Isoalantolactone suppresses gallbladder cancer progression via inhibiting the ERK signalling pathway.

CONTEXT: Gallbladder cancer (GBC) is the most common malignant tumour of the biliary tract. Isoalantolactone (IAL), an active sesquiterpene lactone compound isolated from the roots of Inula helenium L. (Asteraceae), has antitumour effects. OBJECTIVE: This study investigates the effects of IAL on GBC. MATERIALS AND METHODS: In vitro, NOZ and GBC-SD cells were treated with IAL (0, 10, 20 and 40 µM) for 24 h. The DMSO-treated cells were selected as a control. Cell proliferation, migration, invasion and apoptosis were measured by the CCK-8 assay, transwell assay, flow cytometry and western blot. In vivo, subcutaneous tumour xenografts were constructed by injecting nude mice (BALB/C) with 5 × 106 NOZ cells. Mice were divided into the control group (equal amount of DMSO), the IAL group (10 mg/kg/day) and the IAL + Ro 67-7476 group (IAL, 10 mg/kg/day; Ro 67-7476, 4 mg/kg/day). The study duration was 30 days. RESULTS: Compared with the DMSO group, cell proliferation of NOZ (IC50 15.98 µM) and GBC-SD (IC50 20.22 µM) was inhibited by about 70% in the IAL 40 µM group. Migration and invasion were suppressed by about 80%. Cell apoptosis rate was increased about three-fold. The phosphorylation level of ERK was decreased to 30-35%. Tumour volume and weight (about 80% reduction) were suppressed by IAL in vivo. Moreover, the effects of IAL were abolished by Ro 67-7476 in vitro and in vivo. DISCUSSION AND CONCLUSIONS: Our findings indicate that IAL could inhibit GBC progression in vitro and in vivo by inhibiting the ERK signalling pathway.

Managing Pathologic Scars by Injecting Auto-Cross-linked Hyaluronic Acid: A Preliminary Prospective Clinical Study.

BACKGROUND: Pathological scars are unattractive and may significantly impair the patient's quality of life. Current treatments provide inconsistent results, and none may be regarded as definitive. Recently, an auto-cross-linked hyaluronic acid (HA) formulation, IAL-SYSTEM ACP, featuring a long residence time and an enhanced safety profile, has been successfully used to prevent surgical adhesions, treat tendon lesions and rejuvenate the face and the décolletage. This study aims to preliminarily investigate whether IAL-SYSTEM ACP may also be effective in treating pathological scars resulting from burns, trauma or iatrogenic causes. METHODS: Patients presenting one pathological scar were prospectively recruited and treated with two IAL-SYSTEM ACP injections carried out two weeks apart. Scar improvement was measured comparing the patient and observer scar assessment scale (POSAS) scores collected before treatment (T0) and 90 days after the second injection (T90) using nonparametric tests. The effect of age and scar type over score variation was investigated through quantile regression analysis. RESULTS: Forty-one patients, 10 women and 31 men (median age, 34 years) were recruited. No patient dropped out, and no significant adverse event was observed. At T90, the median observer total score decreased by 11 units (- 77.5%) and the median patient total score decreased by 15 units (- 73.7%). The difference was significant (p < 0.001) in both cases. Traumatic injuries and young patient's age were the most significant predictors of a positive treatment outcome. CONCLUSIONS: Treatment of pathological scars by two IAL-SYSTEM ACP injections, carried out two weeks apart, may provide significant clinical benefits. These findings should be the subject of further investigations. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

A novel actinomycete Streptomyces enissocaesilis exhibiting algicidal activity against the toxic cyanobacterium Phormidium angustissimum.

Harmful cyanobacterial blooms that occur in freshwater can produce hazardous cyanotoxins as contaminants that threaten ecosystems, aquatic organisms, and human health. In the present study, the actinobacterium Streptomyces enissocaesilis, strain M35, isolated from soils, exhibited the strongest algicidal activity against the toxic cyanobacterium Phormidium angustissimum TISTR 8247. To improve the P. angustissimum removal efficiency of strain M35, the optimum carbon and nitrogen sources were determined as starch and yeast extract, respectively. Response surface methodology (RSM) using the Box-Behnken design (BBD) revealed that the optimal independent parameters among the culture medium conditions for enhancing the algicidal activity of strain M35 were 21.5 g/L starch, 0.57 g/L yeast extract, and a pH value of 8.00. The Phormidium sp. removal efficiency increased notably from 80.8 to 94.4% under the optimum conditions. In a batch experiment, the removal of P. angustissimum in an internal airlift loop (IAL) bioreactor containing immobilized strain M35 on a plastic medium indicated a high anti-Phormidium activity of 94.8%, whereas in a continuous system, strain M35 exhibited a removal efficiency of 85.5%. This study revealed that this actinobacterium could potentially be utilized to remove the toxic cyanobacterium Phormidium from water.

Efficacy and analysis of modified "three-tube method" in the treatment of intrathoracic anastomotic leakage after esophagectomy.

BACKGROUND: This study aimed to retrospectively evaluate the clinical efficacy of the modified "three-tube method" for the treatment of intrathoracic anastomotic leakage (IAL) after esophagectomy, and to analyze the independent risk factors for prolonging the treatment time of the modified "three-tube method". METHODS: From January 2013 to December 2018, IAL was reported in 22 patients with esophageal cancer who underwent esophagectomy with intrathoracic anastomosis. By reviewing and analyzing the clinical data of the 22 patients, the efficacy of the modified "three-tube method" treatment and the independent risk factors associated with a longer treatment duration of the modified "three-tube method" were evaluated. RESULTS: Of the 22 patients, 19 were male (86.4%). The average age was 65.2 years old. A total of 4 patients (18.2%) underwent preoperative neoadjuvant chemotherapy; 6 patients (27.3%) had a Charlson comorbidity index (CCI) score of 1-3; the average diagnosis time of IAL was 9.5 days; the median intervention time was 4 days; and the average fistula length was 1.5 cm. The average albumin level after surgery was 30.5 g/L, and the average C-reactive protein (CRP) level was 139.4 mg/L. The modified "three-tube method" average treatment time was 19.5 days. One patient (4.5%) died of respiratory failure during treatment. Univariate analysis and multivariate analysis by establishing multiple linear regression model found that the date of intervention and the fistula size were significantly associated with a longer treatment duration of the modified "three-tube method". CONCLUSIONS: The modified "three-tube method" is a safe and effective means for non-surgical treatment of IAL after esophagectomy. The intervention time and the fistula size are independent risk factors for prolonging the treatment time of the modified "three-tube method".

Simulation of the θ' Precipitation Process with Interfacial Anisotropy Effects in Al-Cu Alloys.

The effects of anisotropic interfacial properties and heterogeneous elasticity on the growth and ripening of plate-like θ'-phase (Al2Cu) in Al-1.69 at.% Cu alloy are studied. Multi-phase-field simulations are conducted and discussed in comparison with aging experiments. The precipitate/matrix interface is considered to be anisotropic in terms of its energy and mobility. We find that the additional incorporation of an anisotropic interfacial mobility in conjunction with the elastic anisotropy result in substantially larger aspect ratios of the precipitates closer to the experimental observations. The anisotropy of the interfacial energy shows comparably small effect on the precipitate's aspect ratio but changes the interface's shape at the rim. The effect of the chemo-mechanical coupling, i.e., the composition dependence of the elastic constants, is studied as well. We show that the inverse ripening phenomenon, recently evidenced for δ' precipitates in Al-Li alloys (Park et al. Sci. Rep. 2019, 9, 3981), does not establish for the θ' precipitates. This is because of the anisotropic stress fields built around the θ' precipitates, stemming from the precipitate's shape and the interaction among different variants of the θ' precipitate, that disturb the chemo-mechanical effects. These results show that the chemo-mechanical effects on the precipitation ripening strongly depend on the degree of sphericity and elastic isotropy of the precipitate and matrix phases.

Comparative performances of seven quantitative Reverse-Transcription Polymerase Chain Reaction assays (RT-qPCR) for detecting SARS-CoV-2 infection in samples from individuals suspected of COVID-19 in São Paulo, Brazil.

Introduction: Brazil is the second largest country with COVID-19 positive cases worldwide. Due to the potent spread of the virus and the scarcity of kits and supplies, the Brazilian Ministry of Health has granted authorization for the use of kits available during this emergency, without an accurate evaluation of their performance. This study compared the performance and cost-effectiveness of seven molecular assays/kits available in São Paulo, Brazil, for SARS-CoV-2 diagnosis. Materials and methods: A total of 205 nasopharyngeal/oropharyngeal samples from suspected cases of COVID-19, were tested using the following assays: (i) GeneFinder COVID-19 plus RealAmp kit; (ii) 2019-nCoV RNA PCR-Fluorescence Probing, Da An Gene Co.; (iii) in-house RT-qPCR SARS-CoV-2 IAL; (iv) 2019-nCoV kit, IDT; (v) molecular SARS-CoV-2 (E) kit, Bio-Manguinhos; (vi) Allplex 2019-nCoV modified Assay, Seegene Inc, and (vii) Biomol one-step COVID-19 kit, IBMP. The criteria for determining a SARS-CoV-2 true positive result included the cycle threshold cut-off values, the characteristics of exponential/linear curves, the gene target diversity, and a positive result in at least two assays. Results: The overall sensitivity of the assays listed were GeneFinder 83.6%, Da An Gene 100.0%, IAL 90.4%, IDT 94.6%, Bio-Manguinhos 87.7%, Allplex 97.3%, and IBMP 87.7%. The minor sensitive gene target was RdRP. Although all assays had a Cohen's Kappa index ≥0.893, the best tests used multiplex assays identifying N-gene and/or E-gene targets. Conclusion: All assays tested accurate for diagnosis, but considering cost-effectiveness (cost, time consumption, number of samples tested, and performance), the in-house IAL assay was ideal for COVID-19 diagnosis in São Paulo, Brazil.

Estradiol induces gene proximity and MLL-MLLT3 fusion in an activation-induced cytidine deaminase-mediated pathway.

Epidemiological data have linked birth control formulations to an increased risk of infant acute leukemia involving MLL rearrangements. Reverse transcription polymerase chain reaction (RT-PCR) studies showed that 10 nM estradiol enhanced MLL transcription in addition to its common translocation partners, MLLT2 (AF4) and MLLT3 (AF9). The same concentration of estradiol triggered MLL and MLLT3 co-localization without affecting the interaction of genes located on the same chromosomes. Estradiol also stimulated the generation of MLL-MLLT3 fusion transcripts as seen by RT-PCR. RNAi knockdown of activation-induced cytidine deaminase (AICDA) suppressed the induction of MLL-MLLT3 fusion transcript formation observed with estradiol. Additionally, chromatin immunoprecipitation (ChIP) analysis showed estradiol dependent localization of AICDA in MLL intron 11, upstream of a hotspot for both DNA cleavage and rearrangement, but not downstream within intron 12. Combined, these studies show that levels of estradiol consistent with that observed during pregnancy have the potential to initiate MLL fusions through an AICDA-mediated mechanism.