穗花杉双黄酮对3T3-L1细胞增殖、凋亡及生物钟的影响

目的探讨穗花杉双黄酮对3T3-L1细胞增殖、凋亡及生物钟的影响。方法采用不同浓度穗花杉双黄酮处理正常和过表达昼夜节律运动输出周期故障蛋白(CLOCK)的3T3-L1细胞,CCK-8和EdU检测细胞增殖情况;流式细胞仪检测细胞凋亡情况;real-timeRT-PCR检测生物钟基因CLOCK和BMAL1的表达;Westernblotting检测Cleavedcaspase-3、Bcl-2、CLOCK及BMAL1蛋白的表达。结果低浓度(2.5~10.0mg/L)穗花杉双黄酮作用于3T3-L1细胞后,3T3-L1细胞增殖和凋亡无影响(P>0.05),但是当浓度达20和40mg/L反而抑制增殖(P<0.05);通过上调Cleavedcaspase-3表达,下调Bcl-2表达,促进凋亡的发生。较低浓度穗花杉双黄酮对3T3-L1细胞CLOCK、BMAL1基因和蛋白的表达无影响(P>0.05),而浓度达20和40mg/L则呈抑制作用(P<0.05);过表达CLOCK基因可减弱穗花杉双黄酮对3T3-L1细胞的抑制作用(P<0.05)。结论3T3-L1细胞在培养状态仍保持昼夜节律性,较高浓度穗花杉双黄酮可抑制3T3-L1细胞增殖并促进凋亡,其可作为抑制细胞增殖的药物,其作用机制与生物钟基因表达有关。

Image-guidedcryoablation in unresectable or recurrent advanced colorectal cancer:a retrospective study

Purpose: The present study retrospectively analyzed thesafety and efficacy of computed tomography(CT)-guided cryoablationin the treatment ofunresectable or recurrent advanced colorectal cancer, which did not respond well to or experienced progression with radiotherapy or chemotherapy. Materials and Methods: From January 2013 to April 2015, 31 lesions in 27 patients(16 males, 11 females; mean age of 57.2 years) with pelvic unresectableadvanced or recurrent colorectal cancer were included in the study.The tumor diameter was approximately 3.37 ±1.41 cm. The primary tumor included 25 rectal cancers, 1 sigmoid colon adenocarcinoma, and 1 ileocecal mucinous adenocarcinoma. Cryoablation was performed with 17-gauge cryoprobes and monitored by 64-slice spiral CT. Follow-up was carried out by contrast-enhanced magnetic resonance imaging(MRI). The treatment efficacy was evaluated by symptom palliation, decreased carcinoembryonic antigen(CEA) serum level, and tumor response. Results: The cryoablation procedure was well-tolerated in all patients without major complications or procedure-related mortality. Long-term complications included abscess formation(1 patient), skin frostbite and post-sacrum antrum formation(1 patient). Pain relief was satisfactory in patients with perineal pain(P<0.001), and the median time of pain relief was 3.0 months. Complete ablations were obtained in 22 lesions of 18 patients, while 9 lesions in 9 patients underwent incomplete ablation. The median time to local recurrence for lesions with complete ablations was 15.0 months, and that to the progression of tumors with incomplete ablation was 4.0 months. Conclusion: CT-guided cryoablation is a minimally invasive, safe, and effective therapeutic option for unresectableadvanced or recurrent colorectal cancer. The treatment is well-tolerated by patients, and pain relief is achieved rapidly.

Successful treatment of an enormous rectal mucosa-associated lymphoid tissue lymphoma by endoscopic full-thickness resection:A case report

BACKGROUND Colorectal mucosa-associated lymphoid tissue lymphoma(MALToma),a rare kind of nongastric MALToma,lacks consensus on its endoscopic features and standard therapies.According to previous studies on the clinical characteristics and outcomes of colorectal MALToma,endoscopic resection remains a good therapeutic strategy.CASE SUMMARY A 71-year-old woman suffered intermittent hematochezia for 1 mo,accompanied with abdominal pains but without weight loss,fever,chills or fatigue.Colonoscopy showed a massive hemispheric mass with rough and hyperemic mucosa in the lower rectum.Narrow-band imaging magnifying endoscopy detected some branching abnormal blood vessels and disappearance of glandular structure,which was similar with the tree-like appearance sign in gastric MALToma.Endoscopic ultrasonography revealed the lesion to be hypoechoic,boundary-defined,and echo uniform inside,originating from the muscularis propria.Abdominal enhanced computed tomography(CT)demonstrated a soft tissue mass with defined boundary.No enlarged superficial lymph nodes were detected by B-mode ultrasound.C13-urea breath test and serum Helicobacter pylori antibody were both negative.The patient underwent endoscopic full-thickness resection.Postoperative pathological analysis indicated colorectal MALToma.The patient remained asymptomatic after discharge,and follow-up positron emission tomography–CT and colonoscopy showed no residual lesion,remnants or lymph node metastasis.CONCLUSION This case provides new information on the specific endoscopic features of colorectal MALToma and an alternative treatment for patients.

Core-shell nanoparticle enhanced Raman spectroscopy in situ probing the composition and evolution of interfacial species on PtCo surfaces

The composition and evolution of interfacial species play a key role during electrocatalytic process.Unveiling the structural evolution and intermediate during catalytic process by in situ characterization can shed new light on the electrocatalytic reaction mechanism and develop highly efficient catalyst.However,directly probing the interfacial species is extremely difficult for most spectroscopic techniques due to complicated interfacial environment and ultra-low surface concentration.Herein,electrochemical core-shell nanoparticle enhanced Raman spectroscopy is utilized to probe the composition and evolution processes of interfacial species on Au@Pt,Au@Co,and Au@PtCo core-shell nanoparticle surfaces.The spectral evidences of interfacial intermediates including hydroxide radical(OH*),superoxide ion(O_(2)^(−)),as well as metal oxide species are directly captured by in situ Raman spectroscopy,which are further confirmed by the both isotopic experiment and density functional theory calculation.These results provide a mechanistic guideline for the rational design of highly efficient electrocatalysts.

In situ tracking of the lithiation and sodiation process of disodium terephthalate as anodes for rechargeable batteries by Raman spectroscopy

Organic compounds represent an appealing group of electrode materials for rechargeable batteries due to their merits of biomass,sustainability,environmental friendliness,and processability.Disodium terephthalate(Na_(2)C_(8)H_(4)O_(4),Na_(2)TP),an organic salt with a theoretical capacity of 255 mAh·g^(-1),is electroactive towards both lithium and sodium.However,its electrochemical energy storage(EES)process has not been directly observed via in situ characterization techniques and the underlying mechanisms are still under debate.Herein,in situ Raman spectroscopy was employed to track the de/lithiation and de/sodiation processes of Na2TP.The appearance and then disappearance of the–COOLi Raman band at 1625 cm^(-1) during the de/lithiation,and the increase and then decrease of the–COONa Raman band at 1615 cm^(-1) during the de/sodiation processes of Na2TP elucidate the one-step with the 2Li+or 2Na+transfer mechanism.We also found that the inferior cycling stability of Na2TP as an anode for sodium-ion batteries(SIBs)than lithium-ion batteries(LIBs)could be due to the larger ion radium of Na+than Li+,which results in larger steric resistance and polarization during EES.The Na2TP,therefore,shows greater changes in spectra during de/sodiation than de/lithiation.We expect that our findings could provide a reference for the rational design of organic compounds for EES.

The Masc gene product controls masculinization in the black cutworm, Agrotis ipsilon

Sex determination has been studied in the model lepidopteran species Bombyx mori, but it remains poorly understood in lepidopteran pests. In the present study, we identified and characterized the Masculinizer (Masc) gene in a Noctuidae pest species, Agrotis ipsilon. Sequence analysis revealed that AiMasc encodes a protein of 658 amino acids that has two CCCH-type zinc finger domains and two conserved cysteine residues (Cys-277 and Cys-280). We assessed the masculinizing activity AiMasc in BmN cells and found that/z'Masc induced expression of the male-specific doublesex isoform. Disruption of Masc via clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) in A. ipsilon caused abnormalities in abdominal segments and external genitalia, resulting in male-specific sterility. These results suggest that Masc participates in the process of sex determination in A. ipsilon. Successful identification of sex-determination gene in a pest species may enable the development of novel genetic approaches for pest control.

CRISPR/Cas9-mediated Tyrosine hydroxylase knockout resulting in larval lethality in Agrotis ipsilon

Tyrosine hydroxylase (TH)is involved in insect melanin and the catecholamine biosynthesis pathway.TH as an enzyme catalyzing the conversion of tyrosine to 3,4- dihydroxyphenylalanine is the first step reaction in the pathway.Although TH has been proven to affect the pigmentation of the epidermis and development in many insects,there is no report about physiological function of the THgene inAgrotis ipsilon.Here we cloned the TH gene from A.ipsilon.Semi-quantitative real-time polymerase chain reaction (PCR) analysis showed that AiTH was expressed at all development stages.Moreover,its high expression levels in the head and epidermis suggest that it is mainly related to pigment deposition and insect development.Then,we used the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9system to target the AiTH gene:deletion events were detected at the target sites.Compared with the control group,a few mutants with the phenomenon of narrowing in the egg shell and embryos can develop but cannot hatch;the other hatched embryos were seriously dehydrated after hatching and died within the first day.Quantitative real-time PCR analysis revealed that THwas down-regulated inAiTHmutants.Here,our work demonstrated thatAiTHplays an important role in growth and development of newly hatched larvae;meanwhile,it would be a promising target to explore a control strategy for A.ipsilon.

CRISPR/Cas9-based mutation reveals Argonaute 1 is essential for pigmentation in Ostrinia fumacalis

Ostrinia fumacalis (Lepidoptera: Pyralidae) is one of the most destructive agricultural pests in Asia. Traditional pest-management methods include sex pheromone capture, transgenic crops that produce Bacillus thuringiensis toxin, and pesticides. Although these strategies control pest populations effectively, they also cause negative side effects, including dramatically increased pesticide resistance, severe pollution, and hazards for human health. Recently developed genome editing tools provide new prospects for pest management and have been successfully used in several species. However, few examples have been reported in the agricultural pest O. fumacalis due to a lack in genomic information. In this report, we identified only one transcript of O. fumacalis Argonaute 1 (OfAgo 1) gene from the genome and cloned the open reading frame. OfAgol presented the maximum expression at the embryo stage or in the fat body during the larval stages. To understand its function, an OfAgol mutant was constructed using the Clustered Regularly Interspaced Short Palindromic Repeat/RNA-guided Cas9 nuclease (CRISPR/Cas9). Mutagenesis of OfAgol disrupted cuticle pigmentation by dowregulating micro RNAs and pigmentation-related genes. This is the first report for the cloning and functional analysis of OfAgol, revealing a role of OfAgol in cuticle pigmentation. The current report also established a CRISPR/Cas9 system in O. fumacalisy providing a new insight for pest management.

Disruption of egg-specific protein causes female sterility in Bombyx mori

Yolk proteins are the main source of nutrients during embryonic and early larval development in oviparous animals. Therefore, vitellogenesis is crucial for reproduction. The silkworm, Bombyx mori, is a model lepidopteran insect in which there are three yolk proteins: vitellin, 30-kDa protein, and egg-specific protein (Esp). In this study, we explored the gene function of Esp through transgenic clustered regularly interspaced palindromic repeats (CRISPR) / CRISPR-associated protein 9 technology-mediated mutations in the silkworm. We found that Esp mutation resulted in female sterility but had no effect on male fertility. Female mutants could lay eggs after mating, but the eggs were smaller and lighter colored than those laid by wild-type females. The most important finding is that the eggs laid by female mutants did not hatch. Furthermore, we observed stable inheritance of female sterility caused by Esp mutation through successive generations. Thus, Esp encodes a yolk protein that is crucial for female reproductive success and is a potential target for pest control.