Modulation of proinflammatory mediators by viruses-bacteria synergism in human osteoblasts-an in vitro study.

BACKGROUND/PURPOSE: Viruses-bacteria synergistic interaction is associated with destructive periodontal diseases. However, the underlying mechanism for tissue destruction is not fully elucidated. In this study, lipopolysaccharide from Porphyromonas gingivalis (Pg-LPS) and polyinosinic-polycytidylic acid (poly I:C) were used to simulate bacteria and viruses, respectively. The possible combined effects of both molecular patterns on secretion of interleukin (IL)-6 and prostaglandin E2 (PGE2) from osteoblasts were determined. The effects of povidone-iodine (PVP-I) on the secretion of IL-6 and PGE2 were also examined. METHODS: Viability of treated osteoblastic cells (MG63) was examined by detection the mitochondrial dehydrogenase activity. Secretion of IL-6 and PGE2 was detected using the enzyme-linked immunosorbent assay (ELISA). Mitogen-activated protein kinases (MAPKs) and cyclooxygenase-2 (COX-2) were determined using the Western blotting analysis. RESULTS: Pg-LPS or poly I:C significantly enhanced the production of IL-6 and PGE2 in MG63 cells. The additive/synergistic effects of Pg-LPS/poly I:C on production of IL-6 and PGE2 were evident. The levels of phosphorylation of p38 MAPK and c-Jun N-terminal kinase (JNK) and expression of COX-2 protein were enhanced by Pg-LPS and/or poly I:C. On the other hand, the level of phosphorylation of extracellular signal-regulated kinase (ERK) was reduced by Pg-LPS and/or poly I:C. The stimulatory secretion of PGE2 by poly I:C was significantly reduced by PVP-I. CONCLUSION: Concomitant infection of viruses and bacteria may be potentially harmful to the tooth supporting tissues by production of proinflammatory mediators. The results suggest the potential anti-inflammatory effect of PVP-I on bacterial or viral infection.

E-Z Isomerization Mechanism of the Green Fluorescent Protein Chromophore: Remote Regulation by Proton Dissociation of the Phenol Group.

Dronpa, a GFP (green fluorescent protein)-like fluorescent protein, allows its fluorescent and nonfluorescent states to be switched to each other reversibly by light or heat through E-Z isomerization of the GFP chromophore. In this article, a GFP chromophore (p-HBDI) in water is used as a model to explore this E-Z isomerization mechanism. Based on the experimental solvent isotope effect (kH2O/kD2O = 2.30), the E-Z isomerization of p-HBDI in water is suggested to go through the remote-proton-dissociation-regulated direct mechanism with a proton transfer in the rate-determining step. The fractionation factor (ϕ) of the water-associated phenol proton of p-HBDI in the transition state is found to be 0.43, which is exactly in the range of 0.1-0.6 for the fractionation factor (ϕ) of the transferring proton in the transition state of R2O···H···O+H2 in water. This means that the phenol proton of E-p-HBDI in the transition state is on the way to the associated water oxygen during the E-Z isomerization. The proton dissociation from the phenol group of p-HBDI remotely regulates its E-Z isomerization. Less proton dissociation from the phenol group (pKa = 8.0) at pH = 1-4 results in a modest reduction in the E-Z isomerization rate of p-HBDI, while complete proton dissociation from the phenol group at pH = 11-12 also reduces its E-Z isomerization rate by one order of magnitude because of the larger charge separation in the transition state of the p-HBDI anion. All of these results are consistent with the remote-proton-dissociation-regulated direct mechanism but against the water-assisted addition/elimination mechanism.

Synthesis and properties of the para-trimethylammonium analogues of green fluorescence protein (GFP) chromophore: The mimic of protonated GFP chromophore.

At low pH, protons from the external, bulk solution can protonate the phenoxide group of the p-HBDI chromophore in wild-type green fluorescent protein (wtGFP) and its mutants, and likely continue to tentatively protonate the phenol hydroxyl group of the same chromophores. Because the protonated GFP chromophore is a transient, we prepare the stable p-trimethylammonium analogues (2a and 2b) of the GFP chromophore to mimic it and explore their properties. What we found is that the p-trimethylammonium analogues of the GFP chromophore have the highly electrophilic amidine carbon, blue-shifted electronic absorption, smaller molar absorptivity, smaller fluorescent quantum yield, and faster E-Z thermoisomerization rate. The amidine carbon of the p-trimethylammonium analogue (2b) of the GFP chromophore is the only site that is attacked by very weak nucleophile of water, resulting in ring-opening of the imidazolinone moiety. The half-life of its decay rate in D2O is around 33 days. Actually, acid-catalyzed hydrolysis of p-HBDI also results in ring-opening of the imidazolinone moiety. The ratio of the acid-catalyzed hydrolysis rate constants [kobs(p-HBDI)/kobs(1b)] between p-HBDI and 1b (p-dimethylammonium analogue of the GFP chromophore) is dramatically increased from 0.30 at pH = 2 to 0.63 at pH = 0. This is the evidence that more and more phenol hydroxyl groups of p-HBDI are tentatively protonated in a low-pH aqueous solution and that accelerates hydrolysis of p-HBDI in the way similar to the quaternary ammonium derivatives 2a and 2b in water. With this view point, 2a and 2b still can partially mimic the cationic p-HBDI with the protonated phenol hydroxyl group. Implication of the experiment is that the amidine carbon of the chromophore in wtGFP and its mutants at very low pH should be highly electrophilic. Whether ring-opening of the imidazolinone moiety of the GFP chromophore would occur or not depends on if water molecules can reach the amidine carbon of the chromophore inside wtGFP and its mutants.

Clinical effect of traditional Chinese spinal orthopedic manipulation in treatment of chondromalacia patellae.

OBJECTIVE: To evaluate the clinical effect of traditional Chinese spinal orthopedic manipulation (TCSOM) in treating chondromalacia patellae (CP). METHODS: Sixty cases of CP patients were randomly assigned to a TCSOM group and a Celecoxib group according to the random number table method. All patients in the TCSOM group were treated with a maximum of 10 spinal manipulations and rehabilitation training of quadriceps femoris. The symptoms before and after treatment were assessed with visual analog scale (VAS) and Kujala functional knee scoring system (KFKSS). A symptom improvement rate (SIR) was implemented in order to evaluate the effects of the treatments. RESULTS: The symptoms of 16 patients in the TCSOM group quickly resolved after the first spinal manipulation and 8 cases were significantly improved. The VAS scores in the TCSOM group after 4 weeks of treatment were significantly lower than those in the Celecoxib group. The KFKSS scores in the TCSOM group after 4 weeks of treatment were significantly higher than those in the Celecoxib group. Side effects of the treatment were not reported. Symptom improvement rate based on the VAS in the TCSOM group indicated more significant improvements than the Celecoxib group. CONCLUSION: TCSOM has greater efficacy than Celecoxib capsules for relief of the symptoms of CP.

Genomic analysis, trajectory tracking, and field surveys reveal sources and long-distance dispersal routes of wheat stripe rust pathogen in China.

Identifying sources of phytopathogen inoculum and determining their contributions to disease outbreaks are essential for predicting disease development and establishing control strategies. Puccinia striiformis f. sp. tritici (Pst), the causal agent of wheat stripe rust, is an airborne fungal pathogen with rapid virulence variation that threatens wheat production through its long-distance migration. Because of wide variation in geographic features, climatic conditions, and wheat production systems, Pst sources and related dispersal routes in China are largely unclear. In the present study, we performed genomic analyses of 154 Pst isolates from all major wheat-growing regions in China to determine Pst population structure and diversity. Through trajectory tracking, historical migration studies, genetic introgression analyses, and field surveys, we investigated Pst sources and their contributions to wheat stripe rust epidemics. We identified Longnan, the Himalayan region, and the Guizhou Plateau, which contain the highest population genetic diversities, as the Pst sources in China. Pst from Longnan disseminates mainly to eastern Liupan Mountain, the Sichuan Basin, and eastern Qinghai; that from the Himalayan region spreads mainly to the Sichuan Basin and eastern Qinghai; and that from the Guizhou Plateau migrates mainly to the Sichuan Basin and the Central Plain. These findings improve our current understanding of wheat stripe rust epidemics in China and emphasize the need for managing stripe rust on a national scale.

Potential Targets for CRISPR/Cas Knockdowns to Enhance Genetic Resistance Against Some Diseases in Wheat (Triticum aestivum L.).

Wheat is one of the most important food crops worldwide. Even though wheat yields have increased considerably in recent years, future wheat production is predicted to face enormous challenges due to global climate change and new versions of diseases. CRISPR/Cas technology is a clean gene technology and can be efficiently used to target genes prone to biotic stress in wheat genome. Herein, the published research papers reporting the genetic factors corresponding to stripe rust, leaf rust, stem rust, powdery mildew, fusarium head blight and some insect pests were critically reviewed to identify negative genetic factors (Susceptible, S genes) in bread wheat. Out of all reported genetic factors related to these disease, 33 genetic factors (S genes) were found as negative regulators implying that their down-regulation, deletion or silencing improved disease tolerance/resistance. The results of the published studies provided the concept of proof that these 33 genetic factors are potential targets for CRISPR/Cas knockdowns to improve genetic tolerance/resistance against these diseases in wheat. The sequences of the 33 genes were retrieved and re-mapped on the latest wheat reference genome IWGSC RefSeq v2.1. Phylogenetic analysis revealed that pathogens causing the same type of disease had some common conserved motifs and were closely related. Considering the significance of these disease on wheat yield, the S genes identified in this study are suggested to be disrupted using CRISPR/Cas system in wheat. The knockdown mutants of these S genes will add to genetic resources for improving biotic stress resistance in wheat crop.

Calcium alginate/curdlan/corn starch@calcium alginate macrocapsules for slowly digestible and resistant starch.

This study aimed to enhance the content of slowly digestible starch (SDS) and resistant starch (RS) by designing a type of novel shell-controllable calcium alginate/curdlan/corn starch@calcium alginate (CA/CL/CS@CA) core-shell structured macrocapsule. These macrocapsules were obtained by coating calcium alginate/curdlan/corn starch beads with different concentrations of sodium alginate. The results of the swelling power indicated that as the concentration of sodium alginate in the shell increased from 0% to 0.5%, the swelling degree of the macrocapsules at 95 °C decreased from 5.64 ± 0.09 g/g to 4.30 ± 0.12 g/g. The in vitro digestion experiments indicated that the content of SDS and RS of the macrocapsules at a sodium alginate concentration of 0.5% reached 42.21 ± 1.04% and 28.82 ± 1.05%, respectively. These results suggest that the CA/CL/CS@CA core-shell structured macrocapsules have the potential to be used as functional foods for cardiovascular disease, diabetes, and obesity patients.

Genetic Diversity of Wheat Stripe Rust Fungus Puccinia striiformis f. sp. tritici in Yunnan, China.

The stripe rust of wheat is one of the devastating diseases in China, which is caused by fungus Puccinia striiformis f. sp. tritici (Pst). The Yunnan Province of China is located in the south-western part, and holds distinctive geographical and climate features, while wheat growth and epidemics of stripe rust fungus are fully dissimilar to the major wheat-growing regions of China. It is important to discover its origin and migration to control the disease. In this study, 352 isolates were sampled from 11 spots of the Yunnan Province during the wheat growing season from 2004 to 2015 and analyzed with SNPs markers of housekeeping genes. Results revealed that 220 haplotypes were inferred from the concatenating sequences; among them, 5 haplotypes (viz., 'H86', 'H18', 'H8', 'H15' and 'H23') comprised over 24.5% of the population. The haplotype diversity, nucleotide diversity, mutation rate and recombination events were 0.992, 6.04 × 10-3, 4.46 × 10-3 and 18.0 respectively, which revealed the genetic diversity of Pst populations among all locations. Four grouping methods, such as UPGMA-tree, PCA, PLS-DA and STRUCTURE, were employed for the categorization of the Pst populations conferring to their races and topographical localities. All methods were found significant and mostly had co-linear relations with each other. The analysis of molecular variance (AMOVA) conferred total variation was 9.09%, and 86.20% of variation was within the populations. The current study also exposed a comparatively high genetic multiplicity within the population, while low genetic inconsistency among the populations. Furthermore, the molecular records on the gene pole (Nm = 18.45) established that the migration of the stripe rust pathogen occurred among all locations in Yunnan province. The ancestral haplotype was detected in Yuxi. Based on the trajectories of upper airflow and genetic diversity of Pst populations in different locations, it is suggested that the locations Dehong, Dali, Lincang and Baoshan are probably a major source of Pst in Yunnan.

Effects of Chinese Herbal Medicines on the Risk of Overall Mortality, Readmission, and Reoperation in Hip Fracture Patients.

Hip fracture is a major public health concern, with high incidence rates in the elderly worldwide. Hip fractures are associated with increased medical costs, patient dependency on families, and higher rates of morbidity and mortality. Chinese herbal medicine (CHM) is typically characterized as cost-effective and suitable for long-term use with few side effects. To better understand the effects of CHM on hip fracture patients, we utilized a population-based database to investigate the demographic characteristics, cumulative incidence of overall mortality, readmission, reoperation, and patterns of CHM prescription. We found that CHM usage was associated with a lower risk of overall mortality [P = 0.0009; adjusted hazard ratio (HR): 0.47, 95% confidence interval (CI): 0.30-0.73], readmission (P = 0.0345; adjusted HR: 0.67, 95% CI: 0.46-0.97), and reoperation (P = 0.0009; adjusted HR: 0.57, 95% CI: 0.40-0.79) after adjustment for age, type of hip fracture, surgical treatment type, and comorbidities. We also identified the herbal formulas, single herbs, and prescription patterns for the treatment of hip fracture by using association rule mining and network analysis. For hip fracture patients, the most common CHM coprescription pattern was Du-Zhong (DZ) → Xu-Duan (XD), followed by Du-Huo-Ji-Sheng-Tang (DHJST) → Shu-Jing-Huo-Xue-Tang (SJHXT), and Gu-Sui-Bu (GSB) → Xu-Duan (XD). Furthermore, XD was the core prescription, and DZ, GSB, SJHXT, and DHJST were important prescriptions located in cluster 1 of the prescription patterns. This study provides evidence for clinical CHM use as an adjunctive therapy that offers benefits to hip fracture patients.

A Clinical Observation of Functional Abdominal Pain Syndrome in Patients Treated by Traditional Chinese Spinal Orthopedic Manipulation.

OBJECTIVE: To evaluate the clinical effect of traditional Chinese spinal orthopedic manipulation (TCSOM) in treating patients with functional abdominal pain syndrome (FAPS) in comparison with Pinaverium Bromide (Dicetel, PBD), and to assess a possible cause for FAPS. METHODS: Eighty patients with FAPS were randomly and equally assigned to the TCSOM group and PBD group according to the random number table. All patients in the TCSOM group were treated with a maximum of 5 times of spinal manipulations. Patients in the PBD group were instructed to take 50 mg 3 times a day, consistently for 2 weeks. The symptoms of pre- and post-treatment were assessed on a visual analog scale (VAS) pain score. A symptom improvement rating (SIR) was implemented to evaluate the effects of the treatments. RESULTS: The symptoms of 27 cases of the TCSOM group were relieved soon after the first TCSOM treatment and 9 cases were significantly improved. The VAS pain scores in the TCSOM group were significantly lower than those in the PBD group after 2 weeks treatment. According to the SIR based on VAS, the TCSOM group included 30 cases with excellent results, 7 cases with good, and 3 cases with poor. Adverse events to the treatment were not reported. Based on VAS, the PBD group reported 8 cases with excellent results, 10 cases with good and 22 cases with poor. There was a significant difference between the two groups (P<0.01). CONCLUSIONS: The displacement of intervertebral discs and/or vertebra in the thoracic or lumbar region seems to be a contributing factor in the symptoms of FAPS. TCSOM is an effective treatment for FAPS.