Network pharmacological and molecular docking verification of the mechanism of Osteoking in preventing deep vein thrombosis of lower limb.

OBJECTIVE: The aim of this study was to predict the mechanism of Osteoking in preventing deep vein thrombosis (DVT) of the lower limb by network pharmacology and molecular docking. MATERIALS AND METHODS: The relevant active components and targets of Osteoking were collected through the TCMSP database, and the relevant disease targets of DVT were collected through the GeneCards, OMIM, and DisGeNET databases. The intersecting gene targets of Osteoking and DVT were obtained using Venny 2.1.0 software. PPI network construction and core target selection using Cytoscape 3.9.0 software. The Metascape database was used for GO and KEGG enrichment analysis of relevant targets. Finally, the molecular docking of the main active components and key targets was carried out. RESULTS: There are 361 potential targets and 71 core targets of Osteoking in preventing deep vein thrombosis of the lower limb. Signal pathways are involved in various diseases such as cancer, diabetic complications, atherosclerosis, and more. Some of the most common pathways include AGE-RAGE signaling pathway and Calcium signaling pathway. Molecular docking results showed that the main active components of Osteoking had relatively stable binding activities with the key targets. CONCLUSIONS: Osteoking can play a role through multiple targets and multiple signal pathways to prevent the formation of deep venous thrombosis of the lower limb after fracture.

Risk factors and etiological characteristics of urinary tract infection in hospitalized continuous ambulatory peritoneal dialysis patients.

OBJECTIVE: This study aimed to explore the risk factors and etiological characteristics of urinary tract infection (UTI) in continuous ambulatory peritoneal dialysis (CAPD) patients. PATIENTS AND METHODS: A total of 90 CAPD patients with UTI comprised the infection group, while 32 CAPD patients without UTI constituted the control group. The risk factors and etiological characteristics of UTI were analyzed. RESULTS: Of the 90 bacterial strains isolated, 30 were Gram-positive (33.3%) and 60 were Gram-negative (66.7%). Urinary stones or urinary tract structural changes were more prevalent in the infection group (71.1%) than in the control group (46.9%) (χ² = 6.076, p = 0.018). The proportion of patients with residual diuresis less than 200 ml was higher in the infection group (50%) than in the control group (15.6%) (χ² = 11.533, p = 0.001). The distribution of primary disease differed between the two groups. Patients in the infection group had higher CAPD vintage, levels of triglycerides, fasting blood glucose, blood creatinine, blood phosphorus, and calcium-phosphorus product than those in the control group. Multivariate binary logistic regression analysis indicated that residual diuresis less than 200 ml (OR = 3.519, p = 0.039) and urinary stones or structural changes (OR = 4.727, p = 0.006) were independent risk factors for UTI. CONCLUSIONS: Urine cultures of CAPD patients with UTI contained a complex distribution of pathogenic bacteria. Urinary stones or structural changes and residual diuresis less than 200 ml were independent risk factors for UTI.

[Validation of a hyperkalemia prediction model in chronic kidney disease].

Objective: To validate the accuracy and consistency of a previously established prediction model for the occurrence of hyperkalemia in non-dialytic chronic kidney disease (CKD) patients. Methods: All patients diagnosed with CKD from Outpatient Department of Shanghai Changzheng Hospital during the 4th quarter of 2020 were recruited. Demographic data, clinical characteristics and prediction model-related parameters of the patients were collected and analyzed. Receiver operating characteristic (ROC) curve was drawn to evaluate the effectiveness of the model, and the specificity and sensitivity were calculated based on the cut-off value of 4 obtained from the previous model. The improved Hanley method was used to compare the area under the curve (AUC) between the previously established model and current validation dataset. The calibration curve was drawn to verify the model calibration degree. Results: A total of 434 patients diagnosed with non-dialytic CKD were enrolled, among whom 233 were males and 201 were females, with an average age of (55±16) years. According to the measured serum potassium values, the prevalence of hyperkalemia was 7.6%. And 33 patients were allocated to the hyperkalemia group and 401 patients were to the normal potassium group. There was no significant difference in age and sex between the two groups (both P>0.05). A combination of hyperkalemia and heart failure (27.3% vs 3.7%, P<0.001), diabetes (42.4% vs 19.7%, P=0.002), and acidosis (51.5% vs 7.0%, P<0.001) were more frequently in the hyperkalemia group, compared with the normal serum potassium group. Patients in the hyperkalemia group were more likely to have a past history of serum potassium ≥5.0 mmol/L (48.5% vs 2.5%, P<0.001). For the drugs that could increase serum potassium levels, there was a significant correlation between Chinese herbal medicine and the occurrence of hyperkalemia, while renin-angiotensin-aldosterone system inhibitor (RAASi) and potassium supplementation showed no significant difference between the two groups. The results of ROC curve analysis showed that the AUC was 0.914, with the sensitivity of 84.8% and the specificity of 79.8% with the cut-off value of 4. The difference of AUC between the previously established risk assessment model of hyperkalemia in patients with non-dialytic CKD and current validation dataset was not statistically significant (Z=1.924, P=0.054), indicating the good accuracy and consistency of the prediction model. In the calibration curve, when the predicted risk of patients was below 0.4 or above 0.6, the prediction effect of the model was better. Conclusion: The previously-constructed hyperkalemia prediction model in non-dialytic CKD patients had good accuracy and consistency, and could be used to evaluate the risk of hyperkalemia in all stages of non-dialytic CKD patients.

Alcohol, coffee and tea intake and the risk of cognitive deficits: a dose-response meta-analysis.

AIMS: Lifestyle interventions are an important and viable approach for preventing cognitive deficits. However, the results of studies on alcohol, coffee and tea consumption in relation to cognitive decline have been divergent, likely due to confounds from dose-response effects. This meta-analysis aimed to find the dose-response relationship between alcohol, coffee or tea consumption and cognitive deficits. METHODS: Prospective cohort studies or nested case-control studies in a cohort investigating the risk factors of cognitive deficits were searched in PubMed, Embase, the Cochrane and Web of Science up to 4th June 2020. Two authors searched the databases and extracted the data independently. We also assessed the quality of the studies with the Newcastle-Ottawa scale. Stata 15.0 software was used to perform model estimation and plot the linear or nonlinear dose-response relationship graphs. RESULTS: The search identified 29 prospective studies from America, Japan, China and some European countries. The dose-response relationships showed that compared to non-drinkers, low consumption (<11 g/day) of alcohol could reduce the risk of cognitive deficits or only dementias, but there was no significant effect of heavier drinking (>11 g/day). Low consumption of coffee reduced the risk of any cognitive deficit (<2.8 cups/day) or dementia (<2.3 cups/day). Green tea consumption was a significant protective factor for cognitive health (relative risk, 0.94; 95% confidence intervals, 0.92-0.97), with one cup of tea per day brings a 6% reduction in risk of cognitive deficits. CONCLUSIONS: Light consumption of alcohol (<11 g/day) and coffee (<2.8 cups/day) was associated with reduced risk of cognitive deficits. Cognitive benefits of green tea consumption increased with the daily consumption.

[Screening of drugs that selectively inhibit uveal melanoma cells with SF3B1 mutations].

OBJECTIVE: To screen compounds that can selectively inhibit uveal melanoma cells with splicing factor 3B subunit 1 (SF3B1) mutations in comparison with isogenic SF3B1 wild-type counterparts in a cell model of SF3B1 mutant allele knockout. METHODS: Principal component analysis was used to analyze transcriptome alternative splicing in TCGA cohorts of uveal melanoma with wild-type SF3B1 and SF3B1 mutations, and abnormal alternative splicing events derived from SF3B1 mutations were identified. The SF3B1 mutant allele in Mel202 cells was knocked out using CRISPR-Cas9 technology, and Sanger sequencing was used to verify the edited sequence. MTT and colony formation assays were used to assess the proliferation of Mel202 and Mut-KO cells. RT-PCR agarose electrophoresis combined with Sanger sequencing was used to determine alternative splicing events in Mel202 and Mut-KO cells. MTT assay was performed to screen the compounds that showed selective inhibitory effect against Mel202 cells with SF3B1 mutation. RESULTS: Specific knockout of SF3B1 mutant allele in Mel202 cells obviously promoted the cell proliferation and caused changes in alternative splicing of ZDHHC16 and DYNLL1 transcripts. The screening data showed that 13 compounds had selective inhibitory activity against Mel202 cells with SF3B1 mutation (Fold change≥2), and among them, tetrandrine and lapatinib showed good dose-effect curves. CONCLUSION: This study provides a cell screening model for identification of potential individualized treatment drugs for patients with uveal melanoma with SF3B1 mutation.

[Analysis of follow-up results of chrono-chemotherapy or conventional chemotherapy combined with intensity modulated radiotherapy in locally advanced nasopharyngeal carcinoma].

Objective: To evaluate the long-term effect and safety of chrono-chemotherapy combined with intensity modulated radiotherapy (IMRT) in locally advanced nasopharyngeal carcinoma (NPC). Methods: 160 patients with locally advanced NPC were randomly divided into a chrono group and conventional group according to random number table. In the first stage, all patients underwent two cycles of induced chemotherapy, consisting of docetaxel, cisplatin and 5-Fu every 21 days. Notably, patients received chrono-moduated chemotherapy according to circadian rhythm in the chrono group, and conventional chemotherapy in the conventional group. Then, 21 days after the completion of first stage, three cycles of concurrent cisplatin chemotherapy every 21 days were given to all patients during IMRT. The median follow-up after the completion of radiotherapy was 31 months. Long-term side effects and the survival of patients were observed. Results: Patients in the chrono group had significantly lower rates of hearing loss (22.72%), dysphagia (0) and neck fibrosis (4.54%) compared with those in the conventional group (39.13%、8.69%, 15.94%, respectively, all P<0.05). Meanwhile, the 1- year overall survival rates (97.0% vs 92.8%), 3-year overall survival rates (80.3% vs 81.2%), 1-year progression free survival rates (95.5% vs 87.0%), 3-year progression free survival rates (71.2% vs 73.9%), 1-year locoregional relapse-free survival rates (97.0% vs 95.7%), 1-year locoregional relapse-free survival rates (92.4% vs 92.8%), 1-year distant metastasis-free survival rates (97.0% vs 98.6%) and 3-year distant metastasis-free survival rates (90.9% vs 91.3%) between the chrono group and the conventional group were not statistically significant (all P>0.05). Conclusions: Compared with conventional chemotherapy, chrono-chemotherapy combined with IMRT didn't affect long-term survival, but reducing the incidence of adverse events in patients with locally advanced NPC.

Effects of ${\rm \small L}$-methionine on growth performance, carcass quality, feather traits, and small intestinal morphology of Pekin ducks compared with conventional ${\rm \small {DL}}$-methionine.

The research studied the effects of l-methionine (l-Met) on growth performance, carcass quality, feather traits, and small intestinal morphology of Pekin ducks compared with conventional dl-methionine (dl-Met). A total of 1080, 1-day-old male Pekin ducks were randomly allotted to 9 groups with 6 replicate pens of 20 birds each. During the starter phase (1 to 14 d), ducks were fed a basal diet (Met, 0.30%) or that supplemented with dl-Met or l-Met at 0.05, 0.10, 0.15, or 0.20% of feed. During the grower phase (15 to 35 d), ducks were fed a basal diet (Met, 0.24%) or that supplemented with dl-Met or l-Met at 0.04, 0.08, 0.12, or 0.16% of feed. Compared with ducks fed the basal diet, supplementation with either dl-Met or l-Met increased the body weight (BW) of ducks at days 14 and 35, increased average daily gain (ADG) and average daily feed intake (ADFI), decreased F:G at the starter phase, and increased ADG over the whole 35-d period (P < 0.05). The efficacy of l-Met compared to dl-Met was 140.1% for 14-d BW, 137.6% for ADG and 121.0% for F:G for days 1 to 14. Ducks fed diets supplemented with l-Met had greater proportion of leg muscle, higher than in ducks provided with dl-Met (P < 0.05). The breast muscle proportion was enhanced with dl-Met rather than l-Met supplementation (P < 0.01). The back feathers score and fourth primary wing feather length were increased with dl-Met or l-Met supplementation (P < 0.01), and there was increased efficacy of l-Met relative to dl-Met for back feathers score (153.1%). Dietary dl-Met or l-Met supplementation increased villus height of ileal mucosa of ducks at days 14 and 35 (P < 0.01). Overall, dietary l-Met or dl-Met supplementation affected the growth performance of ducks during the starter phase, and improved the feather traits and small intestinal morphology. The efficacy of l-Met to dl-Met ranged from 120 to 140% for growth performance of young ducks (1 to 14 d) and was 153% for the feather traits of ducks (35 d).

Effects of dietary rapeseed meal inclusion levels on growth performance, organ weight, and serum biochemical parameters in Cherry Valley ducks.

The aim of this study was to investigate the effects of the inclusion levels of different types of rapeseed meal (RSM) on performance, organ weight, and serum biochemical parameters in Cherry Valley ducks in the starter period and grower-finisher period. In Exp. 1, a total of 750 seven-day-old male ducklings were divided into 5 dietary treatments with 6 replicate pens of 25 birds per pen. The starter diets with the inclusion of 0, 5, 10, 15, or 20% of double-low RSM contained 0, 1.37, 2.15, 3.46, or 5.31 µmol glucosinolates (GLS)/g in the finished feed (from day 7 to 21). In Exp. 2, a total of 900 fifteen-day-old male ducklings were divided into 6 dietary treatments with 6 replicate pens of 25 birds per pen. The grower-finisher diets with the inclusion of 0, 5, 10, 15, 20, or 25% of Indian RSM contained 0, 7.67, 15.34, 24.66, 31.21, or 38.44 µmol GLS/g in the finished feed (from day 15 to 42). For ducklings in the starter period (Exp. 1), body weight gain and feed intake decreased linearly as the dietary double-low RSM inclusion level increased at day 7 to 14, while growth rate was not influenced by dietary double-low RSM inclusion levels at day 15 to 21 and day 7 to 21. For ducks in the grower-finisher period (Exp. 2), growth performance decreased linearly as the dietary RSM inclusion level increased from 5 to 20%. In addition, dietary RSM inclusion levels induced liver enlargement in ducklings at day 21 (5 to 20% double-low RSM with 1.37 to 5.31 µmol/g GLS) and thyroid enlargement accompanied by increased serum AST and ALP activities in ducks at day 42 (5 to 15% Indian RSM with 7.67 to 23.66 µmol/g GLS). Therefore, our results indicated that the upper limit of using RSM sources in feed formulation should consider the anti-nutritional factor of GLS content at different stages of duck growth.

Effect of manganese source on manganese absorption and expression of related transporters in the small intestine of broilers.

An experiment was conducted to investigate the effect of manganese (Mn) source on Mn absorption and expressions of Mn, amino acid, and peptide transporters in the small intestine of broilers. A total of 320 Mn-deficient 15-day-old Arbor Acres male broilers were randomly assigned to 5 treatments with 8 replicates/treatment and 8 chicks/replicate and fed an Mn-unsupplemented control diet or the control diet supplemented with 110 mg Mn/kg from either MnSO4, or 1 of 3 organic Mn chelates with weak (OW), moderate (OM), or strong (OS) chelation strength for 14 D. The plasma Mn contents were higher (P < 0.03) in supplemental Mn groups than in the control group, in OS group than in OM group, and in OM group than in OW and MnSO4 groups on day 28. Broilers fed diets supplemented with Mn had higher (P < 0.02) duodenal divalent metal transporter 1 (DMT1) and ferroportin 1 (FPN1) mRNA levels and FPN1 protein level on both days 21 and 28 than those fed the control diet. Duodenal DMT1 mRNA and protein levels were higher (P < 0.05) in OM and OS groups than in OW and MnSO4 groups on day 28. The mRNA levels of amino acid transporters [b0, +-type amino acid transporter 1 (B0AT1) and excitatory amino acid transporter 3 (EAAT3)] were higher (P < 0.0005), and peptide transporter 1 was lower (P < 0.04) in the ileum than in the duodenum and jejunum; however, Mn source did not affect (P > 0.05) mRNA levels of amino acid and peptide transporters in the small intestine of broilers. The results from the present study indicate that both DMT1 and FPN1 facilitated Mn absorption, however, the amino acid and peptide transporters might not be involved in the transport of the organic Mn chelates; organic Mn chelates with moderate and strong chelation strength, especially strong chelation strength, showed higher Mn absorption possibly due to enhanced DMT1 expression in the duodenum of broilers.

Estimation of calcium requirements for optimal productive and reproductive performance, eggshell and tibial quality in egg-type duck breeders.

Optimizing the dietary calcium (Ca) level is essential to maximize the eggshell quality, egg production and bone formation in poultry. This study aimed to establish the Ca requirements of egg-type duck breeders from 23 to 57 weeks of age on egg production, eggshell, incubation, tibial, plasma and ovary-related indices, as well as the expression of matrix protein-related genes. Totally, 450 Longyan duck breeders aged 21 weeks of age were allotted randomly into five treatments, each with six replicates of 15 individually caged birds. The data collection started from 23 weeks of age and continued over the following 35 weeks. The five groups corresponded to five dietary treatments containing either 2.8%, 3.2%, 3.6%, 4.0% or 4.4% Ca. The tested dietary Ca levels increased (linear, P <0.01) egg production and egg mass, and linearly improved (P <0.01) the feed conversion ratio (FCR). Increasing the dietary Ca levels from 2.8% to 4.4% increased (P <0.01) the eggshell thickness and eggshell content. The tested Ca levels showed a quadratic effect on eggshell thickness and ovarian weight (P <0.01); the highest values were obtained with the Ca levels 4.0% and 3.6%, respectively. Dietary Ca levels affected the small yellow follicles (SYF) number and SYF weight/ovarian weight, and the linear response (P <0.01) was significant vis-à-vis SYF number. In addition, dietary Ca levels increased (P <0.05) the tibial dry weight, breaking strength, mineral density and ash content. Plasma and tibial phosphorus concentration exhibited a quadratic (P <0.01) response to dietary Ca levels. Plasma calcitonin concentration linearly (P <0.01) increased as dietary Ca levels increased. The relative expression of carbonic anhydrase 2 in the uterus rose (P <0.01) with the increment of dietary Ca levels, and the highest value was obtained with 3.2% Ca. In conclusion, Longyan duck breeders fed a diet with 4.0% Ca had superior eggshell and tibial quality, while those fed a diet with 3.6% Ca had the heaviest ovarian weights. The regression model indicated that the dietary Ca levels 3.86%, 3.48% and 4.00% are optimal levels to obtain maximum eggshell thickness, ovarian weight and tibial mineral density, respectively.

Effects of metabolic abnormalities, hyperandrogenemia and clomiphene on liver function parameters among Chinese women with polycystic ovary syndrome: results from a randomized controlled trial.

PURPOSE: To investigate the effects of metabolic abnormalities, hyperandrogenemia and ovulation induction by clomiphene/acupuncture on liver function parameters among women with polycystic ovary syndrome (PCOS). METHODS: This is a secondary analysis of a randomized controlled trial. All 1000 subjects were diagnosed as PCOS by modified Rotterdam criteria. Liver function parameters, metabolic panel and hormone profile were measured at baseline and after treatment. The relationship between liver parameters with metabolic, hormonal parameters and ovulation induction was examined. RESULTS: PCOS women with metabolic syndrome had higher liver enzyme levels but lower bilirubin and bile acid levels than without. PCOS women with hyperandrogenemia had higher liver enzyme, bilirubin levels than without. Correlation analyses showed that worsening of metabolic parameters was associated with higher liver enzyme levels but lower bilirubin and bile acid levels, while increased androgen levels were associated with higher liver enzyme, bilirubin and bile acid levels. Ovulation induction with clomiphene citrate could decrease bilirubin and bile acid levels, while acupuncture had no obvious effect on liver function. CONCLUSIONS: Among PCOS women, metabolic abnormalities and hyperandrogenemia impaired different liver function parameters. Clomiphene could decrease the bilirubin and bile acid levels while acupuncture had no obvious effect on liver function.

Dietary supplementation of organic or inorganic chromium modulates the immune responses of broilers vaccinated with Avian Influenza virus vaccine.

Dietary supplementation with the organic chromium (Cr) has been shown to positively affect the immune function of poultry. However, to our knowledge, no experiment has been done to directly compare the impacts of Cr chloride and chromium picolinate (CrPic) on the immune responses of broilers vaccinated with Avian Influenza (AI) virus vaccine. Therefore, the present experiment was conducted to investigate the effects of supplemental Cr sources (Cr chloride and CrPic) and levels on the growth performance and immune responses of broilers vaccinated with AI virus vaccine so as to provide an effective nutritional strategy for improving immune function of broilers. A total of 432 1-day (d)-old male broiler chicks were used in a 1 plus 2×4 design. Chickens were given either a diet without Cr supplementation (control) or diets supplemented with 0.4, 0.8, 1.6, or 3.2 mg Cr/kg as either Cr chloride or CrPic for 42 d. Compared to the control, dietary Cr supplementation had no effect (P>0.05) on average daily gain, average daily feed intake and gain : feed of broilers during the starter and grower phases, but increased (P<0.05) the relative weights of bursa of fabricius on d 21 and thymus, spleen, or bursa of fabricius on d 42, serum antibody titers against AI virus on d 21, 28, 35 and 42, blood T-lymphocyte transformation rate on d 28 and 42, blood T-lymphocyte percentage on d 42, and serum interleukin-2 contents on d 28. Broilers fed the diets supplemented with the inorganic Cr chloride had higher (P<0.05) weights of thymus, spleen and bursa of fabricius than those fed the diets supplemented with the CrPic on d 42. In addition, broilers fed the diets supplemented with the CrPic had higher (P<0.05) antibody titers against AI virus than those fed the diets supplemented with the inorganic Cr chloride on d 21 and 35. These results indicate that dietary Cr supplementation improved immune responses of broilers vaccinated with AI virus, and the inorganic Cr chloride was more effective than the CrPic in increasing the relative weights of lymphoid organs, however, the CrPic was more effective than the inorganic Cr chloride in enhancing the serum antibody titer against AI virus.

Effects of curcumin on performance, antioxidation, intestinal barrier and mitochondrial function in ducks fed corn contaminated with ochratoxin A.

Curcumin has been attributed with antioxidant, anti-inflammatory, antibacterial activities, and has shown highly protective effects against enteropathogenic bacteria and mycotoxins. Ochratoxin A (OTA) is one of the major intestinal pathogenic mycotoxins. The possible effect of curcumin on the alleviation of enterotoxicity induced by OTA is unknown. The effects of dietary curcumin supplementation on OTA-induced oxidative stress, intestinal barrier and mitochondrial dysfunctions were examined in young ducks. A total of 540 mixed-sex 1-day-old White Pekin ducklings with initial BW (43.4±0.1 g) were randomly assigned into controls (fed only the basal diet), a group fed an OTA-contaminated diet (2 mg/kg feed), and a group fed the same OTA-contaminated feed plus 400 mg/kg of curcumin. Each treatment consisted of six replicates, each containing 30 ducklings and treatment lasted for 21 days. There was a significant decrease in average daily gain (ADG) and increased feed : gain caused by OTA (P<0.05); curcumin co-treatment prevented the decrease in BW and ADG compared with the OTA group (P<0.05). Histopathological and ultrastructural examination showed clear signs of enterotoxicity caused by OTA, but these changes were largely prevented by curcumin supplementation. Curcumin decreased the concentrations of interleukin-1ß, tumor necrosis factor-α and malondialdehyde, and increased the activity of glutathione peroxidase induced by OTA in the jejunal mucosa of ducks (P<0.05). Additionally, curcumin increased jejunal mucosa occludin and tight junction protein 1 mRNA and protein levels, and decreased those of ρ-associated protein kinase 1 (P<0.05). Notably, curcumin inhibited the increased expression of apoptosis-related genes, and downregulated mitochondrial transcription factors A, B1 and B2 caused by OTA without any effects on RNA polymerase mitochondrial (P<0.05). These results indicated that curcumin could protect ducks from OTA-induced impairment of intestinal barrier function and mitochondrial integrity.

A novel FGFR1-binding peptide attenuates the degeneration of articular cartilage in adult mice.

OBJECTIVE: We previously reported that genetic ablation of (Fibroblast Growth Factors Receptors) FGFR1 in knee cartilage attenuates the degeneration of articular cartilage in adult mice, which suggests that FGFR1 is a potential targeting molecule for osteoarthritis (OA). Here, we identified R1-P1, an inhibitory peptide for FGFR1 and investigated its effect on the pathogenesis of OA in mice induced by destabilization of medial meniscus (DMM). DESIGN: Binding ability between R1-P1 and FGFR1 protein was evaluated by enzyme-linked immuno sorbent assay (ELISA) and molecular docking. Alterations in cartilage were evaluated histologically. The expression levels of molecules associated with articular cartilage homeostasis and FGFR1 signaling were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting and immunohistochemistry (IHC). The chondrocyte apoptosis was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay. RESULTS: R1-P1 had highly binding affinities to human FGFR1 protein, and efficiently inhibited extracellular signal-regulated kinase (ERK)1/2 pathway in mouse primary chondrocytes. In addition, R1-P1 attenuated the IL-1ß induced significant loss of proteoglycan in full-thickness cartilage tissue from human femur head. Moreover, this peptide can significantly restore the IL-1ß mediated loss of proteoglycan and type II collagen (Col II) and attenuate the expression of matrix metalloproteinase-13 (MMP13) in mouse primary chondrocytes. Finally, intra-articular injection of R1-P1 remarkably attenuated the loss of proteoglycan and the destruction of articular cartilage and decreased the expressions of extracellular matrix (ECM) degrading enzymes and apoptosis in articular chondrocytes of mice underwent DMM surgery. CONCLUSIONS: R1-P1, a novel inhibitory peptide for FGFR1, attenuates the degeneration of articular cartilage in adult mice, which is a potential leading molecule for the treatment of OA.

[Preclinical application of MR and fluorescent dual-modality imaging combined with photothermal therapy in HER-2 positive breast cancer].

Objective: To construct superparamagnetic iron oxide nanoparticles (SPIONs) coated on trastuzumab and indocyanine green (ICG) and then investigate whether the coated nanoparticles (NPs) targeted to human epidermal growth factor receptor-2 (HER-2) receptors on breast cancer cells in vitro and in vivo. Methods: The Fe(3)O(4)-trastuzumab-ICG NPs were constructed. And a series of characteristics of the NPs were evaluated. The uptake ability of SK-BR-3, a HER-2 positive breast cancer cell, was observed by transmission electron microscopy. Then the NPs were injected in the tail veins of SK-BR-3 xenograft tumor-bearing mice to observe the aggregation of NPs in the tumor sites by MRI and fluorescent imaging. Furthermore, when the NPs was gathered at the tumor sites, the near infrared thermal imaging system was used to monitor the tumor temperature after the near infrared radiation. Results: The successfully constructed Fe(3)O(4)-trastuzumab-ICG NPs had the size of (25.93±4.25) nm. The absorption peak was 828 nm, which was as same as the emission wavelength of ICG. The NPs had a high relaxation rate of approximately 107.65 mM(-1)·s(-1). The maximum temperature of NPs solution could reach to 57.8℃ after continuous near infrared laser irradiation. The transmission electron microscopy imaging revealed that the NPs could target and enter into the endoplasmic reticulum of SK-BR-3 cells. MRI analysis showed the lowest T(2) relaxation time in the tumor sites 24 h after tail vein injection of the NPs. The △T(2) of the tumor sites in the Fe(3)O(4)-trastuzumab-ICG group (30.7±4.8) ms was higher compared with that of control group (3.1±1.1) ms, Fe(3)O(4)-IgG-ICG group (4.4±0.9) ms and trastuzumab + Fe(3)O(4)-trastuzumab-ICG group (11.3±3.8) ms., respectively, all showing statistically significant differences (P<0.05). The fluorescence imaging revealed that the NPs was concentrated transiently in the intraperitoneal organs and tumor sites, then excreted into the bladder. After 24 h, there was an obvious aggregation in the tumor sites. The near infrared thermal imaging experiments showed that the temperature of tumor sites in Fe(3)O(4)-trastuzumab-ICG group could go up to 49.4℃ after continuous near infrared light irradiation. Conclusion: The newly constructed Fe(3)O(4)-trastuzumab-ICG NPs have the potential to act as a multifunctional imaging agent and a powerful tool for photothermal therapy for HER-2 positive breast cancer.

Risk of major bleeding in patients with non-valvular atrial fibrillation treated with oral anticoagulants: a systematic review of real-world observational studies.

OBJECTIVE: To conduct a systematic review of real-world (RWD) studies comparing the risk of major bleeding (MB) among patients with non-valvular atrial fibrillation (NVAF) on direct oral anticoagulants (DOACs) or warfarin. METHODS: MEDLINE, Embase, NHS-EED, and EconLit were searched for RWD studies published between January 2003 and November 2016 comparing MB risk among DOACs and warfarin. Proceedings of clinical conferences from 2012 to 2016 were reviewed. RESULTS: A total of 4218 citations were identified, 26 of which met eligibility criteria. Most studies were retrospective analyses of administrative claims databases and patient registries (n = 23 of 26); about half were based in the United States (n = 15). Apixaban showed a significantly lower risk of MB versus warfarin in all eight included studies. MB risk was either significantly lower (n = 9 of 16) or not significantly different (n = 7 of 16) between dabigatran and warfarin; there was no significant difference between rivaroxaban and warfarin in all seven included studies. The risk was significantly lower with apixaban versus rivaroxaban (n = 7 of 7) but not significantly different from dabigatran (n = 6 of 7). MB risk was significantly lower (n = 3 of 4) or not significantly different (n = 1 of 4) with dabigatran versus rivaroxaban. No evidence was identified for edoxaban. CONCLUSION: DOACs were associated with similar or lower risks of MB versus warfarin. A lower MB risk was consistently observed for apixaban, but less consistently for dabigatran; MB risk was similar between rivaroxaban and warfarin. Among DOACs, the risk of MB with apixaban was consistently lower than with rivaroxaban, but similar to dabigatran.

Effects of sodium, 1,25-dihydroxyvitamin D3 and parathyroid hormone fragment on inorganic P absorption and Type IIb sodium-phosphate cotransporter expression in ligated duodenal loops of broilers.

Three experiments were conducted with 22-day-old Arbor Acres male broilers to study the effects of Na+, 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] and parathyroid hormone fragment [PTH (1-34)] on inorganic P absorption and Type IIb sodium-phosphate cotransporter (NaP-IIb) mRNA and protein expression levels in ligated duodenal loops. The duodenal loops were perfused with solutions (pH = 6) containing zero, 50, or 150 mmol/L of Na+ as NaCl in Exp. 1, containing zero, 30, or 300 pmol/L of 1,25-(OH)2D3 in Exp. 2, or containing zero, 65, or 650 pmol/L of PTH (1-34) in Exp. 3, respectively. Compared with the control, additions of 50 and 150 mmol/L of Na+, 30 and 300 pmol/L of 1,25-(OH)2D3, or 65 and 650 pmol/L of PTH (1-34) to the perfusates promoted (P < 0.02) the P absorption percentages and rates, respectively. Additions of the above-mentioned concentrations of Na+ or 1,25-(OH)2D3 to the perfusates increased (P < 0.003) NaP-IIb mRNA level in the duodenum of broilers, and a similar trend (P = 0.08) was observed for PTH (1-34). The Na+, 1,25-(OH)2D3, and PTH (1-34) had no effects (P > 0.15) on NaP-IIb protein level in the duodenum of broilers. The results indicate that increased P absorption due to perfusions of Na+, 1,25-(OH)2D3 or PTH (1-34) might be attributed to enhanced NaP-IIb expression in the duodenum of broilers.

Impact of maternal heat stress in conjunction with dietary zinc supplementation on hatchability, embryonic development, and growth performance in offspring broilers.

The aim of this study was to investigate whether maternal dietary supplementations with different zinc (Zn) sources could reduce the deleterious effect of maternal heat stress on hatchability performance and progeny growth performance. A completely randomized design (n = 6) with 2 maternal environmental temperatures [normal 21 ± 1°C (NT) vs. high 32 ± 1°C (HT)] × 3 maternal dietary supplemental Zn levels [Zn-unsupplemented control diet (CON), the control diet + 110 mg of Zn/kg of diet as either inorganic ZnSO4 (iZn) or organic Zn with a moderate chelation strength (oZn)] was used. HT decreased (P < 0.05) fertility, hatchability, chick hatch weight, and embryonic survival. HT also decreased (P ≤ 0.05) progeny BW, ADG, and ADFI at one to 21, 22 to 28, and 29 to 42 d of age as well as breast muscle ratio and plasma aspartate aminotransferase and creatine kinase activities at 42 d of age. Maternal dietary Zn supplementation with either iZn or oZn increased (P < 0.004) Zn contents in yolk and liver, non-weak chick ratio, as well as progeny BW, ADFI, and survivability at one to 21 d of age. Notably, the addition of oZn increased (P < 0.05) hatchability and progeny thigh meat quality by reducing b* value. The progeny ADG at one to 21 d and 22 to 28 d of age and BW at 28 d of age from maternal NT were not affected (P > 0.46) by maternal dietary Zn supplementation, but the above 3 indices from HT-iZn (P < 0.05) and HT-oZn (P < 0.003) were higher than those from HT-CON. Our results indicate that maternal heat stress impairs hatching performance, embryonic development, and progeny growth performance, inducing metabolic changes, while supplementation of Zn in maternal diets regardless of Zn sources improved hatch chick quality and survivability of offspring and alleviated the negative effect of maternal heat stress on growth performance of offspring during the starter period. In addition, maternal dietary supplementation with the organic Zn improved hatchability and progeny meat quality.

Dietary non-phytate phosphorus requirement of broilers fed a conventional corn-soybean meal diet from 1 to 21 d of age.

An experiment was conducted to investigate the effect of dietary non-phytate phosphorus (NPP) level on growth performance, bone characteristics and phosphorus metabolism-related gene expressions, so as to evaluate the dietary NPP requirement of broiler chicks fed a conventional corn-soybean meal diet from 1 to 21 d of age. A total of 540 day-old Arbor Acres male chicks were randomly allocated to one of nine treatments with six replicate cages of 10 birds per cage in a completely randomized design, and fed a basal corn-soybean meal diet (containing 0.08% of NPP) supplemented with 0.10, 0.15, 0.25, 0.30, 0.35, 0.40, 0.45, or 0.50% of inorganic phosphorus in the form of CaHPO4·2H2O, respectively. Each diet contained the constant calcium content of about 1.0%. The results showed that daily weight gain, serum inorganic P, tibia bone strength, tibia ash percentage, tibia bone mineral content (BMC) and density (BMD), middle toe ash percentage, middle toe BMC and BMD were affected (P < 0.0001) by dietary NPP level, and increased linearly (P < 0.0001) and quadraticly (P < 0.004) as dietary NPP levels increased. The gene expression of type IIb sodium-phosphate cotransporter (NaPi-IIb) in the duodenum was affected (P < 0.03) and decreased linearly (P < 0.002) as dietary NPP levels increased. Dietary NPP requirements estimated based on fitted broken-line models (P < 0.0001) of the sensitive indices including daily weight gain, tibia bone strength, tibia ash percentage, tibia BMC and BMD as well as middle toe ash percentage were 0.34∼0.39%. The results from this study indicate that tibia BMC and BMD might be new, sensitive, and noninvasive criteria to evaluate the dietary NPP requirements of broilers, and the dietary NPP requirement is 0.39% for broiler chicks fed a conventional corn-soybean meal diet from 1 to 21 d of age.

Characterization of RsMYB28 and RsMYB29 transcription factor genes in radish (Raphanus sativus L.).

Glucosinolates (GSLs) are important secondary metabolites in Brassicaceae plants. Previous studies have mainly focused on GSL contents, types, and biosynthesis-related genes, but the molecular characterization patterns of GSL biosynthesis-related transcription factors remain largely unexplored in radish (Raphanus sativus L.). To isolate transcription factor genes regulating the GSL biosynthesis, genomic DNA and cDNA sequences of RsMYB28 and RsMYB29 genes were isolated in radish. Two R2R3-MYB domains were identified in the deduced amino acid sequences. Subcellular localization and yeast-one hybrid assays indicated that both the RsMYB28 and RsMYB29 genes were located in the nucleus and possessed transactivation activity. Reverse transcription quantitative analysis showed that the RsMYB28 and RsMYB29 genes were expressed in seeds, leaves, stems, and roots at the seedling, taproot thickening, and mature stages. Both genes were highly expressed during the seedling and taproot thickening stages. The expression level of RsMYB28 was found to be up-regulated following wounding, glucose, and abscisic acid treatments, whereas RsMYB29 was up-regulated following wounding and methyl jasmonate treatments. These results provide insights into the biological function and characterization of the RsMYB28 and RsMYB29 genes, and facilitate further dissection of the molecular regulatory mechanism underlying the GSL biosynthesis in radish.