Dysambiol, an Anti-inflammatory Secomeroterpenoid from a Dysidea sp. Marine Sponge.

An unusual secomeroterpenoid, dysambiol (1), was isolated from a Dysidea sp. marine sponge collected from the South China Sea. Dysambiol features an unprecedented secomeroterpene scaffold with a rare lactone bridge. The structure of 1 was determined by extensive spectroscopic analysis, Mosher's method, and electronic circular dichroism calculation. Dysambiol displayed potent anti-inflammatory activity in LPS-induced Raw 264.7 macrophages by regulating the NF-κB/MPAK signaling pathway.

Gene Therapy Activates Retinal Pigment Epithelium Cell Proliferation for Age-related Macular Degeneration in a Mouse Model.

OBJECTIVE: Age-related macular degeneration (AMD) is a degenerative retinal disease. The degeneration or death of retinal pigment epithelium (RPE) cells is implicated in the pathogenesis of AMD. This study aimed to activate the proliferation of RPE cells in vivo by using an adeno-associated virus (AAV) vector encoding ß-catenin to treat AMD in a mouse model. METHODS: Mice were intravitreally injected with AAV2/8-Y733F-VMD2-ß-catenin for 2 or 4 weeks, and ß-catenin expression was measured using immunofluorescence staining, real-time quantitative reverse transcription polymerase chain reaction (PCR), and Western blotting. The function of ß-catenin was determined using retinal flat mounts and laser-induced damage models. Finally, the safety of AAV2/8-Y733F-VMD2-ß-catenin was evaluated by multiple intravitreal injections. RESULTS: AAV2/8-Y733F-VMD2-ß-catenin induced the expression of ß-catenin in RPE cells. It activated the proliferation of RPE cells and increased cyclin D1 expression. It was beneficial to the recovery of laser-induced damage by activating the proliferation of RPE cells. Furthermore, it could induce apoptosis of RPE cells by increasing the expression of Trp53, Bax and caspase3 while decreasing the expression of Bcl-2. CONCLUSION: AAV2/8-Y733F-VMD2-ß-catenin increased ß-catenin expression in RPE cells, activated RPE cell proliferation, and helped mice heal from laser-induced eye injury. Furthermore, it could induce the apoptosis of RPE cells. Therefore, it may be a safe approach for AMD treatment.

A Hypoxia-Regulated Retinal Pigment Epithelium-Specific Gene Therapy Vector Reduces Choroidal Neovascularization in a Mouse Model.

BACKGROUND: Wet age-related macular degeneration (wAMD) is characterized by the presence of choroidal neovascularization (CNV). Although there are some clinical drugs targeting vascular endothelial growth factor (VEGF) and inhibiting CNV, two major side effects limit their application, including the excessive activity of anti-VEGF and frequent intraocular injections. To explore better treatment strategies, researchers developed a hypoxic modulator retinal pigment epithelium (RPE)- specific adeno-associated virus (AAV) vector expressing endostatin to inhibit CNV. However, the mechanism of endostatin is complex. Instead, soluble fms-like tyrosine kinase-1 (sFlt-1) can inhibit VEGF-induced angiogenesis through two simple and clear mechanisms, giving rise to sequestration of VEGF and forming an inactive heterodimer with the membrane-spanning isoforms of the VEGF receptor Flt-1 and kinase insert domain-containing receptor. OBJECTIVE: In this study, we chose sFlt-1 as a safer substitute to treat wAMD by inhibiting VEGFinduced angiogenesis. METHODS: The AAV2/8-Y733F-REG-RPE-sFlt-1 vector was delivered by intravitreal injection to the eyes of mice. AAV2/8-Y733F vector is a mutant of the AAV2/8 vector, and the REG-RPE promoter is a hypoxia-regulated RPE-specific promoter. Two animal models were used to evaluate the function of the vector. RESULTS: In the cobalt chloride-induced hypoxia model, the results demonstrated that the AAV2/8- Y733F-REG-RPE-sFlt-1 vector induced the expression of the sFlt-1 gene in RPE cells through hypoxia. In the laser-induced CNV model, the results demonstrated that the AAV2/8-Y733F-REG-RPE-sFlt- 1 vector reduced laser-induced CNV. CONCLUSION: Hypoxia regulated, RPE-specific AAV vector-mediated sFlt-1 gene is a hypoxiaregulated antiangiogenic vector for wAMD.

Evaluation of the predictive performance of Bayesian dosing for warfarin in Chinese patients.

AIM: To evaluate the accuracy and predictive performance of Bayesian dosing for warfarin in Chinese patients. MATERIALS & METHODS: Six multiple linear regression algorithms (Wei, Lou, Miao, Huang, Gage and IWPC) and a Bayesian method implemented in Warfarin Dose Calculator were compared with each other. RESULTS: Six multiple linear regression warfarin dosing algorithms had similar predictive ability, except Miao and Lou. The mean prediction error of Bayesian priori and posteriori method were 0.01 mg/day (95% CI: -0.18 to 0.19) and 0.17 mg/day (95% CI: -0.05 to 0.29), respectively, and Bayesian posteriori method demonstrated better performance in all dose ranges. CONCLUSION: The Bayesian method showed a good potential for warfarin maintenance dose prediction in Chinese patients requiring less than 6 mg/day.

Emodin suppresses angiogenesis and metastasis in anaplastic thyroid cancer by affecting TRAF6­mediated pathways in vivo and in vitro.

Emodin has been recognized to be an anti­cancer agent against a number of types of human cancer. It was demonstrated that TNF receptor­associated factor 6 (TRAF6) was correlated with cancer angiogenesis and metastasis. The present study confirmed the association between TRAF6 and the angiogenesis/metastasis of anaplastic thyroid cancer (ATC). The anti­angiogenesis and metastatic effects of emodin, in addition to its molecular mechanisms in ATC, were investigated. A total of two ATC cell lines, namely 8505c and SW1736, were studied. ATC cells were implanted into nude mice to form xenografts or to establish lung metastasis models. Emodin was used to incubate ATC cells or to treat animals orally. An MTT assay was used to assess cell proliferation. A wound healing assay was employed to evaluate cell migration. ELISA analysis was used to detect the vascular endothelial growth factor (VEGF) content. Western blotting was used to determine the protein expression levels. In the in vivo study, cancer angiogenesis was assessed by micro vascular density measurement. The lung metastatic rate was the criterion for cancer metastasis. The results of the present study demonstrated that the proliferation of ATC was inhibited by emodin. The activation of the TRAF6/hypoxia inducible factor (HIF)­1α/VEGF and TRAF6/basigin (CD147)/matrix metalloproteinase­9 (MMP9) pathways were associated with the angiogenesis and metastasis of ATC. In a concentration­dependent manner, emodin inhibited the TRAF6/HIF­1α/VEGF and TRAF6/CD147/MMP9 signaling pathways to suppress angiogenesis and metastasis. In conclusion, emodin exerted anti­angiogenic and anti­metastatic activities in ATC by affecting TRAF6­mediated pathways.

Dysiarenone, a Dimeric C21 Meroterpenoid with Inhibition of COX-2 Expression from the Marine Sponge Dysidea arenaria.

Dysiarenone (1), a dimeric C21 meroterpenoid featuring an unprecedented 2-oxaspiro[bicyclo[3.3.1]nonane-9,1'-cyclopentane] carbon skeleton, was isolated from the marine sponge Dysidea arenaria. The structure of 1 was determined by HRMS and NMR spectroscopic analyses coupled with ECD calculations. Dysiarenone showed inhibitory activities against COX-2 expression and the production of prostaglandin E2 with an IC50 value of 6.4 µM in LPS-stimulated RAW264.7 macrophages.

Simultaneous measurement of refractive index and temperature based on all-dielectric metasurface.

In this paper, a novel kind of sensors for simultaneous measurement of refractive index and temperature based on all-dielectric metasurfaces is proposed. The metasurfaces are constructed by an array of silicon nanoblocks on top of the bulk fused silica substrate. We used three-dimensional full wave electromagnetic field simulation by finite integral method to accurately calculate the transmission spectrum of the metasurfaces. Two transmission dips corresponding to the electric and magnetic resonances are observed. Both dips shift as the ambient refractive index or the temperature changes. Simulation results show that the sensing sensitivities of two dips to the refractive index are 243.44 nm/RIU and 159.43 nm/RIU, respectively, while the sensitivities to the temperature are 50.47 pm/°C and 75.20 pm/°C, respectively. After introducing four holes into each silicon nanoblock, the electromagnetic field overlap in the surrounding medium can be further promoted, and the sensitivities to the refractive index increase to 306.71 nm/RIU and 204.27 nm/RIU, respectively. Our proposed sensors have advantages of polarization insensitive, small size, and low loss, which offer them high potential applications in physical, biological and chemical sensing fields.

Dysivillosins A-D, Unusual Anti-allergic Meroterpenoids from the Marine Sponge Dysidea villosa.

Four unusual meroterpenoids, dysivillosins A-D (1-4), were isolated from an organic extract of the marine sponge Dysidea villosa collected from the South China Sea. Their planar structures were determined by 1D and 2D NMR and HRESIMS techniques, while the relative and absolute configurations were elucidated by NOESY experiments and comparison between the calculated and experimental ECD spectra. To the best of our knowledge, dysivillosins A-D are the first examples of terpene-polyketide-pyridine hybrid metabolites from the nature. Anti-allergic activity evaluation showed that compounds 1-4 potently inhibited the release of ß-hexosaminidase, a marker of degranulation, in a dose-dependent manner with IC50 values of 8.2-19.9 µM. Additionally, the four meroterpenoids could downregulate the production of lipid mediator leukotrienes B4 (LTB4) and pro-inflammatory cytokine interleukin-4 (IL-4) in the antigen-stimulated RBL-2H3 mast cells. Further biological investigations revealed that dysivillosin A (1) could suppress the phosphorylation of Syk and PLCγ1 in IgE/FcɛRI/Syk signaling pathway, which resulted in the inhibition of degranulation and the downregulation of LTB4 and IL-4 production in mast cells.

An Alternative Strategy for Treatment of Distal Tibiofibular Syndesmotic Disruption: A Technical Note.

Metal screws are the most widely used in treating syndesmotic injuries; however, failure and the rigidity of the screws can threaten the success of the treatment and increase the cost of care. We performed open reduction and internal fixation of a fibula fracture followed by fixation of the tibiofibular joint with an olive wire and one half of an external fixator. We present a review of 1 patient treated with this material. We believe this method to be a physiologic fixation system with the advantage of being safe, inexpensive, adjustable, and effective; however, further clinical investigation and comparison to other methods is needed in order to better understand it clinical utility.

Dysiherbols A-C and Dysideanone E, Cytotoxic and NF-κB Inhibitory Tetracyclic Meroterpenes from a Dysidea sp. Marine Sponge.

Four new tetracyclic meroterpnes, dysiherbols A-C (1-3) and dysideanone E (4), were isolated from a Dysidea sp. marine sponge collected from the South China Sea. Their complete structures and absolute configurations were unambiguously determined by a combination of NMR spectroscopic data, ECD calculations, and single-crystal X-ray diffraction analysis. Within the sesquiterpene quinol structures, dysiherbols A-C possess an intriguing 6/6/5/6-fused tetracyclic carbon skeleton. The NF-κB inhibitory and cytotoxic activity evaluation disclosed that dysiherbol A (1) showed potent activity with respective IC50 values of 0.49 and 0.58 µM, which were about 10-fold and 20-fold more potent than those of dysiherbols B (2) and C (3), which feature hydroxy and ketone carbonyl groups at the C-3 position.

Clinical characteristics and neurologic recovery of patients with cervical spinal tuberculosis: should conservative treatment be preferred? A retrospective follow-up study of 115 cases.

OBJECTIVE: To present the clinical characteristics and prognostic factors of neurologic recovery in patients with cervical spinal tuberculosis (CST). METHODS: General description and multivariate analysis were used to detect possible predictors of the outcome of patients with neurologic deficit. Follow-up data were used to generate a Kaplan-Meier curve of neurologic recovery. RESULTS: Protective factors in neurologic recovery included less involved vertebrae, surgery, and higher Japanese Orthopaedic Association score before treatment; not shorter symptom duration was not a protective factor. Normal neurologic function was present in 30% of patients 6 months after treatment, in 56% of patients 12 months after treatment, and in 93% of patients 28 months after treatment. The cumulative complete neurologic recovery rates at 6 months, 12 months, and 28 months were 44%, 68%, and 91.7% in the surgery group and 16.7%, 38.8%, and 94.4% in the nonsurgery group. CONCLUSIONS: Surgery and Japanese Orthopaedic Association score before treatment are important predictors of neurologic recovery in patients with CST. A neurologic recovery curve could predict neurologic recovery in patients with CST and indicate in patients with neurologic deficit how long it will take to achieve complete neurologic recovery. The effect of surgery is quick, and the effect of conservative treatment is slower, but no difference in neurologic recovery was found in the long-term. Conservative treatment should be tried in every patient with CST with no obvious indication for surgery. In contrast to patients with tuberculosis without cervical spine involvement but with more complications, comprehensive conservative therapy should be preferred for patients with neurologic deficit to avoid unnecessary surgery and overtreatment and to conserve medical resources. Indications for surgical intervention for CST should be carefully selected.

(±)-Quassidines I and J, two pairs of cytotoxic bis-ß-carboline alkaloid enantiomers from Picrasma quassioides.

(±)-Quassidines I (1) and J (2), two pairs of new bis-ß-carboline alkaloid enantiomers, were isolated from the stems of Picrasma quassioides. Their structures were determined by the analysis of spectroscopic data, including HRESIMS and 2D NMR, and confirmed by single-crystal X-ray diffraction analysis. The racemic mixtures of 1 and 2 were resolved into two pairs of enantiomers, (+)-S-1a and (-)-R-1b and (+)-S-2a and (-)-R-2b, by HPLC using a chiral Daicel IB-3 column, respectively, which represents the first successful example to resolve bis-ß-carboline racemic mixtures. The absolute configurations of the two pairs of enantiomers were determined by comparison between the calculated and experimental ECD spectra. The cytotoxicity evaluation revealed that (+)-S-1a and (+)-S-2a showed more potent cytotoxicity against human cervical HeLa and gastric MKN-28 cancer cell lines with IC50 values of 4.03-6.30 µM than their enantiomers with IC50 values of 9.64-12.3 µM; however, the two (+)-S-quassidines showed similar activities to their enantiomers against the mouse melanoma B-16 cancer cell line.

Dysidinoid A, an unusual meroterpenoid with anti-MRSA activity from the South China Sea sponge Dysidea sp.

An unusual meroterpenoid, dysidinoid A (1), was isolated from the South China Sea sponge Dysidea sp. Its structure was elucidated by extensive spectroscopic methods including HRESIMS and 2D NMR, and its absolute configuration was determined by single-crystal X-ray diffraction analysis. Dysidinoid A (1) is the first meroterpenoid from Nature bearing a 9,4-friedodrime skeleton and a 2,5-dionepyrrole unit. Dysidinoid A (1) showed potent antibacterial activity against two strains of pathogenic bacteria methicillin-resistant Staphylococcus aureus (MRSA) with MIC90 values of 8.0 µg/mL against both.

[Determination of chlorogenic acid and baicalin in Yinhuang film-coated tablets by HPLC isocratic elution].

OBJECTIVE: To establish an HPLC method for determination of chlorogenic acid and baicalin in Yinhuang film-coated tablets. METHOD: HPLC isocratic elution was adopted, with the C18 column (4.6 mm x 250 mm, 5 microm) as the chromatographic column, methanol-water-phosphoric acid (45: 55: 0.2) as the mobile phase. The flow rate was set at 1.0 mL x min(-1), and column temperature was set at 30 degrees C. The detection wavelength was 328 nm, and the sample size was 20 microL. RESULT: At concentrations ranging between 2.200-44.00, 24.20-242.0 mg x L(-1), peak area of chlorogenic acid and baicalin showed good linear relationship, the average recoveries were 99.63% (RSD 1.8%, n = 9) and 99.72% (RSD 1.9%, n = 9), respectively. CONCLUSION: The method is simple that it can accurately measure the content of chlorogenic acid and baicalin in Yinhuang film-coated tablets.

[²°¹TI and (99m)Tc-MIBI scintigraphy in evaluation of neoadjuvant chemotherapy for osteosarcoma].

OBJECTIVE: To evaluate the application of ²°¹TI and (99m)Tc-MIBI scintigraphy in assessment of neoadjuvant chemotherapy for osteosarcoma. METHODS: Twenty-two patients with osteosarcoma underwent both ²°¹TI and (99m)Tc-MIBI scintigraphy. According to tumor necrosis rate (TNR), 22 patients were classified into three groups: Group 1(necrosis less than 50%), Group 2(50% ≊ 89% necrosis) and Group 3(necrosis greater than 90%). The uptake ratio(UR) was obtained in images before and after chemotherapy. The alteration ratio(AR) and tumor necrosis ratio (TNR) were calculated. RESULTS: In ²°¹Tl images,UR(pre) and UR(post) (mean ± s.d.) of Group 1 were 2.14 ± 0.67, 2.07 ± 0.71 (P>0.05); UR(pre) and UR(post)of Group 2 were 3.45 ± 1.57 and 2.02 ± 0.97 (P<0.01); UR(pre) and UR(post) of Group 3 were 3.57 ± 0.67 and 1.36 ± 0.20 (P<0.01). In (99m)Tc-MIBI images, UR(pre)and UR(post) of Group 1 were 1.66 ± 0.42 and 1.85 ± 0.70 (P>0.05); UR(pre) and UR(post) of Group 2 were 2.39 ± 1.41 and 1.68 ± 0.72 (P<0.05);UR(pre) and UR(post) of Group 3 were 2.56 ± 0.60 and 1.19 ± 0.14 (P<0.01). The AR value in (201)Tl scintigraphy was -0.03-0.72, the liner regression analysis of AR versus TNR showed a highly significant positive correlation (r=0.95). The AR value in (99m)Tc-MIBI scintigraphy was -1.21-0.64, the liner regression analysis of AR versus TNR showed a highly significant positive correlation (r=0.71). The liner regression analysis of AR in ²°¹TI scintigraphy versus AR in (99m)Tc-MIBI scintigraphy showed a highly significant positive correlation (r=0.70). CONCLUSION: The AR changes significantly after neoadjuvant chemotherapy and is positively correlated with TNR, which indicates that ²°¹TI- and (99m)Tc-MIBI scintigraphy can be used for evaluation of neoadjuvant chemotherapy.

[Value of (99m)Tc-MIBI/(18)F-FDG-dual-isotope simultaneous acquisition in diagnosis and treatment of myocardial infarction].

OBJECTIVE: To investigate the value of(99m)Tc-MIBI/(18)F-FDG-dual-isotope simultaneous acquisition (DISA) in diagnosis and treatment of patients with myocardial infarction. METHODS: Sixty-three patients with myocardial infarction who underwent DISA before and after treatment were enrolled in the study. All cases were divided into subgroups based on different treatment and myocardial viability: Group A1 (n = 16) with coronary revascularization and viable myocardium, A2 (n = 9) same as A1 but no viable myocardium; B1 (n = 6) with coronary revascularization + stem cell transplantation and viable myocardium, B2 (n = 7) same as B1 but no vital myocardium; C1 (n = 8) with stem cell transplantation and viable myocardium, C2 (n = 17) same as C1 but on viable myocardium. The changes of uptake rate of (99m)Tc-MIBI/(18)F-FDG before and after treatment were analyzed with SPSS 13.0 software. RESULT: There were statistical significances in DF value of (99m)Tc-MIBI or (18)F-FDG imaging before and after treatment in all groups (P <0.05), except Group A2 (P>0.05). The improvement of blood perfusion and metabolism in cardiac survival groups was more marked than that in non-cardiac survival groups after treatment (P<0.05). Furthermore, Group B1 was superior to Groups A1 and C1; Group B2 was superior to Group A2 and Group C2 (P<0.05 or<0.01). CONCLUSION: (99m)Tc-MIBI/(18)F-FDG DISA can detect myocardial viability and is of value for patients with myocardial infarction to choose appropriate therapeutic strategies. The degree of cardiac improvement after treatment can be evaluated by DISA.

Fluorescence quenching of CdSe quantum dots by nitroaromatic explosives and their relative compounds.

CdSe quantum dots (QDs) were synthesized in oleic acid and octadecene medium under high-temperature and dispersed in chloroform. Nitroaromatic explosives and their relative compounds, 2,4,6-trinitrotoluene (TNT), 2,4-dinitrotoluene (DNT), nitrobenzene (NB), 2,4-dinitrochlorobenzene (DNBCl) and p-nitrotoluene (NT) can obviously cause the fluorescence quenching of the synthesized QDs. Under the optimum conditions, a nonlinear response was observed over the concentration range of 10(-8) to 10(-5) M for them all. The modified Stern-Volmer quenching equations of ln I(0)/I versus C show a good linear relation in 10(-5) M order of magnitude, and the detection limits approach 10(-6) to 10(-7) M.

[Efficient isolation of chondrocytes from rabbit articular cartilage with three-step enzymatic digestion and observation of their biological characteristics during cultivation in vitro].

OBJECTIVE: To observe the effect of isolating the chondrocytes from articular cartilage with the method of three-step enzymatic digestion, and the biological characteristics of the isolated chondrocytes during cultivation in vitro in order to evaluate their biological activity. METHODS: The method of three-step enzymatic digestion was designed that the articular cartilage was digested one by one with the 1 g/L trypsin and 1 g/L EDTA, 1 g/L hyaluronidase and 2 g/L collagenase I in the culture medium to isolate chondrocytes. The harvesting and viability rate of the primary chondrocytes were detected. During the passage cultivation in vitro, the changes of the chondrocytes shape and growth were observed, the changes of the collagen type I and II and aggrecan in the extracellular matrix were investigated and detected. RESULTS: (1) The extracellular matrix of articular cartilage was completely dissolved by the three-step enzymatic digestion, and the chondrocytes were completely isolated from the solid matrix. The number of the harvested chondrocytes from every gram of wet cartilage was 50.3 x 10(6) on average, and their viability rate was 98.8% on average. (2) The primary and first passage chondrocytes had triangle or multi-angle shape, and became elliptic shape at the growing confluence with the positive immunohistochemical stain of collagen type II and the strong heterochromia to toluidine blue. The content of sulfate glycosaminoglycans (GAG) in the extracellular matrix of the primary passage cells was (92 +/- 10) microg/cm(2). The chondrocytes after the third passaging gradually became spindle shape with the negative stain of collagen type II and the weak heterochromia to toluidine blue. The content of sulfate GAG of the fourth passage cells was (48 +/- 12) microg/cm(2). CONCLUSION: (1) The method of three-step enzymatic digestion can make the extracellular matrix of articular cartilage completely degraded, and has advantages in the high efficiency of harvesting primary chondrocytes, high cellular viability rate and simple manipulation. (2) The primary and first passage chondrocytes have fine biological activity, and the chondrocytes after the third passaging have lost their special biological activity.

Investigating the role of 99mTc-TRODAT-1 SPECT imaging in idiopathic Parkinson's disease.

OBJECTIVE: To investigate the role of 99mTc-TRODAT-1 SPECT in diagnosis and assessing severity of idiopathic Parkinson's disease (PD). METHODS: Thirty-eight patients with primary, tentative diagnosis of PD and eighteen age-matched normal controls were studied with 99mTc-TRODAT-1 SPECT imaging. The regions of interests (ROIs) were drawn manually on cerebellum (CB), occipital cortex (OC) and three transverse plane slice-views of striatums, the semiquantitative BG (background)/[(OC+CB)/2] were then calculated. RESULTS: A lower uptake of 99mTc-TRODAT-1 in striatums were displayed in thirty-six out of thirty-eight PD patients by visual inspection, compared to controls. In twenty-four PD cases with (Hoehn and Yahr scale) HYS stage I, a greater loss of DAT uptake was found in striatum and its subregions contralateral striatum to the affected limbs than in the same regions of the controls, although the striatal uptake was bilaterally reduced. Using Spearman correlation analysis showed that the reduction of the uptake ratios significantly correlated with the UPDRS in striatum and all its subregions in the PD group (P<0.05), a similar change was also found in the putamen by using the rating scale of Hoehn and Yahr (P<0.05). However, analysis of variance (ANOVA) did not show any relationship between the decreasing uptake of 99mTc-TRODAT-1 and increasing severity of PD patients, although the specific uptake of 99mTc-TRODAT-1 was continuously decreased in the striatum by visual inspection with the progress of PD from HYS stage I to III. CONCLUSION: 99mTc-TRODAT-1 SPECT imaging may serve as a useful method for improving the correct diagnosis of PD. In assessing the role of 99mTc-TRODAT-1 SPECT in disease severity of PD, UPDRS can offer a comprehensive index, although the Hoehn and Yahr assessment may be available in part.