Traditional Chinese medicine reverses cancer multidrug resistance and its mechanism.

Chemotherapy is one of the most commonly used clinical treatments among the currently available cancer therapies. However, the phenomenon of Multidrug resistance (MDR) has become a challenge in the treatment process, weakening the impact of chemotherapy. Extensive research on elucidating the development of cancer MDR has identified the following mechanisms that play a critical role in the development of several MDR reversal agents: abnormal expression of cell membrane transporters, adaptation of cancer cells to the microenvironment, regulation of hypoxia, repair of DNA damage and reduction of apoptosis, the enhancement of the EMT process, the existence of cancer stem cells (CSCs), and the abnormal activation of key signaling pathways. However, they failed to demonstrate significant efficacy due to severe side effects during their clinical trials. Traditional Chinese medicines (TCMs) are known to play an important anti-cancer role since they have low toxicity, high efficacy, and safety and can reverse MDR. TCMs reversal agents can be divided into Chinese medicine monomers, synthetic monomers, analogs, or derivatives. Several studies have shown that TCMs can effectively overcome cancer MDR and can be effectively used for treating cancer patients.

Genome-wide scanning for CHD1L gene in papillary thyroid carcinoma complicated with type 2 diabetes mellitus.

PURPOSE: Papillary thyroid carcinoma (PTC) represents the most common subtype of thyroid cancer (TC). This study was set out to explore the potential effect of CHD1L on PTC and type 2 diabetes mellitus (T2DM). METHODS: We searched for T2DM susceptibility genes through the GWAS database and obtained T2DM-related differentially expressed gene from the GEO database. The expression and clinical data of TC and normal samples were collated from the TCGA database. Receiver operating characteristic (ROC) curve analysis was subsequently applied to assess the sensitivity and specificity of the CHD1L for the diagnosis of PTC. The MCP-counter package in R language was then utilized to generate immune cell score to evaluate the relationship between CHD1L expression and immune cells. Then, we performed functional enrichment analysis of co-expressed genes and DEGs to determine significantly enriched GO terms and KEGG to predict the potential functions of CHD1L in PTC samples and T2DM adipose tissue. RESULTS: From two genes (ABCB9, CHD1L) were identified to be DEGs (p < 1 * 10-5) that exerted effects on survival (HR > 1, p < 0.05) in PTC and served as T2DM susceptibility genes. The gene expression matrix-based scoring of immunocytes suggested that PTC samples with high and low CHD1L expression presented with significant differences in the tumor microenvironment (TME). The enrichment analysis of CHD1L co-expressed genes and DEGs suggested that CHD1L was involved in multiple pathways to regulate the development of PTC. Among them, Kaposi sarcoma-associated herpesvirus infection, salmonella infection and TNF signaling pathways were highlighted as the three most relevant pathways. GSEA analysis, employed to analyze the genome dataset of PTC samples and T2DM adipose tissue presenting with high and low expression groups of CHD1L, suggests that these differential genes are related to chemokine signaling pathway, leukocyte transendothelial migration and TCELL receptor signaling pathway. CONCLUSION: CHD1L may potentially serve as an early diagnostic biomarker for PTC, and a target of immunotherapy for PTC and T2DM.

The role of macrophages during breast cancer development and response to chemotherapy.

Macrophages play an important role in the immune system as a key host defense against pathogens. Non-polarized macrophages can differentiate into pro-inflammatory classical pathway-activated macrophages or anti-inflammatory alternative pathway-activated macrophages, both of which play central roles in breast cancer growth and progression in a process called polarization of macrophages. Classical pathway-activated and alternative pathway-activated macrophages can transform into each other and their transformational properties and orientation are determined by cytokines in the tumor microenvironment. Tumor-associated macrophages display many functions, such as tissue reforming, participating in inflammation and tumor growth in breast cancer progression. Some cytokines, such as interleukins and transcriptional activators, reside in the tumor microenvironment and influence tumor-associated macrophages. Chemotherapy is a common treatment for breast cancer and macrophages play an important role in mammary tumor cell migration, cancer invasion, and angiogenesis. This review summarizes the activities of tumor-associated macrophages in the mammary tumor, chemotherapeutic processes and some potential strategies for breast cancer therapy.

25-OH-vitamin D deficiency identifies poor tumor response in hepatocellular carcinoma treated with transarterial chemoembolization.

PURPOSE: Vitamin D is implicated linked to liver cancer and chronic liver diseases, but its association with tumor response in hepatocellular carcinoma (HCC) patients undergoing transarterial chemoembolization (TACE) remains unclear. This study aimed to determine whether vitamin D levels influence tumor response in HCC patients treated with TACE. METHODS: A total of 58 HCC patients undergoing TACE were enrolled in the study. Serum 25-hydroxyvitamin D (25-OHD) levels were determined at baseline and 1 day after TACE using electrochemiluminescence immunoassay. Response to TACE was evaluated after a 4-6 week interval. Univariate and multivariate analyses with Cox regression model were performed to determine the risk factors associated with tumor response. Receiver operating characteristic (ROC) curve analysis was performed to assess the predictive performance of baseline 25-OHD levels on tumor response in HCC patients undergoing TACE. RESULTS: 43.1% of HCC patients showed 25-OHD deficiency. Baseline 25-OHD level was associated with liver cirrhosis (P = 0.025), vascular invasion (P = 0.031), Barcelona Clinic Liver Cancer stage (P = 0.002) and an alanine aminotransferase increase after TACE (P = 0.021). Serum 25-OHD level was significantly decreased 1 day after TACE (P = 0.045). Multiple tumor numbers (P = 0.034) and low baseline 25-OHD levels (P = 0.040) were independently correlated with poor tumor response after TACE. ROC curve analysis showed that baseline 25-OHD levels present better predictive performance for OR in those patients, compared with other current clinical test pointers. CONCLUSION: Our study suggested that 25-OHD deficiency at baseline is a prognostic indicator for a poor tumor response in hepatocellular carcinoma treated with TACE.

Prognostic analysis of stage III gastric cancer after curative surgery according to the newest TNM classification.

AIM: To study the prognostic factors of gastric cancer (GC) patients who were classified with stage III disease according to the newest TNM classification. METHODS: This study retrospectively enrolled 279 patients who underwent radical gastrectomy from January 2012 to December 2014 at our hospital and who were diagnosed with stage III GC according to the new 8th edition of the TNM classification. The patient data that were collected included age, sex, pathological parameters, survival, lymph node ratio, neo-adjuvant chemotherapy with oxaliplatin and S-1, and operation type. The characteristics, survival, and prognostic factors of the patients were analyzed by univariate and multivariate analyses. RESULTS: The median OS of the patients after curative surgery was 19 months, and the 3-year survival rate (3-YSR) was 25.3%. A univariate analysis showed that tumor location (P = 0.01), neo-adjuvant chemotherapy (P = 0.005), pathological T stage (P = 0.002), pathological N stage (P < 0.001), lymph node ratio (LNR) (P < 0.001), and operation type (P = 0.032) were significantly associated with overall survival. A multivariate analysis revealed that neo-adjuvant chemotherapy (P = 0.009), pathological T stage (P = 0.012), and LNR (P < 0.001) were independent prognostic factors. CONCLUSIONS: Neo-adjuvant chemotherapy, pathological T stage, and LNR were independent prognostic factors for the overall survival of patients with stage III GC. The neo-adjuvant chemotherapy with oxaliplatin and S-1 can be used for the patients to improve their survival.

Organic egg production, egg quality, calcium utilization, and digestibility in laying hens fed with mushroom (Flammulina velutipes) stem waste

The possibility of using mushroom (Flammulina velutipes) stem waste (FVW) in layer diets was explored in this study. A total of 180, 40-week-old ISA Brown laying hens were randomly assigned to 5 groups, with 6 replications of 6 birds for each group. Layers were fed a standard basal diet as control; antibiotic (0.05% flavomycin); 2% FVW; 4% FVW; and 6% FVW. No significant differences (p>0.05) were observed on hen daily egg production, egg weight, egg mass, unmarketable eggs, feed intake, feed conversion ratio, and live weight among experimental groups. Haugh unit was found higher (p0.05) by feeding FVW in this study. Yolk color was found higher (p<0.05) in FVW fed groups than in control and antibiotic groups. The shell color was higher (p<0.05) both in 4% FVW and 6% FVW fed groups than in control and antibiotic groups. Calcium retention and calcium in egg shell deposition were higher (p<0.05) both in 4% FVW and 6% FVW fed groups than in control and antibiotic groups respectively. Apparent nutrient retention for dry matter, crude protein, crude fat, and organic matter were higher (p<0.05) in FVW fed groups than in control and antibiotic groups. FVW at 6% level in layer ration can be used for organic eggs production which would be safe and sound for human consumption.

Organic egg production, egg quality, calcium utilization, and digestibility in laying hens fed with mushroom (Flammulina velutipes) stem waste

The possibility of using mushroom (Flammulina velutipes) stem waste (FVW) in layer diets was explored in this study. A total of 180, 40-week-old ISA Brown laying hens were randomly assigned to 5 groups, with 6 replications of 6 birds for each group. Layers were fed a standard basal diet as control; antibiotic (0.05% flavomycin); 2% FVW; 4% FVW; and 6% FVW. No significant differences (p>0.05) were observed on hen daily egg production, egg weight, egg mass, unmarketable eggs, feed intake, feed conversion ratio, and live weight among experimental groups. Haugh unit was found higher (p<0.05) in FVW fed groups than in antibiotic fed groups. Shape index, shell weight, shell thickness, yolk weight, yolk index, yolk weight and albumen weight were not affected (p>0.05) by feeding FVW in this study. Yolk color was found higher (p<0.05) in FVW fed groups than in control and antibiotic groups. The shell color was higher (p<0.05) both in 4% FVW and 6% FVW fed groups than in control and antibiotic groups. Calcium retention and calcium in egg shell deposition were higher (p<0.05) both in 4% FVW and 6% FVW fed groups than in control and antibiotic groups respectively. Apparent nutrient retention for dry matter, crude protein, crude fat, and organic matter were higher (p<0.05) in FVW fed groups than in control and antibiotic groups. FVW at 6% level in layer ration can be used for organic eggs production which would be safe and sound for human consumption.(AU)

Characterization of air-liquid interface culture of A549 alveolar epithelial cells

Alveolar epithelia play an essential role in maintaining the integrity and homeostasis of lungs, in which alveolar epithelial type II cells (AECII) are a cell type with stem cell potential for epithelial injury repair and regeneration. However, mechanisms behind the physiological and pathological roles of alveolar epithelia in human lungs remain largely unknown, partially owing to the difficulty of isolation and culture of primary human AECII cells. In the present study, we aimed to characterize alveolar epithelia generated from A549 lung adenocarcinoma cells that were cultured in an air-liquid interface (ALI) state. Morphological analysis demonstrated that A549 cells could reconstitute epithelial layers in ALI cultures as evaluated by histochemistry staining and electronic microscopy. Immunofluorescent staining further revealed an expression of alveolar epithelial type I cell (AECI) markers aquaporin-5 protein (AQP-5), and AECII cell marker surfactant protein C (SPC) in subpopulations of ALI cultured cells. Importantly, molecular analysis further revealed the expression of AQP-5, SPC, thyroid transcription factor-1, zonula occludens-1 and Mucin 5B in A549 ALI cultures as determined by both immunoblotting and quantitative RT-PCR assay. These results suggest that the ALI culture of A549 cells can partially mimic the property of alveolar epithelia, which may be a feasible and alternative model for investigating roles and mechanisms of alveolar epithelia in vitro.

Characterization of air-liquid interface culture of A549 alveolar epithelial cells.

Alveolar epithelia play an essential role in maintaining the integrity and homeostasis of lungs, in which alveolar epithelial type II cells (AECII) are a cell type with stem cell potential for epithelial injury repair and regeneration. However, mechanisms behind the physiological and pathological roles of alveolar epithelia in human lungs remain largely unknown, partially owing to the difficulty of isolation and culture of primary human AECII cells. In the present study, we aimed to characterize alveolar epithelia generated from A549 lung adenocarcinoma cells that were cultured in an air-liquid interface (ALI) state. Morphological analysis demonstrated that A549 cells could reconstitute epithelial layers in ALI cultures as evaluated by histochemistry staining and electronic microscopy. Immunofluorescent staining further revealed an expression of alveolar epithelial type I cell (AECI) markers aquaporin-5 protein (AQP-5), and AECII cell marker surfactant protein C (SPC) in subpopulations of ALI cultured cells. Importantly, molecular analysis further revealed the expression of AQP-5, SPC, thyroid transcription factor-1, zonula occludens-1 and Mucin 5B in A549 ALI cultures as determined by both immunoblotting and quantitative RT-PCR assay. These results suggest that the ALI culture of A549 cells can partially mimic the property of alveolar epithelia, which may be a feasible and alternative model for investigating roles and mechanisms of alveolar epithelia in vitro.

Development of coding single nucleotide polymorphic markers in the pearl oyster Pinctada fucata based on next-generation sequencing and high-resolution melting analysis.

The pearl oyster Pinctada fucata is an important commercial marine shellfish that is cultured for producing saltwater pearls. In this study, 468 single nucleotide polymorphisms (SNPs) were screened from P. fucata transcriptome data, and 119 polymorphic SNPs were successfully isolated by a two-step small-amplicon high-resolution melting assay. Of these, 88 were annotated with BLAST in the Nr database and 90 were in the open reading frame, including 16 non-synonymous SNPs and 74 synonymous SNPs; 12 SNPs were in the 3'-untranslated region (UTR) and 1 was in the 5'-UTR. Twenty-five SNPs were randomly chosen to test the genetic diversity of 40 wild individuals from Liusha Bay, China. All of the loci had two alleles. The observed and expected heterozygosities ranged from 0.0417 to 0.6042 and from 0.2945 to 0.5053, respectively. Minor allele frequencies ranged from 0.1771 to 0.5000, and the polymorphism information content ranged from 0.2516 to 0.3750. These novel SNP markers can contribute to P. fucata genetics and breeding studies.

Association of ectomycorrhizal fungi with Picea crassifolia (Pinaceae, Piceoidae) from high-altitude stands in Mount Helan Nature Reserve, China.

We investigated the diversity of ectomycorrhiza associated with the endemic Picea crassifolia in Mount Helan National Nature Reserve in Inner Mongolia, China. Toward this objective, we conducted morphological and molecular identification of ectomycorrhizae in soil cubes taken from pure P. crassifolia stands. Eleven types of ectomycorrhizal (ECM) organisms were separated, briefly described, and identified. Nine morphotypes belonged to the phylum Basidiomycotina [Amphinema byssoides, Cortinarius sp (cf. limonius), Cortinarius vernus, Inocybe cf. nitidiscula, Inocybe sp 1, Sebacina incrustans, Sebacina sp, Suillus luteus, and Piceirhiza tuberculata x Picea crassifolia (comb. Nov.)], and two morphotypes to the phylum Ascomycotina (Cenococcum geophilum and Helvella sp). The diversity of ECM organisms in P. crassifolia was lower than that reported by other studies on spruce or pine forests, or on sporocarp diversity in the high-mountain forests of China. Most of the fungi in the rhizosphere did not correspond to species previously recorded as sporocarps above ground. Here, several new ectomycorrhiza morphotypes are proposed and described. We also confirmed the ectomycorrhizal status of the genus Sebacina (order Sebacinales).

Expression and function of PDGF-α in columnar epithelial cells of age-related cataracts patients.

We studied the expression and function of platelet-derived growth factor A (PDGF-α) in the lens epithelial cells of cataracts patients. Ninety age-related cataracts patients were recruited in our hospital between January 2012 and January 2014. The expression levels of platelet-derived growth factor receptor (PDGFR) in the anterior capsule of the lens at different degrees of turbidity, and PDGF-α in the aqueous humor were detected. A human lens epithelium cell line was also cultured and studied. To investigate its function, PDGF-α was used to treat a PDGFR-silenced human lens epithelium cell line to observe changes in the proliferation, transfer, and epithelial mesenchymal transition (EMT). The expression of PDGF-α and its receptor increased in patients with more serious cataracts. Lens epithelium cells stimulated by PDGF-α showed greater proliferation and migration. The degree of EMT was also upregulated in cells stimulated by PDGF-α. However, silencing the expression of PDGFR inhibited the effects. The development and severity of age-related cataracts was related to the secretion and expression of PDGF-α. This may be a new therapeutic target for cataracts treatment.

Sequence and phylogenetic analyses of the M and N genes of porcine epidemic diarrhea virus (PEDV) strains in Anhui Province, China.

To assess the homology and phylogenetic relationship between porcine epidemic diarrhea virus (PEDV) Anhui strains and other PEDV strains, molecular homology and phylogenetic analyses of Anhui PEDV field strains were compared with those of reference strains. The results revealed that the M and N genes of PEDV were 681 and 1326 bp long, respectively. The nucleotide sequences of the N genes of Anhui PEDV strains were 95.9-99.9% homologous with each other, and the deduced amino acid sequences were 92.5-99.8% homologous. Compared with the PEDV reference strains, the Anhui PEDV field strains had 94.1-99.5% nucleotide sequence homology in the N gene and 91.2-97.5% amino acid mutation homology in the N protein. The nucleotide sequences of the M genes of Anhui PEDV were 98.3-100% homologous, and the deduced amino acid sequences were 96.5-99.6% homologous. Compared with the PEDV reference strains, the Anhui PEDV field strains had 96.9-100% nucleotide sequence homology in the M gene and 96.5-99.6% amino acid homology in the M protein. The Anhui strains were genetically similar to USA strains (USA/Iowa/16465/2013 and USA/Indiana/17846/2013) but different from European (CV777;Br1/7), Korean (Chinju99), and Japanese (83p-5) strains.

Quantitative trait locus analysis of leaf morphological characters, yield-related traits, and secondary metabolite contents in Eucommia ulmoides.

Eucommia ulmoides is cultivated for the production of eucommia rubber and Chinese herbal drugs. Molecular breeding methods, such as marker-assisted selection (MAS), have the potential to improve the efficiency of E. ulmoides breeding. Quantitative trait locus (QTL) analysis was applied to identify marker-trait associations for E. ulmoides using an F1 mapping population of 152 individuals derived from a cross between the wild genotype Xiaoye and the cultivar Qinzhong No. 1. A total of 78 QTLs were identified for 12 leaf traits involving morphology, yield, and secondary metabolites. Phenotypic variance explained by individual QTLs ranged from 10.4 to 53.3%. Fifteen QTL clusters, each harboring loci controlling at least two leaf traits, were detected across nine linkage groups. Co-location of these QTLs may be due to pleiotropy or linkage. Three main QTL regions for secondary metabolites were consistently identified each year. QTL information from this study furthers our understanding of the genetic architecture of these economically important traits and of MAS in E. ulmoides breeding.

Comparative transcriptome analysis of Nile tilapia (Oreochromis niloticus) in response to alkalinity stress.

To identify genes of potential importance to alkalinity tolerance, RNA sequencing (RNA-Seq) was performed to survey gill transcriptome profiles from freshwater (FW) and alkaline water (AW) exposed Nile tilapia (Oreochromis niloticus). A total of 22,724,036 (AW)/16,461,040 (FW) single-end reads were generated in which 20,304,348 (AW)/14,681,290 (FW) reads (90.0/89.72%) were aligned to the reference genome. Differential expression analysis revealed 302 up-regulated and 193 down-regulated genes between AW- and FW-exposed fish. These differentially expressed genes were enriched in several Gene Ontology (GO) terms related to "stress response", "heme binding", and "carbonate dehydratase activity". Meanwhile, significant KEGG pathways were enriched in energy metabolism, including nitrogen and sulfur metabolism. These results demonstrate the response of Nile tilapia exposed to alkaline-water and might provide valuable information to further understand the molecular mechanisms of adaptation of fish to alkaline environments.

Circulating autoantibody to CD25 may be a potential biomarker for early diagnosis of esophageal squamous cell carcinoma.

BACKGROUND: The present study was undertaken to develop an in-house enzyme-linked immunosorbent assay (ELISA) using human leukocyte antigen class II restricted epitopes to test circulating autoantibodies to CD25 as a biomarker for esophageal cancer. METHODS: A total of 97 patients with esophageal squamous cell carcinoma (ESCC) and 226 healthy subjects were recruited for this study and their plasma samples were collected for antibody analysis with the ELISA approach. RESULTS: Mann-Whitney U test showed that the IgG anti-CD25 antibody level was significantly higher in the patient group than the control group (P < 0.001) while the IgA antibody level was not significantly different between these two groups (P = 0.361). Spearman correlation analysis failed to reveal a significant correlation between the levels of anti-CD25 IgG and IgA antibodies in either the patient group (r = -0.027, P = 0.797, n = 94) or the control group (r = 0.055, P = 0.429, n = 209). The sensitivity against >90 % specificity was 37.2 % for the IgG assay with an inter-assay deviation of 9.4 %, and 8.2 % for the IgA assay with an inter-assay deviation of 13.0 %. Based on a cut-off value determined by the 99th percentile of control IgG levels, the positive rate was 7.4 % in patients with ESCC, in which patients at stage I had the highest positivity (11.5 %) (χ (2) = 11.10, P = 0.001, OR = 12.12, 95 % CI 1.93-75.94). CONCLUSIONS: This work suggests that circulating IgG autoantibody to CD25 may be a potential biomarker for early diagnosis of esophageal cancer.

Infertility caused by male partners with genetic defects in Sichuan Province of China.

The purpose of this study was to detect chromosomal aberrations and azoospermia factor (AZF) microdeletions in male patients with reproductive problems and to summarize related clinical features to provide reliable information for evaluating prenatal and preimplantation diagnoses. A large cohort of 5083 men with various phenotypes of male infertility was analyzed via G-banding karyotyping, and Origin 8.0 was used to analyze the prevalence of abnormalities. Additionally, patients with azoospermia, oligozoospermia, and oligoasthenozoospermia were analyzed using multiplex polymerase chain reaction to detect microdeletion in the AZF. We identified 387 patients with abnormal karyotypes, and the ratio was 7.61%. Among them were 175 patients with Klinefelter's syndrome, which was the most common numerical chromosomal abnormality and accounted for 45.22% of all chromosomal aberrations. The frequencies of increased satellites, balanced translocations, and Robertsonian translocations were 6.47, 7.00, and 3.62%, respectively. Multiplex polymerase chain reaction performed in 810 cases with azoospermia, oligozoospermia, and oligoasthenozoospermia found a ratio of AZF microdeletions of 4.94%. The finding suggests that chromosomal abnormalities and AZF deletion are main factors that result in male infertility. Detecting these common genetic variations is necessary in infertile men seeking assisted reproductive technology.

A study of circulating anti-CD25 antibodies in non-small cell lung cancer.

PURPOSE: Tumors can trigger specific immune response to tumor-associated antigens but the precise mechanism remains unclear. Since regulatory T-lymphocytes (Treg) play a crucial role in controlling autoimmune responses, the present work was undertaken to test whether dysfunction of Treg cells could be involved in developing autoimmunity in patients with lung cancer. METHODS: In this study, we developed an in-house enzyme-linked immunosorbent assay to test circulating anti-CD25 autoantibodies among 272 patients with non-small cell lung cancer (NSCLC) and 226 control subjects matched in age, gender and smoking history. RESULTS: Mann-Whitney U test showed that the anti-CD25 IgG level was significantly higher in patients with NSCLC than control subjects (Z = -7.48, P < 0.001) while the anti-CD25 IgA level was not significantly changed in the patient group as compared with the control group (Z = -1.34, P = 0.181). Spearman correlation analysis failed to reveal a significant correlation between the levels of anti-CD25 IgG and IgA either in patients with NSCLC (r = -0.034, P = 0.578) or in control subjects (r = 0.055, P = 0.429). ROC analysis showed an AUC of 0.70 for anti-CD25 IgG, in which NSCLC at stage III had the highest AUC (0.75). The sensitivity against a specificity of >90 % was 35.0 % for anti-CD25 IgG assay with an inter-assay deviation of 9.4 %, and 4.0 % for anti-CD25 IgA assay with an inter-assay deviation of 13.0 %. CONCLUSIONS: Circulating anti-CD25 IgG antibody may be a useful biomarker for prognosis of lung cancer.