Mechanism of Gentisic Acid on Rheumatoid Arthritis Based on miR-19b-3p/RAF1 Axis.

OBJECTIVE: To investigate the therapeutic effect of gentisic acid (GA) on rheumatoid arthritis (RA) based on the miR-19b-3p/RAF1 axis. METHODS: The cell counting kit-8 method was used to detect the growth inhibitory effect of different concentrations of GA on MH7A cells, and the drug concentration of GA was determined in the experiment. The quantificational real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-19b-3p and RAF1. RAF1, extracellular regulated protein kinases1/2 (ERK1/2) and phospho-ERK1/2 (p-ERK1/2) were examined by Western blotting. Three methods (dual-luciferase assay, qRT-PCR and Western blot analysis) were used to verify miR-19b-3p targeting RAF1. Flow cytometry was performed to detect MH7A cell apoptosis. Transwell and wound healing assays were used to determine the invasion and migration capacities of MH7A cells. RESULTS: The growth of MH7A cells was gradually inhibited with increasing GA concentration. When the GA concentration exceeded 80 mmol/L, GA was significantly cytotoxic to MH7A cells, so the half maximal inhibitory concentration of GA for MH7A cells was calculated as 67.019 mmol/L. GA upregulated miR-19b-3p expression, downregulated RAF1 expression, inhibited ERK1/2 phosphorylation, induced MH7A cell apoptosis and suppressed MH7A cell invasion and migration (P<0.05 or P<0.01). RAF1 was identified as the target of miR-19b-3p and reversed inhibitory effects on miR-19b-3p expression (P<0.05 or P<0.01). The miR-19b-3p inhibitor upregulated RAF1 expression and ERK1/2 phosphorylation, suppressed MH7A cell apoptosis and induced MH7A cell invasion and migration (P<0.01). CONCLUSION: GA regulated miR-19b-3p/RAF1 axis to mediate ERK pathway and inhibit the development of RA.

The protective effect of gentisic acid on rheumatoid arthritis via the RAF/ERK signaling pathway.

BACKGROUND: RAF and ERK pathways are known to be activated in human rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS), which play an important role in the pathogenesis and destruction of RA. Gentisic acid (GA) was a natural product derived from plants, which has been reported can attenuate pressure overload-induced cardiac hypertrophy and fibrosis in mice through inhibition of the ERK1/2 pathway. Whether GA can inhibit the occurrence and development of RA through RAF/ERK signaling pathway has not been reported. The purpose of this study is to determine whether GA may have a certain therapeutic effect on RA-FLS. METHOD: Bovine type II collagen was used to establish a rat model of rheumatism. Enzyme-linked immunosorbent assay was used to detect inflammatory factors, anti-inflammatory mediators, and rheumatoid factor. Hematoxylin and eosin and TUNEL staining were used to detect the effect of GA on histochemical with rheumatoid arthritis. RAF, ERK, and p-ERK expressions in synovial tissue were measured by western blot and immunohistochemical. Besides, human rheumatoid arthritis fibroblast-like synoviocytes cell line MH7A was used to investigate the biological behavior influenced by GA. Apoptosis assay was performed to detect apoptosis of GA on MH7A cells. Transwell invasion assay was performed to detect the ability of cell migration. RESULT: The result showed that GA could reduce joint swelling and inflammation. At the same time, it can also promote the apoptosis of synovial cells and down-regulate the RAF/ERK pathway. CONCLUSION: GA may ameliorate inflammatory factors' abnormality, synovial hyperplasia, and apoptosis of synovium via inhibiting the RAF/ERK signaling pathway.

Physicochemical and molecular transformation of novel functional peptides from Baijiu.

A novel strategy for screening and identifying peptides present in Baijiu was developed based on magnetic solid-phase extraction with magnetic S-doped graphene (M-G-S) as adsorbent combined with ultrahigh-performance liquid chromatography with high resolution tandem mass spectrometry. In total, 28 peptides consisting of amino acids from 3 to 9 were preliminarily identified, and significantly higher in the number than that of direct concentration and SPE with C18 as the adsorbent. Six peptides were confirmed with their corresponding synthetic reference standards by comparing their retention time, high resolution MS/MS spectra, and NMR spectroscopic studies. Parallel reaction monitoring integrated with the internal standard method was utilized to quantify identified peptides with concentrations ranging from 1.14 to 10.25 ng mL-1, and prediction results of bioactivity comprising antioxidation or ACE inhibitors were obtained. These discoveries were conducive to understanding the versatility benefit of Baijiu and paved the way to study the interaction between peptides and volatile substances.

General recommendation for assessment and management on the risk of glucocorticoid-induced osteonecrosis in patients with COVID-19.

BACKGROUND/OBJECTIVE: Coronavirus disease 2019 (COVID-19) is a disaster in human medical history and glucocorticoids remain the most promising therapy. Osteonecrosis is a disease caused by reduced intraosseous blood flow to bones in the joints, which will rapidly induce joint destruction. Approximately one-third patients with severe acute respiratory syndrome (SARS) who received high cumulative doses and long treatment durations of glucocorticoids occurred osteonecrosis. Considering the similarity of SARS and COVID-19 on their pathogen, clinical characteristics, and therapeutic strategies, it is particularly desirable to investigate whether osteonecrosis will become a common sequela among convalescent COVID-19 patients. METHODS: This multi-strategy study was designed by integrating different research methods, such as meta-analysis, systematic review, and cross-sectional investigations to address above study objectives. At first, two meta-analyses were performed on the osteonecrosis incidence among SARS patients and the clinical data of glucocorticoid exposure among COVID-19 patients. Then, a systematic review of low-dosage glucocorticoid associated osteonecrosis and a cross-sectional investigation of glucocorticoid exposure of COVID-19 patients in Wuhan city of China were also conducted. Moreover, the pathogenesis, diagnosis, prevention, and treatment options for osteonecrosis patients with COVID-19 infection were further presented and discussed. RESULTS: Our meta-analysis showed that 32% of SARS patients had developed osteonecrosis after receiving glucocorticoid treatment with high dose, and our system review supported that low level glucocorticoid exposure might also lead to the occurrence of osteonecrosis. Similarly, 40% of COVID-19 patients had undergone glucocorticoid treatment according to our meta-analysis. The cross-sectional investigation in Wuhan city of China found that the average of cumulative glucocorticoid exposure level was 504 â€‹mg calculated by the dosage of methylprednisolone. Notably, a confirmed osteonecrosis case was identified from 1406 patients with COVID-19 during our cross-sectional investigation, implying that preventive management of osteonecrosis should be better started with regular clinical follow-up observation. CONCLUSION: Growing evidence of the glucocorticoid therapy for COVID-19 patients prompts us to establish risk-classification-based early screening and to introduce early prevention protocol of its associated osteonecrosis that will be of clinical significance in favor of improved prognosis of this disease. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: To establish risk-classification-based early screening and to introduce early prevention protocol of glucocorticoid-induced osteonecrosis will be of clinical significance in favor of improved prognosis of COVID-19.

Administration of mircoRNA-135b-reinforced exosomes derived from MSCs ameliorates glucocorticoid-induced osteonecrosis of femoral head (ONFH) in rats.

Exosomes were found to exert a therapeutic effect in the treatment of osteonecrosis of the femoral head (ONFH), while miR-135b was shown to play an important role in the development of ONFH. In this study, we investigated the effects of concomitant administration of exosomes and miR-135b on the treatment of ONFH. A rat mode of ONFH was established. TEM, Western blotting and nanoparticle analysis were used to characterize the exosomes collected from human-induced pluripotent stem cell-derived mesenchymal stem cells (hiPS-MSC-Exos). Micro-CT was used to observe the trabecular bone structure of the femoral head. Real-time PCR, Western blot analysis, IHC assay, TUNEL assay, MTT assay and flow cytometry were performed to detect the effect of hiPS-MSC-Exos and miR-135b on cell apoptosis and the expression of PDCD4/caspase-3/OCN. Moreover, computational analysis and luciferase assay were conducted to identify the regulatory relationship between PDCD4 mRNA and miR-135b. The hiPS-MSC-Exos collected in this study displayed a spheroidal morphology with sizes ranging from 20 to 100 nm and a mean concentration of 1 × 1012 particles/mL. During the treatment of ONFH, the administration of hiPS-MSC-Exos and miR-135b alleviated the magnitude of bone loss. Furthermore, the treatment of MG-63 and U-2 cells with hiPS-MSC-Exos and miR-135b could promote cell proliferation and inhibit cell apoptosis. Moreover, PDCD4 mRNA was identified as a virtual target gene of miR-135b. HiPS-MSC-Exos exerted positive effects during the treatment of ONFH, and the administration of miR-135b could reinforce the effect of hiPS-MSC-Exos by inhibiting the expression of PDCD4.

Multiscale Time-Sharing Elastography Algorithms and Transfer Learning of Clinicopathological Features of Uterine Cervical Cancer for Medical Intelligent Computing System.

Intelligent medical diagnosis and computing system faces many challenges in complex object recognition, large-scale data imaging and real-time diagnosis, such as poor real-time computing, low efficiency of data storage and low recognition rate of lesions. In order to solve the above problems, this paper proposes a medical intelligent computing system and a series of algorithms for the clinical pathology of cervical cancer based on the multi-scale imaging and transfer learning framework. Firstly, based on data dimensions, imaging errors and other factors, this paper designs a multi-scale time-sharing elastic imaging algorithm based on image reconstruction time and data sample characteristics. Then, taking the burst imaging cohort and the calculation data set of new cervical cancer cases as the objects, based on the difficulties of cervical cancer feature modeling, this paper proposes the transfer learning algorithm of clinical and pathological features of cervical cancer. Finally, a medical intelligent computing system for cervical cancer pathology analysis and calculation with high efficiency and reliability is established. A series of proposed algorithms are compared with single-scale Retinex (SSR), which is based on single-scale Retinex migration learning (SSR-TL). The experimental results show that the proposed algorithm in cervical cancer pathological imaging and scoring, as well as the feature extraction and recognition of lesions, especially the efficiency of system execution, is obviously due to the comparison algorithm.

The implicit beliefs and implicit behavioral tendencies towards smoking-related cues among Chinese male smokers and non-smokers.

BACKGROUND: The dual-process theory is central to several models of addiction, implying the importance of automatic processes in the maintenance and development of addiction. Implicit beliefs are traces of previous experience which relate to the representation in cognition. Implicit behavioral tendencies are traces of previous experience which relate to the representation in behavioral tendencies. In this study, we aim to provide behavioral evidence for implicit beliefs and implicit behavioral tendencies towards smoking-related cues among Chinese male smokers and non-smokers. We also examine the relationships among implicit beliefs, implicit behavioral tendencies and smoking behaviors of smokers. METHODS: In order to achieve these goals, we used an implicit association test (IAT) to measure implicit beliefs and implicit behavioral tendencies simultaneously. Thirty-nine smokers and twenty-five non-smokers were tested, using smoking-related words and images, as well as neutral words and images as stimuli. RESULTS: Our analysis shows significant differences in smokers' and non-smokers' implicit beliefs and behavioral tendencies (t62 = 3.494, p < 0.001; t62 = 5.034, p < 0.001). In the group of smokers, implicit beliefs and implicit behavioral tendencies were positively correlated with each other (r = 0.460, p < 0.01). In addition, smokers' scores for implicit behavioral tendencies are negatively correlated with the number of cigarettes smoked per day (r = - 0.51, p < 0.001). CONCLUSIONS: This study suggests that implicit beliefs and behavioral tendencies toward smoking-related cues vary significantly between Chinese male smokers and non-smokers. In addition, there is a positive correlation between implicit beliefs and behavioral tendencies within smokers. It also shows for the first time that the implicit behavioral tendencies are related to smoking behaviors. Our results may be considered as references for smoking cessation interventions focused on changes at the implicit level, and they provide a new perspective for measuring different dimensions of implicit attitudes by an IAT. This finding might promote the development of the network theory of implicit attitudes.

Effect of Jiawei Fengshining on Synovial Cell Apoptosis and TGF-ß1/Smad Signaling Pathway in Rats with Rheumatoid Arthritis.

BACKGROUND/AIMS: Jiawei Fengshining (JWFSN) is a new formula originated from Fengshining, a classic formula for the treatment of rheumatoid arthritis (RA). The mechanism of JWFSN in the treatment of RA is still unclear. The aim of this study was to evaluate the effect of JWFSN formula on the inflammatory mediator levels in the serum and the TGF-ß1/Smad pathway in the synovium and to explore the underlying mechanisms of JWFSN formula to ameliorate synovial hyperplasia and apoptosis inhibition of synovium in rats with RA. METHOD: SPF female Wistar rats were randomly divided into 6 groups: the blank control group, the model control group, the positive drug group, and the low-, medium-, and high- dose JWFSN groups, with 8 rats in each group. Enzyme-linked immunosorbent assay (ELISA) was used to detect inflammatory mediators, anti-inflammatory mediators, and rheumatoid factor (RF). The pathological condition and apoptosis of the synovial tissue were detected by hematoxylin and eosin (HE) and TUNEL staining, respectively. TGF-ß1, p-Smad2, p-Smad3, and Smad7 protein expressions in synovial tissue were measured by western blot assay. In addition, human rheumatoid arthritis fibroblast-like synoviocytes cell line MH7A was treated with 20% JWFSN-containing serum to obtain in vitro data. RESULT: The administration of JWFSN was found to ameliorate synovial hyperplasia and promote apoptosis; increase the serum contents of anti-inflammatory mediators; reduce inflammatory mediators and RF contents; and inhibit the TGF-ß1/Smad signaling pathway in CIA rats. In vitro JWFSN treatment increased the apoptosis of MH7A cells and decreased cell viability. Additionally, JWFSN treatment inhibited the TGF-ß1/Smad signaling pathway in MH7A cells. Interestingly, kartogenin (TGF-ß1/Smad pathway activator) treament reversed the effects of JWFSN treatment. CONCLUSION: JWFSN may ameliorate inflammatory factors' abnormality, synovial hyperplasia, and apoptosis inhibition of synovium via the TGF-ß1/Smad signaling pathway.

Corrigendum: Expanded CURB-65: a new score system predicts severity of community-acquired pneumonia with superior efficiency.

This corrects the article DOI: 10.1038/srep22911.

Immature monocytes contribute to cardiopulmonary bypass-induced acute lung injury by generating inflammatory descendants.

BACKGROUND: As immune regulatory and effector cells, monocytes play an important role in the blood-extracorporeal circuit contact-related acute lung injury in patients undergoing cardiopulmonary bypass (CPB). However, circulating monocytes are phenotypically and functionally heterogeneous, so we characterised how immature monocytes affect acute lung injury induced by CPB. METHODS: The identification and dynamic changes in monocyte subsets were monitored by flow cytometry in patients undergoing CPB and in a rat model of CPB. The differentiation and migration of monocyte subsets were explored by in vitro cultures and adoptive transfer in the CPB rat model. RESULTS: We observed a dramatic increase of two monocyte subsets in the peripheral blood of patients undergoing CPB, involving tumour necrosis factor (TNF)-α-producing, mature intermediate CD14highCD16+ monocytes and a novel immature CD14lowCD16- subset. The immature CD14lowCD16- monocytes possessed limited ability for TNF-α production, and failed to suppress T-cell proliferation mediated by T-cell receptor signalling. However, these immature cells were highly proliferative and could differentiate into TNF-α producing, mature CD14highCD16+ monocytes. In the rat model of CPB, we further demonstrated that CPB induced migration of immature monocytes into the lungs, either from the bone marrow or from the spleen. Moreover, we confirmed the hypothesis that immature subsets could contribute to CPB-induced acute lung injury by giving rise to TNF-α producing descendants. CONCLUSIONS: The immature CD14lowCD16- monocytes might contribute to blood-circuit contact-induced acute lung injury by generating TNF-α-producing, mature monocytes. New strategies based on monocyte manipulation could be a promising therapeutic approach for minimising CPB-related lung injury.

Expanded CURB-65: a new score system predicts severity of community-acquired pneumonia with superior efficiency.

Aim of this study was to develop a new simpler and more effective severity score for community-acquired pneumonia (CAP) patients. A total of 1640 consecutive hospitalized CAP patients in Second Affiliated Hospital of Zhejiang University were included. The effectiveness of different pneumonia severity scores to predict mortality was compared, and the performance of the new score was validated on an external cohort of 1164 patients with pneumonia admitted to a teaching hospital in Italy. Using age ≥ 65 years, LDH > 230 u/L, albumin < 3.5 g/dL, platelet count < 100 × 10(9)/L, confusion, urea > 7 mmol/L, respiratory rate ≥ 30/min, low blood pressure, we assembled a new severity score named as expanded-CURB-65. The 30-day mortality and length of stay were increased along with increased risk score. The AUCs in the prediction of 30-day mortality in the main cohort were 0.826 (95% CI, 0.807-0.844), 0.801 (95% CI, 0.781-0.820), 0.756 (95% CI, 0.735-0.777), 0.793 (95% CI, 0.773-0.813) and 0.759 (95% CI, 0.737-0.779) for the expanded-CURB-65, PSI, CURB-65, SMART-COP and A-DROP, respectively. The performance of this bedside score was confirmed in CAP patients of the validation cohort although calibration was not successful in patients with health care-associated pneumonia (HCAP). The expanded CURB-65 is objective, simpler and more accurate scoring system for evaluation of CAP severity, and the predictive efficiency was better than other score systems.

[Roof folding and rotary pushing for the treatment of back to back fractures of distal radius and ulna in children].

OBJECTIVE: To evaluate the technique and the clinical effect of folding roof and rotary pushing in treatment of children with distal radius and ulna fracture of "back to back". METHODS: From January 2012 to February 2014,38 children with distal radius and ulna fracture of "back to back" were treated by using the technique of folding roof and rotary pushing to reset and splint fixation including 23 males and 15 females with an average age of 9.5 years old ranging from 6 to 14 years old. Injury time was from 45 min to 3 days (averaged 1.3 days). All cases was unilateral closed fracture without symptoms of nerve injury occurred. The wrist joint anteroposterior and lateral radiographs showed double fracture of radius and ulna, and the broken end of radius was typical "back to back" displacement. The quality of reduction was assessed according to Dienst recommendation on the combination of Aro measurement, and the therapeutic effect was evaluated using standard of Anderson function. RESULTS: All patients were followed up from 3 to 13 months with an average of 6 months. There were no iatrogenic nerve injury. Thirty cases were treated successfully for the first time, 8 cases were again reset successfully; 28 cases were anatomical reduction, 7 cases were near anatomic reduction, 3 cases were functional reduction. At the second day 7 cases with hand and finger swelling appeared in multiple reset patients. Quality results of reduction were excellent in 33 cases, good in 5 cases. According to the standard of Anderson function evaluation, 35 cases were excellent, 3 cases were good. All fractures were healed with of deformity of wrist. CONCLUSION: Using the technique of folding roof and rotary pushing in treatment of children with distal radius and ulna fracture of "back to back" is very successful, the patient's limb function recovered well, the whole operation process is simple.

Submassive hepatic necrosis distinguishes HBV-associated acute on chronic liver failure from cirrhotic patients with acute decompensation.

BACKGROUND & AIMS: Distinguishing between acute on chronic liver failure (ACLF) and decompensated liver cirrhosis is difficult due to a lack of pathological evidence. METHODS: A prospective single-center study investigated 174 patients undergoing liver transplantation due to acute decompensation of hepatitis B virus (HBV)-associated liver cirrhosis. Two groups were distinguished by the presence or absence of submassive hepatic necrosis (SMHN, defined as necrosis of 15-90% of the entire liver on explant). Core clinical features of ACLF were compared between these groups. Disease severity scoring systems were applied to describe liver function and organ failure. Serum cytokine profile assays, gene expression microarrays and immunohistochemical analyzes were used to study systemic and local inflammatory responses. RESULTS: SMHN was identified in 69 of 174 patients proven to have cirrhosis by histological means. Characteristic features of SMHN were extensive necrosis along terminal hepatic veins and spanning multiple adjacent cirrhotic nodules accompanied by various degrees of liver progenitor cell-derived regeneration, cholestasis, and ductular bilirubinostasis. Patients with SMHN presented with more severely impaired hepatic function, a higher prevalence of multiple organ failure (as indicated by higher CLIF-SOFA and SOFA scores) and a shorter interval between acute decompensation and liver transplantation than those without SMHN (p<0.01 for all parameters). Further analyzes based on serum cytokine profile assays, gene expression microarrays and immunohistochemical analyzes revealed higher levels of anti-inflammatory cytokines in patients with SMHN. CONCLUSIONS: SMHN is a critical histological feature of HBV-associated ACLF. Identification of a characteristic pathological feature strongly supports that ACLF is a separate entity in end-stage liver disease.

Imbalance between circulating CD4+ regulatory T and conventional T lymphocytes in patients with HBV-related acute-on-chronic liver failure.

BACKGROUND & AIMS: The important pathophysiological role of immune dysfunction, especially innate immune dysfunction in patients with acute-on-chronic liver failure (ACLF), has been investigated in recent years, but dysregulation of adaptive immunity remains poorly elucidated. The aim of this study was to (i) determine the CD3(+) T-lymphocyte count and the balance between CD4(+) regulatory T (Tregs) and conventional T cells (Tconv) in hepatitis B virus (HBV)-related ACLF patients; (ii) analyse the frequencies of Tregs subpopulations; and (iii) assess the suppressive potency of CD4(+) Tregs and each fraction. METHODS: We enrolled 20 HBV-ACLF patients, 10 septic shock subjects, 20 chronic hepatitis B (CHB) patients and 20 healthy volunteers (HC). Based on flow cytometry, we performed the absolute counting of circulating T lymphocytes and phenotyping of CD4(+) Tregs and quantified the effects of Tregs and each subpopulation on Tconv proliferation by CFSE staining. RESULTS: Compared with CHB patients and HC, we observed an equal reduction in peripheral T subsets in HBV-ACLF and septic shock subjects; the number of CD4(+) Tregs remained unchanged and the Tconv count declined, promoting elevation of the Treg-to-Tconv ratio. The frequencies of Treg-II and -III were elevated in HBV-ACLF. Functional studies showed that the suppressive capacity of Tregs was preserved in the HBV-ACLF group and Treg-II came first. CONCLUSIONS: Similar to septic shock subjects, in HBV-ACLF patients there exists a reduction in CD4(+) T lymphocytes, predominantly CD4(+) Tconv, and the development of suppressive CD4(+) Tregs greatly prevails over Tconv, constituting important characteristics of adaptive immune dysfunction of HBV-ACLF.

[Characterization of a p-nitrophenol degrading bacterium Pseudomonas sp. PDS-7 and cloning of degradation relevant genes].

OBJECTIVE: The aim of this study was to (i) isolate and characterize bacteria capable of degrading p-nitrophenol (PNP); (ii) determine the kinetics of biodegradation, (iii) clone and express the PNP-degrading related genes. METHODS: Enrichment method and serial dilution spread-plate method were employed to isolate PNP-degrading strain. Morphological, physiological & biochemical tests and 16S rDNA sequence analysis were used to identify the isolate. Degradation kinetics was studied by flask test. PNP-degrading related genes were cloned by SEFA-PCR method. Hydroxyquinol 1,2-dioxygenase encoding gene pnpC was cloned into pET29a to construct the recombinant plasmid pETpnpC and expressed in E. coli BL21 (DE3). The activity of the expressed product was determined by spectrophotometric method. RESULTS: Strain PDS-7 capable of utilizing PNP as the sole carbon, nitrogen and energy source was isolated and identified as Pseudomonas sp. It could tolerate the PNP concentration up to 80 mg/L, the optimal temperature for degradation was about 30 degrees C and alkaline pH benefited PNP degradation. Hydroxyquinol 1,2-dioxygenase and maleylacetate reductase encoding gene pnpC and pnpD were cloned and sequenced respectively, the sequence was deposited in GenBank with the accession number EU233791. pnpC was expressed in E. coli BL21 (DE3), the expressed product in cell-free crude extracts showed ortho ring cleavage activity to hydroxyquinol and catechol, with the special activity 0.45 U/mg protein and 0.37 U/mg protein, respectively, indicating pnpC gene encoding hydroxyquinol 1,2-dioxygenase was actively expressed. CONCLUSION: One PNP-degrading strain Pseudomonas sp. PDS-7 was isolated and identified. Its degradation kinetics was studied. Its degradation relevant genes were cloned and expressed.

[Isolation of phenol-degrading bacteria from natural soil and their phylogenetic analysis].

Five bacterial strains capable of utilizing phenol as sole carbon source for growth were isolated from non-contaminated natural soil sample after enrichment in the presence of phenol. They were preliminarily identified according to their phylogenetic analysis and physiological and biochemical characteristics. Strain PHD-2, PHD-4 and PHD-5 belonged to the genera of Ralstonia, Acinetobacter and Microbacterium respectively; strain PHD-1 and PHD-3 were from the genus of Pseudomonas. Homology comparing of their 16S rRNA gene sequences and phylogenetic analysis displayed the high biodiversity of phenol-degrading microorganisms in the natural soil.

Isolation and characterization of a carbofuran-degrading strain Novosphingobium sp. FND-3.

A gram-negative Novosphingobium sp. strain FND-3 capable of degrading carbofuran was isolated and characterized. The carbofuran-degrading ability of strain FND-3 was investigated under various culture conditions. Strain FND-3 showed a high average carbofuran-degrading rate of 28.6 mg L(-1) h(-1) in mineral salts medium with 100 mg L(-1) carbofuran. GC/MS analysis pointed out the presence of several unknown metabolites. One hydrolyzate was identified as 2-hydroxy-3-(3-methypropan-2-ol) phenol via hydrolysis of carbofuran phenol. The appearance of another metabolite with M(+) of 180 m/z indicated that the hydroxylation of carbofuran occurred at the aromatic ring. One novel degrading product with M(+) of 239 m/z was identified as 2-hydroxy-3-(3-methylpropan-2-ol) benzene-N-methylcarbamate via hydrolyzing at the ether bond of furanyl ring of carbofuran. Strain FND-3 was also able to degrade other N-methylcarbamate pesticides.

Suppression of IL-8 gene transcription by resveratrol in phorbol ester treated human monocytic cells.

Resveratrol (3,4',5-trihydroxy-trans-silbene), a natural phytoalexin found in grapes and other food products, has promising anti-inflammatory and anticancer effects. To observe the modulation of interleukin-8 (IL-8) production in human monocytic cells by resveratrol and explore its mechanism at the gene transcription level, U937 cells were stimulated with phorbol 12-myristate 13-acetate (PMA) for 24h. IL-8 protein in supernatants was measured by radioimmunoassay. The cytotoxicity of PMA, dexamethasone and resveratrol was accessed by MTT cell proliferation assay. The RNA level of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and IL-8 were detected by RT-PCR using specific primers. DNA binding activities of NF-kappaB and AP-1 were examined by electrophoretic mobility shift assay (EMSA). 0.01-100 nM PMA could significantly induce IL-8 production in U937 cells; 10 microM Dexamethasone and 10, 1, 0.1 microM resveratrol could inhibit PMA-induced IL-8 protein production and mRNA accumulation. The cytotoxicity did not contribute to their inhibitory effect. The DNA binding activity of AP-1 was inhibited by dexamethasone and resveratrol, but resveratrol has little effect on PMA-induced NF-kappaB activation. Resveratrol could inhibit PMA-induced IL-8 production in U937 cells at protein and mRNA levels. The suppression of IL-8 gene transcription by resveratrol was, at least partly, due to inhibition of AP-1 activation.