low-dose radiation for a long time aged between 23 and 56, whose average age were 27.1 years old.In addition, their lengths of service were from 2 to 34 years (average 7.9 years) and average exposure time was 8 hours a day each year for more than three months. Average annual radiation dose equivalent was from 1.8 to 16.5 mSv (average 7.3 mSv). Total radiation dose equivalent was from 3.8 to 425.0 mSv (average 97.3 mSv).

Dose-dependent expression changes of early response genes to ionizing radiation in human lymphoblastoid cells.

The sensitivity of cancer cells as well as normal cells in response to ionizing radiation (IR) is believed to be associated with the early inducible expression of specific genes. Using cDNA microarray technology, here we explored and compared the global transcriptional changes in human lymphoblastoid AHH-1 cells irradiated with 0.05-, 0.2-, 0.5-, 2.0- and 10-Gy doses of gamma-rays 4 h after exposure. A dose as low as 0.05 Gy was efficient in inducing a transcriptional response including the up-regulation of 25 genes, some of which are involved in signal transduction pathways, e.g. BMPR2, GPR124, MAPK8IP2 and AGGF1, and the down-regulation of 18 genes. Expression of some genes was altered only at a specific dose. Most importantly, we discovered a number of radiation-response genes, e.g. DNA repair gene XPC, tumor protein p53 inducible protein 3 gene (TP53I3), immediate early response 5 gene, whose transcriptional levels were increased or depressed by IR in a dose-dependent trend within the dose range 0.05-10 Gy. The dose-dependent induced expression of TP53I3 and XPC was confirmed by Northern blot analyses. Using quantitative real-time PCR, we further confirmed that XPC gene induction was dose dependent as well as time dependent, reaching a peak 4 h post-2 Gy and 10 h post-0.05 Gy. The maximum induced expression level of the XPC gene was higher after 2 Gy (3.2-fold) than 0.05 Gy (1.93-fold). The identification of these radiation-inducible genes, especially those exhibiting a dose-dependent response, not only expands our knowledge of the mechanisms underlying the diverse biological effects induced by IR, but provides candidates for developing novel biomarkers of radiation injury.

[Expressions of RhoC, CD44v6 and ICAM-1 in human gastric cancer and its clinicopathological significance].

OBJECTIVE: To investigate the expressions of RhoC, CD44v6 and ICAM-1 in gastric cancer and their correlations. METHODS: SABC immunohistochemistry was used to detect the expressions of RhoC, CD44v6 and ICAM-1 in the specimens from 40 cases with gastric cancer. Their correlations were reviewed by clinicopathological data. RESULTS: The expression of RhoC, CD44v6 and ICAM-1 were not correlated with tumor differentiation and invasion depth (P> 0.05), but significantly correlated with lymph metastasis and pTNM stage (P< 0.05). There was a significant correlation between the expression of RhoC and the expression of CD44v6 or ICAM-1 (r=0.355, P=0.006; r=0.354, P=0.003) respectively. If RhoC was positive-expressed, and either of CD44v6 or ICAM-1 was positive-expressed, the sensitivity and the specificity for predicting the lymphatic metastasis was 93.75%, 62.5% respectively. CONCLUSIONS: The positive expressions of RhoC, CD44v6 and ICAM-1 are useful biological markers for predicting the metastatic potential of gastric cancer. The combined detection of three markers is a useful method for predicting lymphatic metastasis of gastric cancer.