Effects of manganese glycine on eggshell quality, eggshell ultrastructure, and elemental deposition in aged laying hens.

Poor eggshell quality of eggs laid by aged laying hens is the major problem affecting the length of the rearing period in the laying hen industry. Trace elements are required and play vital roles in the eggshell quality of laying hens. Appropriate dose of organic microelements is environmentally friendly and sufficient to satisfy the needs of hens because of their greater bioavailability and lower excretion than inorganic forms. The aim of this experiment was to investigate the effects of manganese (Mn) glycine (MG) on eggshell quality, elemental deposition, and eggshell ultrastructure in aged laying hens. A total of 720 Hy-Line Brown hens 70 weeks old were assigned equally to four groups with six replicates of 30 birds each. The hens were fed basal diets (without Mn supplementation) supplemented with 120 mg/kg of Mn from manganese sulfate monohydrate (MSM), or 40, 80, or 120 mg/kg Mn from MG for 12 weeks. Dietary supplementation with 80 mg/kg Mn from MG resulted in the greatest eggshell strength after 6 weeks of treatment (P = 0.047), and in greater eggshell strength than observed in the MSM control after 12 weeks of treatment (P = 0.025). After 12 weeks of treatment, the eggs of hens in the MG groups showed lower mammillary layer thickness in the blunt end, equator, and acute end than observed in the MSM control group (P < 0.001). With the exception of the blunt ends of eggs from hens in the 120 mg/kg MG group, the eggs of hens in the MG groups, compared with the MSM control group, exhibited a lower mammillary layer ratio, and greater palisade layer ratio and effective layer ratio in the blunt end, equator, and acute end (P < 0.001). Dietary supplementation with 80 mg/kg Mn from MG, compared with the MSM control and 40 and 120 mg/kg MG, resulted in the greatest palisade layer thickness and effective layer thickness, and the lowest mammillary layer thickness in the equator (P < 0.001, P = 0.001, P < 0.001, respectively). Furthermore, supplementation with 80 mg/kg Mn from MG exhibited the greatest ratio of the palisade layer and effective layer, and the lowest mammillary layer ratio in the blunt end and equator (all P < 0.001). The Mn content of eggshells in hens-fed diets supplemented with 80 and 120 mg/kg Mn from MG was greater than that in the MSM control and 40 mg/kg MG groups (P = 0.035). Dietary supplementation with 80 or 120 mg/kg Mn from MG resulted in greater tibia Mn content than observed in the 40 mg/kg MG group (P = 0.019), and greater yolk Mn content than observed in the 40 mg/kg MG and MSM control groups (P = 0.018). In conclusion, dietary supplementation with 80 mg/kg Mn from MG, compared with the MSM control (120 mg/kg Mn), may increase the deposition efficiency of Mn, alter eggshell elemental composition, improve eggshell ultrastructure, and enhance eggshell strength in aged laying hens.

Effects of exogenous energy on synthesis of steroid hormones and expression characteristics of the CREB/StAR signaling pathway in theca cells of laying hen.

Energy and the cAMP-response element binding protein (CREB)/steroidogenic acute regulatory protein (StAR) signaling pathway play important roles in steroid hormone production and follicular development in hens. This present study aimed to investigate the effects of exogenous energy on the synthesis of steroid hormones and the expression characteristics of the CREB/StAR signaling pathway in theca cells of laying hen. The primary theca cells of small yellow follicles were randomly divided into 6 treatments and cultured in medium with glucose concentrations of 1, 1.5, 3, 4.5, 6, and 7.5 mg/mL for 48 h. It was found that growth was robust and cell outlines were clear when cells were cultured with 1, 1.5, 3, and 4.5 mg/mL glucose, but cell viability was diminished and cell density decreased after exposure to glucose at 6 and 7.5 mg/mL for 48 h. Cell viability showed an increasing and then decreasing quadratic response to increasing glucose concentration in culture (r2 = 0.688, P < 0.001). The cell viability of theca cells cultured with 4.5 mg/mL glucose was greater than those cultured with 1, 1.5, 6, and 7.5 mg/mL glucose (P < 0.05). The concentration of estradiol in the medium containing 3 mg/mL glucose was higher than in medium containing 1, 1.5, and 6 mg/mL glucose (P < 0.05). There was an increasing and then decreasing quadratic correlation between progesterone concentrations and glucose concentrations (r2 = 0.522, P = 0.002). The concentration of progesterone in medium with 4.5 mg/mL glucose was higher than in medium with 1 and 7.5 mg/mL glucose (P < 0.05). There was an increasing and then decreasing quadratic correlation between the relative expression of CREB1 (r2 = 0.752, P < 0.001), StAR (r2 = 0.456, P = 0.002), CYP1B1 (r2 = 0.568, P < 0.001), and 3ß-HSD (r2 = 0.319, P = 0.018) in theca cells of laying hens and glucose concentrations after treatment with different glucose concentrations for 48 h. After treatment with 4.5 mg/mL glucose, the expression of StAR, CYP1B1, and 3ß-HSD genes were increased compared to treatment with 1, 1.5, 3, 6, and 7.5 mg/mL glucose (P < 0.001). There was an increasing and then decreasing quadratic correlation between glucose concentrations and protein expression of CREB1 (r2 = 0.819, P < 0.001), StAR (r2 = 0.844, P < 0.001), 3ß-HSD (r2 = 0.801, P < 0.001), and CYP11A1 (r2 = 0.800, P < 0.001) in theca cells of laying hens. The protein expression of CREB1, StAR, and 3ß-HSD in theca cells cultured with 4.5 mg/mL glucose was higher than in other groups (P < 0.001). The results indicate that the appropriate glucose concentration (4.5 mg/mL) can improve the synthesis of steroid hormones in theca cells of laying hens through the upregulation of key genes and proteins in the CREB/StAR signaling pathway.

Effects of energy restriction during growing phase on the productive performance of Hyline Brown laying hens aged 6 to 72 wk.

The aim of this study was to assess the effects of energy-restricted feeding during growing phase on the productive performance of Hyline Brown laying hens aged 6 to 72 wk. A total of 720 six-week-old layer chicks were allocated equally to 3 groups with 6 replicates of 40 pullets each, and were fed 1 of 3 diets that were nutritionally similar except for the apparent metabolizable energy corrected for nitrogen (AMEn) content. At the age of 6 to 17 wk, the pullets in the control group were given diet with 2,850 kcal/kg AMEn, and were fed ad libitum. The levels of AMEn in diet of pullets in the experimental groups were 90% (2,565 [2,850 × 90%] kcal/kg) and 80% (2,280 [2,850 × 80%] kcal/kg) of that in control group, and the daily amount of feed was restricted to the absolute quantity of the diet consumed by pullets in control group. At the age of 18 to 72 wk, all the hens were fed with the same diets ad libitum. As energy restriction increased in the growing phase, body weight (BW) dropped at the ages of 12 and 15 to 23 wk (at 23 wk: P = 0.001; at other ages: P < 0.001), but it showed no significant difference at 24 wk (P = 0.071). At 20 wk, restricting energy induced a delay in the development of sexual organs, including the ovary stroma, oviduct, and small yellow follicle (P < 0.05), as well as a delay in sexual maturity (P < 0.05). Consequently, the laying rate in the first and second periods dropped linearly (P = 0.046, 0.030, and 0.038, P < 0.001, respectively). The coefficient of variation (CV) in the BW at 19, 20, and 21 wk (P = 0.040, 0.023, and 0.042, respectively), the CV of age at first egg (P < 0.001), and CV of individual egg number at age 18 to 72 wk (P < 0.001) decreased linearly. There was a linear increase in the laying rate of hens in the later periods (at age 32-72 wk, P < 0.05), as well as in the average total egg number per hen and average laying rate at the age of 18 to 72 wk (P = 0.006). The average egg mass also showed a linear increase with increasing levels of energy restriction (P < 0.001). In summary, although appropriate energy restriction during growing phase delayed sexual maturity and sexual organ development in early-laying Hyline Brown pullets, it improved uniformity of BW, age at first egg laying, and individual egg number, and increased egg number per hen, laying rate, average egg mass, and number of settable eggs from 18 to 72 wk of age.

Postoperative pulmonary complications in older patients undergoing elective surgery with a supraglottic airway device or tracheal intubation.

The two most commonly used airway management techniques during general anaesthesia are supraglottic airway devices and tracheal tubes. In older patients undergoing elective non-cardiothoracic surgery under general anaesthesia with positive pressure ventilation, we hypothesised that a composite measure of in-hospital postoperative pulmonary complications would be less frequent when a supraglottic airway device was used compared with a tracheal tube. We studied patients aged ≥ 70 years in 17 clinical centres. Patients were allocated randomly to airway management with a supraglottic airway device or a tracheal tube. Between August 2016 and April 2020, 2900 patients were studied, of whom 2751 were included in the primary analysis (1387 with supraglottic airway device and 1364 with a tracheal tube). Pre-operatively, 2431 (88.4%) patients were estimated to have a postoperative pulmonary complication risk index of 1-2. Postoperative pulmonary complications, mostly coughing, occurred in 270 of 1387 patients (19.5%) allocated to a supraglottic airway device and 342 of 1364 patients (25.1%) assigned to a tracheal tube (absolute difference -5.6% (95%CI -8.7 to -2.5), risk ratio 0.78 (95%CI 0.67-0.89); p < 0.001). Among otherwise healthy older patients undergoing elective surgery under general anaesthesia with intra-operative positive pressure ventilation of their lungs, there were fewer postoperative pulmonary complications when the airway was managed with a supraglottic airway device compared with a tracheal tube.

[Response characteristics of tislelizumab combined with chemotherapy in first-line treatment of locally advanced or metastatic non-squamous non-small cell lung cancer].

Objective: To investigate the response characteristics of patients with locally advanced/metastatic non-squamous non-small cell lung cancer (nsq-NSCLC) treated with tislelizumab in combination with chemotherapy in the first line. Methods: Patients with nsq-NSCLC who achieved complete or partial remission after treatment with tislelizumab in combination with chemotherapy or chemotherapy alone in the RATIONALE 304 study, as assessed by an independent review board, were selected to analyze the response characteristics and safety profile of the responders. Time to response (TTR) was defined as the time from randomization to the achievement of first objective response. Depth of response (DpR) was defined as the maximum percentage of tumor shrinkage compared with the sum of the baseline target lesion length diameters. Results: As of January 23, 2020, 128 patients treated with tislelizumab in combination with chemotherapy achieved objective tumor response (responders), representing 57.4%(128/223) of the intention-to-treat population, with a TTR of 5.1 to 33.3 weeks and a median TTR of 7.9 weeks. Of the responders (128), 50.8%(65) achieved first remission at the first efficacy assessment (week 6), 31.3%(40) at the second efficacy assessment (week 12), and 18.0%(23) at the third and subsequent tumor assessments. The percentages of responders who achieved a depth of tumor response of 30% to <50%, 50% to <70% and 70% to 100% were 45.3%(58/128), 28.1%(36/128) and 26.6%(34/128), respectively, with median progression-free survival (PFS) of 9.0 months (95% CI: 7.7 to 9.9 months), 11.5 months (95% CI: 7.7 months to not reached) and not reached (95% CI: 11.8 months to not estimable), respectively. Tislelizumab plus chemotherapy were generally well tolerated in responders with similar safety profile to the overall safety population. Conclusion: Among responders to tislelizumab in combination with chemotherapy for nsq-NSCLC, 82.0%(105/128) achieves response within the first two tumor assessments (12 weeks) and 18.0%(23/128) achieves response at later (18 to 33 weeks) assessments, and there is a trend toward prolonged PFS in responders with deeper tumor response.

Effects of energy-restricted feeding during rearing on sexual maturation and reproductive performance of Rugao layer breeders.

The aim of this study was to assess the effects of energy-restricted feeding during rearing on the sexual maturation and reproductive performance of Rugao layer breeders. A total of 2,400 8-wk-old Rugao layer breeders were randomly assigned to one of 5 groups (480 pullets per group) with eight replicates and were fed one of 5 diets that were nutritionally similar with the exception of apparent metabolizable energy corrected for nitrogen (AMEn) content (2,850, 2,750, 2,650, 2,550, and 2,450 kcal AMEn/kg) from 8 to 18 wks of age. The daily amount of feed was restricted to the absolute quantity of the diet consumed by laying hens fed 2,850 kcal AMEn per kg diet ad libitum (control). From 18 to 52 wks of age, all hens were fed basal diets ad libitum. The body weight of layer breeders at 18 wks of age decreased linearly with increasing energy restriction (P < 0.001), but caught up within 3 wks of ad libitum feeding (P = 0.290). The coefficient of variation of the body weight of the hens at 18, 21, and 24 wks of age decreased linearly (P = 0.010, 0.025, and 0.041, respectively) with increasing energy restriction during rearing. Energy-restricted feeding delayed sexual organ development at 18, 20, and 22 wks of age, including the number of large yellow follicles, oviduct length, oviduct length index, oviduct index, and ovary stroma index (P < 0.05), and delayed sexual maturity, including the age at laying the first egg and the age at 5% and 50% egg production (P = 0.042, 0.004, and 0.029, respectively). Consequently, egg number from 5% to 50% egg production decreased linearly as the degree of energy restriction increased (P = 0.001) and egg production of hens in the energy-restricted feeding groups was lower than that of hens in the ad libitum feeding group (6.36, 6.43, 6.4, and 4.61% vs. 14.29%; P < 0.05) from 18 to 20 wks of age. Furthermore, egg weight increased linearly as energy restriction increased (P < 0.001) and laying hens in the most severe energy-restricted feeding group had more setting eggs (normal eggs weighing >40 g) than hens in the ad libitum feeding and lighter energy-restricted feeding groups (149.57 vs. 144.34, 142.66, 143.63, and 141.78; P < 0.05). No significant differences were observed in fertility, hatchability of fertile eggs, and hatchability of setting eggs (P = 0.381, 0.790, and 0.605, respectively). In conclusion, moderate energy restriction (85.97%, 2,450 vs. 2,850 kcal AMEn/kg) from 8 to 18 wks of age increased egg weight as well as the production of setting eggs in native layer breeders throughout the laying period, without adverse effects on productive performance from 18 to 52 wks of age, or fertility and hatchability at 52 wks of age.

Genome-wide association studies for mottled eggs in chickens using a high-density single-nucleotide polymorphism array.

Mottled eggs in layer chickens are gaining increasing attention because of the economic impact on the egg industry caused by the reduced sale value of commodity eggs. However, the genetic architecture underlying mottled eggs is not well understood. The genetic architecture underlying the mottled egg trait was investigated using genome-wide association studies (GWAS) by high-density arrays, using a total of 407 pink eggs and 799 blue eggs from an F2 resource population generated by crossing Dongxiang Blue-shelled and White Leghorn chickens. The mottled egg score in blue eggs was found to be higher than that in pink eggs. The single-nucleotide polymorphism heritability of mottled egg at laying day and storage for 7 days was 0.18 and 0.20, respectively. Bivariate GWAS provided 29 significant loci, mainly located on GGA2, GGA3, GGA8, GGA10, GGA15, GGA17, and GGA23, affecting mottled egg on laying day. Candidate genes RIMS2, SLC25A32, RIMBP2, VPS13B, and RGS3 were obtained for mottled eggshell by bivariate GWAS and gene annotation. Our findings provide new insights into the genetic architecture of mottled egg in hens, and demonstrate that a genomic selection method would be profitable for breeding out the mottled egg trait.

Dietary supplementation of total flavonoids from Rhizoma Drynariae improves bone health in older caged laying hens.

Caged layer osteoporosis (CLO) is a common bone metabolism diseases and poses a great threat to the production of laying hens. So far, there is no effective nutrition intervention to prevent CLO. The objective of this study was to evaluate the effects of dietary total flavonoids from Rhizoma Drynariae (TFRD), a Chinese herbal, on bone health, egg quality, and serum antioxidant capacity of caged laying hens. A total of two hundred sixteen, 54-wk-old Lohmann Pink-shell laying hens at were allocated to 3 groups with 6 replicates of 12 hens per replicate. The control group was fed a basal diet (BD) and 2 treatment groups additionally supplied with 0.5 or 2.0 g/kg TFRD, respectively. Results showed that supplying 2.0 g/kg TFRD enhanced the activities of serum total antioxidant capacity (P < 0.01) and glutathione peroxidase (P < 0.05) and had higher femur and tibia bone mineral density (both P < 0.05) compared with the control group. Dietary 2.0 g/kg TFRD also reduced the activities of serum alkaline phosphatase (P < 0.01), tartrate resistant acid phosphatase (P < 0.01), and the contents of osteocalcin (P < 0.01). Furthermore, tibia histomorphology observation showed that the microstructure of bone tissue was improved after TFRD treatment. Egg quality was not affected by TFRD while the egg weight significantly increased (P < 0.01). These findings suggested that TFRD has beneficial effects on bone health in older caged laying hens.

Effects of high-dose selenium-enriched yeast on laying performance, egg quality, clinical blood parameters, organ development, and selenium deposition in laying hens.

Organic selenium (Se) supplementation from Se-enriched yeast (SY) has been advocated and approved for use in animal feeds by some nutritionists and researchers rather than inorganic Se from sodium selenite. However, there is little available safety data of SY in laying hens. A subchronic study was conducted to determine if high-dose SY affects the safety of hens. A total of 768, 30-wk-old, Hy-Line Brown hens were randomly assigned to 1 of 4 groups (192 laying hens per group) with 6 replicates of 32 birds each. After a 2-wk acclimation period, the birds were fed diets supplemented with 0, 0.3, 1.5, or 3.0 mg/kg Se from SY for 12 wk. Throughout the study period, clinical observations and laying performance were measured. The hematological and chemical parameters of blood samples and the Se concentration in eggs were examined after SY supplementation for 4, 8, and 12 wk, and the egg quality was measured after 12 wk. At the end of the study, full post-mortem examinations were conducted: breast Se concentrations were measured, visceral, and reproductive organs were weighed, and specified tissues were collected for subsequent histological examinations. Although the Se concentrations in the eggs and breast meat from hens fed 3.0 mg/kg of Se from SY were 1036.73% and 2127.93% higher (P < 0.001) than those from hens fed a basal diet after 12 wk, no treatment-related changes of toxicological significance were observed. Therefore, up to 3 mg/kg organic Se from SY can be used to supplement the diets for laying hens without adverse effects following 84-d administration.

Comparison of dynamic change of egg selenium deposition after feeding sodium selenite or selenium-enriched yeast.

The aim of this study was to compare the dynamic change of egg selenium (Se) deposition after sodium selenite (SS) or selenium-enriched yeast (SY) supplementation for 1, 3, 5, 7, 14, 21, 28, 56, and 84 d. A total of 576 32-wk-old Hy-Line Brown laying hens were randomly assigned to 3 groups (192 laying hens per group) with 6 replicates, and fed a basal diet (without Se supplementation) or basal diets with 0.3 mg/kg of Se from SS or 0.3 mg/kg of Se from SY, respectively. The results showed that the Se concentrations in the eggs from hens fed a SY-supplemented diet were significantly higher (P < 0.001) than those from hens fed a SS-supplemented diet or a basal diet after 3 d. And the Se concentrations in the eggs from hens fed a SS-supplemented diet were significantly higher (P < 0.001) than those from hens fed a basal diet after 14 d. There was a positive linear and quadratic correlation between Se concentrations in the eggs from hens fed a SY-supplemented diet (r2 = 0.782, P < 0.001; r2 = 0.837, P < 0.001) or SS-supplemented diet (r2 = 0.355, P < 0.001; r2 = 0.413, P < 0.001) and number of feeding days. The Se concentrations in the breasts from hens fed a SY-supplemented diet were 126.98% higher (P < 0.001) than those from hens fed a SS-supplemented diet, and were 299.44% higher (P < 0.001) than those from hens fed a basal diet after the 84-d feeding period. In conclusion, the dietary Se was gradually transferred into eggs with the extension of the experimental duration. The deposition rate of Se in the eggs from hens fed a SY-supplemented diet was much more rapid than that from hens fed a SS-supplemented diet, and the organic Se from SY had higher bioavailability as compared to inorganic Se from SS.

Safety evaluation of daidzein in laying hens: Effects on laying performance, hatchability, egg quality, clinical blood parameters, and organ development.

Daidzein has become increasingly popular as a dietary supplement, particularly for postpeak-estrus animals, as a safe and natural alternative estrogen-like compound. However, there is little available safety data of daidzein in laying hens. A study was conducted to examine if high-dose daidzein affected the safety of hens, including mortality, laying performance, egg quality, hematological parameters, clinical chemical parameters, organ development parameters, and hatchability. A total of 2,448 42-wk-old Rugao laying hens were randomly assigned to 4 groups with 6 replicates of 102 birds each (612 laying hens per group). After a 2-wk acclimation period, the birds were fed diets supplemented with 0, 10, 100, or 200 mg/kg of daidzein for 12 wk. The hatchability of setting eggs increased linearly with increasing dietary daidzein supplementation (P = 0.034), while the hatchability of fertile eggs also tended to increase linearly (P = 0.069). The red cell distribution width (RCDW) and coefficient variation of RCDW showed an increasing and then decreasing quadratic response to increasing dietary daidzein supplementation (P = 0.001 and 0.002, respectively). No statistically significant changes were observed in mortality, laying performance, egg quality, clinical chemistry parameters, or organ development parameters (P > 0.05). The magnitude of these hematological changes was such that they were considered to be of no toxicological significance. Therefore, a nominal daidzein concentration of 200 mg/kg is not expected to cause adverse effects following daily administration to laying hens for 84 d.

Tumorigenic potential is restored during differentiation in fusion-reprogrammed cancer cells.

Detailed understanding of the mechanistic steps underlying tumor initiation and malignant progression is critical for insights of potentially novel therapeutic modalities. Cellular reprogramming is an approach of particular interest because it can provide a means to reset the differentiation state of the cancer cells and to revert these cells to a state of non-malignancy. Here, we investigated the relationship between cellular differentiation and malignant progression by the fusion of four independent mouse cancer cell lines from different tissues, each with differing developmental potentials, to pluripotent mouse embryonic stem (ES) cells. Fusion was accompanied by loss of differentiated properties of the four parental cancer cell lines and concomitant emergence of pluripotency, demonstrating the feasibility to reprogram the malignant and differentiative properties of cancer cells. However, the original malignant and differentiative phenotypes re-emerge upon withdrawal of the fused cells from the embryonic environment in which they were maintained. cDNA array analysis of the malignant hepatoma progression implicated a role for Foxa1, and silencing Foxa1 prevented the re-emergence of malignant and differentiation-associated gene expression. Our findings support the hypothesis that tumor progression results from deregulation of stem cells, and our approach provides a strategy to analyze possible mechanisms in the cancer initiation.

Left ventricular longitudinal rotation changes in primary hypertension patients with normal left ventricular ejection fraction detected by two-dimensional speckle tracking imaging.

In order to assess the left ventricular (LV) longitudinal rotation (LR) in primary hypertension (PH) patients with a normal LV ejection fraction. Conventional echocardiography was performed in 61 healthy subjects and 64 PH patients. The apical four-chamber views in these patients were acquired by GE-Vivid7 or E9, then the peak radial strain in the systolic period and the strain rate in systole, in early and late diastolic periods, were measured. Segmental LR and global LR were assessed by using two-dimensional speckle tracking imaging (2D-STI). The peak radial strain rate in the early diastolic period in PH patients was significantly lower than that in healthy subjects. The rotational degrees of the middle and base lateral, the apex and the middle septum walls in PH patients were significantly different from those of the healthy subjects. The healthy subjects had prominent counter-clockwise LR (0.29°±2.86°) and the PH patients had prominent clockwise LR (-2.13°±2.93°) in non-LV wall hypertrophy and (-2.43°±2.66°) in LV wall hypertrophy. The time delay between the LV lateral wall and the septum wall in PH patients correlated to the peak LR. We concluded that 2D-STI can assess the time delay between the LV lateral wall and the septum wall to the peak LR and clockwise LR in patients with PH, and prove that PH patients have a clockwise LR. By this, we conclude that in PH patients, the LV early systolic function have changed.

Non-viral FoxM1 gene delivery to hepatocytes enhances liver repopulation.

Hepatocyte transplantation as a substitute strategy of orthotopic liver transplantation is being studied for treating end-stage liver diseases. Several technical hurdles must be overcome in order to achieve the therapeutic liver repopulation, such as the problem of insufficient expansion of the transplanted hepatocytes in recipient livers. In this study, we analyzed the application of FoxM1, a cell-cycle regulator, to enhance the proliferation capacity of hepatocytes. The non-viral sleeping beauty (SB) transposon vector carrying FoxM1 gene was constructed for delivering FoxM1 into the hepatocytes. The proliferation capacities of hepatocytes with FoxM1 expression were examined both in vivo and in vitro. Results indicated that the hepatocytes with FoxM1 expression had a higher proliferation rate than wild-type (WT) hepatocytes in vitro. In comparison with WT hepatocytes, the hepatocytes with FoxM1 expression had an enhanced level of liver repopulation in the recipient livers at both sub-acute injury (fumaryl acetoacetate hydrolase (Fah)(-/-) mice model) and acute injury (2/3 partial hepatectomy mice model). Importantly, there was no increased risk of tumorigenicity with FoxM1 expression in recipients even after serial transplantation. In conclusion, expression of FoxM1 in hepatocytes enhanced the capacity of liver repopulation without inducing tumorigenesis. FoxM1 gene delivered by non-viral SB vector into hepatocytes may be a viable approach to promote therapeutic repopulation after hepatocyte transplantation.

Cuprous oxide nanoparticles inhibit the growth and metastasis of melanoma by targeting mitochondria.

Metal and its oxide nanoparticles show ideal pharmacological activity, especially in anti-tumor therapy. Our previous study demonstrated that cuprous oxide nanoparticles (CONPs) selectively induce apoptosis of tumor cells in vitro. To explore the anti-tumor properties of CONPs in vivo, we used the particles to treat mouse subcutaneous melanoma and metastatic lung tumors, based on B16-F10 mouse melanoma cells, by intratumoral and systemic injections, respectively. The results showed that CONPs significantly reduced the growth of melanoma, inhibited the metastasis of B16-F10 cells and increased the survival rate of tumor-bearing mice. Importantly, the results also indicated that CONPs were rapidly cleared from the organs and that these particles exhibited little systemic toxicity. Furthermore, we observed that CONPs targeted the mitochondria, which resulted in the release of cytochrome C from the mitochondria and the activation of caspase-3 and caspase-9 after the CONPs entered the cells. In conclusion, CONPs can induce the apoptosis of cancer cells through a mitochondrion-mediated apoptosis pathway, which raises the possibility that CONPs could be used to cure melanoma and other cancers.

Differential levels of elastin fibers and TGF-ß signaling in the skin of Caucasians and African Americans.

BACKGROUND: While skin color is the most notable difference among ethnic skins the current knowledge on skin physiological and aging properties are based mainly on Caucasian skin studies. OBJECTIVE: To evaluate histological differences in elastin fiber network and differential responsiveness to TGF-ß in skin of Caucasians and African Americans. METHODS: These studies were undertaken using human skin biopsies, primary dermal fibroblasts, Western blot analyses, immunofluorescence microscopy, cDNA array and quantitative real-time PCR. RESULTS: In Caucasian subjects, tropoelastin expression and elastin fibers in photoprotected skin was substantially less than in age-matched African American skin. Expression of tropoelastin in photoexposed skin of African American was similar to their photoprotected skin, suggesting that photoexposure did not affect elastin fibers in African American skin to the same extent as Caucasian skin. An elevated level of TGF-ß1 present in media from dermal fibroblasts derived from African American skins correlated well with the higher levels of TGF-ß mRNA in African American skins analyzed by cDNA array. Treatment with TGF-ß1 resulted in a considerably higher induction of elastin mRNA in dermal fibroblasts from African Americans than from Caucasian fibroblasts, indicative of enhanced TGF-ß signaling in African American skins. Furthermore, UVA exposure decreased levels of elastin mRNA in Caucasian fibroblasts compared to African Americans fibroblasts. CONCLUSION: These results suggest that there are ethnic differences in the elastin fiber network and in TGF-ß signaling in African American and Caucasian skin, and that African American have less UV dependent loss of elastin than Caucasian which may contribute to the different perceived aging phenotypes.

Genetic background of Citrus nobilis Lour. 'Gonggan' based on the chloroplast trnL gene.

The genetic backgrounds of many Citrus varieties are quite complex. Classifications and phylogenetic relationships of Citrus species have become the focus of researchs. Some conserved genes of chloroplast genome's research have been proven effective in determining the biosources of hybrids and phylogenetic analysis. Thus, we studied variations among the chloroplast trnL gene sequences of 10 Citrus species, including C. nobilis Lour. 'Gonggan'. The amplification results of different trnL target genes and identification of the double-enzyme cut after cloning show that lengths of all trnL sequences were within 895 to 935 bp and a total of 24 variation sites were detected among the 10 material samples. Clustering analysis revealed differences in trnL genes caused by systematic evolution and allowed the determination of variations among Citrus varieties. Variation sites of trnL sequences can be used in the phylogenetic classification and species identification of Citrus, and the results agreed with random amplified polymorphic DNA marker results. C. nobilis Lour. 'Gonggan' is closely associated with the other two varieties in Zhaoqing area, and C. nobilis Lour. 'Gonggan' and C. haniana Hort. ex Tseng 'Sihuihanggan' can be classified into the same category. C. nobilis Lour. 'Gonggan' as a natural hybrid is probably a hybrid with C. haniana Hort. ex Tseng 'Sihuihanggan' as its female parent.

A novel anti-ageing mechanism for retinol: induction of dermal elastin synthesis and elastin fibre formation.

Dermal elastic fibres are extracellular matrix protein complexes produced by fibroblasts and involved in skin elasticity. Elastin fibres decrease with age as a result of reduced synthesis and increased degradation, resulting in skin sagging and reduced skin elasticity. In this study, we show that retinol (ROL), known to enhance dermal collagen production, is also enhancing elastin fibre formation. ROL induced elastin gene expression and elastin fibre formation in cultured human dermal fibroblasts. Topical treatment of cultured human skin explants with a low dose (0.04%) of ROL increased mRNA and protein levels of tropoelastin and of fibrillin-1, an elastin accessory protein, as documented by QPCR and immunohistochemistry staining. Luna staining confirmed the increased elastin fibre network in the ROL-treated skin explants, as compared with untreated controls. These data demonstrate that ROL exerts its anti-ageing benefits not only via enhanced epidermal proliferation and increased collagen production, but also through an increase in elastin production and assembly.