CircDDX17 reduces 5-fluorouracil resistance and hinders tumorigenesis in colorectal cancer by regulating miR-31-5p/KANK1 axis.

OBJECTIVE: Colorectal cancer (CRC) is one of the most common malignancies worldwide. Chemotherapy resistance is a considerable obstacle to CRC treatment. Circular RNAs (circRNAs) are involved in the pathogenesis of many cancers. This study aimed to investigate the role and molecular basis of DEAD-box helicase 17 circRNA (circDDX17) in 5-fluorouracil (5-Fu) sensitivity and CRC progression. MATERIALS AND METHODS: The levels of circDDX17, microRNA-31-5p (miR-31-5p) and kidney ankyrin repeat-containing protein 1 (KANK1) were detected by quantitative real-time PCR or western blot assay. Cell viability was assessed by Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis rate was monitored by flow cytometry. Cell invasion capacity was evaluated by transwell assay. Western blot assay was conducted to measure the expression of matrix metallopeptidase 9 (MMP9) and E-cadherin. The interaction among circDDX17, miR-31-5p and KANK1 was indicated by bioinformatics analysis and dual-luciferase reporter assay. Xenograft assay was performed to analyze tumor growth and 5-Fu sensitivity in vivo. RESULTS: CircDDX17 and KANK1 were down-regulated, while miR-31-5p was upregulated in CRC tissues and cells. Upregulation of circDDX17 enhanced 5-Fu sensitivity and impeded CRC development. CircDDX17 inhibited 5-Fu resistance and CRC progression via sponging miR-31-5p. Besides, KANK1 depletion attenuated the effect of circDDX17 upregulation on chemosensitivity and CRC progression. CircDDX17 regulated KANK1 expression by binding to miR-31-5p. Moreover, circDDX17 overexpression blocked tumor growth and elevated 5-Fu sensitivity in vivo. CONCLUSIONS: Upregulation of circDDX17 strengthened chemosensitivity of CRC to 5-Fu and blocked CRC progression by regulating miR-31-5p/KANK1 axis, which might provide an effective treatment strategy for CRC patients.

Effect of propofol and sevoflurane anesthesia on postoperative cognitive function and levels of Aß-42 and Tau in patients undergoing hepatectomy.

OBJECTIVE: To investigate the effect of propofol and sevoflurane anesthesia on postoperative cognitive function. PATIENTS AND METHODS: Medical records of 280 patients who underwent hepatectomy in Jiangxi Provincial People's Hospital from April 2012 to July 2016 were retrospectively analyzed. Among those patients, 135 patients underwent propofol anesthesia (propofol group), and 145 patients under sevoflurane combined anesthesia (sevoflurane group). Hemodynamics was recorded 5 min before the induction of anesthesia (T0), after the induction of anesthesia (T1), at the beginning of the incision (T2), immediately after the incision (T3) and after the end of the surgery (T4). According to the Mini-Mental State Examination (MMSE), patients' cognitive function was evaluated before surgery. The levels of Aß-42 and Tau proteins in the patient's serum were measured. RESULTS: The stability of the mean arterial pressure after induction of anesthesia in the propofol group was higher than that of the sevoflurane group (p<0.05). MMSE scores in the propofol group were higher than those in the sevoflurane group (p<0.05). MMSE scores of patients in both groups 7 days after surgery were higher than those at 3 days after surgery (p<0.05). At 3 and 7 days after surgery, the levels of Aß-42 in the propofol group were lower than those in the sevoflurane group (p<0.05) and the levels of Tau protein in the propofol group were higher than those in the sevoflurane group. The levels of Aß-42 and Tau protein on the 3rd day after surgery in both groups were significantly higher than those before surgery (p<0.05). The Aß-42 levels decreased at 7 days after surgery in both groups (p<0.05). The level of Tau protein on the 7th day after surgery was higher than that before surgery and 3 days after operation (p<0.05). CONCLUSIONS: Compared with sevoflurane anesthesia, propofol may improve postoperative Aß-42 and Tau protein levels in patients with hepatocellular carcinoma, and ameliorate postoperative cognitive function.

Bone characteristics, histopathology, and chondrocyte apoptosis in femoral head necrosis induced by glucocorticoid in broilers.

Femoral head necrosis (FHN) is a common disorder in fast-growing broilers in the poultry industry, but the pathogenesis of FHN has not been clarified completely. In the present study, glucocorticoid (GC) administration was used to induce FHN in broilers. Compared with normal birds, histopathology showed that the length of the articular cartilage of GC-induced FHN broilers was thicker while the proliferative zone and prehypertrophic zone were obviously thinner. Moreover, hematoxylin and eosin (HE) staining showed the apoptotic chondrocyte in the growth plate of the femoral head in FHN-affected birds. Bone parameters also decreased significantly in GC-induced FHN broilers. In addition, as for the mRNA expression, GC-induced FHN broilers had an apparent reduction in Col-II, Col-X, and Bcl-2 but a significant promotion of Caspase-3, Caspase-9, ASK-1, and JNK-1 when compared with the normal birds. It showed glucocorticoid induced FHN in broilers by affecting the proliferation, differentiation, and apoptosis of chondrocytes accompanying the retarding of bone growth.

The expression of osteopontin in breast cancer tissue and its relationship with p21ras and CD44V6 expression.

OBJECTIVE: This study aimed to investigate the expression of osteopontin (OPN), p2lras, and CD44V6 in breast cancer tissues, and to analyze the relationships between their expression and a patient's clinicopathological characteristics and five-year survival rate. MATERIALS AND METHODS: Streptavidin-peroxidase immunohistochemistry was used to detect the expression of OPN, p2lras, and CD44V6 in tissue samples from 96 breast cancer patients, and the multivariate Cox proportional hazards model (mCOX-PHM) was used to analyze the factors that affect prognosis. RESULTS: Among the 96 breast cancer patients studied, positive staining for OPN, CD44V6, and p21ras was observed in 54.2%, 58.3%, and 43.8% of samples, respectively. The expression of OPN and CD44V6 were positively correlated (r = 0.58), and the expression of OPN and p21ras were also positively correlated (r = 0.25). Coexpression OPN, CD44V6, and p21ras was negatively correlated with a patient's five-year survival rate (p < 0.05). Kaplan-Meier analysis indicated that a patient without OPN, CD44V6, or p21ras expression had an improved survival (p < 0.05). Results from the mCOX-PHM analysis indicated that CD44V6 expression, the degree of tumor differentiation, and lymph node metastasis were all independent factors that indicate prognosis. The combined detection of OPN, CD44V6, and p21ras could contribute to a more accurate assessment of the biological behavior of breast cancers, and could help to indicate the prognosis of breast cancer patients.

Acute pulmonary embolism caused by highly aggregated intravenous immunoglobulin.

BACKGROUND AND OBJECTIVES: Six patients died and one patient survived following infusion of a specific lot of intravenous immunoglobulin (IVIG) within half an hour in May 2008. This study elucidated the underlying pathogenesis. MATERIALS AND METHODS: A variety of protein fractionation and identification approaches were employed to determine the abnormal components in IVIG products obtained from the hospital where the patients were treated. Animal studies using mice and monkeys were conducted to elucidate the pathophysiological mechanisms. In animal experiments, the effect and distribution of immunoglobulin was investigated using HE staining and immunohistochemistry (IHC) separately, while platelets and fibrinogen depletion were utilized to determine a possible link between thromboembolism formation in animals and the lethal effect of the IVIG. The size and distribution of the protein aggregates were determined with Coulter Counter Multisizer-3 after the dilution of the IVIG with plasma, and the lethal effect of the protein aggregates was simulated with artificial microparticles. RESULTS: The IVIG retrieved from the hospital was found to have striking similarities to the heat-treated IVIG in terms of protein aggregation profiles and lethal effects. Post-mortem examination indicated that immunoglobulin aggregates were mainly found in the lung of the animals, while depletion of platelets and fibrinogen from the IVIG preparations failed to prevent the death of the animals. Similar amount of artificial microparticles caused animal death in similar fashion. CONCLUSIONS: Our findings indicate that the retrieved IVIG exerted its lethal effects by blocking the pulmonary circulation without markedly altering the coagulation cascade or immunological events.

Downregulation of basic fibroblast growth factor is associated with femoral head necrosis in broilers.

Femoral head necrosis (FHN) is a metabolic cartilage disease of rapidly growing broilers. The aim of the present study was to investigate the role of basic fibroblast growth factor (bFGF) in the apoptotic processes associated with FHN. Broilers were selected and categorized based on clinical examination in 3 groups: healthy, femoral head separation, or femoral head separation with growth plate lacerations. Hematoxylin and eosin staining showed fewer chondrocytes in the resting zone of the growth plates when FHN occurred. Moreover, the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay revealed a significant increase in chondrocyte apoptosis. Furthermore, immunohistochemical assays and real-time quantitative PCR analysis demonstrated a decline in bFGF expression. In addition, reduced Bcl-2 mRNA expression was observed along with a corresponding increase in Bax and caspase-3 mRNA expression in FHN samples. There was a correlation between bFGF protein expression and the proportion of TUNEL-positive cells and a correlation between bFGF mRNA expression and expression of Bax, and caspase-3. The results of the study suggested that the expression of bFGF was reduced in the process of chondrocyte apoptosis, which could play an important role in the pathogenesis of FHN in chickens.

Effects of age and dietary soybean oil level on eggshell quality, bone strength and blood biochemistry in laying hens.

The objective of the study was to investigate the differences in eggshell quality, bone quality and serum bone biochemistry markers associated with changes in age and dietary soybean oil levels in laying hens. A total of 54, 19-week-old Hy-Line Brown laying hens were housed in 18 battery cages (3 birds/cage) and randomly divided into three diet treatments for 90 d: control-fat (CF, 1.9% soybean oil), moderate-fat (MF, 7% soybean oil) and high-fat (HF, 10% soybean oil). The hens' body weights (BW), egg production, egg weights, eggshell thickness and femoral diameter were higher at d 90 than at d 60 or d 30. Meanwhile, feed intake, relative bone weights, all bone strength parameters and serum Ca were lower at d 90 or 60 than at d 30. Compared to the CF hens, the feed intake, BW, abdominal fat pad weights and serum alkaline phosphatase activity were elevated in MF or HF hens. The eggshell thickness, relative femoral and tibial weight, femoral stiffness, femoral modulus, tibial mixed force and serum calcium and phosphorus levels were lower in MF or HF hens than CF hens. These findings suggest that bone loss in caged hens starts from an early stage of the laying period, and dietary oil (particularly with diets over 10% soybean oil) has harmful effects on eggshell quality, bone strength and bone mineralisation from an early stage of the laying period.

Expression and identification of recombinant chicken vascular endothelial growth factor in Pichia pastoris and its role in the pathogenesis of tibial dyschondroplasia.

Vascular endothelial growth factor (VEGF) is an essential mediator of angiogenesis and endochondral ossification. To explore the role of VEGF in avian diseases such as tibial dyschondroplasia (TD), a typical disorder of endochondral ossification, we expressed and identified recombinant chicken VEGF (chVEGF) protein in Pichia pastoris and evaluated its effects on thiram-induced TD in broiler chickens. The SDS-PAGE showed that 2 recombinant proteins, with molecular weights of ~46 and ~70 kDa, were obtained. Western blot analysis indicated that the 2 proteins were recognized by rabbit anti-chicken and goat anti-human VEGF polyclonal antibodies. Moreover, the mixture of the proteins significantly stimulated angiogenesis in the chick chorioallantoic membrane. In 21-d-old broilers that had been fed a thiram-enriched diet (100 mg/kg of thiram for 2 d at 8 d old) to induce TD, intramuscular injection of the chVEGF proteins (at a dosage of 10 or 30 µg/kg) significantly reduced the severity of TD but had no effect on TD incidence or BW; decreased serum Ca and P concentrations and tartrate-resistant acid phosphatase activity and elevated serum alkaline phosphatase activity; enhanced the total antioxidant capacity, superoxide dismutase, and glutathione peroxidase activities in the liver and kidney; upregulated the expression of type X collagen, matrix metalloproteinase (MMP)-13, and Runx2; and downregulated the Bcl-2 expression in the growth plates. In thiram-treated broilers at 15 d old, the chVEGF proteins upregulated the expression of MMP-13 and Runx2, and had different effects on type X collagen and Bcl-2 expression at different dosages. Our results indicate that exogenous chVEGF proteins promoted the recovery of TD-affected growth plates by improving the antioxidant capacity in the liver and kidney and by regulating differential expression of genes relating to endochondral ossification at different stages of TD development; VEGF deficiency in the growth plates was involved in the pathogenesis of TD.

The Ihh signal is essential for regulating proliferation and hypertrophy of cultured chicken chondrocytes.

The Indian hedgehog (Ihh) signal plays a vital role in regulating proliferation and hypertrophy of chondrocytes. To investigate its function in postnatal chicken (Gallus gallus) chondrocytes, cyclopamine was used to inhibit Ihh signaling. The MTT and ALP assays revealed the downgrade-proliferation and upgrade-differentiation of chondrocytes. To further elucidate the mechanism, the mRNA expression levels of Ihh, parathyroid hormone related protein (PTHrP), Gli-2, Bcl-2, Bone Morphogenetic Protein 6 (BMP-6), type X collagen (Col X) and type II collagen (Col II) were detected by quantitative real-time RT-PCR analysis, and the protein expressions of Ihh, Col X, and Col II were determined using Western blot analysis. After the Ihh signal was blocked, chondrocytes demonstrated high expression levels of PTHrP and Col X and low levels of Gli-2, BMP-6, Bcl-2 and Col II although Ihh expression was increased. Based on these results, the Ihh signal is essential for balancing chicken chondrocyte proliferation and hypertrophy, and the regulatory function of PTHrP acts in an Ihh-dependent manner. Furthermore, BMP-6 and Bcl-2 played roles in maintaining the development of chondrocytes and may be downstream regulatory factors of Ihh signaling.

Development of an enzyme-linked immunosorbent assay for detection of chicken osteocalcin and its use in evaluation of perch effects on bone remodeling in caged White Leghorns.

Osteocalcin (OC) is a sensitive biochemical marker for evaluating bone turnover in mammals. The role of avian OC is less clear because of the need for a chicken assay. Our objectives were to develop an assay using indirect competitive ELISA for detecting chicken serum OC and use the assay to examine the effects of perches on bone remodeling in caged hens. Anti-chicken OC polyclonal antibody was produced by immunization of rabbits with a recombinant OC from Escherichia coli. Chicken OC extracted from bone was used as a coated protein, and purified chicken OC was used for calibration. The limit of detection of the developed OC ELISA was 0.13 ng/mL. The intra- and interassay CV were <7 and <12%, respectively. The sensitivity of the developed OC ELISA was compared with a commercial Rat-Mid OC ELISA in laying hens housed in conventional cages with or without perches. Serum samples were collected from 71-wk-old White Leghorn hens subjected to 4 treatments. Treatment 1 was control chickens that never had access to perches during their life cycle. Treatment 2 chickens had perches during the pullet phase (0 to 16.9 wk of age), whereas treatment 3 chickens had perches only during the egg-laying phase of the life cycle (17 to 71 wk of age). Treatment 4 chickens always had access to perches (0 to 71 wk of age). Correlation between the 2 assays was 0.62 (P < 0.0001). Levels of serum OC using the developed chicken ELISA were higher than that detected using the Rat-Mid ELISA (P < 0.0001). Results from the chicken ELISA assay showed that hens with perch access had higher concentrations of serum OC than hens without perches during egg laying (P = 0.04). Pullet access to perches did not affect serum OC levels in 71-wk-old hens (P = 0.15). In conclusion, a chicken OC ELISA has been validated that is sensitive and accurate with adequate discriminatory power for measuring bone remodeling in chickens.

Expression of TRPV6 and CaBP-D28k in the egg shell gland (uterus) during the oviposition cycle of the laying hen.

1. The aim of this study was to investigate the localisation of the transient receptor potential vanilloid channel type 6 (TRPV6) in egg shell gland (ESG) and examine the dynamic expression of TRPV6 and Calbindin-d28k (CaBP-D28k), as well as the changes in concentration of total calcium (Ca), total inorganic phosphorus (P), alkaline phosphatase (ALP), parathyroid hormone (PTH) and calcitonin (CT) in plasma during the oviposition cycle. 2. The plasma ALP activity was notably increased at 8 h. In addition, plasma CT was highest at 0 h and significantly lower at 8 h. The change of plasma PTH concentration increased slightly post-oviposition and reached a maximum at 16 h. 3. Immunohistochemical analysis indicated that TRPV6 was strongly localised to the apical luminal epithelium of the mucosa. The mRNA levels of TRPV6 and CaBP-D28k in the ESG remained very low from 0 to 4.5 h, but were significantly increased at 16 h. Furthermore, Western blotting analysis showed that the expression of TRPV6 and CaBP-D28k also reached a maximum at 16 h and was different from the concentration of CaBP-D28k. 4. In conclusion, the epithelial Ca(2+) channel TRPV6 is strongly expressed in the epithelial cells of the eggshell gland, and the increase of TRPV6 and CaBP-D28k mRNA and protein expression during eggshell formation suggests that active Ca(2+) transcellular transport exerts significant effects in delivering active calcium in the ESG.

Changes of blood parameters associated with bone remodeling following experimentally induced fatty liver disorder in laying hens.

Studies have demonstrated that obesity and osteoporosis are linked disorders in humans. This study examined the hypothesis that excessive lipid consumption affects bone metabolism in laying hens. A total of one hundred 63-wk-old laying hens were randomly divided into 2 treatments and fed either a regular layer diet (control) or a high energy and low protein diet (HE-LP; experimental treatment) for 80 d. Egg production, feed intake, and BW were recorded at various days during the treatment. At d 80, ten randomly chosen birds per treatment group were killed. Abdominal fat weight, liver weight, and liver fat content were determined. Serum levels of total calcium, inorganic phosphate, and alkaline phosphatase were measured using a biochemical analyzer. Serum concentrations of osteocalcin, leptin-like protein, and estrogen were measured by enzyme-linked immunosorbent assay. Tibia length and width were measured using a vernier caliper; density of the right tibias was determined using an x-ray scanner; and mechanical properties of the left tibias were analyzed using a material testing machine. The expression of osteocalcin and osteoprotegerin mRNA in the keel bone was analyzed by real-time PCR. The concentration of osteocalcin protein in the keels was measured using western blot. Compared with control hens, hens fed the HE-LP diet had lower egg production, lower feed intake, greater liver fat content, and greater abdominal fat pad mass (P < 0.05). Feeding the HE-LP diet increased serum alkaline phosphatase activity, osteocalcin, leptin-like protein, and estrogen concentrations (P < 0.05), and decreased the keel osteocalcin concentrations (P < 0.05). There were significant positive correlations between the serum concentrations of leptin-like protein, estrogen, and osteocalcin regardless of treatment (P < 0.05). The results indicated that HE-LP diet induced a fatty liver disorder in laying hens with an upregulation in bone turnover and exacerbated skeletal damage. The data supported a role for lipid metabolism in skeletal heath of laying hens.

Bone response of broiler chickens (Gallus gallus domesticus) induced by corticosterone.

Experiments were conducted with chickens exposed to corticosterone (CORT), with the aim of determining its effects on bone characteristics. At 7 d of age, the experimental birds were injected daily with CORT (4 mg/kg of body mass) for 1 week. CORT administration significantly decreased the body weight while increasing relative liver weight of the chickens and the bone parameters were also decreased. Histology and immunohistochemistry of type X collagen revealed that CORT reduced the lengths of proliferative and prehypertrophic zone in growth plate and the number of positive chondrocytes in the prehypertrophic zone. In conclusion exposure to CORT depressed the growth performance and retarded the longitudinal growth of the long bones by inhibiting the proliferation and differentiation of chondrocytes in growth plate in broilers.

Localisation and expression of TRPV6 in all intestinal segments and kidney of laying hens.

1. The aim of this study was to investigate the localisation and expression of the epithelial Ca2+ channel TRPV6 (transient receptor potential vanilloid channel type 6) in different intestinal segments and kidney of laying hens during peak lay. 2. Immunohistochemical analysis of the intestine indicated that TRPV6 was localised to the brush-border membranes of the duodenum, jejunum, ileum, caecum, and rectum. Expression was weaker in the rectum, and little or no expression was found in crypt and goblet cells. In addition, TRPV6 mRNA was quantified amongst different intestinal segments, and expression was highest in the duodenum and jejunum. Furthermore, Western blotting indicated that the duodenum expressed the greatest amount of TRPV6 and the rectum the least with the other segments expressing intermediate levels. 3. In the kidney, distinct immunopositive staining for TRPV6 was detected at the apical domain of the distal convoluted tubules (DCT) and medullary connecting tubules (CNT). Interestingly, distribution of TRPV6 extended to the proximal convoluted tubules (PCT). Furthermore, the kidney expressed lower TRPV6 mRNA and protein levels compared with that in the duodenum. 4. In conclusion, the epithelial Ca2+ channel TRPV6 is strongly expressed in the apical cells of the entire intestine and the renal tubules, suggesting that active Ca2+ transcellular transport plays a crucial role in dietary calcium (re)absorption in laying hens.

Letrozole inhibits the osteogenesis of medullary bone in prelay pullets.

This study was performed to investigate the effect of letrozole, an aromatase inhibitor, on osteogenesis of medullary bone in prelay pullets. Three hundred fifteen 95-d-old ISA prelay pullets were used. After 10 d of adaptation in the cages, 15 pullets were selected randomly to collect the serum and bone samples and the rest were randomly assigned to 2 groups with 3 replicates each. One group was control and the other was letrozole-treated, fed 0.5 mg of letrozole per prelay pullet per day for 18 d. The serum and bone samples from these birds were collected during the experiment. Estradiol and testosterone in serum were assayed using commercial RIA kits. The serum alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP), Ca, and inorganic P were measured by an automatic biochemistry analyzer with commercial kits. The periosteum perimeter, endosteum perimeter, cortical bone index, cortical width, cortical bone area, and cortical area ratios of tibia were measured by transmitted scanner and a computer-assisted image analyzer. Our results showed that relative to the control-fed pullet, letrozole-fed pullets had reduced serum estrogen (57.5%), Ca (33.2%), ALP (33.6%), and TRAP (24.2%) and that values of serum estrogen, Ca, estrogen receptor expression, tibia radiographic density, serum ALP, and TRAP were all reduced (P < 0.05) and the serum P had a degressive trend in letrozole-treated groups. By contrast, the serum androgen and the tibia cortical bone index values were higher in the letrozole-treated group (P < 0.05). No differences were observed in the periosteum perimeter, endosteum perimeter, cortical width, and cortical area ratios of tibia between the 2 groups. The results showed that letrozole can inhibit the development of bone and medullary osteogenesis by inhibiting the synthesis of estrogen and its receptor in prelay pullets.

Effects of Gushukang, a Chinese herbal medicine, on bone characteristics and osteoporosis in laying hens.

In this study, we evaluated the effects of the herb medicine formula Gushukang (GSK) on bone characteristics and osteoporosis in end-of-lay hens. One thousand 55-wk-old ISA caged layers were allotted randomly to 2 groups. The control group was given the basal diet, and the GSK group was given the basal diet supplemented with additional GSK (1 g/kg) for 10 wk. Egg production, shell quality, bone radiographic density, and biochemical markers of bone turnover were determined. The results showed that GSK significantly increased the egg laying rate and decreased the percentage of cracked eggs (P < 0.05).The serum calcium, phosphate, and alkaline phosphatase were decreased (P < 0.05) in the GSK-treated group compared with the control group, whereas bone characteristics were significantly improved (P < 0.05). The results suggested that GSK can improve egg production and prevent bone loss by inhibiting bone turnover.

Chicken receptor activator of nuclear factor-kappaB ligand induces formation of chicken osteoclasts from bone marrow cells and also directly activates mature osteoclasts.

Receptor activator of nuclear factor-kappaB ligand (RANKL), which functions as a major determinant of osteoclast differentiation and activation, is a type II transmembrane protein and is expressed in osteoblasts-stromal cells. The aim of this study was to clarify the role of chicken RANKL (chRANKL) in chicken osteoclast differentiation and to determine its effect on mature chicken osteoclasts. In the present study, chRANKL protein was first cloned and expressed in Escherichia coli. We then treated chicken bone marrow cells with chRANKL protein and found that it induced the formation of chicken osteoclast-like multinucleated cells in a dose-dependent manner in the presence of human macrophage colony-stimulating factor. Moreover, the addition of chicken osteoprotegerin could block the effect of chRANKL with regard to osteoclast-like multi-nucleated cell formation and bone resorption. Using primary cultures of chicken osteoclasts on bone slices, we also found that bone resorption pits per cell increased with chRANKL concentration in a dose-dependent manner. The chRANKL-treated hens exhibited increased blood Ca(++) levels within 2 h after injection, showing that chRANKL also activates osteoclasts in vivo. These results clearly indicate that the expressed protein is functional and may also be a critical factor for chicken osteoclastogenesis and bone resorption.

Effects of ipriflavone on caged layer bone metabolism in vitro and in vivo.

The effects of ipriflavone on caged layer bone metabolism were examined in vitro and in vivo. Ipriflavone at 10(-8) M stimulated the activity of osteoblasts cultured from embryonic chick calvariae, and 10(-9) to 10(-7) M inhibited osteoclasts from chick tibias and humeri. Ipriflavone concentrations of 10(-4) and 10(-5) M inhibited osteoblast activity. These results suggest that ipriflavone influences bone metabolism by regulating the functional balance between osteoblasts and osteoclasts. Based on these in vitro experiments, in vivo studies were conducted to further clarify the effects of ipriflavone. Five hundred 58-wk-old ISA caged layers were divided into 5 groups that were fed diets containing 0, 15, 25, 50, and 100 ppm of ipriflavone. The experiment lasted 70 d. Egg production increased in hens fed 25 ppm and decreased in hens fed 50 and 100 ppm when compared with the controls and hens fed 15 ppm (P < 0.05). Egg weight, shell quality, BW, and serum P, Ca, estrogen, and bone mineral content were not affected by inclusion of ipriflavone in the diet. Hens consuming 25 ppm of ipriflavone had greater serum alkaline phosphatase and bone gla-protein levels than controls. Adding 25 ppm of ipriflavone to the feed appears to be close to an ideal level for clinical treatment of osteoporosis because of improved egg production while maintaining bone mineral content.

[Effects of Cistanche deserticola Y.C. Ma on serum creatine kinase and ultrastructures of skeletal muscles in mice].

The results showed that after mice were administrated with Cistanche deserticola, the duration of swimming was prolonged, and the increase of serum creative kinase was inhibited after exercises. Moreover, in the skeletal muscle ultrastructures it was found that after loaded swimming the glycogen became rich, and hyperplasia and hypertrophy of mitochondria occurred without any injury to myofibril.