A bacterial effector manipulates plant metabolism, cell death, and immune responses via independent mechanisms.

Bacterial pathogens inject effector proteins inside plant cells to manipulate cellular functions and achieve a successful infection. The soil-borne pathogen Ralstonia solanacearum (Smith), the causal agent of bacterial wilt disease, secretes > 70 different effectors inside plant cells, although only a handful of them have been thoroughly characterized. One of these effectors, named RipI, is required for full R. solanacearum pathogenicity. RipI associates with plant glutamate decarboxylases (GADs) to promote the accumulation of gamma-aminobutyric acid (GABA), which serves as bacterial nutrient. In this work, we found that RipI can also suppress plant immune responses to bacterial elicitors, which seems to be unrelated to the ability of RipI to induce GABA accumulation and plant cell death. A detailed characterization of the RipI features that contribute to its virulence activities identified two residues at the C-terminal domain that mediate RipI interaction with plant GADs and the subsequent promotion of GABA accumulation. These residues are also required for the appropriate homeostasis of RipI in plant cells and the induction of cell death, although they are partially dispensable for the suppression of plant immune responses. Altogether, we decipher and uncouple the virulence activities of an important bacterial effector at the biochemical level.

A bacterial effector protein prevents MAPK-mediated phosphorylation of SGT1 to suppress plant immunity.

Nucleotide-binding domain and leucine-rich repeat-containing (NLR) proteins function as sensors that perceive pathogen molecules and activate immunity. In plants, the accumulation and activation of NLRs is regulated by SUPPRESSOR OF G2 ALLELE OF skp1 (SGT1). In this work, we found that an effector protein named RipAC, secreted by the plant pathogen Ralstonia solanacearum, associates with SGT1 to suppress NLR-mediated SGT1-dependent immune responses, including those triggered by another R. solanacearum effector, RipE1. RipAC does not affect the accumulation of SGT1 or NLRs, or their interaction. However, RipAC inhibits the interaction between SGT1 and MAP kinases, and the phosphorylation of a MAPK target motif in the C-terminal domain of SGT1. Such phosphorylation is enhanced upon activation of immune signaling and contributes to the activation of immune responses mediated by the NLR RPS2. Additionally, SGT1 phosphorylation contributes to resistance against R. solanacearum. Our results shed light onto the mechanism of activation of NLR-mediated immunity, and suggest a positive feedback loop between MAPK activation and SGT1-dependent NLR activation.

miR-144 negatively regulates the effect of CCNB1 protein on the biological behavior of hepatoma cells.

Liver cancer poses a great threat to the life safety of patients, which is a common malignant tumor worldwide. This study aims to explore the effect of miR-144 negatively regulating CCNB1 on the biological behavior of liver cancer cells, including the proliferation, apoptosis and migration of liver cancer cells, so as to provide a sufficient biological basis for the treatment of liver cancer. A 3 armour hospital at the records of 100 patients with liver cancer in 2015-2019 as the research object, and resection of the liver cancer cells and tissue adjacent to carcinoma as the research samples, using polymerase chain reaction (PCR) for the organization of miR-144 gene and detect CCNB1 protein expression level, and by using a technique called RNA interference to silence the CCNB1 gene, and try to transfer by transfection CCNB1 protein, thus all kinds of biological behaviour of hepatocellular carcinoma cells. The liver tissue of miR-144 is low, the level of gene expression CCNB1 protein expression level is higher, the expression level in liver cancer cells directly influences the curative effect of hepatocellular carcinoma patients, the miR-144 gene can negative regulation CCNB1 protein, through this kind of negative adjustment to the biological behavior of liver cancer cells have a profound impact.

Enhancing nitrate and phosphorus removal from stormwater in a fold-flow bioretention system with saturated zones.

The traditional bioretention systems possess a remarkably low nitrogen and phosphorus removal effect. The removal rate fluctuates greatly, and even appears as negative removal of nitrogen and phosphorus. Four simulated bioretention experimental columns with different bilayer media, packing composition and structure were constructed. Based on the traditional fillers, the modified composite fillers with hydroxy-aluminum and modified vermiculite sludge particle (HAVSP) were added. The traditional filler (C1) and the modified composite filler (C2) were added respectively, moreover saturated zones were set up to enhance the effect of nitrogen and phosphorus removal. Removal of nutrients from experimental columns by simulated runoff efficiency was evaluated and compared. In addition, the effect of media depth on phosphorus retention and denitrifying enzyme activity in bioretention columns was also evaluated. The experimental column #2 filled with C2 had the optimum removal effect on TP (93.70%), however, the removal effect of TP by filling C1 experimental columns was insufficient (57.36%). Designed to remove nitrate (NO3--N) and total nitrogen, the experimental column #4 showed the best performance (83.54% and 92.15%, respectively). In this study, we propose a fold-flow bioretention system by filling HAVSP in combination with saturated zones. The runoff water quality can be effectively improved, and a new bioretention cell configuration can be provided for efficient stormwater treatment.

Molecular cloning and functional analysis of lotus salt-induced NnDREB2C, NnPIP1-2 and NnPIP2-1 in Arabidopsis thaliana.

Dehydration-responsive element bindings transcription factor (DREBs) and plasma membrane intrinsic proteins (PIPs) have been characterized multi-functions in plant growth and metabolism, as well as in the adaptation to various stresses. In this study, we cloned the full-length cDNA of NnDREB2C from a salt-tolerated lotus species with RT-PCR methods. Analysis of qRT-PCR demonstrated that NnDREB2C mRNA in the leaf dramatically increased after the treatments of NaCl, abscisic acid, low temperature and mannitol. Next, NnDREB2C was cloned into constitutive expression vector pSN1301, which in turn transformed into Arabidopsis thaliana to investigate its function in plants. NnDREB2C overexpression significantly elevated Arabidopsis tolerance against salt and drought stresses, showing higher survival rates, lower conductivity and more chlorophyll content than those of wild-type plants. Moreover, higher germination rates were observed in the NnDREB2C overproducing plants when subjected into the stresses of NaCl and mannitol. Furthermore, we investigate the potential down-stream genes regulated by NnDREB2C and observed a significant increase in expressions of several genes belonging to PIPs family, including PIP1-1, PIP1-2, PIP1-3, PIP1-4 and PIP1-5. Consistently, overexpressed NnPIP1-2 and NnPIP2-1 conferred Arabidopsis the tolerance to stresses. Taken together, we concluded that overexpression of NnDREB2C enhanced the tolerance of salt and drought stresses in plants, which might probably be derived from the increased expression of the genes belonging to PIPs family.

Castleman Disease of the Parotid Gland: A Case Report.

Castleman disease is a rare lymphoproliferative disorder characterized by the formation of painless hyperplastic lymph nodes. It can affect any lymph nodes of the body but rarely affects the salivary gland. We describe a patient with unicentric Castleman disease presenting as a parotid tumor and discuss the diagnosis and treatment of Castleman disease. A 39-year-old man had a painless lump on the right side of his face for 1 year. A computed tomography scan showed a well-defined, homogeneous mass in the right parotid gland. The patient underwent a right-sided superficial parotidectomy with preservation of the facial nerve. Histopathologic analysis of the excisional biopsy specimen confirmed the diagnosis of Castleman disease. No further treatment was administered, and the patient recovered well; no recurrence was present at the 1-year follow-up. Castleman disease is a lymphoproliferative disease that rarely affects the parotid gland. Histopathologic examination is regarded as the gold standard for diagnosis of Castleman disease. The current treatment for patients with unicentric Castleman disease is surgical resection. For patients with multicentric Castleman disease, more complex therapies are required.

A bacterial type III effector targets plant vesicle-associated membrane proteins.

The soilborne bacterial pathogen Ralstonia solanacearum is one of the most destructive plant pathogens worldwide, and its infection process involves the manipulation of numerous plant cellular functions. In this work, we found that the R. solanacearum effector protein RipD partially suppressed different levels of plant immunity triggered by R. solanacearum elicitors, including specific responses triggered by pathogen-associated molecular patterns and secreted effectors. RipD localized in different subcellular compartments in plant cells, including vesicles, and its vesicular localization was enriched in cells undergoing R. solanacearum infection, suggesting that this specific localization may be particularly relevant during infection. Among RipD-interacting proteins, we identified plant vesicle-associated membrane proteins (VAMPs). We also found that overexpression of Arabidopsis thaliana VAMP721 and VAMP722 in Nicotiana benthamiana leaves promoted resistance to R. solanacearum, and this was abolished by the simultaneous expression of RipD, suggesting that RipD targets VAMPs to contribute to R. solanacearum virulence. Among proteins secreted in VAMP721/722-containing vesicles, CCOAOMT1 is an enzyme required for lignin biosynthesis, and mutation of CCOAOMT1 enhanced plant susceptibility to R. solanacearum. Altogether our results reveal the contribution of VAMPs to plant resistance against R. solanacearum and their targeting by a bacterial effector as a pathogen virulence strategy.

Screening for oesophageal cancer.

BACKGROUND: Oesophageal cancer is a global heath problem. The prognosis for advanced oesophageal cancer is generally unfavourable, but early-stage asymptomatic oesophageal cancer is basically curable and could achieve better survival rates. The two most commonly used tests are cytologic examination and endoscopy with mucosal iodine staining. The efficacy of the screening tests is controversial, and the true benefit and efficacy of screening remains uncertain because of the potential lead-time and length-time biases. This review was conducted to examine the evidence for the efficacy of screening for oesophageal cancer (squamous cell carcinoma and adenocarcinoma). OBJECTIVES: To determine the efficacy of early screening, using endoscopy with iodine staining or cytologic examination, in reducing mortality from oesophageal cancer in asymptomatic individuals from high-risk and general populations. SEARCH METHODS: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (2012, Issue 8), The Cochrane Library (2012, Issue 8), MEDLINE (1950 to August 2012), EMBASE (1980 to August 2012), Allied and Complementary Medicine (AMED) (1985 to August 2012), Chinese Biomedical Database (CBM) (January 1975 to August 2012), VIP Database (January 1989 to August 2012), China National Knowledge Infrastructure (CNKI) (January 1979 to August 2012), and the Internet. We also searched reference lists, conference proceedings, and databases of ongoing trials. There was no restriction on language or publication status in the search for trials. SELECTION CRITERIA: We included only randomised controlled trials (RCT) of screening versus no screening for oesophageal cancer. Randomisation of groups or clusters of individuals was acceptable. DATA COLLECTION AND ANALYSIS: Two review authors independently scanned the titles and abstracts from the initial search for potential trials for inclusion. We did not find any trials that met the inclusion criteria. MAIN RESULTS: The electronic search identified 3482 studies. Two authors independently reviewed the references. The reports of 18 studies were retrieved for further investigation. None met the eligibility criteria for a RCT investigation of the effects of screening versus no screening for oesophageal cancer. AUTHORS' CONCLUSIONS: There were no RCTs that determined the efficacy of screening for oesophageal cancer. Non-RCTs showed a high incidence and the reported better survival after screening could be caused by selection bias, lead-time and length-time biases. RCTs are needed to determine the efficacy of screening for oesophageal cancer.

The GABA transaminase GabT is required for full virulence of Ralstonia solanacearum in tomato.

The utilization of plant gamma-aminobutyric acid (GABA) is essential for the pathogenicity of the bacterial plant pathogen Ralstonia solanacearum. A knockout mutant in the GABA transaminase-encoding gene gabT is unable to utilize GABA as a nutrient and its ability to cause disease in plants is strongly compromised. However, the proximity of the gabD gene (encoding a succinate semialdehyde dehydrogenase) in the same operon raises the question of whether an impact on the gabD gene underlies or contributes to the virulence attenuation of the ΔgabT mutant. In this work, we use genetic complementation to show that the expression of the gabT gene is able to rescue the impaired virulence of the ΔgabT knockout mutant in tomato plants, confirming that the gabT-encoded GABA transaminase is indeed required for full virulence of R. solanacearum in a natural host plant.

Tomato Stem Injection for the Precise Assessment of Ralstonia solanacearum Fitness in Planta.

Ralstonia solanacearum is a soil-borne pathogen with worldwide distribution that causes bacterial wilt disease in more than 250 plant species. R. solanacearum invades plants through the roots, reaches the vascular system, and colonizes the whole plant by moving through the xylem, where it eventually replicates rapidly, causing plant death. Usual assays to measure the virulence of R. solanacearum under laboratory conditions rely on soil-drenching inoculation followed by observation and scoring of disease symptoms. Here, we describe a protocol to assess the replication of R. solanacearum following injection into tomato stems. This protocol includes four major steps: 1) growth of tomato plants; 2) R. solanacearum injection into tomato stems; 3) collection of tomato xylem samples and bacterial quantitation; and 4) data analysis and representation. This method bypasses the natural penetration process of the pathogen, thus minimizing variation associated with stochastic events during bacterial invasion, and provides a sensitive and accurate measurement of bacterial fitness inside xylem vessels.

SGT1 is not required for plant LRR-RLK-mediated immunity.

Plant immune signalling activated by the perception of pathogen-associated molecular patterns (PAMPs) or effector proteins is mediated by pattern-recognition receptors (PRRs) and nucleotide-binding and leucine-rich repeat domain-containing receptors (NLRs), which often share cellular components and downstream responses. Many PRRs are leucine-rich repeat receptor-like kinases (LRR-RLKs), which mostly perceive proteinaceous PAMPs. The suppressor of the G2 allele of skp1 (SGT1) is a core immune regulator required for the activation of NLR-mediated immunity. In this work, we examined the requirement of SGT1 for immune responses mediated by several LRR-RLKs in both Nicotiana benthamiana and Arabidopsis. Using complementary genetic approaches, we found that SGT1 is not limiting for early PRR-dependent responses or antibacterial immunity. We therefore conclude that SGT1 does not play a significant role in bacterial PAMP-triggered immunity.

Epidemiological pattern of maxillofacial fractures in northern China: A retrospective study of 829 cases.

This study was performed to examine the epidemiological features of maxillofacial fracture, including the incidence, causes, age and sex distribution, methods of treatment, and prognosis, in a local area.A retrospective study was performed to investigate the epidemiological characteristics of 829 patients with maxillofacial fractures treated in a hospital in northern China from August 2011 to July 2019. Sex, age, etiology, fracture site, and treatment method were obtained from the medical records.The average age of all 829 patients was 36.1 years, and most patients were in the 20- to 29-year age group. The male to female ratio was 3.04:1.00. Traffic accidents were the main cause of the maxillofacial fractures. The mandible was the most commonly fractured bone, and the parasymphysis was the most frequently affected site. Head injury was the most common associated injury. Open surgery with internal fixation was the first-choice treatment for most cases.Traffic accidents were the main cause of maxillofacial fractures, followed by falling. Open surgery with internal fixation was the leading treatment choice. Both functional and esthetic outcomes should be considered in the treatment of maxillofacial fractures.

A Bacterial Effector Protein Hijacks Plant Metabolism to Support Pathogen Nutrition.

Many bacterial plant pathogens employ a type III secretion system to inject effector proteins within plant cells to suppress plant immunity. Whether and how effector proteins also co-opt plant metabolism to support extensive bacterial replication remains an open question. Here, we show that Ralstonia solanacearum, the causal agent of bacterial wilt disease, secretes the effector protein RipI, which interacts with plant glutamate decarboxylases (GADs) to alter plant metabolism and support bacterial growth. GADs are activated by calmodulin and catalyze the biosynthesis of gamma-aminobutyric acid (GABA), an important signaling molecule in plants and animals. RipI promotes the interaction of GADs with calmodulin, enhancing the production of GABA. R. solanacearum is able to replicate efficiently using GABA as a nutrient, and both RipI and plant GABA contribute to a successful infection. This work reveals a pathogenic strategy to hijack plant metabolism for the biosynthesis of nutrients that support microbial growth during plant colonization.

Effect of citrus flavonoids and tocotrienols on serum cholesterol levels in hypercholesterolemic subjects.

CONTEXT: Preliminary studies have suggested that both citrus flavonoids and palm tocotrienols reduce cholesterol levels in laboratory animals. OBJECTIVE: To examine the effect of these nutrients in combination on blood levels of cholesterol and related cardiovascular disease risk factors. DESIGN: Two open-label studies and 1 double-blind study are reported. SETTING: Outpatient clinical research setting. PATIENTS: Three groups (n=10, n=10, n=120) of hypercholesterolemic men and women (cholesterol levels >230 mg/dL) between the ages of 19 and 65 years were recruited. INTERVENTION: Subjects were randomized to consume either 270 mg citrus flavonoids plus 30 mg tocotrienols (S) or placebo (P) daily for a period of 4 weeks (group 1 [G1] and group 2 [G2]) or 12 weeks (group 3 [G3]). MAIN OUTCOME MEASURES: Measurements of fasting levels of blood cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), and triglycerides were made at baseline and 4 weeks (all groups) and at 8 weeks and 12 weeks (G3). RESULTS: Daily treatment with S significantly improved cardiovascular parameters compared to P in all groups. Significant reductions were shown in total cholesterol (20%-30%), LDL (19%-27%), apolipoprotein B (21%), and triglycerides (24%-34%). HDL levels remained unchanged in G1 and G2 but increased 4% (nonsignificant) in G3 and was accompanied by a significant increase in apolipoprotein A1 (5%).

Recent Advances in Antischistosomal Drugs and Agents.

Schistosomiasis is a notable neglected tropical disease caused by trematodes that inflame mainly the intestines, bladder, and liver. Because of the unavailability of a schistosomiasis vaccine, control of the disease depends mainly on chemotherapy. Praziquantel (PZQ), which is active against all schistosome species and the recommended drug by the World Health Organization for schistosomiasis treatment at either the community or individual level, has become the exclusive drug because of its low cost and efficacy against the adult form of all schistosome species. In view of rapid re-infection following treatment and concern about the development of tolerance and/or resistance to praziquantel, there is an urgent need for research and development of novel drugs for the prevention and treatment of schistosomiasis. This comprehensive review shall attempt to briefly review the recent advances in the chemotherapy of schistosomiasis in the literature from 1990s to now, particularly focusing on the context of potential development of antischistosomal agents. Their antischistosomal properties, advantages, and disadvantages as well as structure-activity relationships are reviewed in a simple manner. It shall be of interest for both the synthetic chemist as well as the pharmacologist.