An improved quantitative real-time polymerase chain reaction technology for Helicobacter pylori detection in stomach tissue and its application value in clinical precision testing.

BACKGROUND: Helicobacter pylori (H. pylori) infection is a serious human health threat. The empiric H. pylori treatment paradigm guided by traditional testing technologies has led to antibiotic resistance. Here, we improved the qPCR method to provide technical support for precision H. pylori diagnosis and treatment. METHODS: Two pairs of primers and probes targeting the glmM gene were designed to detect H. pylori, and a multiplex qPCR method was established for virulence factor detection. Then, a rapid urease test (RUT), culturing and qPCR were performed on 141 specimens collected from Xinqiao Hospital of China in 2017 to evaluate the qPCR detection capability. Finally, the H. pylori infectious amount and virulence genes were detected by qPCR. RESULTS: 1. The improved qPCR method which used two pairs of primers had a higher detection rate (100%) and better accuracy (p = 0.000), compared with the qPCR using a pair of primers. It also had better consistency with the bacterial culture than with RUT (Kappa =0.440, p < 0.001). 2. The H. pylori infectious amount was significantly positively associated with gastritis in corpus (p = 0.003) and gastric erosion (p = 0.043). The H. pylori infectious amount in gastric precancerous patients was significantly lower than that in H. pylori-positive patients (p < 0.05), and the infectious H. pylori-vacA s1+ amount was significantly greater than that of H. pylori-vacA s1- (p < 0.05). 3. The vacA s1 frequency was significantly higher than that of vacA m1/cagA+/babA2+ in chronic superficial gastritis (p = 0.000), peptic ulcer (p = 0.037) and gastric erosion (p = 0.009). The H. pylori-vacA+/cagA+/babA2+ frequency showed a significant positive correlation (p < 0.05). CONCLUSIONS: The H. pylori infectious amount and presence of H. pylori virulence factors showed complex correlations with gastric disease occurrence and development. The improved qPCR with good detection performance can be used for quantitative H. pylori detection and testing for the virulence genes vacA s1, vacA m1, cagA and babA2 simultaneously. These findings will provide valuable information for disease diagnosis and treatment.

[Promoting tanshinone synthesis of Salvia miltiorrhiza root by a seed endophytic fungus, Phoma herbarum D603].

The interaction of endophytes and host plant is an effective mean to regulate the growth and secondary metabolism of medicinal plants. Here we want to elucidate the effects and mechanism of Phoma herbarum D603 on the root development and tanshinone synthesis in root of Salvia miltiorrhiza by endophyte-plant coculture system. The mycelium of P. herbarum D603 was colonized in the root tissue space, and formed a stable symbiotic relationship with host plant. The in vitro activities analysis showed that the concentration of IAA produced by D603 can reach(6.45±0.23) µg·mL~(-1), and this strain had some abilities of phosphorus solubilization and siderophore production activities. The coculture experiment showed that strain D603 can significantly promote the synthesis and accumulation of tanshinones in the root of S. miltiorrhiza, in which after 8 weeks of treatment with D603, the content of tanshinone Ⅱ_A in the roots reached up to(1.42±0.59) mg·g~(-1). By the qRT-PCR analysis results, we found that D603 could improve the expression levels of some key genes(DXR, DXS, GGPP, HMGR, CPS) of tanshinone biosynthesis pathway in host plant S. miltiorrhiza, but the promoting effect mainly occurred in the early stage of the interaction, and the enzyme activity level decreased in varying degrees of the later stage. In summary, seed-associated endophyte P. herbarum D603 can promote the growth and root development of S. miltiorrhiza by producing hormones, promoting nutrient absorption and siderophore production, and promote the synthesis and accumulation of tanshinones by regulating the expression level of key genes in the synthetic pathway in S. miltiorrhiza.

Peri-operative application of intra-aortic balloon pumping reduced in-hospital mortality of patients with coronary artery disease and left ventricular dysfunction.

BACKGROUND: There are few reports of peri-operative application of intra-aortic balloon pumping (IABP) in patients with coronary artery disease (CAD) and different grades of left ventricular dysfunction. This study aimed to analyze the early outcomes of peri-operative application of IABP in coronary artery bypass grafting (CABG) among patients with CAD and left ventricular dysfunction, and to provide a clinical basis for the peri-operative use of IABP. METHODS: A retrospective analysis of 612 patients who received CABG in the General Hospital of People's Liberation Army between May 1995 and June 2014. Patients were assigned to an IABP or non-IABP group according to their treatments. Logistic regression analysis was performed to investigate the influence of peri-operative IABP implantation on in-hospital mortality. Further subgroup analysis was performed on patients with severe (ejection fraction [EF] ≤ 35%) and mild (EF = 36%-50%) left ventricular dysfunction. RESULTS: Out of 612 included subjects, 78 belonged to the IABP group (12.7%) and 534 to the non-IABP group. Pre-operative left ventricular EF (LVEF) and EuroSCOREII predicted mortality was higher in the IABP group compared with the non-IABP group (P < 0.001 in both cases), yet the two did not differ significantly in terms of post-operative in-hospital mortality (P = 0.833). Regression analysis showed that IABP implantation, recent myocardial infarction, critical status, non-elective operation, and post-operative ventricular fibrillation were risk factors affecting in-hospital mortality (P < 0.01 in all cases). Peri-operative IABP implantation was a protective factor against in-hospital mortality (P = 0.0010). In both the severe and mild left ventricular dysfunction subgroups, peri-operative IABP implantation also exerted a protective role against mortality (P = 0.0303 and P = 0.0101, respectively). CONCLUSIONS: Peri-operative IABP implantation could reduce the in-hospital mortality and improve the surgical outcomes of patients with CAD with both severe and mild left ventricular dysfunction.

Effects of Resveratrol on the Mechanisms of Antioxidants and Estrogen in Alzheimer's Disease.

OBJECTIVE: To observe the effects of resveratrol (Res) on the antioxidative function and estrogen level in an Alzheimer's disease (AD) mouse model. METHODS: First, we examined the effects of Res on an AD mice model. SAMP8 mice were selected as the model, and normal-aging SAMR1 mice were used as the control group. The model mice were randomly divided into three groups: a model group, high-dose Res group (40mg/kg, intraperitoneal (ip)), and low-dose Res group (20mg/kg, ip). After receiving medication for 15 days, the mice were subjected to the water maze test to assess their spatial discrimination. The spectrophotometric method was used to detect the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) as well as the malondialdehyde (MDA) content. Quantitative PCR (q-PCR) was used to detect SOD, GSH-Px, CAT, and heme oxygenase-1 (HO-1) mRNA level changes. Western blot analysis detected HO-1 and Nrf2 protein expression. Second, we researched the effect of Res on the estrogen level in the SAMP8 model mice. The model mice were randomly divided into four groups: a model group, estrogen replacement group (0.28 mg/kg, intramuscular (im), estradiol benzoate), high-dose Res group (5 mg/kg, im), and low-dose Res group (2.5 mg/kg, im). The mice were injected, once every three days, for 5 weeks. Q-PCR was used to detect brain tissue mRNA expression changes. Western blot analysis detected ERα, ERß, and ChAT protein expression. An enzyme-linked immunosorbent assay (ELISA) kit was used to detect the expression of E2 and amyloid ß protein (Aß) in brain tissue. RESULTS: Compared with the control treatment, Res could improve the spatial abilities of the mice to a certain extent and also increase the expression of SOD, GSH-Px, CAT, and HO-1 at the mRNA level (P<0.05). In addition, enhanced SOD, GSH-Px, and CAT activities and HO-1 protein levels and decreased MDA content (P<0.05) were detected in the brain tissue of the Res-treated mice. The cytoplasmic Nrf2 content in the Res-treated mice was also decreased while the nuclear Nrf2 content and the nuclear translation rate of Nrf2 were increased (P<0.05). Res could decrease the expression of ERß in the brain tissue at the mRNA and protein levels and the expression of Aß in the brain tissue at the protein level. Res could also increase the mRNA and protein expression of ERα and ChAT and the protein expression of estradiol in the brain tissue. CONCLUSION: Res can increase the antioxidant capacity of AD models through the Nrf2/HO-1 signaling pathway. In addition, Res can enhance estrogen levels in an AD model. These findings provide a new idea for the treatment of AD.

[Well-to-Wheels Fossil Energy Consumption and CO2 Emissions of Hydrogen Fuel Cell Vehicles in China].

Hydrogen fuel cell vehicles (FCVs) have the advantage of high energy efficiency and zero tailpipe emissions. They have been progressively commercialized in recent years. Hydrogen production has diversified technological pathways, which vary greatly in terms of energy and environmental impacts. In this study, the life cycle assessment (LCA) method was applied to evaluate well-to-wheels (WTW) fossil energy consumption and carbon dioxide (CO2) emissions of FCVs using various hydrogen production pathways. The greenhouse gases, regulated emissions, and energy use in transportation (GREET) model, developed by the Argonne National Laboratory, was applied as the assessment tool, and a China-specific database was investigated and developed to evaluate typical hydrogen production pathways. Then, we compared the WTW fossil energy consumption and CO2 emissions of FCVs with those of gasoline vehicles (GVs), hybrid electric vehicles (HEVs), and battery electric vehicles (BEVs). The results indicated that renewable-energy-based electrolysis of water and biomass gasification are two prospective hydrogen production pathways with significant WTW energy and climate benefits which can help FCVs reduce fossil energy consumption and CO2 emissions by approximately 90% more than GVs. Among the current pathways with mass adoption, hydrogen production from coke oven gas (COG) has substantial energy and CO2 mitigation benefits, which enables FCVs to achieve a lower WTW fossil energy consumption than HEVs and lower WTW CO2 emissions than HEVs and BEVs. Considering the resource reserves and technological maturity in China, hydrogen production from COG and other industrial by-products is recommended for hydrogen energy and FCV development in the short term. In the medium and long terms, utilization of renewable energy to produce hydrogen should be promoted.

Interleukin-33 Predicts Poor Prognosis and Promotes Renal Cell Carcinoma Cell Growth Through its Receptor ST2 and the JNK Signaling Pathway.

BACKGROUND/AIMS: Renal cell carcinoma (RCC) is currently the ninth most common cancer in men. Interleukin (IL)-33 expression has previously been associated with a number of cancers; however, its biological role in RCC is poorly understood. In this study, we sought to elucidate the role of IL-33 in RCC. METHODS: Serum IL-33 levels were measured by ELISA. IL-33 expression in clinical RCC samples was examined by immunocytochemistry. The proliferation and apoptosis rate of RCC were determined by CCK8 and flow cytometry. Mcl1 and Bcl-2 expression were measured by quantitative real-time PCR and western blotting. JNK expression were measured by western blotting and flow cytometry. The in vivo role of IL-33 in RCC tumorigenesis was examined by animal models. RESULTS: We found that increased expression of IL-33 in RCC was associated with tumor-lymph node-metastasis (TNM) stage and inversely correlated with prognosis. IL-33 enhances RCC cell growth in vivo and stimulates RCC cell proliferation and prevents chemotherapy-induced tumor apoptosis in vitro. Furthermore, we demonstrated that IL-33 promotes RCC cell proliferation and chemotherapy resistance via its receptor ST2 and the JNK signaling activation in tumor cells. CONCLUSION: Our findings suggest that targeting IL-33/ST2 and JNK signaling may have potential value in the treatment of RCC.