Papillary thyroid cancer organoids harboring BRAFV600E mutation reveal potentially beneficial effects of BRAF inhibitor-based combination therapies.

BACKGROUNDS: Papillary thyroid cancer (PTC), which is often driven by acquired somatic mutations in BRAF genes, is the most common pathologic type of thyroid cancer. PTC has an excellent prognosis after treatment with conventional therapies such as surgical resection, thyroid hormone therapy and adjuvant radioactive iodine therapy. Unfortunately, about 20% of patients develop regional recurrence or distant metastasis, making targeted therapeutics an important treatment option. Current in vitro PTC models are limited in representing the cellular and mutational characteristics of parental tumors. A clinically relevant tool that predicts the efficacy of therapy for individuals is urgently needed. METHODS: Surgically removed PTC tissue samples were dissociated, plated into Matrigel, and cultured to generate organoids. PTC organoids were subsequently subjected to histological analysis, DNA sequencing, and drug sensitivity assays, respectively. RESULTS: We established 9 patient-derived PTC organoid models, 5 of which harbor BRAFV600E mutation. These organoids have been cultured stably for more than 3 months and closely recapitulated the histological architectures as well as mutational landscapes of the respective primary tumors. Drug sensitivity assays of PTC organoid cultures demonstrated the intra- and inter-patient specific drug responses. BRAFV600E inhibitors, vemurafenib and dabrafenib monotherapy was mildly effective in treating BRAFV600E-mutant PTC organoids. Nevertheless, BRAF inhibitors in combination with MEK inhibitors, RTK inhibitors, or chemotherapeutic agents demonstrated improved efficacy compared to BRAF inhibition alone. CONCLUSIONS: These data indicate that patient-derived PTC organoids may be a powerful research tool to investigate tumor biology and drug responsiveness, thus being useful to validate or discover targeted drug combinations.

Clinical Efficacy of Topical Tacrolimus on Conjunctival Hyperemia Caused by Prostaglandin Analogues.

Objective: To evaluate the clinical efficacy of tacrolimus ophthalmic solution on conjunctival hyperemia caused by prostaglandin analogues. Methods: A retrospective analysis was performed on 120 patients diagnosed with bilateral primary open-angle glaucoma (POAG). The enrolled patients developed symptoms of conjunctival hyperemia during the administration of travoprost ophthalmic solution. The patients were divided into two groups: 0.004% travoprost solution was administered in the control group. A combination of 0.004% travoprost solution with tacrolimus was administered in the experimental group. Clinopathological parameters including intraocular pressure (IOP), subjective dry eye symptom score (SDES), hyperemia score, and noninvasive tear break-up time (NIBUT) were recorded at week 0, 1, 2, and 4. Two-way ANOVA for repeated measurement was employed for statistical analysis using SPSS 22.0 software. Results: At week 1, 2, and 4, the IOP and SDES of both the control and experimental groups were significantly lower when compared the values at week 0 (before treatment). No significant differences in the IOP values between the two groups were observed at all time points. At week 2, the SDES and hyperemia score were lower in the experimental group than those in the control group, and the NIBUT was significantly longer in the experimental group. The above parameters showed no significant difference at week 4 between the two groups, although the average SDES and hyperemia score were slightly lower in the experimental group. Conclusion: Tacrolimus ophthalmic solution can relieve conjunctival hyperemia, improve ocular surface conditions, and reduce discomfort caused by prostaglandin analogues.

Tris(1,3-dichloro-2-propyl) phosphate reduces longevity through a specific microRNA-mediated DAF-16/FoxO in an unconventional insulin/insulin-like growth factor­1 signaling pathway.

Tris(1,3-dichloro-2-propyl) phosphate (TDCPP) has received concerns due to its frequent detection in environmental media and biological samples. Our previous study has indicated TDCPP reduced the lifespan of Caenorhabditis elegans (C. elegans) by triggering an unconventional insulin/insulin-like growth factor signaling (IIS) pathway. This study continued to investigate the possible deleterious effects of TDCPP relating to longevity regulation signal pathways and biological processes. Specifically, this study uniquely performed small RNA transcriptome sequencing (RNA-seq), focusing on the underlying mechanisms of TDCPP-reduced the longevity of C. elegans in-depth in microRNAs (miRNAs). Based on Small RNA-seq results and transcript levels of mRNA involved in the unconventional IIS pathway, a small interaction network of miRNAs-mRNAs following TDCPP exposure in C. elegans was preliminarily established. Among them, up-regulated miR-48 and miR-84 (let-7 family members) silence the mRNA of daf-16 (the crucial member of the FoxO family and pivotal regulator in longevity) via post-transcription and translation dampening abilities, further inhibit its downstream target metallothionein-1 (mtl-1), and ultimately contributed to the reduction of nematode longevity and locomotion behaviors. Meanwhile, the high binding affinities of TDCPP with miRNAs cel-miR-48-5p and cel-miR-84-5p strongly support their participation in the regulation of nematode mobility and longevity. These findings provide a comprehensive analysis of TDCPP-reduced longevity from the perspective of miRNAs.

Impact of ocean waves on transport of underwater spilled oil in the Bohai Sea.

To investigate the impact of ocean waves on transport of underwater spilled oil, a wave action module is developed and embedded into an underwater oil spill model. Combined with a validated hydrodynamic background data, the model is applied to a short-term numerical simulation of the 2011 Penglai 19-3 oil spill event in the Bohai Sea. Numerical result shows that the model result is rationally consistent with the field observation. In addition, waves can influence surface oil slick in both horizontal and vertical directions: in horizontal direction, Stokes drift tends to make oil slick spread more widely under the action of wind-waves; in vertical direction, breaking waves tend to entrain the surface oil into water column, and then affects the vertical distribution of spilled oil. Furthermore, sensitivity study shows that using different parameterizations of wave entrainment can make some difference to the model output, but the effect is limited. Besides, the impact of input wave errors varies on different model outputs. This study is expected to provide offshore oil spill emergency responders and researchers with useful information on oil spill monitoring, emergency plans and environmental damage assessment.

Ameliorative Effect of Parishin C Against Cerebral Ischemia-Induced Brain Tissue Injury by Reducing Oxidative Stress and Inflammatory Responses in Rat Model.

BACKGROUND: Gastrodia elata Blume (Orchidaceae) is a widely used traditional Chinese herbal medicine in the clinical practice of China, to treat nervous headache, convulsions, dizziness, neurasthenia, and so on. Parishin C (Par C), one of the major bioactive components of Gastrodia elata Blume, is known to exert many different biological activities, including antipsychotic and neuroprotective effects. However, there is little research about its neuroprotective effect in an ischemic stroke model. The objective of the present study is thus to investigate the neuroprotective effects of Par C against cerebral ischemia damage. METHODS: Rats were pretreated with Par C (25, 50, or 100 mg/kg/day, i.p.) for 21 days, then subjected to 2 h of middle cerebral artery occlusion (MCAO) and 22 h of reperfusion. Neurological deficient scores, brain water content, histopathology, TCC staining were performed to assess the neuroprotective effects of Par C. Meanwhile, the oxidative stress, inflammation and apoptosis-related markers of brain tissue were evaluated by corresponding assay kits. Besides, the antioxidant and pro-inflammatory expression was measured by real-time quantification PCR (RT-qPCR). RESULTS: Our findings indicated that the pre-treatment with Par C improved nerve function, suppressed oxidative stress, and pro-inflammatory factors release in rats with cerebral ischemia damage. Besides, Par C significantly increased antioxidant expression and declined pro-inflammatory cytokines expression. CONCLUSION: Par C is shown to exert neuroprotective effects partly via inhibiting oxidative stress and inflammation in a rat model of MCAO.

Tris(1,3-dichloro-2-propyl)phosphate Reduces the Lifespan via Activation of an Unconventional Insulin/Insulin-Like Growth Factor-1 Signaling Pathway.

Tris(1,3-dichloro-2-propyl)phosphate (TDCPP) is an environmental contaminant that has attracted increasing concern due to its presence in environmental media and biological samples. Our previous study demonstrated that exposure to TDCPP reduced the lifespan of Caenorhabditis elegans, but the mechanisms, including the relevant signaling pathways, are unclear. The current study found that TDCPP exposure triggers an unconventional insulin/insulin-like growth factor signaling (IIS) pathway, not by disrupting the insulin-like growth factor-1 receptor DAF-2/IGF1R but by inhibiting the downstream tumor-suppressor factor DAF-18/PTEN. This inhibition reduces PI(3,4,5)P3 (PIP3) dephosphorylation, causing buildup that increases the activation of the Akt/Protein Kinase B (PKB) family of serine/threonine kinases. This activation induces DAF-16/FoxO phosphorylation and promotes the sequestration of DAF-16/FoxO in the cytoplasm, reducing the lifespan of nematodes. Our results have important diagnostic and therapeutic implications for controlling TDCPP-related diseases, especially those originating with IIS pathway components.

Huang-Qi San improves glucose and lipid metabolism and exerts protective effects against hepatic steatosis in high fat diet-fed rats.

Numerous researches supported that non-alcoholic fatty liver disease (NAFLD) was an emerging problem associated with increased visceral adiposity (obesity), diabetes and related metabolic disorders. Huang-Qi San (HQS) is composed of three traditional Chinese medicines (Astragali Radix, Pueraria Radix and Cortex Mori Radicis) with a weight ratio of 1:2:1. HQS has been reported to be effective in improving glucose-lipid metabolism, but its underlying mechanism on NAFLD has not been fully understood. The purpose of the present study was to assess the protective effects of HQS on obesity-induced hepatic steatosis in rats fed with high fat diet (HFD). Our data revealed that administration of HQS (1.2 and 2.4 g/kg body weight) resulted in significant reduction in body weight (BW) and organs coefficients of visceral fat. The full-Body CT scan demonstrated that HQS reduced liver fat ratio, visceral and subcutaneous fat mass in a dose-dependent manner. Additionally, HQS decreased plasma TC, TG, FFA and FABP4 levels, normalized glucose and insulin levels, and improved the glucose tolerance. Pathological examination showed that HQS alleviated hepatic steatosis and reduced the cell size of epididymal visceral adipose tissue. Hepatic lipid accumulation was also reduced by HQS treatment compared with HFD fed rats. RNA-Seq analysis combining with qPCR demonstrated that the mRNA expression of some important glucose and lipid metabolism-related genes including Acat2, Apoc4, Bhmt, Cyp3a62, Cyp51, Egln3 (Phd3), Fads1, Fads2, Gnmt, Hmgcs1 and Pemt, were significantly changed by HQS treatment. Taken together, these results suggested that HQS had beneficial effects on glucose-lipid metabolism and hepatic steatosis, and its mechanism might be related to the functions of the genes in regulating glucose and lipid metabolism.

[A controlled study on the therapeutic effect of acupuncture and acupuncture combined with drugs on peripheral facial paralysis with normal result of facial nerve magnetic resonance examination].

OBJECTIVE: To explore whether a combination of drugs is needed when acupuncture is used to treat peripheral facial paralysis with normal result of facial nerve magnetic resonance examination. METHODS: A total of 48 normal facial nerve magnetic resonance examination of peripheral facial paralysis patients were randomly divided into an acupuncture combined with drugs group and an acupuncture group, 24 cases in each group. Acupuncture combined with prednisone were applied in the acupuncture combined with drugs group, and simple acupuncture was given in the acupuncture group. The acupuncture treatment methods were the same. Acupuncture mainly applied at Yifeng (TE 17), Quanliao (SI 18), Taiyang (EX-HN 5), Sibai (ST 2), Jiache (ST 6), Dicang (ST 4) on the affected side, Hegu (LI 4, opposite side) for 30 min a day, once every other day. Electroacupuncture was added at Yifeng (TE 17) and Quanliao (SI 18), Jiache (ST 6) and Dicang (ST 4) on the affected side after the acute phase until it was cured. Prednisone was taken orally once a day, with an initial dose of 30 mg, decrease 10 mg every 3 days for 9 days. After 3 and 8 weeks of treatment, the effect was observed by Portmann score. RESULTS: After 3 and 8 weeks of treatment, the cured rates were 50.0% (12/24) and 83.3% (20/24) in the acupuncture group, which were slightly lower than 58.3% (14/24) and 87.5% (21/24) in the acupuncture combined with drugs group，there was no significant difference between the two groups (both P>0.05); the course of treatment in the acupuncture group was (31.2±17.0) d, slightly longer than (29.5±12.8) d in the acupuncture combined with drugs group , but there was no significant difference between the two groups (P>0.05). CONCLUSION: Simple acupuncture treatment on peripheral facial paralysis with normal result of facial nerve magnetic resonance examination can achieve the same effect of acupuncture combined with drugs, and avoid drug-related adverse reactions.

Comparison of pharmacokinetics and drug release in tissues after transarterial chemoembolization with doxorubicin using diverse lipiodol emulsions and CalliSpheres Beads in rabbit livers.

CalliSpheres® Beads (CB) is the first drug-eluting bead (DEB) product in China. Our aim was to compare the effect on the pharmacokinetics of doxorubicin (DOX) and its local concentration between lipiodol emulsions and CB in the process of TACE in rabbit livers. Twenty-five rabbits were distributed into two groups; Group 1 received lipiodol emulsions with DOX, and Group 2 received CB loaded with DOX (CBDOX). DOX was measured in the peripheral blood at different times after treatment. Livers were sampled at 1 week and 1 month for Group 2 after embolization. DOX concentration and distribution were measured in the liver. The administration of DOX by TACE with CBDOX resulted in peripheral blood DOX concentrations of 39.85 ± 13.86 ng/mL at 5 min, with a gradual decrease to 6.89 ± 1.62 ng/mL at 24 h, after treatment. Plasma concentration of DOX after chemoembolization with lipiodol was 225.91 ± 64.88 ng/mL at 5 min and decreased with time by 24 h to 5.06 ± 0.48 ng/mL. In CBDOX group, the drug impregnated an area as far as 200 µm from the bead edge. The tissue concentration of doxorubicin (tissCDOX) ranged from 40.27 µg/mL to 245.70 µg/mL at 1 week and from 5.64 µg/mL to 28.09 µg/mL at 1 month. Plasma concentrations of DOX resulting from CBDOX embolization were significantly lower than that for cTACE. CB could deliver relatively high concentrations of DOX to an area as far as 200 µm from the bead edge for at least 1 month.

Preparative separation and purification of two highly polar alkaloids derived from Semen Strychni extracted with dichloromethane by high-speed countercurrent chromatography.

Brucine chloromethochloride and strychnine chloromethochloride, the two chloromethochloride derivatives formed during the extraction of Semen Strychni in which dichloromethane was used as the extracting solvent, were isolated and purified by high-speed countercurrent chromatography for the first time. The two-phase solvent system composed of chloroform/methanol/0.3 mol/L hydrochloric acid (4:3:2, v/v/v) was selected for separation. From 300 mg of the crude extracts, 56.2 mg of brucine chloromethochloride and 60.2 mg of strychnine chloromethochloride were obtained with the purity of 99.78 and 96.99%, respectively, and the structures were confirmed by mass spectrometry, 1 H, 13 C, and two-dimensional NMR spectroscopy. The results indicated that the present method is a powerful technology for large-scale isolation of alkaloids from Semen Strychni.

[The effective parts of liangxue tongyu prescription on cooling-blood and activating-blood and analysis of chemical constituents by HPLC-MS and GC-MS].

In order to clarify material basis of effective parts of liangxue tongyu prescription, blood-heat and blood-stasis rat model induced by dry yeast was established. The changes of rectal temperature, blood viscosity and plasma viscosity were used to evaluate the cooling-blood and activating-blood effects of liangxue tongyu prescription and its parts. Compared with the model group, the extract from liangxue tongyu prescription, its volatile oil and n-butanol part could significantly reduce rectal temperature (P<0.01), and also reduce blood viscosity and plasma viscosity to various degrees (P<0.01 or P<0.05). So volatile oil and n-butanol part were primarily identified as effective parts of liangxue tongyu prescription. By using GC-MS with normalization method of area to analyze volatile oil of liangxue tongyu prescription, 70 compounds were identified, accounting for about 92.54%, mainly as ß-asarone, paeonol, α-asarone and shyobunone. 42 compounds such as peony glycosides, tannins, and iridoid glycosides were identified by HPLC-MS techniques and standard comparison. The study determined the effective parts of liangxue tongyu prescription and clarified the chemical composition providing the foundation for further studies on material basis of liangxue tongyu prescription.

Effect of DGPS failures on dynamic positioning of mobile drilling units in the North Sea.

Basic features of differential global positioning system (DGPS), and its operational configuration on dynamically positioned (DP) mobile offshore drilling units in the North Sea are described. Generic failure modes of DGPS are discussed, and a critical DGPS failure which has the potential to cause drive-off for mobile drilling units is identified. It is the simultaneous erroneous position data from two DGPS's. Barrier method is used to analyze this critical DGPS failure. Barrier elements to prevent this failure are identified. Deficiencies of each barrier element are revealed based on the incidents and operational experiences in the North Sea. Recommendations to strengthen these barrier elements, i.e. to prevent erroneous position data from DGPS, are proposed. These recommendations contribute to the safety of DP operations of mobile offshore drilling units.