Wagyu-Angus cross improves meat tenderness compared to Angus cattle but unaffected by mild protein restriction during late gestation.

For the spring-calving beef herds, late gestation coincides with winter and early spring, when cows are dependent on feed supplements with low quality hay, which is hard to meet their nutrient requirements. However, the effects of deficiencies of metabolizable protein intake during late gestation on offspring beef quality are unclear. Wagyu cattle have excellent marbling, and insemination with Wagyu vs Angus semen is a practical option for beef producers to improve beef quality. To test, Angus cows (621 ±â€¯73 kg) were selected and randomly separated into two groups. Each group was inseminated with either Angus or Wagyu semen. During the last 90 days of gestation, cows in each group were further separated and received either a low protein diet (85% of the NRC metabolizable protein requirement), which was a low quality hay-based diet common in northwestern region of the U.S., or an adequate protein diet (108% NRC requirement). All progeny was managed together and harvested at a final BW of 576.5 ±â€¯16.6 kg. Wagyu-sired offspring had higher marbling scores and quality grades than Angus (P < 0.01). Protein supplementation did not affect Slice Shear Force (SSF) in either breeds (P = 0.60). However, Wagyu-sired cattle had lower SSF than Angus-sired (P < 0.01). In addition, Wagyu-sired cattle had higher intramuscular fat (P < 0.05) and total collagen content (P < 0.05), but Angus-sired had greater mature collagen cross-links, as shown by higher contents of Pyridinoline (P < 0.01) and Ehrlich Chromogen (P < 0.01). Consistently, the mRNA expression of enzymes catalyzing collagen cross-linking was higher in Angus-sired offspring, including Plod 1 (P < 0.05), Plod 2 (P = 0.08), and P4Hα 2 (P < 0.01). In conclusion, Wagyu-sired cattle had greater tenderness and marbling score compared to Angus-sired, which was associated with lower collagen cross-links. Feeding mature grass hay-based diet with relatively low protein content during late gestation had no major effect on beef quality of subsequent cattle.

[Effect of an oral nutritional supplementation on nutritional status and quality of life in patients with colorectal cancer and postoperative adjuvant chemotherapy: A multi-center prospective randomized control trial].

Objective: To evaluate the effect of oral nutritional supplementation (ONS) on the nutritional status and quality of life in patients with colorectal cancer and postoperative adjuvant chemotherapy. Methods: This study was registered in the Chinese Clinical Trial Registry (ChiCTR-TRC-13003798). A multi-center randomized controlled trial was conducted. Colorectal cancer patients who underwent radical surgery and postoperative adjuvant chemotherapy, and had nutritional risk (nutrition risk screening 2002 score ≥3) when discharge from hospital in six hospitals (Beijing Hospital, Peking University Third Hospital, Guangzhou Nanfang Hospital, Shanghai Xinhua Hospital, Shanghai Ruijin Hospital, and Shanghai The Sixth People's Hospital) from June 2013 to August 2015 were prospectively enrolled. These patients were randomly divided into the ONS group and control group. Patients in the ONS group received dietary guidance and oral nutritional supplements (2092 kJ/day, whole protein enteral nutrition) for 90 days after discharge from hospital, while patients in the control group only received dietary guidance. Anthropometric measurements (body weight, body mass index [BMI], upper arm circumference, gripping power of the dominant hand, triceps skin fold), nutrition-related laboratory tests (hemoglobin, albumin, prealbumin, total cholesterol, triglyceride), gastrointestinal function scores and quality of life (evaluated by EuroQol five dimensions questionnaire) were collected and compared at baseline (at discharge), and at 30-day, 60-day and 90-day after discharge. Results: A total of 90 patients were included into this multi-center study, of whom 5 patients dropped out, 43 patients were assigned to the ONS group and 42 patients to the control group. Compared with baseline, the body weight of patients in the ONS group increased by (1.523±0.525) kg at 60-day and (1.967±0.661) kg at 90-day, which were significantly higher than those of patients in the control group [60-day: (-0.325±0.518) kg, P=0.015; 90-day: (-0.224±0.705) kg, P=0.027, respectively]. A similar pattern was observed for BMI, the ONS group increased by (0.552±0.203) kg/m(2) at 60-day and (0.765±0.205) kg/m(2) at 90-day, which were significantly higher than those of patients in control group [60-day: (-0.067±0.202) kg/m(2), P=0.034; 90-day: (0.022±0.210) kg/m(2), P=0.013]. No significant differences of other anthropometric measurements and nutrition-related laboratory tests were found between the two groups (all P>0.05). Furthermore, there were no significant differences of improvement in gastrointestinal function and quality of life between two groups (all P>0.05). Conclusion: Oral nutritional supplements can improve the body weight and BMI of colorectal cancer patients with nutritional risk receiving postoperative adjuvant chemotherapy, though it does not improve the quality of life.

Excess dietary fluoride affects laying performance, egg quality, tissue retention, serum biochemical indices, and reproductive hormones of laying hens.

The present study aimed to evaluate the effects of excess dietary fluoride (F) on laying performance, egg quality, tissue retention, serum biochemical indices, and serum reproductive hormones of laying hens. A total of 384 Hy-Line Gray hens, 37 wk old, were treated with sodium fluoride added to a corn-soybean meal basal diet at 0, 400, 800, and 1200 mg fluorine/kg feed. The results showed that dietary F levels at 800 and 1200 mg/kg markedly decreased ADFI, laying rate, average egg weight, and increased feed conversion ratio (FCR) (P < 0.05). Dietary F levels at 800 and 1200 mg/kg dramatically decreased the egg quality of albumen height, yolk color, eggshell strength, and eggshell thickness, and on the 49th D, 400 mg/kg F group significantly decreased the eggshell strength, compared to those of control group. Fluoride residues in tissues of hens were increased significantly with the increase of dietary F supplemental levels (P < 0.05). Fluoride concentrations were generally high in feces, eggshell, tibia, kidney, and ovary, and the highest in feces, following with eggshell and tibia, lower in kidney and ovary, and the lowest in serum. Serum uric acid levels and alanine aminotransferase activity increased significantly (P < 0.05), and glucose, triglycerides, and phosphorus decreased significantly (P < 0.05) in response to dietary F concentration, compared to those of the control group, respectively. Dietary F supplementation at 1200 mg/kg significantly decreased (P < 0.05) the estrogen concentrations in serum, compared to those of the control group. Concentrations of progesterone in the fluoride-treated groups were significantly (P < 0.05) decreased relative to those of the control group. In conclusion, these results indicated that the excessive ingestion of F has had a detrimental effect on egg laying rate and quality of eggs by damaging the function of the liver, kidney, and ovary of laying hens.

Effects of in ovo feeding of L-lysine on hatchability, hatching time, and early post-hatch development in domestic pigeons (Columba livia)1.

This study was conducted to evaluate the effects of in ovo feeding (IOF) of L-lysine (Lys) on hatchability and development in pigeon neonates. At day 13 of incubation, fertile eggs were injected with 200 µL of sterilized saline (0.75%, SC group), 1% Conc. Lys solution (Lys group, 2.11 mg Lys dissolved in 200 µL of sterilized saline), with controls not injected (NC group). The results showed that IOF of Lys decreased the hatchability (P < 0.05) compared with other groups, whereas the hatching time was not affected among groups. On day 14 of post-hatch (D14), the body weight (BW) of squabs received Lys IOF was increased relative to the NC group (P < 0.05). Squabs in Lys group exhibited higher (P < 0.05) body weight gain (BWG) than other groups from D14 to day of hatch (DOH). Meanwhile, IOF of Lys increased the brain relative weight on DOH (P < 0.05), and organ index of heart, legs, and gizzard on day 7 of post-hatch (D7) compared with other groups. In addition, the length index of duodenum, jejunum, or ileum had no difference between groups, as well as the weight index except the weight index of duodenum in Lys group was higher than that of SC group on DOH. However, squabs received Lys IOF showed lower villus height, crypt depth, and villus surface area of jejunum than NC group (P < 0.05) on DOH, and lower crypt depth than NC and SC groups on D7. The situation was improved by D14, although there was no significant difference in morphometric trait of jejunum between Lys group and NC group, squabs received Lys IOF showed higher villus height, crypt depth, and villus surface area of jejunum than NC group. In conclusion, the present study demonstrates that IOF of Lys has negative effects on hatchability and development of early post-hatch squabs, but the situation will be improved with the growth of age.

[Staining effect of bovine incisors in vitro by tea solutions].

OBJECTIVE: To determine the optimum staining condition of tea solutions on bovine incisors in vitro, by comparing the color stability of tooth surface of different concentrations of tea solutions and methods on bovine incisors in vitro. METHODS: Twenty bovine incisors with color surface A1 were chosen, then randomly divided into 4 groups (n=10). Group 1: soaked with 2% tea solution continuously for 6 days; group 2: soaked with 2% tea solution for 6 days, but changed fresh tea solution everyday; group 3: soaked with 1% tea solution continuously for 6 days; group 4: soaked with 1% tea solution for 6 days but fresh tea solution changed every day. After 6 days of staining, the surface color (Δ E value) of all the samples were measured with crystal eye. After brushing 30 times with toothbrushes, the color of bovine incisors were measured again. Then the samples were soaked in artificial saliva at 37 ° C, and Δ E value was measured for 14 days. RESULTS: After staining for 6 days, the Δ E values of the 2% tea solution groups were better than those of the 1% groups (20.21 vs. 16.44, 24.09 vs. 19.22, P<0.05); the groups with the same tea solution concentration, a better result was observed for the group soaked with daily fresh tea solution than for the group that experienced continuous staining (24.09 vs. 20.21, 19.22 vs. 16.44, P<0.05). Groups 1 and 2 were selected for subsequent brushing experiments. The color of both groups became lighter after brushing, and a better result was observed for the continuous staining group than for the group stained in daily fresh solution (3.06 vs. 9.51, P<0.05). The samples with better coloring effect soaked with 2% tea solution continuously for 6 days were put into artificial saliva for 14 days. There was not any significant change in coloring at the end of the first two days (1.51 vs. 1.51, P>0.05), and the color was visibly lighter after the third day (1.51 vs. 5.89, P<0.05), and no further significant change was observed until the 14th day (5.81 vs.5.89, P>0.05), which was darker coloring than that of the pre-staining group. CONCLUSION: Continuous staining on bovine incisors with 2% tea solution with subsequent soaking in artificial saliva resulted in consistent coloring from day 3 to day 14, and this method could be used as an ideal model for teeth staining in vitro.

Xuebijing enhances neuroprotective effects of ulinastatin on transient cerebral ischemia via Nrf2-are signal pathways in the hippocampus.

Prior studies have demonstrated that ulinastatin (UTI) plays a beneficial role in regulating cerebral ischemic injury evoked by cardiac arrest (CA). It is noteworthy to find interventions that can enhance effects of this drug and thereby increase its clinical application. Xuebijing (XBJ) is comprised of extracts from Chinese herbs and has been widely used in China as an anti-endotoxicity drug for the treatment of sepsis and ischemic disorders associated with multiple organ dysfunction syndrome. Thus, in this study we examined the effects of a combination of UTI and XBJ to improve neural injury in the process of neurological functions after transient cerebral ischemia. Our results show that CA impaired Nrf2- antioxidant response element (Nrf2-ARE) and superoxide dismutase (SOD) in the hippocampus CA1 region. This process further amplified products of oxidative stress, namely 8-isoprostaglandin F2α (8-iso PGF2α) and 8-hydroxy-2'-deoxyguanosine (8-OHdG). A lower dose of UTI failed to restore Nrf2-ARE and attenuate 8-iso PGF2α and 8-OHdG SOD following CA; however, systemic administration of XBJ amplified the effects of this dose of UTI on antioxidative signal pathway of the hippocampus. Overall, the results of this study have implications for the enhanced neuroprotective role played by a combination of XBJ and UTI in improving neural injury observed in transient cerebral ischemia; and Nrf2-ARE signal is a part of key mechanisms that are involved in neuroprotective effects of XBJ and UTI.

Proteomic analysis of hypothalamus and liver proteins affected by dietary l-arginine supplementation in laying hens.

The goal of this study was to investigate the influence of l-arginine (l-Arg) supplementation on diet-induced changes in hypothalamus and liver proteome of laying hens. Layers were fed either an isonitrogenous control diet or a l-Arg diet. The test included a 2-week acclimation period and a 12-week experimental period. Eight layers per group were sacrificed at terminal of the experiment underwent 12 fasting. Blood and tissue samples of hypothalamus and liver were collected for further analysis. The levels of serum nitric oxide and hypothalamus neuropeptide Y of layers in l-Arg group were increased in comparison with those in control group. Quantitative proteomic analyses showed that a total of 3,715 hypothalamus proteins (235 differentially expressed) and 3797 liver proteins (373 differentially expressed) were detected between control and l-Arg-fed groups. A further enriched Gene Ontology term analysis of proteins found that 17 hypothalamus proteins (11 upregulated and six downregulated) and 29 liver proteins (14 upregulated and 15 downregulated) were altered differentially between the two groups. Our findings revealed the changes in metabolic and hormonal signals in central nervous system and peripheral tissues by responding to l-Arg feeding, which provides a possible way to gain a better understanding of l-Arg function in laying hens.

Research and analysis of 74 bloodstream infection cases of Acinetobacter baumannii and drug resistance.

OBJECTIVE: We aimed at investigating the clinical and bacteriological features and drug resistance of bloodstream infection of Acinetobacter baumannii, so as to provide new evidence for treatment of bloodstream infection of Acinetobacter baumannii. PATIENTS AND METHODS: Statistical analysis was carried out for the clinical and bacteriological features and drug sensitivity of 74 bloodstream infection cases of Acinetobacter baumannii who were admitted to this hospital between July 2016 and June 2017. RESULTS: Among 74 patients, about 72.0% of them were admitted to the ICU and Respiratory Department; the average age of these patients was 63 years old. Among 74 patients, 62 patients stayed in the hospital for over 2 weeks (83.8%), and 35 for over 1 month (47.3%); 72.0% of patients experienced intrusive operation, in which 28.0% of patients dead. The experiment of drug sensitivity showed that tigecycline had the highest sensitivity (100%), sequentially followed by amikacin (over 90.0%) and other anti-bacterial drugs (less than 40.0%). Pan-drug resistance was identified in 42 patients, accounting for 56.8%. Comparison between the pan-drug resistant and non-pan-drug resistant patients showed that in the first two weeks before the positive blood culture, there were statistically significant differences in administration of carbapenem antibiotics and intrusive operation (p<0.05). Among the patients, the lowest resistance to carbapenem antibiotics was 8.16%, while the rate of resistance to other 12 antibiotics was more than 40.00%. Multiple-resistant strain mainly originated from the ICU and the burn center. CONCLUSIONS: In patients with bloodstream infection of Acinetobacter baumannii, the pan-drug resistant strains account for a vast majority with a high mortality rate. Age, intrusive operation and length of stay in hospital longer than 2 weeks are the common susceptible factors, while the administration of carbapenem antibiotics and intrusive operations might be the high-risk factors leading to pan-drug resistant cases.

[Analysis of infection and drug-resistance in 6 107 cases of extrapulmonary tuberculosis in Chengdu area].

Objective: To analyze the infection and drug resistance of extrapulmonary tuberculosis, in order to provide useful references for prevention and treatment of tuberculosis (TB). Methods: A total of 6 107 suspected cases of extrapulmonary tuberculosis, including lymph node, pleural, peritoneal, meningeal, urinary tract, and bone tuberculosis, were collected retrospectively from January 2013 to December 2015 in Public Health Clinical Center of Chengdu. There were 4 033 male and 2 074 female patients, and the average age was 37 years.The epidemiological data, infection and drug resistance were analyzed statistically. Results: There were 21 036 cases of mycobacterial culture from January 2013 to December 2015, of which 7 119 were positive(33.8%). Extrapulmonary tuberculosis was found in 6 107 cases, of which 896 were positive, accounting for 12.6% of the positive cases(896/7 119). Culture-positive cases of extra-pulmonary tuberculosis showed a rising trend from 2013 to 2015 (n=174, 310, 421, respectively), and the annual average increasing rate was over 50%.Cases were distributed at younger ages (21-40 years), with an average age of 34 years.There were more male patients, with a male to female ratio of 2.1∶1. The most common site of extrapulmonary tuberculosis was the bone(62/243, 25.5%), followed by lymph node(283/1 297, 21.8%), pleural(289/1 840, 15.7%), urinary tract(63/452, 13.9%), peritoneum(41/303, 13.5%), meningeal(156/1 915, 8.1%)and other types of tuberculosis(2/57, 3.5%). Drug sensitivity test of 896 extrapulmonary tuberculosis strains showed that the resistance rate (from high to low) was in the order of isoniazid, rifampicin, streptomycin, ofloxacin, ciprofloxacin, amikacin, capreomycin and ethambutol. Multidrug-resistant(MDR)and extremely-drug resistant (XDR) rate was 9.7 %(87/896)and 2.0%(17/896)respectively. Conclusions: Cases of extrapulmonary tuberculosis sent for bacterial culture and the positive rate showed a rising trend by years in Chengdu area.The drug resistance level was higher than that reported both in China and abroad.The prevention and treatment of extrapulmonary tuberculosis should be strengthened in this area.

Inhibitory effects of grape seed extract on growth, quorum sensing, and virulence factors of CDC "top-six" non-O157 Shiga toxin producing E. coli.

Non-O157 Shiga toxin producing Escherichia coli (STECs) have become a growing concern to the food industry. Grape seed extract (GSE), a byproduct of wine industry, is abundant in polyphenols that are known to be beneficial to health. The objective of this study was to evaluate the effect of GSE on the growth, quorum sensing, and virulence factors of Centers for Disease Control and Prevention (CDC) "top-six" non-O157 STECs. Minimal inhibitory concentration (MIC) of GSE was 2mg/ml against E. coli O26:H11, and 4mg/ml against the other non-O157 STECs tested. Minimal bactericidal concentration (MBC) was the same as MIC for all six non-O157 STECs tested. At 5×10(5)CFU/ml inoculation level, 4mg/ml GSE effectively inhibited the growth of all tested strains, while 0.25-2mg/ml GSE delayed bacterial growth. At a higher inoculation level (1×10(7)CFU/ml), GSE had less efficacy against the growth of the selected six non-O157 STECs. Its impact on bacterial virulence was then assessed at this inoculation level. Autoinducer-2 (AI-2) is a universal signal molecule mediating quorum sensing (QS). GSE at concentration as low as 0.5mg/ml dramatically reduced AI-2 production of all non-O157 STECs tested, with the inhibitory effect proportional to GSE levels. Consistent with diminished QS, GSE at concentration of 0.125mg/ml caused marked reduction of swimming motility of all motile non-O157 STECs tested. In agreement, GSE treatment reduced the production of flagella protein FliC and its regulator FliA in E. coli O103:H2 and E. coli O111:H2. Furthermore, 4mg/ml GSE inhibited the production of Shiga toxin, a major virulence factor, in E. coli O103:H2 and E. coli O111:H2. In summary, GSE inhibits the growth of "top-six" non-O157 STECs at the population level relevant to food contamination. At higher initial population, GSE suppresses QS with concomitant decrease in motility, flagella protein expression and Shiga toxin production. Thus, GSE has the potential to be used in food industry to control non-O157 STEC.

1,25(OH)2D3 downregulates the Toll-like receptor 4-mediated inflammatory pathway and ameliorates liver injury in diabetic rats.

BACKGROUND: Fatty acid deposition in the liver can activate a number of pro-inflammatory signaling pathways such as the Toll-like receptor 4 (TLR4) pathway, which may be important in the pathogenesis of nonalcoholic steatohepatitis. 1,25(OH)2D3 downregulates the expression of TLR4 and may represent a novel treatment strategy for reducing hepatocyte injury. Therefore, in this study, we investigated the protective effects of 1,25(OH)2D3 on diabetic liver injury in vivo. METHODS: Streptozotocin (STZ)-induced diabetic rats were randomly divided into five groups and treated with low-dose 1,25(OH)2D3 (0.025 µg/kg/day), medium-dose 1,25(OH)2D3 (0.15 µg/kg/day), high-dose 1,25(OH)2D3 (0.3 µg/kg/day), insulin (protamine zinc insulin 16 U/kg/day, subcutaneous injection), or no intervention (the control group). Sixteen weeks later, the rats were killed, and blood samples were obtained to test lipid profiles and hepatic function. The infiltration of inflammatory cells, the level of fibrosis, and the expression levels of TLR4, nuclear factor-kappa B (NF-κB), and tumor necrosis factor-α (TNF-α) in the liver were analyzed. The hepatocytes were treated with vehicle control, LPS (100 ng), high fat [DMEM + FFA (0.1 mM: palmitic acid, oleic acid, 1:2)], LPS + high fat, vehicle + 1,25(OH)2D3 (10(-7) M), LPS + 1,25(OH)2D3, high fat + 1,25(OH)2D3, or LPS + high fat + 1,25(OH)2D3. RNA and protein were extracted to detect the expression of TLR4 and downstream inflammatory factors such as NF-ΚB, TNF-α, and IL-6. Groups of data were compared by single factor variance analysis. RESULTS: High-dose 1,25(OH)2D3 administration for 16 weeks downregulated the expression of TLR4, NF-κB, and TNF-α in the liver tissue of diabetic rats and attenuated hepatic inflammation and fibrosis, as shown by immunohistochemical staining, hematoxylin and eosin staining, Masson's trichrome staining, reverse transcription polymerase chain reaction (RT-PCR), and western blotting. In vitro, hepatocytes treated with high fat or LPS exhibited significantly increased expression of TLR4, NF-κB, and downstream inflammatory factors (P < 0.05). Intervention with 1,25(OH)2D3 decreased the expression of TLR4, NF-κB, and inflammatory factors (P < 0.05). CONCLUSIONS: 1,25(OH)2D3 exhibited protective effects against diabetes-related liver injury, possibly through downregulation of components of the TLR4 signaling pathway.

Pharmacokinetics and pharmacodynamics of rilotumumab: a decade of experience in preclinical and clinical cancer research.

Rilotumumab is a fully human monoclonal antibody against hepatocyte growth factor, the only known ligand of the MET receptor. Over the last decade, rilotumumab has been extensively tested in preclinical studies and in clinical studies in a variety of cancer types. In this review, we examine the pharmacokinetic and pharmacodynamic data that have been collected in the rilotumumab programme to date, and discuss retrospectively how the knowledge acquired in this programme can be applied to a number of key issues in oncology drug development, including: (i) using preclinical data to inform first-in-human study design; (ii) the role of biomarkers in the identification of a target patient population; (iii) the potential for drug interactions between therapeutic proteins and other anticancer agents; and (iv) pharmacokinetic and pharmacodynamic considerations in phase 3 study design.

Sorafenib combined with percutaneous radiofrequency ablation for the treatment of medium-sized hepatocellular carcinoma.

OBJECTIVE: Sorafenib, an oral multikinase inhibitor, is the proved therapy method for patients with advanced hepatocellular carcinoma (HCC). Based on heat delivery, Radiofrequency ablation (RFA) has been found to achieve complete neoplasm necrosis. It is the most widely performed percutaneous therapy for HCC. However, Study associated combined Sorafenib with RFA therapy for patients with advanced HCC has never been reported. The aim of present study is to explore the efficacy and safety of sorafenib combined with RFA therapy for the patients with medium-sized HCC. PATIENTS AND METHODS: A total of 62 patients diagnosed as HCC were involved in this study. All patients were randomly assigned to sorafenib and RFA (n=30) or RFA-alone (n=32) treatment groups. Treatment outcomes, including recurrence rates, time to progression (TTP) and adverse reactions induced by sorafenib were observed and recorded to assess the efficacy and safety of the combination method. RESULTS: During the overall follow-up period, the recurrence rate of the combination subgroup was 56.7% (17/30), and that of the RFA-alone subgroup was 87.5% (28/32) (p < 0.01). The median TTP was 17.0 months in the combination therapy vs. 6.1 months in the RFA-alone (p < 0.05). Hand-foot skin reactions were reported by 83.3% (25/30) of patients and 46.7% (14/30) reported diarrhea while the most adverse events (AEs) were mild to moderate in the combination subgroup. CONCLUSIONS: Sorafenib combined with RFA significantly decreased recurrence rates and prolonged the survival time of medium-sized HCC patients. The combination therapy is safer and more effective than the control without unexpected side effects. Furthermore, the earlier application, the better results were.

Outbreak of primary inoculation tuberculosis in an acupuncture clinic in southeastern China.

Outbreak of Mycobacterium tuberculosis infections associated with acupuncture has not been reported. Thirteen patients with a painful swollen lump were referred to our hospital. The index patient received acupuncture and paraspinal muscular injection at a local acupuncture clinic in April 2011 and was diagnosed with M. tuberculosis 1 month later. From May 2011 to August 2011, 12 more patients with a swollen lump on the nuchal region or in the lower back or the buttocks region were referred to our hospital. Tuberculin skin test (TST), T-SPOT.TB, acid-fast stain, M. tuberculosis culture, chest radiograph, and lump magnetic resonance imaging (MRI) were performed and the patients were diagnosed with tuberculous abscess of the lump. All 13 patients received intramuscular injection at the paraspinal muscle by two acupuncturists at a local clinic and reported a swollen lump at the injection site. The needles and syringes were reused after autoclave sterilization. The TST was positive in all patients. Twelve patients had positive acid-fast stains. Mycobacterial cultures of abscess specimens were positive in all 13 patients. T-SPOT.TB tests were positive in all patients who underwent the test. The lesions and biopsies were subjected to polymerase chain reaction (PCR) and gene sequencing by the Disease Control Center of Zhejiang Province, China and the causative agent was identified as M. tuberculosis, Beijing type. In conclusion, physicians should consider the possibility of mycobacterial infections, apart from other bacterial agents, in patients with a swollen paraspinal lump following intramuscular injection.

Genetic diversity of the Chinese traditional herb Blumea balsamifera (Asteraceae) based on AFLP markers.

Blumea balsamifera is a commercially important medicinal herb in China and other parts of Asia. It is used to produce borneol. This plant grows in the wild, but resources have diminished greatly in recent years. We examined the genetic diversity of this species to help develop conservation strategies; 35 plants from five provinces were analyzed using AFLPs. Eight AFLP primer combinations generated 1367 fragments, giving a mean of 172 fragments per primer combination. Polymorphism in the germplasm analysis was found for 1360 (99.48%) of the fragments, of which 264 (19.27%) fragments were unique (accession specific) and 423 (25.33%) of the fragments were rare (present in less than 10% of the accessions). The polymorphic fragments were used to group the accessions in a UPGMA phenogram. Most grouping was geographical. In general, accessions coming from Guizhou and Guangxi showed higher diversities as these accessions were scattered in different groups. The genetic distance estimated by Jaccard similarity coefficient index showed low variability among genotypes (coefficient value ranged from 0.60 to 0.95). More attention should be given to the study and conservation of the biodiversity of this economically important genus.

Effects of maternal plane of nutrition and increased dietary selenium in first-parity ewes on inflammatory response in the ovine neonatal gut.

Many areas of the western United States have soils that have increased Se content, and ruminants grazing these rangelands may ingest increased quantities of Se. In addition, high-energy diets or increased Se intake may induce gut inflammation. The objective of this study was to evaluate the effects of maternal plane of nutrition and increased dietary Se during gestation on inflammatory responses in neonatal lamb ileal tissue, a major immune organ. Rambouillet ewes (age = 240 ± 17 d; initial BW = 52.1 ± 6.2 kg) were allocated to 4 treatments arranged in a 2 × 2 factorial. Factors included Se [adequate Se (ASe, 11.5 µg/kg of BW) or high Se (HSe, 77.0 µg/kg of BW)] initiated at breeding, and nutritional plane [100% (CON) or 140% (HIH) of requirements] initiated at d 40 of gestation. Ewes were fed individually from d 40, and lambs were removed at parturition and fed artificial colostrum and milk replacer. Lambs were necropsied at 20 d of age, and ileal tissues were sampled for immunoblotting and real-time quantitative reverse-transcription PCR analyses. The ASe-HIH and HSe-CON treatments had no effect (P = 0.179) on inflammatory signaling compared with ASe-CON. However, greater inflammatory signaling was detected in the HSe-HIH group, as shown by increased (P < 0.05) mRNA expression of tumor necrosis factor-α and chemotaxis IL-8. Consistently, phosphorylation of c-Jun N-terminal kinase, a primary inflammatory signaling mediator, was greater (P < 0.05) in the HSe-HIH group compared with other treatments. Consistent with cytokine expression, mast cell density was less in the HSe-CON group than in other treatments. The expression of transforming growth factor ß mRNA was greater (P < 0.05) in the HSe-HIH group; consistently, collagen content was increased in the HSe-HIH group compared with the ASe-CON group (P < 0.05). In conclusion, independently, neither HSe nor HIH had major effects on inflammation, but in combination, these maternal treatments induced an inflammatory response in the neonatal intestine.

Predictors of relapse in chronic hepatitis B after discontinuation of anti-viral therapy.

BACKGROUND: Optimal duration of anti-viral therapy in chronic hepatitis B virus (HBV) infection remains unclear. AIM: To investigate factors that could predict relapse after stopping anti-viral agents. METHODS: Chronic hepatitis B patients who were treated with anti-viral agents (lamivudine, adefovir, entecavir) and have stopped the treatment were recruited. Anti-viral agents were stopped according to the recommendations of the Asian Pacific Association for the Study of the Liver. Virological relapse was defined as an increase in serum HBV DNA to >1000 copies/mL after discontinuation of treatment. RESULTS: Eighty-four (69 treatment naïve and 15 lamivudine resistant) patients were eligible for this study. Thirty-seven patients developed virological relapse at 4.3 ± 2.9 (range 1-11) months after discontinuation of therapy. The 1-year cumulative probability of virological relapse was 42% and 47% in HBeAg (hepatitis B e antigen)-positive (n = 41) and HBeAg (hepatitis B e antigen)-negative (n = 43) patients, respectively. On multivariate analysis by Cox proportional hazard model, pre-existing lamivudine resistance, delayed suppression of HBV DNA to undetectable level during anti-viral therapy and to a higher HBsAg (hepatitis B surface antigen) level at the end of treatment were associated with virological relapse. Twelve of the 15 (80%) lamivudine resistant patients developed virological relapse. Among the 11 treatment naïve patients who had HBsAg ≤ 2 log IU/mL at the end of treatment, 1 (9%) of them had virological relapse. CONCLUSIONS: Treatment cessation among lamivudine resistant patients is associated with high risk of virological relapse. Serum HBsAg level at the end of treatment and rate of HBV DNA suppression can provide supplementary information to guide the timing of stopping anti-viral drugs.

Isolation and characterization of a Kunitz-type trypsin inhibitor with antiproliferative activity from Gymnocladus chinensis (Yunnan bean) seeds.

A 20-kDa Kunitz-type trypsin inhibitor was isolated from Gymnocladus chinensis (Yunnan bean) seeds. The isolation procedure involved ion exchange chromatography on diethylaminoethyl cellulose (DEAE-cellulose), affinity chromatography on Affi-gel blue gel, ion exchange chromatography on sulfopropyl sepharose (SP-sepharose), and gel filtration by FPLC on Superdex 75. The trypsin inhibitor was adsorbed on DEAE-cellulose, unadsorbed on Affi-gel blue gel, and adsorbed on SP-Sepharose. It dose-dependently inhibited trypsin with an IC(50) value of 0.4 µM. Dithiothreitol reduced its trypsin inhibitory activity, suggesting that an intact disulfide bond is indispensable to the activity. It suppressed [methyl-(3)H] thymidine incorporation by leukemia L1210 cells and lymphoma MBL2 cells with an IC(50) value of 4.7 and 9.4 µM, respectively. There was no effect on human immunodeficiency virus(4)-1 reverse transcriptase activity and fungal growth when the trypsin inhibitor was tested up to 100 µM.

In vitro characterization and pharmacokinetics of dapagliflozin (BMS-512148), a potent sodium-glucose cotransporter type II inhibitor, in animals and humans.

(2S,3R,4R,5S,6R)-2-(3-(4-Ethoxybenzyl)-4-chlorophenyl)-6-hydroxymethyl-tetrahydro-2H-pyran-3,4,5-triol (dapagliflozin; BMS-512148) is a potent sodium-glucose cotransporter type II inhibitor in animals and humans and is currently under development for the treatment of type 2 diabetes. The preclinical characterization of dapagliflozin, to allow compound selection and prediction of pharmacological and dispositional behavior in the clinic, involved Caco-2 cell permeability studies, cytochrome P450 (P450) inhibition and induction studies, P450 reaction phenotyping, metabolite identification in hepatocytes, and pharmacokinetics in rats, dogs, and monkeys. Dapagliflozin was found to have good permeability across Caco-2 cell membranes. It was found to be a substrate for P-glycoprotein (P-gp) but not a significant P-gp inhibitor. Dapagliflozin was not found to be an inhibitor or an inducer of human P450 enzymes. The in vitro metabolic profiles of dapagliflozin after incubation with hepatocytes from mice, rats, dogs, monkeys, and humans were qualitatively similar. Rat hepatocyte incubations showed the highest turnover, and dapagliflozin was most stable in human hepatocytes. Prominent in vitro metabolic pathways observed were glucuronidation, hydroxylation, and O-deethylation. Pharmacokinetic parameters for dapagliflozin in preclinical species revealed a compound with adequate oral exposure, clearance, and elimination half-life, consistent with the potential for single daily dosing in humans. The pharmacokinetics in humans after a single dose of 50 mg of [(14)C]dapagliflozin showed good exposure, low clearance, adequate half-life, and no metabolites with significant pharmacological activity or toxicological concern.