Metabolic Composition and Quality Traits of Polygonatum cyrtonema Hua from Different Germplasms and Age Sections Based on Widely Targeted Metabolomics Analysis.

Polygonatum rhizomes are rich in various compounds with many biological activities and are widely used in functional foods and pharmaceutical products. In order to screen for superior Polygonatum cyrtonema Hua (P. cyrtonema) germplasm and also to elucidate the nutritional and medicinal values of rhizomes, the metabolic composition and quality traits of rhizomes from different germplasms and age sections of P. cyrtonema were analysed by widely targeted metabolomics, and the molecular mechanism of triacylglycerol synthesis was explored. The results showed that the different germplasms and age sections of P. cyrtonema were rich in different nutritional and medicinal components. Of these, the broad-leaved green stem (GK) germplasm is rich in polysaccharides, alkaloids, and lipids; the pointed-leaved green stem (JL) germplasm is rich in flavonoids, steroids, and amino acids, while the pointed-leaved purple stem (JZ) germplasm contains more phenolic acids. The one-year (AT) age section is rich in polysaccharides, steroids, organic acids, and lipids; the three years (CT) age section contains more flavonoids, alkaloids, and amino acid metabolites. Lipids were significantly enriched in the broad-leaved green stem germplasm and the one-year age section. Interestingly, the highest accumulation of triacylglycerols, an important component of lipids, was also found in the GK germplasm and the AT age section. Nineteen, 14, and 13 members of the glycerol-3-phosphate acyltransferase (GPAT), lysophosphatidic acid acyltransferase (LPAT), and diacylglycerol acyltransferase (DGAT) gene families, respectively, involved in triacylglycerol synthesis were also identified. The quantitative real-time PCR (qRT-PCR) results further suggested that the differentially expressed PcDGAT1, PcDGAT2.4, PcGPAT9.1, PcLPAT2.9, and PcLPAT4.3 genes may play important roles in triacylglycerol synthesis in P. cyrtonema. Therefore, this study provides a new theoretical reference for product development and the breeding of new varieties of Polygonatum species.

Reduced JKAP correlates with advanced disease features, inflammation, as well as increased exacerbation risk and severity in asthmatic children.

BACKGROUND: This study aimed to investigate the correlation of JNK pathway-associated phosphatase (JKAP) with clinical features, inflammation, exacerbation risk, and severity in asthmatic children. METHODS: Asthmatic exacerbation children (N = 90), asthmatic remission children (N = 90), and healthy controls (N = 90) were enrolled in this case-control study, whose venous blood samples were collected after enrollment for routine blood test, JKAP, and inflammatory cytokines detection by enzyme-linked immune sorbent assay. The clinical features included demographic data, family history of asthma, and pulmonary ventilation function. RESULTS: JKAP level was the lowest in asthmatic exacerbation children, followed by asthmatic remission children and healthy controls. ROC curve revealed good ability of JKAP in distinguishing three groups from each other, especially in telling asthmatic exacerbation children from healthy controls (AUC: 0.926; 95%CI: 0.887-0.965). In addition, JKAP was negatively correlated with eosinophil count, immunoglobulin E (IgE), tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1ß), interleukin-6 (IL-6), and interleukin-17 (IL-17), positively correlated with forced expiratory volume in 1 sec/forced vital capacity (FEV1/FVC) and FEV1 (%predicted) in asthmatic exacerbation children. Whereas in asthmatic remission children, JKAP was negatively correlated with eosinophil count, TNF-α, IL-1ß, IL-6, and IL-17 and positively correlated with FEV1 (%predicted), but not with IgE or FEV1/FVC. In healthy controls, the correlation of JKAP with clinical features and inflammatory cytokines was non-obvious. For exacerbation severity, JKAP was the highest in mild exacerbation children, followed by moderate exacerbation children, and severe exacerbation children. CONCLUSION: JKAP serves as a potential biomarker for asthmatic susceptibility, inflammation, exacerbation risk, and severity in children.

Quercetin improves endothelial insulin sensitivity in obese mice by inhibiting Drp1 phosphorylation at serine 616 and mitochondrial fragmentation.

Studies have shown that endothelial insulin resistance induced by oxidative stress contributes to vascular dysfunction in metabolic disorders. Quercetin, a natural antioxidant, has been recently shown to exert protective effects on endothelial function. However, the effects of quercetin on endothelial insulin resistance and its underlying mechanism are unclear. Here, we found that chronic oral treatment of obese mice with quercetin increased vascular endothelial insulin sensitivity, accompanied by alleviated mitochondrial fragmentation as revealed by confocal imaging. In addition, western blot analysis showed that quercetin treatment suppressed the levels of dynamin-related protein 1 (Drp1) and phosphorylation at serine 616 in endothelial cells of obese mice. Mechanistically, quercetin specifically suppressed Drp1 phosphorylation at serine 616, whereas it showed little effects on the Drp1 level and its phosphorylation at serine 637 in cultured endothelial cells under oxidative stress. Furthermore, our results also showed that quercetin suppressed Drp1 phosphorylation at serine 616 by inhibiting PKCδ as revealed by western blot analysis. Knockdown of PKCδ with siRNA alleviated the protective effects of quercetin on endothelial-mitochondrial dynamics and insulin sensitivity. These results suggest that chronic oral treatment with quercetin exerts endothelial protective effects through inhibition of PKCδ and the resultant mitochondrial fragmentation.

Treating ischaemic stroke with intravenous tPA beyond 4.5 hours under the guidance of a MRI DWI/T2WI mismatch was safe and effective.

Purpose: Clinical trials have provided evidence that treating patients with acute ischaemic stroke (AIS) beyond 4.5 hours was feasible. Among them using MRI diffusion-weighted imaging/fluid attenuation inversion response (DWI/FLAIR) mismatch to guide intravenous tissue plasminogen activator (tPA) was successful. Our study explored the outcome and safety of using DWI/T2-weighted imaging (T2WI) mismatch to guide intravenous tPA therapy for patients with AIS between 4.5 hours and 12 hours of onset. Method: This was a retrospective study. Records of 1462 AIS patients with the time of onset of <12 hours were reviewed. Those had MRI rapid sequence study and had hyperintense signal on DWI but normal T2WI and received intravenous tPA up to 12 hours of onset were included in the analysis. Their demographics, risk factors, post-tPA complications, National Institutes of Health Stroke Scale (NIHSS) scores and outcome were recorded and analyse. χ2 was used to compare the intergroup variables. SAS was used to perform statistical calculation. A p<0.05 was considered statistically significant. Results: Of 1462 identified, 601 (41%) patients were entered into the final analysis. Among them, 327 (54%) had intravenous tPA within 4.5 hours of onset and 274 (46%) were treated between 4.5-12 hours. After intravenous tPA, 426 cases (71%) had >4 pints of improvement on NIHSS score within 24 hours. Postintravenous tPA, 32 (5.32%) cases had haemorrhagic transformation. 26 (4.33%) were asymptomatic ICH and 4 (0.67%) died. At 90 days, 523 (87%) achieved a modified Rankin scale of 0-2. Conclusion: Using MRI DWI/T2WI mismatch to identify patients with AIS for intravenous tPA between 4.5 hours and 12 hours was safe and effective. The outcome was similar to those used DWI/PWI or DWI/FLAIR mismatch as the screening tool. However, obtaining DWI/T2WI was faster and avoided the need of contrast material.

Molybdenum sulfide Co-catalytic Fenton reaction for rapid and efficient inactivation of Escherichia coli.

As a typical advanced oxidation technology, the Fenton reaction has been employed for the disinfection, owing to the strong oxidizability of hydroxyl radicals (·OH). However, the conventional Fenton system always exhibits a low H2O2 decomposition efficiency, leading to a low production yield of ·OH, which makes the disinfection effect unsatisfactory. Herein, we develop a molybdenum sulfide (MoS2) co-catalytic Fenton reaction for rapid and highly efficient inactivation of Escherichia coli K-12 (E. coli) and Staphylococcus aureus (S. aureus). As a co-catalyst in the Fe(II)/H2O2 Fenton system, MoS2 can greatly facilitate the Fe(III)/Fe(II) cycle reaction by the exposed Mo4+ active sites, which significantly improves the H2O2 decomposition efficiency for the ·OH production. As a result, the MoS2 co-catalytic Fenton system can reach up to 83.37% of inactivation rate of E. coli just in 1 min and 100% of inactivation rate within 30 min, which increased by 2.5 times than that of the conventional Fenton reaction. Furthermore, the ·OH as the primary reactive oxygen species (ROS) in MoS2 co-catalytic Fenton reaction was measured and verified by electron paramagnetic resonance (EPR) and photoluminescence (PL). It is demonstrated an increased amount of ·OH generated from the decomposition of H2O2 in the presence of MoS2, which is responsible for the rapid and efficient inactivation of E. coli and S. aureus. This study provides a new perspective for rapid and highly efficient inactivation of bacteria in environmental remediation.

[Determination of ascitic bacterial 16S rRNA by quantitative PCR-microarray in the diagnosis of spontaneous bacterial peritonitis].

OBJECTIVE: To evaluate the significance of determining ascitic bacterial 16S rRNA by quantitative PCR combined with microarray (PCR-microarray) in the diagnosis of spontaneous bacterial peritonitis (SBP). METHODS: Ascitic bacterial 16SrRNA was determined by real time fluorescent quantitative PCR-microarray in 76 cases of suspected SBP and 6 cases of non-infectious ascites with chronic liver diseases. The results were compared with ascitic bacterial culture simultaneously. RESULTS: Of 76 ascitic samples, 17 were detected bacteria positive by PCR-microarray, including 8 Grams positive(G+) and 9 Grams negative(G-), which was higher than that by bacterial culture which had only 6 ascitic samples detected positive (all G-); the positive rates were 22.4% vs 7.9%, respectively (P < 0.01). The bacterial strains detected by both methods in 6 cases had a consistency with each other. No bacteria were detected in another 6 cases of non-infectious ascites with chronic liver diseases. CONCLUSIONS: Determination of ascitic bacteria 16S rRNA by PCR-microarray has a higher specificity and sensitivity in the diagnosis of SBP as compared with the bacteria culture. Application of this novel method can not only accelerate SBP diagnosis but also stratify the different pathogens.

[Determination of ascitic bacterial 16S rRNA gene in the rapid diagnosis of spontaneous bacterial peritonitis].

OBJECTIVE: To evaluate the value of ascitic bacterial 16S rRNA gene determination in the rapid diagnosis of spontaneous bacterial peritonitis (SBP). METHODS: 16S rRNA gene from bacterial DNA in ascites was determined by quantitative fluorescent polymerase chain reaction (PCR) in 76 patients with suspected SBP and 6 patients with non-infectious ascites. The results were compared with those obtained from bacterial culture. RESULTS: The positive rate of SBP was 22.4% among patients detected with ascitic bacterial 16S rRNA gene determination-based quantitative fluorescent PCR, which was significantly higher than that (7.9%) in patients only received bacterial culture (P<0.05). In addition,in 6 patients with non-infectious ascites,both the 16S rRNA gene determination-based quantitative fluorescent PCR and bacterial culture showed negative results. CONCLUSIONS: 16S rRNA gene determination-based quantitative fluorescent PCR can be an effective tool for the rapid diagnosis of SBP. It is more sensitive than the bacterial culture.

[The roles of saliva testing for preventing hepatitis B virus spreading].

OBJECTIVE: To discuss the significance of testing hepatitis B virus (HBV) from saliva in HBV patients. METHODS: HBV DNA content in serum and saliva of 200 HBV patients and 20 healthy subjects were detected by fluorescence quantitative polymerase chain reaction. According to the serum level of HBV content, four groups were divided: control group A, group B negative, low virus C (1 x 10(3) - 1 x 10(5) copies/ml) and high-group D ( > 1 x 10(5) copies/ml). The relationship of serum and virus content in saliva was analysed. RESULTS: Of 200 HBV cases, 180 were found HBV DNA in serum with positive rate of 90.0%; while 145 were found HBV DNA in saliva with positive rate of 72.5%, and there was no significant difference (chi2 = 1.35, P > 0.05). The significant difference was observed in testing serum and saliva in Group C (100.0% vs. 38.5%; Z = 14.11, P < 0.01). In group D, there was no significant difference found either (100.0% vs. 83.8%; chi2 = 1.05, P > 0.05). Group D virus serum had a high average level of (6.63 +/- 1.55) log copies/ml virus and in the saliva had an average level of (5.21 +/- 1.85) log copies/ml; saliva had serum viral load lower than an order of magnitude average. No HBV DNA was found in serum or saliva from 20 health subjects. CONCLUSION: When the serum contains a high content of HBV DNA virus, the content of saliva HBV DNA virus should be likely high, which might pose a threat of source of infection. A precise quantitative detection of HBV DNA in saliva might be used as evaluation of the level of virus in the body copy for judgment of infection.

[Clinical significance of intrahepatic hepatitis B core antigen (+) in patients with chronic hepatitis B].

OBJECTIVE: This study aimed to assess the clinical significance of intrahepatic hepatitis B core antigen (HBcAg) (+) in patients with chronic hepatitis B (CHB). METHODS: 200 CHB patients were prospectively studied using fluorescence quantitative PCR (FQ-PCR), combined PCR with fluorescence probe hybridization technique, to determine serum HBV DNA. Serum HBeAg was measured quantitatively. Liver biopsies were performed and immunohistochemistry stained liver slides were examined in all the cases. Correlation analyses were performed. RESULTS: Based on the HBV DNA levels, the patients were divided into 5 groups: group A (<3 log10 copies/ml) n=20, group B (>or=3 log10 copies/ml-<5 log10 copies/ml) n=13, group C (>or=5 log10 copies/ml-<6 log10 copies/ml) n=24, group D (>or=6 log10 copies/ml-<8 log10 copies/ml) n=116, and group E (>or=8 log10 copies/ml) n=27, and 87.5% of the CHB patients were intrahepatic HBcAg (+). The rate of HBcAg (+) was 55.0% (11/20) in group A, 53.8% (7/13) in group B, 75.0% (19/24) in group C, 96.6% (112/116) in group D, and 100% (27/27) in group E. A strong correlation was found between the rate of HBcAg (+) and the level of serum HBV DNA (r=0.80). This type of association also appeared between serum HBV DNA levels and HBeAg (+) (r=0.47). Of 20 CHB patients who were serum HBV DNA negative, 25% (5) were HBeAg (+), and 55% (11) were HBcAg (+), whereas 15 patients were both HBV DNA (-) and HBeAg (-), and 46.7% (7) were HBcAg (+). CONCLUSIONS: Intrahepatic HBcAg (+) in CHB patients might be more reliable in reflecting HBV replication. Determination of HBcAg (+) may have clinical significance for evaluating the efficacy of antiviral therapy and for predicting the therapeutic responses to different antiviral agents.