First report on establishment and characterization of the extrahepatic cholangiocarcinoma sarcoma cell line CBC2T-2.

BACKGROUND: Extrahepatic cholangiocarcinoma sarcoma is extremely rare in clinical practice. These cells consist of both epithelial and mesenchymal cells. Patient-derived cell lines that maintain tumor characteristics are valuable tools for studying the molecular mechanisms associated with carcinosarcoma. However, cholangiocarcinoma sarcoma cell lines are not available in cell banks. AIM: To establish and characterize a new extrahepatic cholangiocarcinoma sarcoma cell line, namely CBC2T-2. METHODS: We conducted a short tandem repeat (STR) test to confirm the identity of the CBC2T-2 cell line. Furthermore, we assessed the migratory and invasive properties of the cells and performed clonogenicity assay to evaluate the ability of individual cells to form colonies. The tumorigenic potential of CBC2T-2 cells was tested in vivo using non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. The cells were injected subcutaneously and tumor formation was observed. In addition, immunohistochemical analysis was carried out to examine the expression of epithelial marker CK19 and mesenchymal marker vimentin in both CBC2T-2 cells and xenografts. The CBC2T-2 cell line was used to screen the potential therapeutic effects of various clinical agents in patients with cholangiocarcinoma sarcoma. Lastly, whole-exome sequencing was performed to identify genetic alterations and screen for somatic mutations in the CBC2T-2 cell line. RESULTS: The STR test showed that there was no cross-contamination and the results were identical to those of the original tissue. The cells showed round or oval-shaped epithelioid cells and mesenchymal cells with spindle-shaped or elongated morphology. The cells exhibited a high proliferation ratio with a doubling time of 47.11 h. This cell line has migratory, invasive, and clonogenic abilities. The chromosomes in the CBC2T-2 cells were polyploidy, with numbers ranging from 69 to 79. The subcutaneous tumorigenic assay confirmed the in vivo tumorigenic ability of CBC2T-2 cells in NOD/SCID mice. CBC2T-2 cells and xenografts were positive for both the epithelial marker, CK19, and the mesenchymal marker, vimentin. These results suggest that CBC2T-2 cells may have both epithelial and mesenchymal characteristics. The cells were also used to screen clinical agents in patients with cholangiocarcinoma sarcoma, and a combination of paclitaxel and gemcitabine was found to be the most effective treatment option. CONCLUSION: We established the first human cholangiocarcinoma sarcoma cell line, CBC2T-2, with stable biogenetic traits. This cell line, as a research model, has a high clinical value and would facilitate the understanding of the pathogenesis of cholangiocarcinoma sarcoma.

[In-situ Remediation Effect of Cadmium-polluted Agriculture Land Using Different Amendments Under Rice-wheat Rotation].

Exploring the effects of one-time amendment treatments on cadmium (Cd)-contaminated farmland soils is beneficial for providing a theoretical basis to effectively prevent Cd pollution in farmland soils and ensure the safe production of crops. Five amendments, including straw biochar, fly ash, sepiolite, white marble powder, and shale (particle size <0.2 mm, application rate 2.25 kg·m-2), were applied to the Cd-contaminated farmland soils. The soil nutrients, pH, soil available Cd, and Cd chemical forms in the soils and grain Cd concentration in the planted crops were determined to investigate the effects and persistence of one-time applications of the five amendments. The results showed that:① the application of the five amendments had little effect on soil nutrient content, but all of them could increase soil pH. Amendment treatments improved the transfer of Cd from the acid extraction fraction to residue fraction and further reduced the Cd availability in the soil. The decreasing amplitudes of straw biochar and white marble powder soil conditioner were 20.42%-22.53%, which was higher than those in the other treatments. ② The grain Cd concentrations in rice and wheat were significantly decreased under the amendment treatments with the decreasing amplitudes of 19.88%-48.77% and 5.06%-24.00%, respectively. The Cd concentrations in rice grains under the treatments of straw biochar, fly ash, and white marble powder soil conditioner were 0.195, 0.196, and 0.223 mg·kg-1, respectively, which were lower than those under the other treatments and were close to or approached the National Standard of Food Safety(GB 2762-2017)(0.2 mg·kg-1). ③ The immobilization effects on Cd in farmland soils were decreasing with time under one-time application of the amendments. The available Cd concentrations in the soil and Cd concentrations in crop grains were still lower than those in the control after three rounds of rice-wheat rotation. The straw biochar and white marble powder soil conditioner had a good and long-term effect on reducing Cd availability in soils and Cd concentrations in crop grain, making them ideal materials for safe production in Cd-contaminated soils.

[Stabilization Characteristics of Exogenous Cd in Different Types of Soil].

To clarify the characteristics of stabilization and availability of exogenous cadmium (Cd) in different types of soils, six main agricultural soils in Sichuan province, including acidic purple soil, neutral purple soil, calcareous purple soil, gray fluvo-aquic soil, typical yellow soil, and rinsed yellow soil, were used in this study. A soil culture experiment was conducted to explore the differences in stabilization time, chemical form, and effective Cd content between the six types of soils. Additionally, the effects of exogenous Cd on the growth and Cd accumulation of cabbage growing in different soils was discussed. The results showed the following: 1 with exogenous Cd treatment, the available Cd content of the six soils decreased sharply within 15 d after Cd addition and then tended to be flat. After 30 d, there was no significant difference. The available Cd contents of rinsed yellow soil and acidic purple soil were significantly higher than that of the other four soils, and the content of calcareous purple soil was significantly lower (1.01 mg·kg-1). 2 After stabilization, exchangeable Cd was the main Cd form in the six soils, up to 42.51%-56.07%. The relative proportions of other Cd forms differed greatly between different soils. The proportion of iron and manganese oxides in the rinsed yellow soil and typical yellow soil was higher, whereas organic complex Cd, residual Cd, and carbonate-bound Cd were higher in the gray fluvo-aquic soil, neutral purple soil, and calcareous purple soil, respectively. 3 The characteristics of availability and bioavailability of Cd among different soils were significantly different with different exogenous Cd doses. With the lowest Cd dose (0.5 mg·kg-1), compared with that in the control, there was no significant decrease in Cd content in the edible parts of the cabbage, but Cd accumulation in the edible parts differed. The Cd contents of edible parts of the cabbage growing in typical yellow soil and rinsed yellow soil were significantly higher than that of the other soils. With the highest Cd dose (2.0 mg·kg-1), the growths of cabbage growing in all six soils were significantly inhibited. The available Cd contents of rinsed yellow soil and acidic purple soil were significantly higher than that of the other four soils, which is consistent with the Cd content in the edible parts of the cabbage. The available Cd contents and the Cd content in the edible parts of the cabbage in calcareous purple soil were the lowest, and acidic soils, especially in rinsed yellow soil, had the highest.

The inflammatory cytokine profiles and ocular biometric characteristics of primary angle-closure glaucoma.

OBJECTIVE: To investigate the pathogenesis of primary angle-closure disease (PACG) by measuring the anatomical structures of the anterior and posterior segments of the eye and inflammatory markers in the peripheral blood. METHODS: This case-control study enrolled patients diagnosed with acute PACG (APACG) and chronic PACG (CPACG). It also enrolled control subjects without PACG. The anterior and posterior anatomical features were measured in all study participants. The levels of interleukin (IL)-6, tumour necrosis factor-α and the neutrophil-to-lymphocyte ratio (NLR) in the peripheral blood were measured. RESULTS: This study analysed a total of 99 eyes: 34 eyes from 34 patients with APACG, 28 eyes from 28 patients with CPACG and 37 eyes from 37 control patients with senile cataract. The axis length, corneal diameter, anterior chamber depth and anterior chamber volume were significantly smaller in the APACG and CPACG groups compared with the controls. The level of IL-6 in the peripheral blood of patients with PACG was significantly lower than that of the controls. The NLR in the peripheral blood of patients with PACG was significantly greater than that of the controls. CONCLUSIONS: Changes in the ocular anatomy and some inflammatory markers might be involved in the pathogenesis of PACG.

[Effects of Different Amendments on Cadmium Accumulation in Rice Safety in Cadmium-Contaminated Farmland Under Two Flooding Treatments].

Cadmium (Cd) accumulation in rice has become a serious public concern; thus, it is important to find an effective approach to reducing Cd accumulation in rice grains to ensure food safety. To investigate the effects of different amendments on Cd accumulation in rice in Cd-contaminated farmland under different flooding treatments, a field experiment with three amendments (jade powder, biochar, and fly ash) and two flooding treatments (intermittent flooding and flooding throughout the whole growth period) was conducted. The results showed that:① without amendment application, the soil pH significantly increased, whereas the soil available Cd concentration decreased by 3.81%-17.27% and 2.25%-6.74% with the treatments of flooding throughout the whole growth period and intermittent flooding, respectively. Additionally, the immobilizing efficiency of the treatment of flooding throughout the whole growth period was better than that of intermittent flooding; ② under different flooding treatments, amendment application improved soil pH, resulting in a decrease in the soil available Cd concentration along with an increase in the residual Cd concentration. Under the treatment of intermittent flooding, the soil pH increased by 0.19-2.20 units, and the soil available Cd concentration and Cd concentration in rice grains decreased by 4.72%-22.68% and 2.60%-75.75%, respectively, with the application of different amendments. Under the treatment of flooding throughout the whole growth period, the application of different amendments decreased the soil available Cd concentration and Cd concentration in rice grains by 5.06%-36.63% and 13.28%-77.01%, respectively. The immobilizing efficiency in both flooding treatments was jade powder > biochar > fly ash. ③ Under different flooding treatments, the application of amendments significantly reduced the soil available Cd concentration and Cd concentration in rice grains. Among the three amendments, jade powder showed the best capacity of immobilizing efficiency with the treatment of flooding throughout the whole growth period; the soil Cd reduction rates were 36.63% and 25.16%, and the Cd concentrations in rice grains were 0.058 and 0.170 mg·kg-1 in 2019 and 2020, respectively. The Cd concentrations in rice grains were within the limitation of the National Food Hygienic Standard of China. Therefore, combining flooding throughout the whole growth period with jade powder can be considered as an ideal strategy for ensuring rice safety in Cd-contaminated farmland.

Early Life Antibiotic Exposure and the Subsequent Risk of Autism Spectrum Disorder and Attention Deficit Hyperactivity Disorder: A Systematic Review and Meta-Analysis.

This study was conducted to assess this association between early life antibiotic exposure and the risk of autism spectrum disorder (ASD) and attention deficit hyperactivity disorder (ADHD) in later life. The results showed that early life antibiotic exposure was associated with an increased risk of ASD (OR = 1.13, 95% confidence interval (CI): 1.07-1.21) or ADHD (OR = 1.18, 95% CI: 1.1-1.27). However, this association for ASD (OR = 1.04, 95% CI: 0.97-1.11) or ADHD (OR = 0.98, 95% CI: 0.94-1.02) disappeared when data from sibling-matched studies were pooled. The statistically significant association between early life antibiotic exposure and ASD or ADHD in later life can be partially explained by unmeasured genetic and familial confounding factors.

A fatal case of Exophiala dermatitidis meningoencephalitis in an immunocompetent host: A case report and literature review.

BACKGROUND: Central nervous system (CNS) infection due to Exophiala dermatitidis is rare and fatal, and primarily reported in immunocompromised patients or those with caspase recruitment domain-containing protein 9 deficiency. Herein, we describe a case of an otherwise healthy person (without underlying disease or gene deficiency) diagnosed with Exophiala dermatitidis meningoencephalitis. The patient achieved clinical remission under high-dose antifungal therapy in the first 14 months but died after 2 years of the therapy. CASE PRESENTATION: A 15-year-old student with headache and fever was admitted to our department. Lumbar puncture showed increased cerebrospinal fluid (CSF) pressure, moderately high CSF protein levels and cell counts, and a remarkable decrease in CSF glucose and chloride. Magnetic resonance imaging of the brain revealed multiple lesions and cerebral pia mater enhancement. CSF culture confirmed E. dermatitidis infection. We administered 4-week antifungal therapy of amphotericin B, but his CSF culture remained positive. After receiving the 12-week standard dose of voriconazole (200 mg q12h), the patient's CSF culture became negative, but his condition deteriorated with intracranial lesion enlargement. We administered a high-dose voriconazole therapy (600-800 mg per day) for 12 months, which led to clinical remission. The voriconazole dose was reduced due to adverse effects including hepatic dysfunction and hypokalemia, and the disease progressed with high intracranial pressure and epileptic seizures. CONCLUSIONS: CNS infection caused by E. dermatitidis is fatal and the most serious form of fungal infection. Initially, high-dose and long-term antifungal therapy could be effective. Gene defect and related antifungal immunodeficiency may be the most important pathogenic and lethal factor.

Enlightening the taxonomy darkness of human gut microbiomes with a cultured biobank.

BACKGROUND: In gut microbiome studies, the cultured gut microbial resource plays essential roles, such as helping to unravel gut microbial functions and host-microbe interactions. Although several major studies have been performed to elucidate the cultured human gut microbiota, up to 70% of the Unified Human Gastrointestinal Genome species have not been cultured to date. Large-scale gut microbial isolation and identification as well as availability to the public are imperative for gut microbial studies and further characterizing human gut microbial functions. RESULTS: In this study, we constructed a human Gut Microbial Biobank (hGMB; homepage: hgmb.nmdc.cn ) through the cultivation of 10,558 isolates from 31 sample mixtures of 239 fresh fecal samples from healthy Chinese volunteers, and deposited 1170 strains representing 400 different species in culture collections of the International Depository Authority for long-term preservation and public access worldwide. Following the rules of the International Code of Nomenclature of Prokaryotes, 102 new species were characterized and denominated, while 28 new genera and 3 new families were proposed. hGMB represented over 80% of the common and dominant human gut microbial genera and species characterized from global human gut 16S rRNA gene amplicon data (n = 11,647) and cultured 24 "most-wanted" and "medium priority" taxa proposed by the Human Microbiome Project. We in total sequenced 115 genomes representing 102 novel taxa and 13 previously known species. Further in silico analysis revealed that the newly sequenced hGMB genomes represented 22 previously uncultured species in the Unified Human Gastrointestinal Genome (UHGG) and contributed 24 representatives of potentially "dark taxa" that had not been discovered by UHGG. The nonredundant gene catalogs generated from the hGMB genomes covered over 50% of the functionally known genes (KEGG orthologs) in the largest global human gut gene catalogs and approximately 10% of the "most wanted" functionally unknown proteins in the FUnkFams database. CONCLUSIONS: A publicly accessible human Gut Microbial Biobank (hGMB) was established that contained 1170 strains and represents 400 human gut microbial species. hGMB expands the gut microbial resources and genomic repository by adding 102 novel species, 28 new genera, 3 new families, and 115 new genomes of human gut microbes. Video abstract.

A case report of pyopneumopericardium following bungee jumping in a patient with tuberculosis.

RATIONALE: Pyopneumopericardium related to bungee jumping is a rare occurrence in the current antibiotic era. We present a case of esophagus-seeded Streptococcus sanguinis pyopneumopericardium in a young man with tuberculosis who had just completed bungee jumping. PATIENT CONCERN: A 27-year-old man was hospitalized with a 1-day history of fever, chest tightness, and intermittent sharp chest pain after bungee jumping for the first time. DIAGNOSES: Clinical examinations, thoracentesis, and pericardiocentesis revealed pyopneumopericardium, pyopneumomediastinum, and suppurative pleurisy secondary to bungee-jumping-related traumas. Pericardial fluid cultures were positive for S sanguinis, and Mycobacterium tuberculosis complex genetic test was positive in both sputum and pleural effusion. INTERVENTIONS: The patient improved with drainage and comprehensive antimicrobial therapy. OUTCOMES: The patient developed constrictive pericarditis and underwent pericardiectomy after 6 months of anti-tuberculosis treatment. During the 6-month follow-up after surgery, he recovered uneventfully. LESSONS: This case adds to the long list of bungee-jumping complications. Early diagnosis to initiate appropriate therapy is critical for pyopneumopericardium patients to achieve good outcomes.

The Mouse Gut Microbial Biobank expands the coverage of cultured bacteria.

Mice are widely used as experimental models for gut microbiome (GM) studies, yet the majority of mouse GM members remain uncharacterized. Here, we report the construction of a mouse gut microbial biobank (mGMB) that contains 126 species, represented by 244 strains that have been deposited in the China General Microorganism Culture Collection. We sequence and phenotypically characterize 77 potential new species and propose their nomenclatures. The mGMB includes 22 and 17 species that are significantly enriched in ob/ob and wild-type C57BL/6J mouse cecal samples, respectively. The genomes of the 126 species in the mGMB cover 52% of the metagenomic nonredundant gene catalog (sequence identity ≥ 60%) and represent 93-95% of the KEGG-Orthology-annotated functions of the sampled mouse GMs. The microbial and genome data assembled in the mGMB enlarges the taxonomic characterization of mouse GMs and represents a useful resource for studies of host-microbe interactions and of GM functions associated with host health and diseases.

[Advances of sequencing and assembling technologies for complex genomes].

Complex genomes are noted to be extremely difficult to sequence or assemble by using ordinary methods. Complex genomes are typically characterized as being highly repetitive, highly heterozygous, extremely GC biased, or naturally contaminated, i.e., contaminations which cannot be removed before sequencing. To solve these problems with sequencing and assembling complex genomes, three major techniques include: (1) DNA extraction experiments, (2) Sequencing technologies and platforms, and (3) Algorithms and strategies for assembling. In this review, we summarize these state-of-the-art technologies and strategies used in these directions. We also review the representative projects of complex genome sequencing and address the development of these technologies and strategies for solving the challenges when sequencing or assembling complex genomes.

Patients with chronic hepatitis C receiving sofosbuvir and ribavirin-based treatment, with or without interferon in Zhejiang, China: An observational study.

Hepatitis C virus (HCV) is one of the most important virus as the cause of liver disease in China. The aim of the present study was to explore whether sofosbuvir and ribavirin-based treatment can cure patients with chronic hepatitis C in eastern China. We examined a cohort of HCV-monoinfected patients and 9 patients agreed to participate in our treatment and research. The patients were diagnosed with chronic hepatitis C with or without cirrhosis. Nine patients including 4 female and 5 male met the requirements for selection and were willing to participate in this experiment. Sofosbuvir and ribavirin-based treatment with or without interferon was given to the patients. Viral loads, cytokines, and chemokines were recorded during treatment and after treatment. After 2 weeks of sofosbuvir and ribavirin-based treatment, the viral load of patients decreased to limits of detection. Eight patients were cured. Patients had rapid virological response (RVR) with undetectable viral load at week 4 and sustained virological response (SVR). The interferon-inducible protein-10 (IP-10) decreased after the treatment. However, the patient with cirrhosis failed, as the virus reappeared during SVR4. At the same time, the IP-10 dramatically increased as the relapse of the HCV virus. In summary, the IP-10 has the potential to be the biomarker for the prognostic of HCV.

Simultaneous analysis of 23 illegal adulterated aphrodisiac chemical ingredients in health foods and Chinese traditional patent medicines by ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.

This paper presents an application of ultra high-performance liquid-chromatography-quadrupole-TOF high resolution mass spectrometry (UHPLC-Q-TOF HRMS) for simultaneous analysis of 23 illegal adulterated aphrodisiac type chemical ingredients in health foods and Chinese Traditional Patent Medicines (CTPMs). The mass spectrometer was operated in Information Dependent Acquisition (IDA) mode, which provides crucial information for the elemental composition analysis, structure elucidation and quantitative analysis simultaneously. Quantitative analysis was performed using the peak areas of the precursor ions in the XICs. The method validation included assessment of selectivity, sensitivity, calibration curve, accuracy, precision, recovery, matrix effect and stability. The results show good linear relationship with the concentrations of the analytes over wide concentration ranges (e.g., 0.05-10 µg/g for sildenafil) as all the fitting coefficients of determination r2 are >0.9984. The detection limits (LODs) were in the range of 0.002-0.1 µg/g. The recoveries were able to reach 82.5-103.6%, while the matrix effects ranged from 87.7 to 109.3%. The intra- and inter-day accuracies were in the range of 82.3-113.8%, while the intra- and inter-day precision ranged from 0.4 to 13.6%. Among 40 batches of health foods and 32 batches of CTPMs (including 28 capsules, 32 tablets, 10 liquid and 2 pills) samples, 28 batches of heath foods were positive. The detected chemical ingredients involved sildenafil, tadalafil, aildenafil and sulfoaildenafil. This method can be used for the screening, identification and quantification of illegal adulterated aphrodisiac chemical ingredients in health foods and CTPMs. Moreover, the LC-Q-TOF MS is very useful to structural elucidation of unknown compound.

Oral Microbiota Community Dynamics Associated With Oral Squamous Cell Carcinoma Staging.

Oral squamous cell carcinoma (OSCC) is a highly aggressive cancer and the fourth leading malignancy among males in Taiwan. Some pathogenic bacteria are associated with periodontitis and oral cancer. However, the comprehensive profile of the oral microbiome during the cancer's progression from the early stage to the late stage is still unclear. We profiled the oral microbiota and identified bacteria biomarkers associated with OSCC. The microbiota of an oral rinse from 51 healthy individuals and 197 OSCC patients at different stages were investigated using 16S rRNA V3V4 amplicon sequencing, followed by bioinformatics and statistical analyses. The oral microbiota communities from stage 4 patients showed significantly higher complexity than those from healthy controls. The populations also dynamically changed with the cancer's progression from stage 1 to stage 4. The predominant phyla in the oral samples showed variation in the relative abundance of Fusobacteria, Bacteroidetes, and Actinobacteria. The abundance of Fusobacteria increased significantly with the progression of oral cancer from the healthy controls (2.98%) to OSCC stage 1 (4.35%) through stage 4 (7.92%). At the genus level, the abundance of Fusobacterium increased, while the number of Streptococcus, Haemophilus, Porphyromonas, and Actinomyces decreased with cancer progression. Fusobacterium periodonticum, Parvimonas micra, Streptococcus constellatus, Haemophilus influenza, and Filifactor alocis were associated with OSCC, and they progressively increased in abundance from stage 1 to stage 4. The abundances of Streptococcus mitis, Haemophilus parainfluenzae, and Porphyromonas pasteri were inversely associated with OSCC progression. We selected a bacterial marker panel of three bacteria (upregulated F. periodonticum, down-regulated S. mitis, and P. pasteri), which had an AUC of 0.956 (95% CI = 0.925-0.986) in discriminating OSCC stage 4 from the healthy controls. Furthermore, the functional prediction of oral bacterial communities showed that genes involved in carbohydrate-related metabolism, such as methane metabolism, and energy-metabolism-related parameters, such as oxidative phosphorylation and carbon fixation in photosynthetic organisms, were enriched in late-stage OSCC, while those responsible for amino acid metabolism, such as folate biosynthesis and valine, leucine, and isoleucine biosynthesis, were significantly associated with the healthy controls. In conclusion, our results provided evidence of oral bacteria community changes during oral cancer progression and suggested the possibility of using bacteria as OSCC diagnostic markers.

Immune tolerance of vector beetle to its partner plant parasitic nematode modulated by its insect parasitic nematode.

Immune response of insect vectors to transmitted pathogens or insect hosts against parasites are well studied, whereas the mechanism of tripartite interactions remains elusive. In this study, we investigated the immune interactions of the vector beetle Monochamus alternatus ( Ma) to the devastating plant parasitic nematode Bursaphelenchus xylophilus ( Bx) and the insect parasitic nematode Howardula phyllotretae ( Hp). We report the unique immune mechanism by which the vector beetle tolerates many devastating Bx in its trachea, yet that immune tolerance is compromised by the parasitic nematode Hp. Contact with either nematode species triggers epithelial reactive oxygen species (ROS) production in Ma. Only the entry of Bx, not Hp, infection, induces increased expression of antioxidative genes, through which the ROS levels are balanced in the trachea of beetles. Furthermore, we found that up-regulation of antioxidative genes was induced by the interaction of Toll receptors. In contrast, beetles infected by Hp retain high levels of oxidative stress and melanization in trachea, and as a result, decrease Bx loading. This study highlights the role of Toll receptors in mediating the activation of antioxidative genes in immune tolerance to plant parasitic nematodes, and suggests the use of insect parasites as a biologic control.-Zhou, J., Zhao, L.-L., Yu, H.-Y., Wang, Y.-H., Zhang, W., Hu, S.-N., Zou, Z., Sun, J.-H. Immune tolerance of vector beetle to its partner plant parasitic nematode modulated by its insect parasitic nematode.

Synthesis and characterization of Tamoxifen citrate modified reduced graphene oxide nano sheets for breast cancer therapy.

Theranostic agents are of immense consideration in the current generation nanomedicine. In this study, we have developed a facile approach for the fabrication of Tamoxifen citrate modified nanosized reduced graphene oxide (nano-rGO) with more stability and low cytotoxicity. The prepared nano-rGO sheets were characterized using HR-TEM and AFM imaging techniques. Further, the cytotoxicity was assessed using MTT assay on female BALB/c nude mice MCF-7 cell lines. In addition, by means of continuous-wave near-infrared laser, cancer cells in vivo were significantly ablated because of the photothermal effect stimulated by tamoxifen modified nano-rGO. These results indicated that the prepared tamoxifen modified nano-rGO has the ability to apply in the photothermal therapy of breast cancers. Consequently, further exploration of photothermal therapeutics is desirable for the synthesis of novel nano materials with additional functionalities.

Comparative analysis of the Monochamus alternatus immune system.

The pine sawyer beetle, Monochamus alternatus, is regarded as a notorious forest pest in Asia, vectoring an invasive pathogenic nematode, Bursaphelenchus xylophilus, which is known to cause pine wilt disease. However, little sequence information is available for this vector beetle. This hampered the research on its immune system. Based on the transcriptome of M. alternatus, we have identified and characterized 194 immunity-related genes in M. alternatus, and compared them with homologues molecules from other species known to exhibit immune responses against invading microbes. The lower number of putative immunity-related genes in M. alternatus were attributed to fewer C-type lectin, serine protease (SP) and anti-microbial peptide (AMP) genes. Phylogenetic analysis revealed that M. alternatus had a unique recognition gene, galectin3, orthologues of which were not identified in Tribolium castaneum, Drosophila melanogastor, Anopheles gambiae and Apis mellifera. This suggested a lineage-specific gene evolution for coleopteran insects. Our study provides the comprehensive sequence resources of the immunity-related genes of M. alternatus, presenting valuable information for better understanding of the molecular mechanism of innate immunity processes in M. alternatus against B. xylophilus.

Stevioside induced cytotoxicity in colon cancer cells via reactive oxygen species and mitogen-activated protein kinase signaling pathways-mediated apoptosis.

The role of mitogen-activated protein kinase (MAPK) signaling pathways in cell growth and differentiation has been well established. The present study aimed to investigate the anti-proliferative effect of stevioside on human colon cancer HT-29 cells. Additionally, the effect of stevioside on cell cycle arrest and MAPK signaling pathways in HT-29 cells was explored. Stevioside was observed to significantly inhibit cancer cell growth at a dose of 5 µM at 48 and 72 h. A dose-dependent increase in the apoptosis rate was observed with cell cycle arrest at G2/M phase. In addition, caspase-9 and caspase-3 activity also increased. An increase in reactive oxygen species (ROS) production and a decrease in the mitochondrial membrane potential indicated that the mitochondrial-mediated intrinsic pathway is responsible for apoptotic activity. These results were additionally verified by the elevated expression level of phosphorylated p38 and extracellular signal-regulated kinase mitogen-activated protein kinases (MAPKs). Additionally, by inhibiting ROS production and MAPK activation, the antiproliferative effect of stevioside was suppressed, confirming the hypothesis that ROS and MAPK proteins induce apoptosis in human colon cancer HT-29 cells.

Complete Sequence of a Novel IncR-F33:A-:B- Plasmid, pKP1034, Harboring fosA3, blaKPC-2, blaCTX-M-65, blaSHV-12, and rmtB from an Epidemic Klebsiella pneumoniae Sequence Type 11 Strain in China.

A high fosfomycin resistance rate was observed in Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae (KPC-KP) in our previous study, but little is known about its mechanisms. In this study, we explored the prevalence of plasmid-mediated fosfomycin resistance determinants among fosfomycin-resistant KPC-KP strains from a Chinese university hospital and determined the complete sequence of a novel fosA3-carrying plasmid isolated from an epidemic K. pneumoniae sequence type (ST) 11 strain. A total of 97 KPC-KP strains were studied, of which 57 (58.8%) were resistant to fosfomycin, including 44 (45.4%) harboring fosA3 and 1 harboring fosA. All fosA3-positive strains belonged to the dominant ST11-pulse type (PT) A clone according to multilocus sequence typing and pulsed-field gel electrophoresis, suggesting clonal dissemination. The fosA-positive isolate belonged to ST11-PTE. The fosA3-carrying plasmid pKP1034 is 136,848 bp in length and is not self-transmissible. It is a multireplicon plasmid belonging to IncR-F33:A-: B-. Besides fosA3, a variety of other resistance determinants, including blaKPC-2, rmtB, blaCTX-M-65, and blaSHV-12, are identified in pKP1034, which would allow for coselection of fosA3 by most ß-lactams and/or aminoglycosides and facilitate its dissemination despite limited use of fosfomycin in China. Detailed comparisons with related plasmids revealed that pKP1034 is highly mosaic and might have evolved from alarming recombination of the blaKPC-2-carrying plasmid pKPC-LK30 from Taiwan and the epidemic fosA3-carrying plasmid pHN7A8 from mainland China.