Development of A Nomogram for Progression-free Survival in Patients with Stage II/T3N0 Nasopharyngeal Carcinoma to Explore Different Treatment Modalities.

Purpose To explore the prognostic value of clinical and serological risk factors for progression-free survival (PFS) in stage II and T3N0 nasopharyngeal carcinoma (NPC) and construct a nomogram based on these factors. Additionally, to investigate the long-term survival and short-term toxic reactions of patients in different risk stratification under different treatment modalities. Methods The patients were randomly divided into training and validation cohorts in a 7:3 ratio. Independent prognostic factors were identified using Cox regression analysis, and a nomogram was constructed by combining these predictive factors with the TNM staging system. The nomogram was then validated in the validation cohort, and patients were classified into different risk groups based on the nomogram. The PFS, overall survival (OS), and acute toxicities were compared among different treatment modalities after balancing baseline characteristics. Results Multivariate Cox regression analysis indicated that pathological type, alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) were independent prognostic factors(p<0.05) in this study. The nomogram showed good prognostic accuracy in both the training and validation cohorts (C-index of 0.73 and 0.70, respectively). In the different risk subgroups, there were no statistically significant differences in PFS and OS between radiotherapy and chemoradiotherapy groups(p>0.05). The treatment modality of combined chemotherapy was associated with more acute toxic reactions. Conclusion We established and validated a nomogram for predicting PFS in patients with stage II/T3N0 NPC. Intensity-modulated radiation therapy (IMRT) combined with chemotherapy did not provide additional survival benefits for these patients and was associated with more chemotherapy-related side effects.

The Abnormal Proliferation of Hepatocytes is Associated with MC-LR and C-Terminal Truncated HBX Synergistic Disturbance of the Redox Balance.

Background: Microcystin-LR (MC-LR) and hepatitis B virus (HBV) are associated with hepatocellular carcinoma (HCC). However, the concentrations of MC-LR in drinking water and the synergistic effect of MC-LR and HBV on hepatocellular carcinogenesis through their disturbance of redox balance have not been fully elucidated. Methods: We measured the MC-LR concentrations in 168 drinking water samples of areas with a high incidence of HCC. The relationships between MC-LR and both redox status and liver diseases in 177 local residents were analyzed. The hepatoma cell line HepG2 transfected with C-terminal truncated hepatitis B virus X gene (Ct-HBX) were treated with MC-LR. Reactive oxygen species (ROS), superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) were measured. Cell proliferation, migration, invasion, and apoptosis were assessed with cell activity assays, scratch and transwell assays, and flow cytometry, respectively. The mRNA and protein expression-related redox status genes were analyzed with qPCR and Western blotting. Results: The average concentration of MC-LR in well water, river water and reservoir water were 57.55 ng/L, 76.74 ng/L and 132.86 ng/L respectively, and the differences were statistically significant (P < 0.05). The MC-LR levels in drinking water were correlated with liver health status, including hepatitis, clonorchiasis, glutamic pyruvic transaminase abnormalities and hepatitis B surface antigen carriage (all P values < 0.05). The serum MDA increased in subjects who drank reservoir water and were infected with HBV (P < 0.05). In the cell experiment, ROS increased when Ct-HBX-transfected HepG2 cells were treated with MC-LR, followed by a decrease in SOD and GSH and an increase in MDA. MC-LR combined with Ct-HBX promoted the proliferation, migration and invasion of HepG2 cells, upregulated the mRNA and protein expression of MAOA gene, and downregulated UCP2 and GPX1 genes. Conclusion: MC-LR and HBV may synergistically affect redox status and play an important role in hepatocarcinoma genesis.

Discovery and exploration of widespread infection of mycoviruses in Phomopsis vexans, the causal agent of phomopsis blight of eggplant in China.

Phomopsis vexans, which causes Phomopsis blight of eggplant, has been reported worldwide. To study the biocontrol of this disease, 162 leaf and fruit samples of eggplant Phomopsis blight were collected from Hunan, Hubei, Jiangxi, Sichuan, Zhejiang, Fujian, Guangdong and Anhui Provinces from 2017 to 2019. Eighty-seven pathogenic fungus isolates were identified as P. vexans. The following studies were conducted: screening of sporulation medium, spore morphology analysis, mycovirus detection and identification of novel mycoviruses in these isolates. The results showed that eggplant tissue medium was the most suitable medium for rapid sporulation, and all isolates had mycoviruses consisting of mainly mixed infections. The genome of these mycoviruses varied from 1-15 kb. Five novel mycoviruses infecting P. vexans were obtained, including "Phomopsis vexans fusarivirus 1" (PvFV1), "Phomopsis vexans ourmia-like virus 1" (PvOLV1), "Phomopsis vexans endornavirus 2" (PvEV2), "Phomopsis vexans partitivirus 1" (PvPV1) and "Phomopsis vexans victorivirus L1" (PvVVL1). Thus, PvVVL1 displays a unique genome structure, and this is the first report of a victorivirus consisting of two segments and of a deltapartitivirus infecting the fungus host.

BRCC36 promotes intestinal mucosal barrier injury caused by BMP2 after ischemia reperfusion via inhibiting PPARγ signaling.

As one of the most common pathological changes in trauma and surgery practice, intestinal ischemia-reperfusion (I/R) injury is regarded as a major precipitating factor in the occurrence and development of fatal diseases. BRCA1-BRCA2-containing complex subunit 36 (BRCC36), a deubiquitinase, has been proved important in a variety of pathophysiological processes such as DNA repair, cell cycle regulation, tumorigenesis, and inflammatory response. However, the effect of BRCC36 on intestinal mucosal barrier injury after I/R has not been fully elucidated. Our research found that BRCC36 aggravated intestinal mucosal barrier injury caused by bone morphogenetic protein 2 after I/R by downregulating peroxisome proliferator-activated receptor-γ (PPARγ) signaling. These results suggested that BRCC36/PPARγ axis might serve as a potential therapeutic target for preventing intestinal mucosal barrier injury after I/R.

Complete nucleotide sequence of a novel partitivirus infecting the plant-pathogenic fungus Phomopsis vexans.

Here, we report the molecular characterization of a novel partitivirus from Phomopsis vexans strain PvHZ002, a plant-pathogenic fungus infecting eggplant. The virus was designated "Phomopsis vexans partitivirus 1" (PvPV1). PvPV1 contains two dsRNA segments, dsRNA1 and dsRNA2, which are 1,662 bp and 1,628 bp long, respectively. Each segment contains a single open reading frame, putatively encoding RNA-dependent RNA polymerase (dsRNA 1) and capsid protein (dsRNA 2). A homology search and phylogenetic analysis showed that PvPV1 clustered with viruses of the genus Deltapartitivirus of the family Partitiviridae.

Molecular characterization of a putative gammapartitivirus in the phytopathogenic fungus Nigrospora oryzae.

We report the molecular attributes of a novel bisegmented double-stranded RNA (dsRNA) virus, designated "Nigrospora oryzae partitivirus 1" (NoPV1), from a phytopathogenic fungus Nigrospora oryzae. The genome of NoPV1 contains two dsRNA segments (dsRNA1 and 2), 1875 bp and 1601 bp in length, respectively. dsRNA1 and -2 both have a single open reading frame encoding the RNA-dependent RNA polymerase and coat protein, respectively. NoPV1 has a high degree of sequence identity to members of genus Gammapartitivirus in the family Partitiviridae. This is the first report of a mycovirus in the family Partitiviridae that infects N. oryzae.

The Gastric Microbiome Is Perturbed in Advanced Gastric Adenocarcinoma Identified Through Shotgun Metagenomics.

Objective: Dysbiosis of gastric microbiota such as Helicobacter pylori plays a significant role in pathogenesis and progression of gastric cancer. Our aim was to evaluate the composition and functional effects of gastric microbiota in superficial gastritis (SG) and advanced gastric adenocarcinoma (GC). Methods: We carried out shotgun metagenomic sequencing on gastric wash samples from 6 patients with GC and 5 patients with SG. The taxonomic composition was profiled using MetaPhlAn2 and functional gene pathway was profiled using HUMAnN2. Differences in microbial composition and pathways between the two patient groups were assessed via LEfSe. Results: The gastric microbiota in GC patients was characterized by reduced species richness, enrichment of 13 bacterial taxa and depletion of 31 taxa (q < 0.05). The most representative taxa which were abundant in GC corresponded to the commensals or opportunistic pathogens that usually colonize the oral cavity, including genera Neisseria, Alloprevotella, and Aggregatibacter, species Streptococcus_mitis_oralis_pneumoniae and strain Porphyromonas_endodontalis.t_GCF_000174815. Each of the three GC-associated genera could separate GC from SG completely. In particular, Sphingobium yanoikuyae, a bacterium capable of degrading carcinogenic compounds, was depleted in GC. Functionally, pathways associated with the biosynthesis of lipopolysaccharide (LPS) and L-arginine were enriched in GC, whereas pathways involved in the fermentation of short chain fatty acids (SCFAs) and branched amino acid metabolism were more abundant in SG. Conclusions: Our results present new alterations in the gastric microbiome in patients with GC from a whole-genome perspective, suggesting that microbiome composition and function can be used for prognosis and diagnosis of GC.

[Effects of Neurotrophin Receptor-interacting MAGE Homolog on Apoptosis of Intestinal Epithelial Cells].

OBJECTIVE: To explore whether neurotrophin receptor-interacting MAGE homolog (NRAGE) is involved in the intestinal ischemia-reperfusion (I/R) and its effect on the apoptosis of intestinal epithelial cells and the expression of occludin protein. METHODS: The level of NRAGE protein after the rat small intestine I/R was detected by immunohistochemical (IHC) In vivo. The level of NRAGE protein and mRNA in IEC-6 cells after hypoxia and reoxygenation were tested by Western blot and RT-PCR respectively in vitro. The IEC-6 cells were divided into four groups, including NRAGE overexpression by lentivirus infection (Lv-NRAGE group), interference (sh-NRAGE group), lentivirus control (Lv-control group), and normal control group without lentivirus infection (NC group). The apoptosis of IEC-6 cells after infection was analyzed by flow cytometry. The level of the tight junction protein occludin was detected by Western blot. RESULTS: The expression of NRAGE were highly increased in intestinal mucosa epithelial cells after I/R (P<0.01). The proteins and mRNA levels of NRAGE were increased after 6 h of hypoxia in IEC-6 cellsin vitro. Compared with the Lv-control group, the early apoptosis rate was raised (P<0.01) and the level of occludin was reduced (P<0.01) in Lv-NRAGE group; while the early apoptosis rate was reduced (P<0.01) and the level of occludin was raised in sh-NRAGE group(P<0.001). CONCLUSION: NRAGE may be involved in intestinal I/R and promote the apoptosis and decrease occludin expression of intestinal epithelial cells.

Prognostic value of PPM1D in 800 gastric cancer patients.

Protein phosphatase magnesium­dependent 1 delta (PPM1D) has recently been associated with tumor biology. However, the expression pattern and clinical significance of PPM1D in gastric cancer (GC) have yet to be elucidated. The present study aimed to investigate the clinical and prognostic significance of PPM1D in GC. PPM1D expression was assessed in 800 patients with GC using immunohistochemistry and tissue samples were divided into a PPM1D­positive and ­negative group. The correlation between PPM1D expression and clinicopathological parameters or prognosis was investigated. PPM1D expression was significantly higher in GC tissue than in adjacent normal tissue (48 versus 9.5%; P<0.001). PPM1D positivity was significantly correlated with nodal status, distant metastasis and vascular invasion. Survival analysis indicated that the five­year survival rate in the PPM1D­positive group was significantly lower than that in the PPM1D­negative group (41 versus 72%; p=0.0012). Furthermore, the association between PPM1D positivity and survival rate was still significant following regulation of other prognostic markers in a multivariate analysis [hazard ratio (HR), 6.572; 95% confidence interval (CI), 3.108­13.471; P=0.0018]. In conclusion, the present study suggested that PPM1D positivity is associated with GC invasion and metastasis, and proposed PPM1D positivity as an indicator of unfavorable prognosis in patients with GC.

Etanercept in the treatment of intestinal Behcet's disease.

Behcet's disease (BD) accompanied by intestinal involvement is called intestinal BD. Although recent studies have attained positive feedback with the administration of anti-TNF-α agents in patients with BD, only a few reports on the study of etanercept in intestinal BD have been found. In this study, 35 cases of intestinal BD were treated with conventional therapy (prednisone or methotrexate) for a minimum period of 3 months (group 1). Another 19 patients who failed to respond to conventional therapy were then treated with etanercept (25 mg twice a week for 3 months). During each subsequent relapse, the patients were given the same treatment. The main outcome measures were the four criteria for diagnosis of BD (buccal ulcers, genital ulcers, ocular lesions, and skin lesions), the manifestation of intestinal involvement (abdominal symptoms, double-balloon enteroscopy), laboratory examinations of the acute phase reactants (erythrocyte sedimentation rate) and C-reactive protein, and relapses. As a result of the administered therapy, the healing rate of buccal and genital ulcers, the remission rate of ocular lesions, skin lesions, and abdominal symptoms, the healing rate of intestinal ulcers, and the recovery rate of ESR and CRP were significantly higher in group 2 than those of group 1. The relapse rate in the etanercept therapy was reduced significantly when compared with conventional therapy group. In conclusion, etanercept treatment, in contrast to the conventional therapy, can result in better curative effect and less adverse reactions in intestinal BD.

A fasciclin-like arabinogalactan protein, GhFLA1, is involved in fiber initiation and elongation of cotton.

Arabinogalactan proteins (AGPs) are involved in many aspects of plant development. In this study, biochemical and genetic approaches demonstrated that AGPs are abundant in developing fibers and may be involved in fiber initiation and elongation. To further investigate the role of AGPs during fiber development, a fasciclin-like arabinogalactan protein gene (GhFLA1) was identified in cotton (Gossypium hirsutum). Overexpression of GhFLA1 in cotton promoted fiber elongation, leading to an increase in fiber length. In contrast, suppression of GhFLA1 expression in cotton slowed down fiber initiation and elongation. As a result, the mature fibers of the transgenic plants were significantly shorter than those of the wild type. In addition, expression levels of GhFLAs and the genes related to primary cell wall biosynthesis were remarkably enhanced in the GhFLA1 overexpression transgenic fibers, whereas the transcripts of these genes were dramatically reduced in the fibers of GhFLA1 RNA interference plants. An immunostaining assay indicated that both AGP composition and primary cell wall composition were changed in the transgenic fibers. The levels of glucose, arabinose, and galactose were also altered in the primary cell wall of the transgenic fibers compared with those of the wild type. Together, our results suggested that GhFLA1 may function in fiber initiation and elongation by affecting AGP composition and the integrity of the primary cell wall matrix.

[Value of the laparoscopy combined with double-balloon enteroscopy in diagnosis and treatment of intestinal diseases].

OBJECTIVE: To evaluate the value of laparoscopy combined with double-balloon enteroscopy (DBE) for the diagnosis and treatment of intestinal diseases. METHODS: Clinical data of 69 cases with suspected small bowel diseases undergoing laparoscopic and DBE for the diagnosis and treatment were retrospectively analyzed. RESULTS: The lesions were found in 48 cases by laparoscopy. DBE was required in the remaining 21 patients to identify the underlying condition. All the operations were successfully completed using the laparoscopic approach, including totally laparoscopic bowel resection (n=11), and laparoscopic-assisted bowel resection (n=58). There were no anastomotic leakage, postoperative bleeding, intestinal obstruction, or wound infection. All the patients were discharged within 7 to 9 days after surgery. Postoperative pathological examination showed vascular abnormally (n=10), gastrointestinal stromal tumor (n=20), intestinal adenocarcinoma (n=5), intestinal neurofibroma (n=2), diverticulum (n=5), intestinal mucosal ulceration (n=8), intestinal tuberculosis (n=3), postoperative pouch bleeding (n=1), intestinal polyp (n=6), Crohn's disease (n=5), Meckel diverticulum (n=2), metastatic kidney cancer (n=1), and metastatic lung cancer (n=1). Length of follow up ranged from 3 months to 4 years, during which no re-bleeding occurred, 2 patients with gastrointestinal stromal tumor died of local recurrence and liver metastasis, 1 patient with adenocarcinoma died of local recurrence involving pancreatic head, duodenum, and mesenteric vessels, 2 patients with metastatic disease died of peritoneal recurrence and liver metastasis. CONCLUSION: Laparoscopic combined with DBE has a high detection rate for small intestinal disease with accurate localization, less trauma, and quicker recovery.

Construction and analysis of cotton (Gossypium arboreum L.) drought-related cDNA library.

BACKGROUND: Drought is one of the most important environmental factors causing water stress for cotton, and it greatly limits cotton growth and crop productivity. So far only a few drought-tolerance genes have been functionally characterized in details, and most efforts on this topic have been made in model organisms. Therefore, to identify more drought-related genes in cotton plays a crucial role in elucidating the underlying mechanisms of drought tolerance as well as utilizing bioengineering techniques to improve the tolerance in this organism. FINDINGS: Here we constructed a subtractive drought-tolerance cDNA library using suppressive subtractive hybridization (SSH). Through differential screening and bioinformatics analysis, we identified 392 positive clones with differential expression, corresponding 265 unique genes. By BLAST search against Genbank, we found that more than half of these EST sequences were homologous to those previously known drought-related genes and that there were 57 sequences with unknown functions, suggesting that many more genes are involved in this complex trait. Moreover, using RT-PCR, we examined the expression of nine representative candidate genes and confirmed that their expression levels were increased at different levels under drought stress. CONCLUSION: Our results show that drought tolerance is a complex trait in cotton, which involves the coordination of many genes and multiple metabolism pathways. The candidate EST sequences we identified here would facilitate further functional studies of drought-related genes and provide important insights into the molecular mechanisms of drought-stress tolerance and genetic breeding in cotton.

Histological and ultrastructural observation reveals significant cellular differences between Agrobacterium transformed embryogenic and non-embryogenic calli of cotton.

Over the past few decades genetic engineering has been applied to improve cotton breeding. Agrobacterium medicated transformation is nowadays widely used as an efficient approach to introduce exogenous genes into cotton for genetically modified organisms. However, it still needs to be improved for better transformation efficiency and higher embryogenic callus induction ratios. To research further the difference of mechanisms for morphogenesis between embryogenic callus and non-embryogenic callus, we carried out a systematical study on the histological and cellular ultrastructure of Agrobacterium transformed calli. Results showed that the embryogenic callus developed nodule-like structures, which were formed by small, tightly packed, hemispherical cells. The surface of some embryogenic callus was covered with a fibrilar-like structure named extracellular matrix. The cells of embryogenic calli had similar morphological characteristics. Organelles of embryogenic callus cells were located near the nucleus, and chloroplasts degraded to proplastid-like structures with some starch grains. In contrast, the non-embryogenic calli were covered by oval or sphere cells or small clusters of cells. It was observed that cells had vacuolation of cytoplasm and plastids with a well organized endomembrane system. This study aims to understand the mechanisms of embryogenic callus morphogenesis and to improve the efficiency of cotton transformation in future.