Intestinal microbiota of broilers submitted to feeding restriction and its relationship to hepatic metabolism and fat mass: Fast‐growing strain

The present study was conducted to verify how feed restriction affects gut microbiota and gene hepatic expression in broiler chickens and how these variables are related to body weight gain. For the experiment, 21‐d‐old Cobb500^TM birds were distributed in a completely randomized experimental design with three treatments: T1. Control (ad libitum—3.176 Mcal/kg ME—metabolizable energy—and 19% CP—crude protein); T2. Energetic restriction (2.224 Mcal/kg ME and 19% CP) from 22 to 42 days with consumption equivalent to control; T3. Quantitative restriction (70% restriction, i.e., restricted broilers ingested only 30% of the quantity consumed by the control group—3.176 Mcal/kg ME and 19% CP) for 7 days, followed by refeeding ad libitum from 28 to 42 days. Ileum and caecum microbiota collections were made at 21, 28 and 42 days of age. Hepatic tissue was collected at 28 and 42 days old for relative gene expression analyses. At 43‐d‐old, body composition was quantified by DXA (Dual‐energy X‐ray Absorptiometry). Both feed restriction programmes decreased Lactobacillus and increased Enterococcus and Enterobacteriaceae counts. No differences were found in the refeeding period. Energetic restriction induced the expression of CPT1‐A (Carnitine palmitoyltransferase 1A) gene, and decreased body fat mass. Quantitative feed restriction increased lipogenic and decreased lipolytic gene expression. In the refeeding period, CPT1‐A gene expression was induced, without changing the broilers body composition. Positive associations were found between BWG (Body Weight Gain) and Lactobacillus and Clostridium cluster IV groups, and negatively associations with Enterobacteriaceae and Enterococcus bacterial groups. In conclusion, differences found in microbiota were similar between the two feed restriction programmes, however, hepatic gene expression differences were only found in quantitative restriction. Higher counts of Lactobacillus and Clostridium cluster IV groups in ileum are likely to be related to better broiler performance and low expression of lipogenic genes.     

Calreticulin promotes migration and invasion of nasopharyngeal carcinoma cells by inducing cell EMT

AIM:To observe the expression of calreticulin (CRT) in nasopharyngeal carcinoma tissues, analyze the significance of clinical pathology and the influence on epithelial-mesencymal transition (EMT) of CNE2 cells. METHODS:The expression of calreticulin was detected by immunohistochemistry in 52 nasopharyngeal carcinoma and 57 nasopharyngeal benign tissues, and the significance of clinical pathology was evaluated. The calreticulin gene-specific small interfering RNA was constructed, and then was transfected into the NPC cell line CNE2 using the cationic liposome method. The effect of CRT on the morphological changes of the CNE2 cells was observed under light microscope. The effect of CRT on the cell migration and invasion abilities of the CNE2 cells was detected by Transwell migration and invasion assays. The expression of EMT-related proteins E-cadherin, vimentin, transforming growth factor (TGF)-β and matrix metalloproteinase (MMP)-9 in the CNE2 cells was determined by Western blot. RESULTS:The positive expression rate of CRT in the benign lesion tissues was 19.29% (11/57), which was significantly increased in the nasopharyngeal carcinoma tissues as 82.69% (43/52). The expression rate of CRT was positively correlated with the stage of nasopharyngeal carcinoma and lymph node metastasis (P<0.05). Knockdown of CRT expression made the CNE2 cells showing a spindle shape to a flat, cobblestone-like epithelial state change, arranged more compact, and the migration and invasion abilities were significantly decreased (P<0.05). Knockdown of CRT expression resulted in significant increase in the protein expression of E-cadhe-rin, and the decreases in the protein expression of vimentin, TGF-β and MMP-9 in the CNE2 cells (P<0.05). CONCLUSION:Calreticulin expression in nasopharyngeal carcinoma is significantly higher and positively correlated with nasopharyngeal carcinoma stage and lymph node metastasis. Calreticulin promotes cell migration and invasion of nasopharyngeal carcinoma CNE2 cells by inducing EMT.     

Progress on genomic aberrations in esophageal squamous-cell carcinoma

Esophageal squamous-cell carcinoma (ESCC) is one of the most common malignancies with poor prognosis in China. Since most clinical confirmation of the patients with ESCC is diagnosed at an advanced stage or with lymph node metastasis, the treatment effect was very poor. With the applications of high-throughput microarray and whole-genome sequencing or whole-exome sequencing, several novel cancer-related genes have been identified and revealed, and these genes may become new biomarkers or therapeutic targets. This review is to summarize the research progress in ESCC genome reported recently.     

Establishment of patient-derived esophageal squamous-cell carcinoma xenograft in mice and characteristics of signaling pathways related to proliferation in SCID mice

AIM: To establish and characterize the patient-derived esophageal squamous-cell carcinoma xenograft (PDECX) in mice. METHODS: The samples of human esophageal cancer were grafted into severe combined immunodeficient (SCID) mice. The xenografts were transferred to SCID mice when the first passage of xenografts grew up. The growth of tumors in the first, second and third passages was observed. HE staining was performed. The expression of CK5/6, p63 and p40 in the patient samples, and the first and third passages of the xenografts were detected by immunohistochemical analysis. The expression of mTOR, p-mTOR, p70S6K, p-p70S6K, Akt1, p-Akt (Ser473), Erk1/2 and p-Erk1/2 were determined by Western blot.RESULTS: The PDECX was successfully established. The positive expression of CK5/6, p63 and p40 in the xenografts was consistent with that in the patients' samples. The levels of phosphorylated and total proteins of proliferation-related signaling pathways were different in the xenografts from different patients.CONCLUSION: The PDECX model adequately reflects the tumal heterogeneity that is observed in the patients.     

Liver histology and histomorphometry in hybrid sorubim (Pseudoplatystoma reticulatum × Pseudoplatystoma corruscans) reared on intensive fish farming

This study aimed to characterize the liver histology and histomorphometry in sorubim hybrid of different categories (nursery, growth and grow‐out) reared on fish farming. The categories were defined considering body weight (BW): nursery category (n = 5): BW = 37.06 ± 6.00 g (31.6–45.3 g); growth category (n = 5): BW = 310.40 ± 53.80 g (242.1–376.4 g) and grow‐out category (n = 5): BW = 874.28 ± 27.59 g (846.2–913.1 g). Liver fragments were processed to paraffin inclusion, and sections were stained by haematoxylin and eosin (H&E), PAS (Periodic Acid Schiff) and Perl's staining to histology, histomorphometry and density volumetric of liver structures; glycogen analysis and to detect ferric irons (Fe³⁺) respectively. The hepatosomatic index decreased between the categories (P < 0.01). The percentage of PAS‐positive hepatocytes in the nursery category was higher (P < 0.05) in relation to the growth and grow‐out categories. The hepatocytes from all fish were positive to Perl's staining. The density volumetric of liver structures did not differ among categories except to blood vessels were higher (P < 0.01) in the nursery and growth. The area (μm²) and perimeter (μm) of hepatocytes, and the area (μm²), perimeter (μm) and volume (μm³) of the nuclei from grow‐out fish were lower (P < 0.01) than those from the nursery and growth categories. Changes in morphometric characteristics of hepatocytes may result from metabolic changes associated with body growth surubins; therefore, these morphometric characteristics of liver tissue can be used as functional biomarkers for the assessment of fish health and nutrition status.     

Low protein provision during the first year of life,but not during foetal life,affects metabolic traits,organ mass development and growth in male mink (Neovison vison)

Low protein provision in utero and post‐partum may induce metabolic disorders in adulthood. Studies in mink have mainly focused on short‐term consequences of low protein provision in utero whereas the long‐term responses to low protein (LP) provision in metabolically programmed mink are unknown. We investigated whether low protein provision in utero affects the long‐term response to adequate (AP) or LP provision after weaning in male mink. Eighty‐six male mink were exposed to low (19% of ME from CP; crude protein) or adequate (31% of ME from CP) protein provision in utero, and to LP (~20% of ME from CP) or AP (30–42% of ME from CP) provision post‐weaning. Being metabolically programmed by low protein provision in utero did not affect the response to post‐weaning diets. Dietary protein content in the LP feed after weaning was below requirements; evidenced by lower nitrogen retention (p < 0.001) preventing LP mink from attaining their growth potential (p < 0.02). LP mink had a lower liver, pancreas and kidney weight (p < 0.05) as well as lower plasma IGF‐1 concentrations at 8 and 25 (p < 0.05) weeks, and a higher incidence of hepatic lipidosis at 25 weeks (p < 0.05). Furthermore, LP mink had a higher body fat (p < 0.05) and lower body CP content (p < 0.05) at 50 weeks of age. It is concluded that some effects of low protein provision in utero can be alleviated by an adequate nutrient supply post‐partum. However, long‐term exposure to low protein provision in mink reduces their growth potential and induces transient hepatic lipidosis and modified body composition.     

人类表皮生长因子受体2蛋白在宫颈癌组织中的表达及意义

目的观察宫颈癌组织中人类表皮生长因子受体2（HER-2/neu）蛋白表达及其在宫颈癌发生发展中的作用。方法应用免疫组织化学方法检测20例正常宫颈组织、30例宫颈上皮内瘤样病变（CINⅠ-Ⅲ）和50例宫颈癌组织中HER-2/neu蛋白表达。结果宫颈癌组织中HER-2/neu蛋白表达显著高于宫颈上皮内瘤样病变（P〈0.01）,正常宫颈组织未见阳性表达;宫颈癌组织中HER-2/neu蛋白表达与肿瘤浸润深度、淋巴结转移、临床分期密切相关（P〈0.05）。结论 HER-2/neu蛋白表达上调可能在宫颈癌发生发展中起重要作用。     

Effect of gold nanoparticles on reversing adriamycin resistance of human hepatocellular carcinoma HepG2/ADM cells

AIM: To investigate whether gold nanoparticles (GNPs) reverses adriamycin (ADM), resistance of human hepatocellular carcinoma drug-resistant cell line HepG2/ADM and to explore the potential mechanism. METHODS: The sensitivities of HepG2 cells and HepG2/ADM cells to ADM were tested by MTT assay before and after GNPs pretreatment. The apoptotic rate was examined by flow cytometry. The concentration of ADM in HepG2/ADM or HepG2 cells was determined by ultraviolet-visible spectrophotometer. The content of glutathione (GSH) in HepG2/ADM or HepG2 cells by DTNB method. RESULTS: The half maximal inhibitory concentrations (IC₅₀) of ADM for HepG2/ADM cells were(29.46±1.73) mg/L and (15.18±0.85) mg/L before and after GNPs pretreatment,respectively. The IC₅₀ of ADM for HepG2 cells was (9.16±2.03) mg/L before pretreatment. The apoptotic rate in GNPs+ADM group was higher than that in ADM group (P<0.05). The concentration of ADM in HepG2/ADM group was lower than that in HepG2 group (P<0.01). After GNPs pretreatment, the concentration of ADM in HepG2/ADM cells was higher than that before pretreatment. The content of GSH in HepG2/ADM group was higher than that in HepG2 group (P<0.01). After GNPs pretreatment, the content of GSH in the HepG2/ADM cells was lower than that before pretreatment. CONCLUSION: Gold nanoparticles can reverse ADM resistance of human hepatocellular carcinoma drug-resistant cell line HepG2/ADM, reduce the content of GSH and increase the concentration of ADM in HepG2/ADM cells.