Lysophospholipid and lipase in broilers: combined lysophospholipid and lipase supplementation maintains production performance and modulates metabolic and antioxidant responses under reduced energy feeding regimens

The effects of formic acid or herbal mixture on growth performance, carcass quality, blood chemistry, and gut microbial load in broiler chickens: Formic Acid & Herbal Mixture in Broiler Diets.

A total of 420 day-old male Ross chicks were weighed at d 1 of life and assigned to test diets to assess the efficacy of a new Buttiauxella spp. phytase expressed in Trichoderma reesei. Diets were: positive control (PC) adequate in nutrients and negative control (NC) diet (40% and 17% less available phosphorous (P) and calcium (Ca), respectively) supplemented with 6 levels of phytase 0, 250, 500, 750, 1,000, and 2,000 phytase units (FTU)/kg of diet. All diets had titanium dioxide as digestibility marker and each diet was allocated to ten cages (6 birds/cage). Diets were fed for 3 wk to measure growth performance, apparent retention (AR) on d 17 to 21 and bone ash and ileal digestibility (AID) on d 22. Growth performance and nutrient utilization was lower (p<0.05) for NC vs PC birds. Phytase response in NC birds was linear (p<0.05) with 2,000 FTU showing the greatest improvement on body weight gain (20%), feed conversion (7.4%), tibia ash (18%), AR of Ca (38%), AR of P (51%) and apparent metabolizable energy corrected for nitrogen (5.1%) relative to NC. Furthermore, phytase at ≥750 FTU resulted in AID of total AA commensurate to that of PC fed birds and at ≥1,000 FTU improved (p<0.05) AR of P, dry matter, and N beyond that of the lower doses of phytase and PC diet. In conclusion, the result from this study showed that in addition to increased P and Ca utilization, the new Buttiauxella phytase enhanced growth performance and utilization of other nutrients in broiler chickens in a dose-dependent manner.

An experiment was conducted to investigate the effects of replacing antibiotic growth promoter (AGP) with a combination of essential oil and organic acids (EO + OA) on growth performance, gut microbiota and immune response in broiler chickens. In Experiment 1, 320 day-old broiler chicks were randomly distributed to 32 pens with 10 birds in each pen and the pens were equally allotted to four treatment groups. In Experiment 2, 120 day-old chicks were divided into the same four treatment groups, each group containing three replicated pens with 10 birds in each pen. The groups were (1) Negative Control (NC) without AGP or other growth-promoting feed additives; (2) AGP (NC + enramycin; 125 mg/kg feed; (3) OA (NC + OA; 500 mg/kg feed); and (4) EO + OA (NC + a combination of cinnamaldehyde and calcium formate; 500 mg/kg feed). Experiment 1 lasted for 40 days, whereas Experiment 2 continued for 28 days. In Experiment 2, all birds were orally challenged with Escherichia coli (108 bacteria/bird) on Day 14. Overall intake, growth and feed conversion ratio (FCR) on Day 40 had no difference (P &amp;gt; 0.05) among the groups in Experiment 1. In Experiment 2, growth, feed intake and FCR were not affected by any dietary treatments until Day 14, but after being challenged with E. coli, bodyweight gain and FCR improved (P &amp;lt; 0.05) for AGP and EO + OA compared with NC. Mortality rate was also lower (P &amp;lt; 0.05) for AGP and EO + OA than NC in Experiment 2. Villi height was higher (P &amp;lt; 0.001) in OA and EO + OA groups compared with NC and AGP groups. Any treatment did not affect (P &amp;gt; 0.05) the counts of total bacteria, E. coli and Lactobacillus in the contents of ileum and caecum. However, Salmonella counts in the ileal and caecal contents decreased (P &amp;lt; 0.001) for AGP, OA and EO + OA compared with NC group. Clostridium counts were lower for EO + OA group than for NC and AGP groups in the ileum, and for AGP, OA and EO + OA groups than for NC in the caeca (P &amp;lt; 0.05). Antibody titer on Day 35 against Newcastle disease vaccine was higher in EO + OA group than in NC, AGP and OA groups (P &amp;lt; 0.001). In conclusion, EO + OA did not affect growth and FCR in broilers. However, AGP and EO + AO improved growth performance and FCR after being challenged with E. coli. Moreover, EO + OA was effective in reducing the Clostridium count in the small intestine and caecum and increasing the villus height and antibody titer against Newcastle disease vaccine.

The purpose of this study was to evaluate the outcome of sodium stearoyl-2-lactylate (SSL) supplementation to energy reduced diets on broilers’ growth performance, apparent nutrient digestibility, excreta bacterial counts, and blood lipid profile. A total of 700 one-day-old male (feather sexed) Ross 308 broiler chickens having an average body weight of 43 ± 1.2 g were randomly allotted to five (5) dietary treatments (7 pens/treatment). The treatments consisted of: (1) positive control (PC) (basal diet with metabolisable energy, ME = 3000 kcal/kg starter; 3100 kcal/kg grower; 3200 kcal/kg finisher), (2) negative control (NC) (PC − 50 kcal/kg), (3) SSL1, NC + 0.03% SSL, (4) SSL2, NC + 0.04% SSL, (5) SSL3, NC + 0.05% SSL. The PC group showed higher BWG (body weight gain) and lower FCR (feed conversion ration) than the NC group during days 1 to 7, and 7 to 21. SSL supplemented groups showed higher BWG and better FCR than the NC group, along with a linear increase in BWG and linear decrease in FCR during days 1 to 7 and 7 to 21. Furthermore, a linear increase in overall BWG was observed with SSL doses. Excreta microbial counts were not influenced by energy levels or SSL doses. In blood serum parameters, LDL (low density lipoprotein) cholesterol was reduced with a linear tendency by increasing levels of SSL supplementation. In conclusion, sodium stearoyl-2-lactylate supplementation showed better growth and nutrient digestibility in broilers’ at an early age. Highlights Emulsifiers can help broilers to utilise dietary energy more efficiently Sodium Stearoyl-2- Lactylate helped to increase energy and fat digestibility in broilers As a result, growth performance was improved through Sodium Stearoyl-2-Lactylate supplementation in broilers

Background/Aims: Nasopharyngeal carcinoma (NPC) is rare worldwide but remains highly prevalent in endemic regions, notably in southern China. Radiotherapy remains the treatment of choice for NPC, but radioresistance has been identified as a major cause of therapeutic failure. The Wnt/β-catenin signaling has been found to be involved in NPC radioresistance; however, the effect of β-catenin overexpression on radioresistance remains unknown in NPC until now. This study aimed to examine the impact of β-catenin overexpression on the radiosensitivity of human NPC CNE-2 cells. Methods: Immunohistochemistry was performed to detect the β-catenin expression in normal nasopharyngeal specimens and NPC specimens. The human NPC CNE-2 cell line overexpressing β-catenin was modeled by transfection with the pcDNA3.1/Hygro(+)/β-catenin recombinant vector (transfection group), while cells transfected with the pcDNA3.1/Hygro(+) vector served as negative controls and non-transfected cells served as blank controls. The expression of key molecules of the Wnt/β-catenin signaling pathway was determined using Western blotting and qPCR assays, and the changes of radiation sensitivity were measured with a colony-formation assay. Cell viability was measured by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 -diphenyltetrazolium bromide) assay. In addition, the cell cycle and apoptosis was detected using flow cytometry and the TCF/LEF transcriptional activity was measured with a Dual Luciferase Reporter Assay System. Results: Immunohistochemical staining showed high β-catenin expression in radioresistant NPC specimens, and low expression in radiosensitive NPC specimens and normal nasopharyngeal specimens. Western blotting and qPCR assays detected higher β-catenin expression in the transfection group than in the negative and blank controls (P < 0.01). Down-regulation of GSK-3β expression (P < 0.05) and up-regulation of Cyclin D1 expression (P < 0.01) was detected in β-catenin overexpressing NPC cells exposed to X-ray radiation relative to negative and blank controls. Colony-formation assay revealed higher D₀, D_q and SF in the transfection group than in the negative and blank control groups post-radiation, and the SER in the transfection group was 0.75-fold and 0.68-fold greater than that in the blank and negative control groups, respectively. MTT assay revealed that the viability of CNE-2 cells was significantly higher in the transfection group (96% ± 8.72%) than in the negative control group (74.67 ± 7.05%) and the blank control group (75.33% ± 7.02%) 24 h post-exposure to 6 Gy X-ray radiation (P < 0.05). X-ray radiation led to a lower proportion of CNE-2 cells at the G2/M phase and a lower apoptotic rate in the transfection group than in the negative and blank control groups (P < 0.05). In addition, the TCF/LEF transcriptional activity was higher in the transfection group than in the negative and blank control groups (P < 0.01), and 6 Gy X-ray radiation elevated the TCF/LEF transcriptional activity relative to 0 Gy radiation in the transfection group (P < 0.01). Conclusion: β-catenin overexpression may decrease the radiation sensitivity in NPC CNE-2 cells through activating the downstream transcriptional factors of β-catenin, and reducing G2/M arrest and cell apoptosis.

Objective To investigate the effect of microRNA-1224-5p (miR-1224-5p) on tumor endothelial cells (TECs) of human hepatocellular carcinoma (HCC). Methods Oligonucleotides were chemically synthesized and transfected into TECs using Lipofectamine 2000. TECs were divided into 3 groups: a control (CON) group consisted of TECs without transfection, a negative control (NC) group consisted of TECs transfected with negative control oligonucleotides and Green Fluorescent Protein (GFP), and a micro-up (MU) group consisted of TECs transfected with miR-1224-5p mimic and GFP. The expression of miR-1224-5p was quantified via quantitative reverse transcription polymerase chain reaction (RT-qPCR). The proliferation of TECs was detected using MTT (Thiazolyl Blue Tetrazolium Bromide) assay. Apoptosis and cell cycle were detected via flow cytometry. The migration and invasion of TECs were detected using Ttranswell assay. The ability of tube fromation of TECs was detected using tube formation assay. RESULTS Oligonucleotides were successfully transduced into TECs, and specifically up-regulated the expression of miR-1224-5p. Results The results of RT-qPCR showed that the expression of miR-1224-5p was significantly increased in the MU group (2-ΔΔCt=3.27±0.15) compared with the CON group (2-ΔΔCt=1.00) and the NC group (2-ΔΔCt=1.08±0.11) (P<0.01). The results of MTT assay showed that the cell proliferation was all significantly inhibited in the MU group for the 4 time points compared with the CON group and the NC group (P<0.01). The results of flow cytometry showed that the apoptosis was significantly increased in the MU group (19.29±0.95)% compared with the CON group (8.73±0.64)% and the NC group (9.51±0.56)% (P<0.01). The results of cell cycle detection showed that percentage of G1 phase cells in the MU group (73.3±2.0)% was significantly higher than the CON group (50.7±1.3)% and the NC group (51.7±1.9)% and the percentage of S phase cells in the MU group (23.0±3.0)% was significantly decreased compared with the CON group (31.4±2.7)%, and the NC group (29.1±1.6)%. (P<0.05 or P<0.01). The ability of migration was significantly inhibited in the MU group (51.0±3.6) compared with the CON group (77.7±2.5), and the NC group (79.2±3.5) (P<0.01). And the ability of invasion of TECs was also significantly inhibited in the MU group (9.8±1.3) compared with the CON group (15.8±0.8) and the NC group (15.4±0.9) (P<0.01). The ability of tube formation of TECs in the MU group was inhibited absolutely, but TECs in the CON group and the NC group were not affected. Conclusion Up-regulation of miR-1224-5p could decrease proliferation, induce apoptosis, inhibit migration and invasion, and supress tube formation of TECs of human HCC. Key words: Hepatocellular carcinoma; Vascular endothelial cells of human hepatocellular carcinoma; MicroRNA-1224-5p

To investigate the role of NF-κB/survivin signal pathway in the intima hyperplasia of rat carotid balloon injury restenosis model. NF-κB siRNA lentivirus vector (titer was 1 × 10⁸ TU/ml) was established. Carotid balloon injury restenosis model was made in 33 SD rats. The rats were divided into 4 groups according to different processing methods, including negative control (NC) group (n = 11), NF-κB siRNA group (n =11), NF-κB siRNA+YM155 (survivin inhibitor) (n = 11), the uninjured carotid artery served as the normal control group (n = 11). After 7 days, the carotid sample (n = 5 each group) were harvested to detect the NF-κB and survivin mRNA expression by RT-PCR.The carotid sample were harvested on 28 days (n = 6 each group) for HE staining and measuring intima hyperplasia. Immunohistochemical method was also used to detect the expression of intima proliferation cell nuclear antigen (PCNA) and media α-SM-actin. (1) After 7 days, NF-κB and survivin mRNA expression was significant higher in NC group than in normal control group (P<0.05), the NF-κB mRNA expression was significantly lower in NF-κB siRNA group than in NC group (P<0.05) and similar between NF-κB siRNA group and NF-κB siRNA+YM155 group. The survivin mRNA expression was significantly lower in NF-κB siRNA group compared to NC group (P<0.05) and significantly higher in NF-κB siRNA group than in NF-κB siRNA+YM155 group (P<0.05). (2) After 28 days, intima hyperplasia was observed in NC (0.13 ± 0.01), NF-κB siRNA (0.11 ± 0.01) and NF-κB siRNA+YM155 group (0.09 ± 0.01) mm² (P<0.05). Media area was similar among NC group, NF-κB siRNA group and NF-κB siRNA+YM155 group (P>0.05). I/M ratio was gradually reduced among NC group (1.55 ± 0.07), NF-κB siRNA group (0.92 ± 0.08), NF-κB siRNA+YM155 group (0.76 ± 0.06, all P<0.05). Similar results were found in the residual restenosis rate: NC group (58.71 ± 0.02) %, NF-κB siRNA group (32.13 ± 0.05) %, NF-κB siRNA+YM155 group (26.42 ± 0.03) % (all P<0.05) and expression of vascular smooth muscle cell PCNA: NC group (45.32 ± 7.21) %, NF-κB siRNA group (36.54 ± 6.42) %, NF-κB siRNA+YM155 group (28.57 ± 6.31) % (all P<0.05). On the contrary, the IOD of α-SM-actin in media increased gradually: NC group (0.055 ± 0.006), NF-κB siRNA group (0.072 ± 0.011), NF-κB siRNA+YM155 group (0.084 ± 0.008, all P<0.05). Inhibiting NF-κB expression can significant decrease intima hyperplasia in this model, and this effect may be mediated by inhibiting survivin and reducing the proliferation of vascular smooth muscle cells.

Objective To observe the preventive effect of type Ⅱ collagen on experimental rat articular cartilage damage induced by T-2 toxin,to explore molecular biomarkers of articular cartilage damage and repair,and to provide a theoretical basis for control of articular cartilage damage.Methods Eighty Wistar rats were randomly divided into 4 groups according to their body weights:negative control,positive control,high-dose intervention,and low-dose intervention groups,20 rats in each group.Animals in negative control group were fed with standard rat chow,and animals in other three groups were fed with T-2-toxin-contaminated chow( 100 ng/kgfeed).Animals in negative and positive control groups drank distilled water,animals in high-dose intervention and low-dose intervention groups drank water containing type Ⅱ collagen(0.5,5.0 g/L,respectively).These rats were sacrificed after 3 and 5 months,respectively,and bilateral knee joints were collected.Histopathologic changes in hyaline cartilage were examined by light microscope,serum levels of type Ⅱ collagen carboxyl terminal peptide (CTX-Ⅱ ),cartilage oligomeric matrix protein (COMP) and urinary deoxypyridinoline (DPD) were determined by enzyme-linked immunosorbent assay(ELISA).Results HE staining showed,that the positive control articular chondrocytes were disarranged,deformated,degenerated,with necrosis and extensive areas of chondrocyte loss;but the two intervention groups only showed fibril formation and swelling and surface cartilage cells became round,flat cartilage cells decreased in number,and cartilage cells clustered and so on early pathological changes of osteoarthritis.At the ends of 3 month and 5 month experiment,the levels of serum CTX- Ⅱ in different groups were,negative control[(18.77 ± 4.61),(25.07 ± 9.17)μg/L],high-dose intervention[ (21.11 ± 5.02),(33.20 ± 9.74)μg/L ],low-dose intervention [ ( 19.87 ± 4.53 ),( 29.73 ± 9.32 ) μg/L ] and positive control [ ( 24.43 ± 5.23 ),( 39.17 ±10.49 ) μg/L ] ; the levels of serum COMP were,negative control group [ (5.43 ± 2.75 ),( 6.38 ± 2.23 ) μg/L ],highdose intervention group[ (17.27 ± 4.77),(20.32 ± 4.74)μg/L],low-dose intervention group[(20.13 ± 5.07),(19.44 ± 4.92)μg/L] and positive control group[ (21.37 ± 4.72),(24.52 ± 4.26)μg/L].At the end of 3 month,compared with negative control group,the level of serum CTX- Ⅱ in other three groups increased,but only positive control group increased significantly(P ＜ 0.05) ; at the end of 5 month,compared with negative control group,the level of serum CTX-Ⅱ in other three groups increased significantly,and the difference was statistically significant (all P ＜ 0.05),and the level of CTX-Ⅱ in the two intervention groups was significantly lower compared with that of positive control group(all P ＜ 0.05).Compared with negative control group,the level of serum COMP in other groups increased significantly at the end of 3 month (all P ＜ 0.05) and only the level of serum COMP in high-dose intervention group was significantly lower compared with that of positive control group(P ＜ 0.05).At the end of 5 month,compared with negative control group,the level of serum COMP in other three groups increased significantly,the difference were statistically significant (all P ＜ 0.05) ; the levels of serum COMP in the two intervention groups were significantly lower than that of positive control group(all P ＜ 0.05).At the ends of 3 month and 5 month,the content of urinary DPD in negative control group were[ (3.47 ± 2.20),(4.14 ± 1.06)μg/L],positive control group[ (4.09 ± 2.48),(4.33 ± 3.43)μg/L],high-dose intervention group[ (3.86 ± 2.31 ),(5.72 ± 3.89)μg/L] and low-dose intervention group[ (3.58 ± 2.77),(4.23 ± 2.90)μg/L].The difference between the 4 groups were not statistically significant (F =2.608,2.436,all P ＞ 0.05).Conclusions Type Ⅱ collagen could effectively reduce the level of serum CTX-Ⅱ and COMP in experimental rats and delay the process of articular cartilage damage induced by T-2 toxin. Key words: Cartilage, articular; Collagen type Ⅱ; T-2 toxin

The cytogenetic effects of taking oral contraceptive pills OCPs that contain steroid hormones (estrogen and progesterone) during pregnancy on some aspects of fetus and newborn have been previously reviewed. The objectives of this study are to detect the effect of using different doses of progestin and estrogen by pregnant female mice on the chromosomal aberration CAs and mitotic index MI in their male fetuses. Thirty female mice were divided equally into three major groups: two experimental G1 and G2 and control group according to the level of the dose. Administration of COCPs (either 0.034 or 0.068 mg/kg b. wt. /d dissolved with 0.1ml distilled water) orally once a day, were started at the first day of gestation and continued till day 14 for the experimental groups G1and G2 respectively. When the new pups in each experimental and control group reach two weeks of age, those male offspring 10animals/ group were used for chromosome preparation, while the control group divided into two groups: ten animals represented a negative control given distilled water only, while the other ten animals injected intra peritoneal with mitomycin–C 2mg/Kg represented positive control (once dose). The statistical analysis of mice bone marrow cells have no significant decrease in the mitotic index of the pups belongs to G1 mothers in comparison with negative control group, but it shows a highly significant differences P&lt;0.01 as compared with that of positive control group . Although the pups belong to G2 mothers showed a slight decrease in the mitotic index in comparison with negative control group. Differences were non- significant with negative control and highly significant P&lt;0.01 with positive control group. In addition, the mitotic index of spleen cells of the pups belongs to G1 mothers noticed no significant decrease in comparison with negative control group, however, it shows a significant differences P&lt; 0.05 as compared with positive control group. In the pups belongs to G2 mothers there were significant decrease P&lt; 0.05 in mitotic index in comparison with negative and positive control groups. The results showed that the COCPs caused an increase in chromosomal aberrations CAs with increasing of steroid hormones concentration. In G1 group the CA equal to 19.91%. This reducing in CA was not significant in comparison with those of the negative control which gave 14.33%; this percentage was increased in the result of CAs of the mice treated with mitomycin –C (MMC) which was 48.58%. Treatment with 0.068 mg/kg B wt. of COCPs gave CA equal to 17.18%. This value was not significant in comparison with those of the negative control which was 19.91%. Higher doses of OCPs may cause cytogenetic effects in chromosomal aberrations CAs and decrease the mitotic index (MI) in the bone marrow cells and spleen cells of offspring belonged to mothers treated with oral contraceptive during their pregnancy for 14 days.

Simple SummaryHeat stress is a serious issue in commercial broiler production in hot and humid countries, including Malaysia. Exposure of broilers to heat stress affects their health and productivity. In this context, antibiotics are widely used at sub-therapeutic levels as growth promoters to reduce stress and infectious diseases in order to sustain productivity in commercial broiler farms. However, the extensive use of antibiotics as growth promoters for a long time leads to the development of antibiotic-resistant bacteria and the possibility of antibiotic-resistant genes being transferred among organisms. Recently, postbiotics produced by Lactobacillus plantarum have been widely studied as a feed additive in order to replace in-feed antibiotics. However, to date, no studies have investigated the role of postbiotics in feed for broilers under heat stress.The effects of feeding different postbiotics on growth performance, carcass yield, intestinal morphology, gut microbiota, immune status, and growth hormone receptor (GHR) and insulin-like growth factor 1 (IGF-1) gene expression in broilers under heat stress were assessed in this study. A total of 252 one-day-old male broiler chicks (Cobb 500) were randomly assigned in cages in identical environmentally controlled chambers. During the starter period from 1 to 21 days, all the birds were fed the same basal diet. On day 22, the birds were weighed and randomly divided into six treatment groups and exposed to cyclic high temperature at 36 ± 1 °C for 3 h per day from 11:00 to 14:00 until the end of the experiment. From day 22 to 42 (finisher period), an equal number of birds were subjected to one of the following diets: NC (negative control) basal diet; PC (positive control) basal diet + 0.02% oxytetracycline; or AA (ascorbic acid) basal diet + 0.02% ascorbic acid. The other three groups (RI11, RS5 and UL4) were basal diet + 0.3% different postbiotics (produced from different Lactobacillus plantarum strains, and defined as RI11, RS5 and UL4, respectively). The results demonstrated that birds fed RI11 diets had significantly higher final body weight, total weight gain and average daily gain than the birds that received the NC, PC and AA treatments. The feed conversion ratio was significantly higher in the RI11 group compared with the other groups. Carcass parameters were not affected by the postbiotic-supplemented diet. Postbiotic supplementation improved villi height significantly in the duodenum, jejunum and ileum compared to the NC, PC and AA treatments. The crypt depth of the duodenum and ileum was significantly higher in NC group compared to other treatment groups except RI11 in duodenum, and UL4 in ileum was not different with NC groups. The villus height to crypt depth ratio of duodenum and ileum was significantly higher for the postbiotic treatment groups and AA than the PC and NC treatment groups. The postbiotic RI11 group recorded significantly higher caecum total bacteria and Lactobacillus count and lower Salmonella count compared to the NC and PC treatment groups. The Bifidobacterium population in the NC group was significantly lower compared to the other treatment groups. The postbiotic (RI11, RS5 and UL4) and AA treatment groups showed lower Enterobacteriaceae and E. coli counts and caecal pH than the NC and PC treatment groups. The plasma immunoglobulin M (IgM) level was significantly higher in the birds receiving postbiotic RI11 than those receiving other treatments. The plasma immunoglobulin G (IgG) level was higher in the RI11 treatment group than in the NC, AA and RS5 groups. The plasma immunoglobulin A (IgA) level was not affected by postbiotic supplements. The hepatic GHR mRNA expression level was significantly increased in birds fed postbiotics RI11, RS5 and UL4, AA and PC compared to the NC-fed birds. Postbiotic RI11 led to significantly higher hepatic IGF-1 mRNA expression level compared to the NC, PC, and AA treatments. Mortality was numerically lesser in the postbiotic treatment groups, but not significantly different among all the treatments. In conclusion, among the postbiotics applied in the current study as compared with NC, PC and AA, RI11 could be used as a potential alternative antibiotic growth promoter and anti-stress treatment in the poultry industry.

Response of broiler chickens fed wheat-based diets to xylanase supplementation

Objective To explore the effects of over-expression of Lin28A on glycolysis of gastric cancer cells and the underlying mechanism. Methods Human gastric cancer BGC-823 cells in vitro were stably transfected with lentivirus-mediated over-expression of Lin28A. Lactic acid levels of cell culture medium were detected with biochemical method, phosphofructose kinase(PFK) levels in cells with ELISA, the expression levels of Lin28A, hypoxia-inducible factor (HIF)-1α, glucose transporter (GLUT)-1, and pyruvate kinase M2 (PKM2) were detected using Western blotting, and let-7a RNA using reverse transcription-PCR. Annexin V-allophycocyanin (APC) staining method was used to assess cell apoptosis. Results Compared with the normal control(CON) group and the negative control(NC) group, Lin28A protein level in the over-expression (OE) group was significantly higher (both P<0.001). RNA expression level of let-7a was significantly lower in the OE group than in the NC group (0.602±0.017 vs. 1.001±0.063, P=0.005 2). The levels of the HIF-1α, GLUT-1, PKM2 proteins were all significantly lower in the OE group than in the CON and the NC groups (all P<0.001). The lactic acid levels of cell culture medium in the CON, the NC, and the OE groups were (1.71±0.13), (1.53±0.11), (1.24±0.04)mmol/L, which was significantly lower in the OE group than in the CON group and the NC group (P=0.017 0, 0.031 0). Intracellular PFK level in the OE group was also significantly lower than in the CON group and the NC group [(1.79±0.05)mmol/L vs. (3.71±0.13)mmol/L, P=0.014 0; (1.79±0.05)mmol/L vs. (3.49±0.14)mmol/L, P=0.036 0]. Apoptosis rates of the CON, the NC, and the OE groups were 5.02%±0.14%, 7.16%±0.21%, 10.39%±0.37%, respectively, which was significantly higher in the OE group than in the CON group and the NC group (P=0.000 5, 0.000 7). Conclusion Over-expression of Lin28A can inhibit the expression of let-7a, and induce apoptosis of human gastric cancer cells by suppressing glycolysis of the cells. Key words: Lin28A; Gastric cancer; Glycolysis; Apoptosis

To investigate the expression of miR-155 in patients with diffuse large B-cell lymphoma (DLBCL), and to explore the effect of transfection of miR-155 inhibitor on the biological characteristics of DLBCL cells. A total of 76 patients with DLBCL treated in our hospital were selected from April 2013 to December 2017. In the same time, 40 cases of lymph node reactive hyperplasia (LNRH) were selected as control group. DB cells were cultured and divided into miR-155 inhibitor, negative control and blank groups. The expressions of miR-155 in DLBCL, negative and blank control groups were detected by using real-time PCR, the cell proliferation was detected by MTT assay, the apoptosis was detected by flow cytometry, and the cell migration and invasion were detected by Transwell assay. The relative expression level of miR-155 in tissues of DLBCL patients was significantly higher than that in tissne of controls (1.93±0.16 vs 1.01±0.09) (t=33.991, P=0.000). The expression level of miR-155 increased (P＜0.05) in DLBCL patients with LDH level abnormarity, BCL-2+, MUM1+, Ki-67≥50%, non-GC type, Ann Arbor stage III-IV, extranodal lesion number≥2 and IPI score 3-5. The relative expression level of miR-155 in the miR-155 inhibitor group was lower than that in the negative control group and the blank group (P＜0.05). The absorbance (A) values at 24, 48, 72 and 96 h of culture in the miR-155 inhibitor group were lower than those in the negative control group and the blank group (P＜0.05), while the apoptotic rate was higher than that in the negative control group and the blank group (P＜0.05). Both the migrating cells and invading cell number in the miR-155 inhibitor group were lower than those in the negative control group and the blank group (P＜0.05). The miR-155 highly expresses in DLBCL tissue, which relates with tumor malignancy and invasion progression. The specific inhibition of miR-155 expression in DB cells can reduce cell proliferation, accelerate cell apoptosis, and inhibit cell migration and invasion.

Objective To investigate the effect of estrogen receptor β (ERβ) gene silencing mediated by small interference RNA (siRNA) on apoptosis of human lung cancer LTEP-a2 cells overexpressing ERβ and the sensitivity to 17β-estradiol. Methods siRNA was designed and synthesized as expression vector and transfected into human lung cancer LTEP-a2 cells. After 48 h, the collected cells were randomly divided into the negative control group, the blank control group and the transfected group. Transcription-polymerase chain reaction (RT-PCR) was used to detect ERβ mRNA, and Western blotting was used to detect the expression of ERβ protein. Cellular growth was tested by methylthiazoltetrazolium colorimetry (MTT) assay and the median inhibitory rate (IC50) of each group was calculated. The cell apoptosis rate after treatment by 17β-estradiol was detected by flow cytometry. Results The expression intensities of ERβ mRNA in negative control, blank contro1 and siRNA group were 0.32±0.06, 0.28±0.05 and 0.05±0.02 respectively. The expression of ERβ mRNA was decreased significantly in siRNA group as compared with negative control group (P=0.032) and blank control group (P=0.015). The expression intensities of ERβ protein in negative control, blank contro1 group and siRNA group were 0.18±0.04, 0.28±0.03 and 0.06±0.01, respectively. The expression of ERβ protein was decreased significantly in siRNA group as compared with negative control group (P=0.038) and blank control group(P=0.000). After intervention with different concentrations of 17β-estradiol, the survival rate was decreased significantly in transfected group as compared with negative control group and blank control group ; The value of half maximal inhibitory concentration (IC50) in negative control , blank contro1 and siRNA group were (28.34±4.32)%, (27.66±3.04)% and (8.34±1.22)%, respectively. the growth of the lung cancer cell LTEP-a2 was significantly inhibited in siRNA group as compared with negative control group and blank control group ; the apoptosis rate in the transfected group was significantly higher than that in the negative control group and blank control group [(35.68±1.58)%, (3.55±0.33)% and (2.34±0.58)% respectively]. Conclusion ERβ gene silencing mediated by siRNA might inhibit growth and promote the apoptosis of non-small lung cancer with estrogen intervention. ERβ could play a significant role in screening female sex hormone-dependent non-small lung cancer. Key words: Lung cancer; Sex hormone; Small interference RNA; Estrogen receptor β gene; Apoptosis

ObjectivesTo evaluate the cytocompatibility, bioactivity, and anti-inflammatory potential of the new pre-mixed calcium silicate cement-based sealers Bioroot Flow (BrF) and AH Plus Bioceramic Sealer (AHPbcs) on human periodontal ligament stem cells (hPDLSCs) compared to the epoxy resin-based sealer AH Plus (AHP).Materials and methodsStandardized discs and 1:1, 1:2, and 1:4 eluates of BrF, AHPbcs and AHP after setting were prepared. The following assays were performed: cell attachment and morphology via SEM, cell viability via a MTT assay, cell migration/proliferation via a wound-healing assay, cytoskeleton organization via immunofluorescence staining; cytokine release via ELISA; osteo/cemento/odontogenic marker expression via RT-qPCR, and cell mineralized nodule formation via Alizarin Red S staining. HPDLSCs were isolated from extracted third molars from healthy patients. Comparisons were made with hPDLSCs cultured in unconditioned (negative control) or osteogenic (positive control) culture media. Statistical significance was established at p < 0.05.ResultsBoth BrF and AHPbcs showed significantly positive results in the cytocompatibility assays (cell metabolic activity, migration, attachment, morphology, and cytoskeleton organization) compared with a negative control group, while AHP showed significant negative results. BrF exhibited an upregulation of at least one osteo/cementogenic marker compared to the negative and positive control groups. BrF showed a significantly higher calcified nodule formation than AHPbcs, the negative and positive control groups, while AHPbcs was higher than the negative control group. Both were also significantly higher than AHP group.ConclusionBrF and AHPbcs exhibit adequate and comparable cytocompatibility on hPDLSCs. BrF also promoted the osteo/cementogenic differentiation of hPDLSCs. Both calcium silicate-based sealers favored the downregulation of the inflammatory cytokine IL-6 and the calcified nodule formation from hPDLSCs. BrF exerted a significantly higher influence on cell mineralization than AHPbcs.Clinical relevanceThis is the first study to elucidate the biological properties and immunomodulatory potential of Bioroot Flow and AH Plus Bioceramic Sealer. The results act as supporting evidence for their use in root canal treatment.