However, basal serum iron concentrations and area under the iron curves significantly decreased in the experimental group (p < 0.0001) and remained at the same level throughout the 210-min study. The values obtained for hemoglobin, mean corpuscular volume, ferritin, transferrin, transferrin saturation, ceruloplasmin and total protein were within normal reference ranges. In conclusion, the decrease in serum iron was likely due to the effects of chronic zinc administration, and the decrease in serum iron was not sufficient to cause anemia.The recognized antagonistic actions between zinc and iron prompted us to study this subject in children. A convenience sample was used. Thirty healthy children between 8 and 9 years of age were studied with the aim of establishing the effect of a 3-mo oral zinc supplementation on iron status. Fifteen individuals were given a placebo (control group), and 15 were given 10 mg Zn/day . Blood samples were collected at 0, 60, 120, 180 and 210 min after a 12-h overnight fast, before and after placebo or zinc supplementation. This supplementation was associated with significant improvements in energy, protein, fat, carbohydrate, fiber, calcium, iron, and zinc intake in accordance with the recommendations for age and sex. The basal serum zinc concentration significantly increased after oral zinc supplementation ( Zinc is essential for many biological functions in all living organisms. Zinc deficiency, considered a nutritional disorder, has been associated with a variety of symptoms and diseases. This deficiency may be due to inadequate intake, malabsorption, excessive loss, increased demand, or other pathologies compromising its bioavailability .Like zinc, iron is essential for humans, mainly for oxygen transport, hematopoiesis and central nervous system development. Serum iron is not an adequate parameter to diagnose iron deficiency because it exhibits analytical and biological limitations in assessing iron status. Inadequate dietary iron, physiological states and various pathologies can influence iron status testing .viceversa) may be due to the antagonistic action of zinc during the process of iron absorption from the gastrointestinal tract [The literature reveals that zinc and iron have been studied in children with respect to a number of clinical outcomes such as growth, infection, diarrhea, anemia, cognition and behavior ,4. Combial tract ,7. In thal tract . Howeveral tract .et al. (2010) recently demonstrated that zinc supplementation reduced iron absorption but did not promote anemia in healthy children [Many tests are available for the determination of iron status. However, due to specimen integrity issues, analytical problems, biological and diurnal variations, and interpretation limitations, no gold standard exists . Certainchildren .Accordingly, a need exists for more studies to establish the antagonistic effects of zinc on iron and the onset of anemia to evaluate the benefits in intervention programs. Additionally, zinc is administered in high doses and for long periods in many disorders, particularly in healthy children with growth deficiency, and in these conditions, attention to the onset of anemia is necessary. The aim of this study was to investigate whether a dose of 10 mg Zn/day for 3-mo might affect iron status in healthy children.A total of 30 healthy children of both sexes, aged 8–9 years, from four municipal schools in the northeastern city of Natal, Brazil, were studied. We studied healthy children because there are only rare studies in the literature regarding the effects of zinc on iron metabolism in this population. Many healthy children use oral zinc for growth and development. The published articles have only focused on debilitated children and those with zinc deficiency. Here, the inclusion criteria were healthy schoolchildren, as assessed clinically, anthropometrically and by laboratory assessment. The exclusion criteria included early pubarche, thelarche or menarche; acute, chronic, infectious or inflammatory diseases; and children who had undergone surgery or were using vitamin and mineral supplements.The protocol was approved by the University Hospital Research Ethics Committee of the Federal University of Rio Grande do Norte (UFRN) (no. 542/11). All schoolchildren agreed to participate in the study and informed consent was obtained from their parents and guardians.The volunteers were studied between 2011 and 2012. The study was characterized as a randomized, controlled, triple-blind trial with groups matched by age, formed by a process of non-probability sampling (convenience sample) in which the volunteers were divided into control and experimental groups. The study evaluated 15 schoolchildren as the control group to observe the serum zinc and iron profiles before and after the 3-mo oral placebo. The 3-mo period was established because it is already considered sufficient the study of zinc metabolism. The 15 schoolchildren in the experimental group were administered oral zinc, and serum zinc and iron were observed before and after the 3-mo zinc supplementation. The study began at 7:00 am and ended at 12:00 pm after a 12-h overnight fast . The subAll the schoolchildren were measured for body weight (kg) and height (cm) using an electronic balance  and a stadiometer , respectively. For weight measurement, the child stood on the scale wearing light clothing and without shoes. To measure height, the child remained standing without shoes or head gear and with heels together, arms extended along the body and with the body as upright as possible. The heels, buttocks, shoulders and head were aligned against the wall or vertical surface of the measuring equipment. The height and weight measurements were obtained three consecutive times by the same observer and then averaged. The observer was specifically trained for this purpose. The analysis of nutritional status was also based on the BMI-for-age ,13.Information on food intake was obtained at the beginning of the study and after 3-mo using food records from 3 nonconsecutive days (2 weekdays and 1 weekend day). Mothers were instructed to record each of the following about their child’s food intake: the time of each meal, the foods consumed, and their respective amounts. Energy, macronutrient, fiber, calcium, iron and zinc intake were calculated using NutWin software version 1.5  . Foods nThe experimental group received 10 drops of zinc solution (10 mg Zn/day) added to milk or juice every morning at breakfast. The control group received an oral placebo (sorbitol 10%) under the same conditions. This 10 mg/day dose was considered physiologic and without risk of toxicity in children 8–9 years of age, mainly because it did not exceed the estimated average requirement (EAR) and the UL values.4·H2O) was purchased from Merck . The oral zinc solution (152.97 μmol Zn/day in the form of ZnSO4·7H2O) was prepared at the Pharmacotechnical Laboratory of the Department of Pharmacy, UFRN. Each drop contained 1 mg of elemental zinc. The placebo consisted of the same solution used to prepare the zinc solution. All the material used for the zinc and iron collections, separation, and storage was composed of metal-free propylene plastic. The tubes were purchased from BD-Becton , and the pipettes were purchased from Bio-Rad .Zinc sulfate heptahydrate  according to the manufacturer’s instructions. The zinc sensitivity was 0.01 μg/mL, the intra-assay coefficient of variation was 2.09%, and the normal reference range was 0.7–1.2 μg/mL. The iron sensitivity was 0.01 μg/mL, the intra-assay coefficient of variation was 2.79%, and the normal reference range was between 0.5 and 1.5 μg/mL.The standard zinc solution (1000 mg/mL) was obtained by diluting Titrisol Zinc Standard  in ultra-pure water. The standard iron solution  was obtained in the same manner. The wavelength was 213.9 nm and 248.3 nm for zinc and iron, respectively. The lamp current was 10 mA, and all other procedures, such as calibrations and measurements, were performed in accordance with the manufacturer’s instructions.The hematologic analyses, hematocrit, hemoglobin, and MCV were measured using standard clinical laboratory methods .Biochemical parameters, such as transferrin, transferrin saturation and total protein, were measured by the colorimetric method in a biochemical analyzer . Ferritin was measured by chemiluminescence , and ceruloplasmin was measured by nephelometry .t test was used to compare the data obtained in iron profiles. Paired and unpaired Student’s t tests were used to analyze anthropometric index, energy and nutrient intake, basal serum zinc, area under the iron curves, and biochemical parameters. The Wilcoxon test was used for paired nonparametric testing or the Mann-Whitney test for unpaired nonparametric testing. Tukey’s multiple comparisons test was used to compare iron concentrations. Subsequently, to verify whether the results were true for the population studied, we used the sample size calculation for comparing two means (paired samples) as follows: n = (Z α + Z β)2 σ2D/δ2. Concentrations were expressed as the mean ± SEM. All comparisons were considered to be statistically significant at the 5% significance level. Analyses were performed using GraphPad Prism version 6.0 .Statistical analyses included the Shapiro-Wilk test to analyze the normality of all study data. A paired Student’s 2 = 0.105034 and δ = −0.09, the sample size required would be n ≤ 15.Thirty healthy schoolchildren between 8 and 9 years of age were randomly assigned to the control and the experimental groups, and all completed the supplementation trial. The sample size of 30 schoolchildren was adequate for the conclusions obtained in this study given that for any value of α = 0.05, σAccording to BMI-for-age classification, all schoolchildren were eutrophic during the 3-mo study. The BMI was significantly different at the end of the study for both groups .Energy, protein, fat, carbohydrate, fiber, calcium, iron and zinc were normal according to the recommendations for age and sex, and showed a significant increase after oral zinc supplementation, indicating increased food consumption after the 3-mo study. Although fiber and calcium concentrations increased after oral zinc supplementation, they remained low in comparison to the recommendations by age and sex .i.e., 6.51 mg/day from zinc intake plus 10 mg/day from zinc supplementation. This was significantly different from the control group before and after placebo and the experimental group before zinc supplementation  suggested that Zip14 stimulates the cellular uptake of zinc and nontransferrin-bound iron [et al. (2012) provided evidence that Zip14 appears to be the ideal transporter or site for iron and zinc, which provides a mechanistic view of the interactions between both ions during uptake in the enterocyte [In the presence of organic substances, zinc and iron are absorbed by different mechanisms, particularly when administered as supplements . Emphasiund iron . Iyengarterocyte .Although serum iron is considered an iron-status indicator, it is important to recognize other indicators such as hematologic and biochemical parameters. As such, we analyzed hematocrit, hemoglobin, MCV, ferritin, transferrin, transferrin saturation, ceruloplasmin and total protein. To date, no single publication has provided a comprehensive quantitative analysis of the parameters used to evaluate iron nutritional status. Although zinc supplementation decreased serum iron, it did not affect the aforementioned indicators of iron deficiency. Given that these indicators were within normal reference ranges, iron deficiency and iron-deficiency anemia were ruled out . A likelet al. (2010), in a sample of 11 children, found that serum zinc had a potential inhibitory effect on the serum iron profile, but the deficiency was not sufficient to cause anemia in the children studied [Furthermore, no impairment of the total plasma protein concentration was detected, which, when diminished, might alter serum zinc, iron, and transferrin. Corroborating our results, Antunes  studied . Moreove studied .By contrast, a number of investigators reported a significant decline in hemoglobin in infants supplemented with zinc for 6 mo ,34. Addi.Additionally, zinc appears to interact with copper during absorption, which could affect the status of this micronutrient . For exaThe main limitation of our study was the difficulty in increasing the sample number of schoolchildren, hence the choice of the non-probability sampling method. Other limitations were (i) the extended time period in selecting the schoolchildren; (ii) standardization of the tests; (iii) the performance of 60 analyses; (iv) extensive dietary assessment and biochemical analysis; and (v) logistical complexity.Our study suggests that chronic oral zinc supplementation in physiologic doses most likely inhibited iron absorption in healthy schoolchildren. The negative effect of zinc on serum iron did not promote anemia, although the duration of supplementation and the dose should be considered. These findings are preliminary and warrant additional study.


Pseudomonas spp. Nosample was positive for Staphylococcus aureus andSalmonella spp. On the other hand,Listeria spp. was detected in only one sample of handmadediets. Contamination was significantly higher in the handmade preparations (p< 0.05). Nonconformities were detected with respect to good handlingpractices, which may compromise the diet safety. The results indicate that thesanitary quality of the enteral diets is unsatisfactory, especially handmadediets. Contamination by Pseudomonas spp. is significant becauseit is often involved in infection episodes. With regard to aseptic practices, itwas observed the need of implementing new procedures for handling enteraldiets.Contamination of enteral diets represents a high risk of compromising thepatient's medical condition. To assess the microbiological quality and asepticconditions in the preparation and administration of handmade and industrializedenteral diets offered in a hospital in the Valley of Jequitinhonha, MG, Brazil,we performed a microbiological analysis of 50 samples of diets and 27 samples ofsurfaces, utensils, and water used in the preparation of the diets. In addition,we assessed the good handling practices of enteral diets according to therequirements specified by the Brazilian legislation. Both kinds of enteral dietsshowed contamination by coliforms and  Enteral Nutrition Therapy (ENT) involves therapeutic actions intended to recoverand/or maintain the patient's nutritional status satisfactorily, and is indicatedfor patients who are not able to receive at least two thirds of their daily energyneeds orally . Controlcan be achieved by adopting sanitation and hygiene practices for the environment,foods, and hands of handlers. The latter is of crucial importance, because EnteralNutrition (EN) handling at the bedside is a critical point for meals contaminationby pathogenic bacteria  and industrialized diets, and the aseptic conditions ofpreparation and use of such feeds, produced in a hospital in the JequitinhonhaValley, Minas Gerais, Brazil.This study was conducted in a hospital in the Jequitinhonha Valley, Brazil, MinasGerais and in the Laboratory of Food Hygiene of the Federal University ofJequitinhonha and Mucuri Valleys (UFVJM), Brazil, from July 2010 to November2011.®, Nestlé, Brazil), pureed fruits, andvegetable soup . The industrializeddiets were of the type Isosource 1.5 cal® and Fibersource 1.2Kcal®. Samples from T0 were placed into previously sterilizedplastic containers, and T1 samples were collected directly from the originalcontainers of enteral diets. Then, the samples were transported in styrofoamboxes under refrigeration and analyzed not later than 6 h after beingcollected.Twenty-five samples of enteral diets were analyzed, 13 of them beingnon-industrialized diets  and 12industrialized diets, collected in two different times, immediately after thecompletion of preparation (T0) and after administration to the patient (T1),totalizing 50 sample units, each of them containing at least 100 mL of the diet.The "handmade" preparations consisted of: UHT (or long-life) whole milk, proteinsupplement . In a recipientpreviously sterilized in an autoclave, 250 mL of water were collected.Escherichia coli, viable aerobic mesophilic bacteria,Salmonella spp., Listeriaspp., Listeria monocytogenes,coagulase-positive Staphylococcus, andPseudomonas spp. For countertops and utensils, total andthermo-tolerant coliforms, E. coli, S. aureus,coagulase-positive Staphylococcus, and viable aerobicmesophilic bacteria were analyzed. Analyses of water and EN administrationdevices were conducted to detect Pseudomonas spp. besides themicroorganisms cited for evaluation of countertops and utensils.The diets were analyzed for total coliforms (35 °C) and thermo-tolerant coliforms(45 °C), ®), in 225 mL of watersaline solution (NaCl 0.85%). From this homogenate, decimal dilutions of up to10−6 were made for the handmade diets and 10−3 for theindustrialized diets and water, using tubes with 9 mL of water saline solution(NaCl 0.85%). The analyses of the samples of countertops, utensils, andadministration devices were conducted in tubes containing 10 mL of this samesolution (NaCl 0.85%).Twenty-five grams of each sample were weighed and homogenized in a Stomarcsamples homogenizer model MA440/CF (MarconiE. coli broth (EC)  for detectionof thermo-tolerant coliforms. In addition to the Most Probable Number (MPN)method, was used Petrifilm EC plates  forthe detection of coliforms and E. coli.One mL of each dilution was transferred to three test tubes containing LaurylSulfate Tryptose (LST)  with Durhan tubes in its interior. It wasused the Brilliant Green Bile Broth 2%  for confirmation of totalcoliforms and E. coli incubation was performed at 35°C for 24–48 h. The analyses of thermo-tolerant coliforms followed incubation at45 °C for 24 h.For total coliforms and 2 of surface and cfu/administration devices.The results were expressed in Colony-Forming Units per milliliter of diet orwater (cfu/mL), Most Probable Number per milliliter of diet or water (MPN/mL),cfu/cm2 of surface and cfu/administration equipment.One mL of the dilutions described above was transferred to a Petri plate by thepour plate method, to which standard Plate Count Agar  was added.After the agar homogenization and solidification, the plates were incubated at35 °C for 24–48 h. The results were expressed in cfu/mL of diet or water,cfu/cmL. monocytogenes was initially performed byadding 225 mL of Listeria Enrichment Broth  in 25 g ofsample, followed with incubation at 35 °C for 24 h. Immediatly after, it wasmade an inoculation of 0.1 mL of the previous step in tubes containing 10 mL ofFraser broth , and incubated at 35 °C for 48 h. Furthermore, 0.1mL of the same sample incubated for 24 h in Listeria Enrichment Broth wastransferred to Oxford agar plates  and 0.1 mL to chromogenic agarplates , and incubated at 35 °C for 24–48 h, and checked forgrowth of typical colonies. With the tubes with Broth Fraser it was made thesame plating mentioned, in Oxford agar plates and chromogenic agar plates, tocheck for growth of L. monocytogenes bacteria. In addition,analysis to detect Listeria spp. was conducted, using the kitTECRA Listeria VIA, AOAC 995.22, 2002.09 , incubated at 35 °C for 22–24 h. After this period, the primary selectiveenrichment was accomplished: 1.0 mL of the pre-enrichment medium was transferredto tubes containing 9 mL of Selenite Cystine broth , andincubated at 35 °C for 6–18 h. Next, the secondary selective enrichment wasperformed: 1 mL of the primary enrichment medium was transferred to tubescontaining 9 mL of M broth , and incubated at 35 °C for 6–8 h.After that, the kit TECRA Salmonella VIA, AOAC 989.14, 998.09, 4 related to 10−1dilution and two to 10−2 and 10−3 dilutions, and incubatedat 35 °C for 48 h. After checking for typical colonies growth , count and selectionof five typical colonies were performed for the coagulase test. The result wasexpressed in cfu/mL of diet.The analysis for Staphylococcus Petrifilm™ Staph Express plates were used, and after growth of the colonies, the referred coagulase test was carried out.In the analyses of countertops, utensils, water, and administration devices, forthe counts of Pseudomonas spp. the same dilutions, as alreadydescribed, were used, up to 10−3, and then 0.1 mL was poured ontoPetri plates with Pseudomonas agar for Piocianines , and then incubated at 25 °C for 48 h. As colonies grew , a confirmation test was performed in inclined tubes with TripleSugar Iron - TSI  and then incubated at 25 °C for 24 h. Theisolates growing without change in color were confirmed asPseudomonas spp.To count The assessment of good practices of diets preparation and handling was accordingto the Brazilian technical regulation for enteral nutrition therapy.This regulation contains guidelines for inspection of EN-related activities,including the following: MTNT activities, general conditions, receiving of dietprescriptions, storage, water, preparation, cleaning and sanitation, clothing,handling and packaging, storage and transport, quality assurance, qualitycontrol, and EN administration.Each item had questions, classified as essential (E), when it would impactcritically the EN quality and safety; necessary (N), which would impact lesscritically the EN quality and safety; and recommendable (R), which would notinterfere critically on the EN safety and quality.For the interpretation of the results, each question was considered "conform"(C), when it met the requirements; "nonconform", when the item beinginvestigated did not comply with the requirements; and "not applicable" (NA),when the hospital did not provide the service.The results were analyzed by means of the difference between the values ofmedians using Mann Whitney and Wilcoxon Signed Rank nonparametric testing,correlation and regression analysis using the Minitab software, version 15. Thestatistical significance level was 5% of probability.Pseudomonas spp.Both industrialized and non-industrialized enteral diets showed contamination bycoliforms and thermo-tolerant coliforms, aerobic mesophilic bacteria, andConsidering the microbiological standards under Brazilian law , the perThese results corroborate those found by Pseudomonas spp. (T1) and after administration to the patient (T1), beingconsidered significant results (p < 0.05). No difference was observed in thecounts of fecal coliform (T0 and T1) and pp. (T1).Taking into account the difference between the two instants of collection (T0 andT1), regardless of the type of diet, an increased number of microorganisms  were fouPseudomonas spp., not significant for thecounts of other bacteria analyzed (p > 0.05). The regression equation that bestrepresents the increased amount of Pseudomonas spp. as a functionof time is:There was a significant correlation (p < 0.05) between the time elapsed from thepreparation of the diet until completion of administration to the patients and theincreased population of Pseudomonas spp. population (p = 0.017).So, we can infer that at every minute there is an increase of 0.0158 log (cfu/mL) ofPseudomonas spp., it is worth notingthat the Brazilian legislation does not specify standards for this pathogen, but itis important to consider that its occurrence has been often observed in nosocomialinfections or in isolated clinical materials sent to culture , E.coli (up to 102 cfu/device), aerobic mesophilicmicroorganisms (up to 104 cfu/device), coagulase-positiveStaphylococcus (102 cfu/device).With respect to the water samples used to cleanse the tubes and to dilute thehandmade diets, it was observed absence of total coliforms andWith respect to the water quality, we considered that it is not a source ofcontamination, once it met the standards defined in the RDC N°. 12, of January 12,2001 .The contamination of catheters was considered an important risk factor forcontamination of enteral feeding, including by pathogens, allowing the occurrence ofinfectious exacerbations in patients receiving this type of nutritional support.The questionnaire included 208 topics for investigation, and The following topics did not show nonconformities: receiving of the dietprescription, water, and clothing.Among the questions related to MTNT activities, the hospital in questionsucceeded in some topics considered essential, such as the existence of a formalact to constitute the team, formal record of meetings, and records of medicalprescriptions. However, the following were considered nonconformities: lack ofprocedural protocols for the professionals involved in the therapy, lack oftraining programs, and lack of ENT quality indicators.Regarding the activities related to preparation, general conditions, cleaning andsanitation, handling, storage, packaging and transport, the main nonconformityfound in the category of essential was the relative lack of a separate area forpreparation of EN. The other failures detected were among items considerednecessary and recommended, and among them we can cite: lack of sealed openingsand wire-protected windows, allowing access of insects and rodents into the ENhandling sector, unrestrictive circulation of people in the preparation sector,and the floor of the preparation area was difficult to clean and had cracks.Among the conformities found for all categories , it is worth noting that preparation was made only upon medicalprescription, the staff was trained to perform this activity and such trainingwas recorded, the diets were kept in an exclusive refrigerator, and the dietsproduction facility of this healthcare institution performs sanitation and pestcontrol every semester.Concerning the enteral feeding process, the nonconformities found among the itemsindicated as essential include: the diets were not stored in the refrigeratorwhen not used at the specified time; the feeding devices were not exclusivelyused for feeding, being also used for administration of medications; sanitationof tube connections was not performed during the change of these devices.Among the items considered essential, the conformities found among the ENadministration activities, we can cite the clinical and laboratorial control ofthe patient receiving the enteral diet, and the diet was administered in itsoriginal container. In addition, among the topics considered necessary andrecommended, we can cite pump-assisted administration of the diet; trainednursing staff to use the pumps; and existence of a manual of procedures for ENadministration.Situations not applied to the reality of this healthcare unit are mainly relatedto the fact that MTNT has not established Standard Operational Procedures (SOP)and therefore there are no control records of essential qualities to assure ENsafety.The microbiological quality of the enteral diets analyzed was not satisfactory, andthe aseptic conditions in the investigated hospital with respect to preparation andhandling of enteral diets were favorable to risks of cross contamination.


Using phylogenetic and syntenic tools, we analyzed the Adam genes in various vertebrates ranging from fishes to placental mammals. Our analyses reveal duplication and loss of some sAdams in certain vertebrate species. In particular, there exists an Adam9-like gene in non-mammalian vertebrates but not mammals. We also identified putative group I and group II tAdams in all amniote species that have been examined. These tAdam homologues are more closely related to Adams 9 and 9-like than to other sAdams. In all amniote species examined, group II tAdams lie in close vicinity to Adam9 and hence likely arose from tandem duplication, whereas group I tAdams likely originated through retroposition because of their lack of introns. Clusters of multiple group I tAdams are also common, suggesting tandem duplication after retroposition. Therefore, Adam9/9-like and some of the derived tAdam loci are likely preferred targets for tandem duplication and/or retroposition. Consistent with this hypothesis, we identified a young retroposed gene that duplicated recently from Adam9 in the opossum. As a result of gene duplication, some tAdams were pseudogenized in certain species, whereas others acquired new expression patterns and functions. The rapid duplication of Adam genes has a major contribution to the diversity of ADAMs in various vertebrate species.Members of the disintegrin metalloproteinase (ADAM) family have important functions in regulating cell-cell and cell-matrix interactions as well as cell signaling. There are two major types of ADAMs: the somatic ADAMs (sADAMs) that have a significant presence in somatic tissues, and the testicular ADAMs (tADAMs) that are expressed predominantly in the testis. Genes encoding tADAMs can be further divided into two groups: group I (intronless) and group II (intron-containing). To date,  Adams”  to reflect these important features [i.e., the sADAMs) have a significant presence in somatic tissues and play important roles in embryonic development [Adam genes have been identified in vertebrate and invertebrate animals, and Adam-like sequences have also been found in the yeast species Schizosaccharomyces pombe [The first two members of the ADAM family were originally identified as guinea pig sperm-surface antigens that were recognized by the monoclonal antibody PH-30, which strongly inhibits sperm-egg fusion . Sequencfeatures . It soonelopment ,6. To daes pombe ,8. Functes pombe ,10. ADAMes pombe ,8.A typical ADAM is a type I transmembrane protein, although secreted soluble proteins can also be generated through alternative splicing. The unprocessed form of these transmembrane proteins contains a signal peptide, a pro-domain, a metalloproteinase domain, a disintegrin domain, a cysteine-rich domain, a transmembrane region and a cytoplasmic tail, from N- to C-terminus ,8. Only Adam genes, while 27 Adam loci have been identified to date in the human genome [Adam genes are expressed exclusively or predominantly in the testis, and are referred to as tAdams. These tAdams are further classified into two groups: group I genes , which have no or few introns; and group II genes , which have multiple introns. None of the group II tADAMs contains the consensus zinc-binding motif in the metalloproteinase domain, and are hence likely non-proteolytic. By contrast, group I tADAMs, except for ADAMs 4, 6 and 29, are predicted to be proteolytically active. It should be noted that some tADAMs, including the presumably proteolytic ADAM1, undergo further processing to remove the metalloproteinase domain during sperm maturation [The mouse genome contains at least 34 n genome ,13. Abouturation ,14. Anotturation . Such coturation ,15. One turation ,17.tAdam genes have only been identified in eutherians, and it is not clear how they originated during evolution and if there are any tAdams in other animals. Most of our current knowledge on the functions of tADAMs comes from studies in mice, which show that several tAdam genes are important for fertilization in this rodent species. For example, Adam1a-/-, Adam2-/- and Adam3-/- male mice are all infertile. Detailed analyses indicate that sperms lacking any of these tADAMs are defective in their migratory activities in the female reproductive tract, as well as in in vitro sperm-egg interaction [Adam24-/- mice: these mice displayed reduced fertility with high level of polyspermic fertilization [Adams 1a and 3, appear to have been pseudogenized in the human genome [tAdam genes in various animal species will not only help us understand how these genes evolved, but also provide important clues on which of these genes may be expressed and functional in individual species. Given the importance of tADAMs in fertilization, the latter information may be of particular interest to veterinary and agricultural research.Thus far, eraction –21. A dilization . Howevern genome ,24, suggtAdams in all amniote species that have been examined. Phylogenetic and syntenic (gene linkage) analyses suggest that these tAdams originally derived from ancient Adam9, a somatic Adam that is conserved in vertebrates, and/or Adam9-like, which we identified here as a close paralogue of Adam9. Two mechanisms, tandem duplication and retroposition, were found to underlie the expansion of the tAdam genes during vertebrate evolution. These results provide important insight into the evolution of tAdams, and form the basis for future functional studies of these important reproductive genes in various vertebrate species.In the current study we identified putative group I and group II Adam genes. Specifically, we performed consistent mining of representative vertebrate species  to understand the broader evolution of the ADAM genes. This was augmented with data obtained from additional species, such as rat, rabbit, pig, dog, platypus, zebra finch, flycatcher, coelacanth, Japanese medaka, cave fish, spotted gar, elephant shark and lamprey, where a more specific understanding of synteny/phylogeny was needed to provide clarity on the data. Potential retroposed Adam genes were identified as Adam genes with no or few (≤ 2) introns by UCSC Genome Browser BLAT search [Adam genes, along with surrounding genome sequences that are 1 kb upstream and downstream, were analyzed with the CENSOR program [The sequences of annotated ADAM proteins from humans and mice were BLASTed against the EST databases and genome sequences of other vertebrate species to identify T search . To iden program .Adams. Phylogenetic and syntenic analyses were carried out as described below. To determine the exon/intron boundaries of Adam genes, deduced amino acid sequences were searched against the UCSC Genome Browser using BLAT [Three main criteria, phylogeny, synteny, and conservation of exon/intron boundaries, were used to establish the orthology with existing human and mouse ing BLAT .Adam genes from various species  alignmeThe Bayesian analyses were conducted with MrBayes version 3.2.2 . Each anNeighbor joining and maximum likelihood trees were generated by using the corresponding algorithms included in the MEGA 6.0 software . For neiAdam gene was entered into the search box of Genomicus [Adam gene in a specific species was validated by searching the sequences of nearby genomic regions that include the syntenic anchor genes.For syntenic analyses, an individual enomicus , Metazomenomicus , and NCBet al. identified 4 and 22 Adam genes in Ciona and zebrafish, respectively, and we have also reported the presence of 15 homologues of mammalian sAdam genes in Xenopus tropicalis [C. elegans, Drosophila and Ciona [Drosophila and Ciona, which do not belong to the ADAM10/17 clade. These ADAMs cluster with the rest of mammalian sADAMs, but no distinct orthology could be established [A previous study by Huxley-Jones opicalis ,36. In aopicalis –38. Thesnd Ciona ,37. One ablished .Xenopus [sAdam genes during early vertebrate evolution. However, these published studies failed to identify an orthologue of ADAM7 in zebrafish and Xenopus, and ADAM8 seems to be missing in Xenopus [Adam genes and, particularly, the origins of tAdams, we performed a search for Adam genes in various vertebrate species, including X. tropicalis, Anolis, chick, opossum, mouse and human. Phylogenetic trees of deduced protein sequences encoded by these genes were generated by three methods: Bayesian  and ADAM7 form a cluster, likely as a result of tandem duplication . Similarn tandem . Four anal to it ,46.Xenopus ADAM13, which plays an important role in neural crest induction and migration as well as eye formation, is an orthologue of mammalian ADAM33 [et al. reported the presence of two Adam19 loci (Adam19a and Adam19b) but not Adam13/33 in zebrafish [Adam13/33 only exists in tetrapods. The two Adam19 loci in zebrafish are unlikely the result of TGD, as both are also present in the spotted gar  gene that has been reported to be expressed exclusively in the testis of Xenopus laevis [Adams 7 and 28 are sometimes classified together with the tAdams as “reproductive” Adams, as Adams 7 and 28 are predominantly expressed in the mouse epididymis [Adam28 is expressed in a wide variety of somatic tissues, and our phylogenetic analyses strongly indicate that ADAMs 7 and 28 do not belong to the subfamily that includes ADAMs 9, 9-like and tADAMs. Thus ADAMs 7 and 28 were not included in the subsequent analyses.In an attempt to find potential  opossum , but notd 9-like  Figs. Als genome , there is laevis . Interesididymis . HowevertAdams shows a unique feature: in all amniote species that were examined, group II tAdams cluster together in a genomic region that also includes Adam9  that may serve as evidence for the retroposition events they underwent. We therefore started out to search for young retroposed genes that derived from the Adam9/9-like/tAdam subfamily, in which the characteristic features of retroposition may still be well preserved.Unlike group II position . HoweverAdam9 that has only one intron  in nucleotide sequence. The Adam9a gene contains 24 introns and conserved exon/intron boundaries that are found in Adam9 genes of other vertebrate species (data not shown), and is hence the opossum orthologue of Adam9. The lack of introns and very high sequence homology to Adam9a suggest that Adam9b was likely retroposed recently from Adam9a. This is further supported by the presence of a polyA tail in the Adam9b gene . Retroposed genes were thought to be non-functional because they were not expected to carry any regulatory elements required for transcription [tAdam genes, which likely derived from retroposition, are predominantly expressed in the testis.A common consequence of gene duplication is the loss of function of duplicated genes. The HUGO Gene Nomenclature Committee website listed 11 human II genes . The psecription . Howevercription . Direct cription . Interescription ,55. Thiscription ,58. TherAdam2 promoter was reported to bind Mrf/Gm98, a transcriptional activator in the central nervous system [tAdams are also expressed in some somatic tissues in mice and humans Click here for additional data file.S2 Fig(DOCX)Click here for additional data file.S3 Fig(DOCX)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.


Cardiovascular disease is the leading cause of death in the Chinese population. Although general prevalence estimates of cardiovascular risk factors (CVRFs) are available for Chinese adults, prevalence estimates covering all adult age groups by race/ethnicity have not been reported. The aim of this study is to estimate the current prevalence and clustering of major CVRFs in Chinese adults, including a plurality of ethnic minorities.A cross-sectional survey was conducted in a nationally representative sample of 23,010 adults aged 18 years and older from 2007 to 2011. Questionnaires and physical examinations were performed, and fasting blood was collected for laboratory measurements. The prevalence of traditional CVRFs, including hypertension, diabetes, dyslipidemia, overweight, and current smoking, were determined.The prevalence of the major CVRFs, including hypertension, diabetes, dyslipidemia, overweight, and current smoking were 24.3%, 4.3%, 49.3%, 32.0%, and 21.7%, respectively. These risk factors were significantly associated with sex, age, region, ethnicity, and education levels. Overall, 70.3%, 40.3%, and 16.7% of Chinese adults had ≥1, ≥2, or ≥3 CVRFs, respectively. Men, northern and rural residents were more likely to have clustered CVRFs compared with women, southern and urban residents, respectively. Compared with Han residents, Hui and Mongolian residents were more likely, and Tujia and Miao residents were less likely, to have ≥1, ≥2, or ≥3 risk factors. The prevalence of Chinese women having ≥1, ≥2, or ≥3 CVRFs decreased with increasing levels of education.The prevalence and clustering of CVRFs is still high in Chinese adults ≥18 years old, especially in men and in individuals living in the northern and rural areas. Of note, there are differences in cardiovascular risk among different ethnic groups. Therefore, targeted and enhanced intervention measures are required to reduce the risk of cardiovascular disease and the corresponding economic burden of disease in China. China is a multiethnic country with 56 ethnic groups. The Han-Chinese account for >90% of the total population, whereas the other 55 ethnic minorities account for <10% of the total population. Among these minorities, 18 ethnic groups, including the Mongolian, Hui, Tibetan, Miao, Yi, Tujia, Korean, Zhuang, Manchu, Uygur, Buyi, Dong, Yao, Bai, Hani, Kazakhs, Dai, and Li populations, comprise 1 million individuals. The geographic distribution of Chinese minorities is extremely uneven, and minority populations living in the northwest and southwest regions of China account for 30.1% and 29.4% of the total minority population, respectively. In China, the minority population is mainly distributed in Inner Mongolia, Xinjiang, Ningxia, Guangxi, Tibet, Yunnan, Heilongjiang, Jilin, Liaoning, Qinghai, Sichuan, Guizhou, Gansu, Hunan, Hubei, and Hainan, and most minorities have specific lifestyles. In addition, racial and ethnic differences are known to influence the prevalence and risks of CVD.16 Therefore, there is a need to understand the risk factors involved in CVD and to establish rational prevention strategies to decrease morbidity related to CVD in a representative population sample of Chinese multiethnic adults. Currently, no information is available in the literature regarding the ethnic differences in the prevalence of CVD risk factors among Chinese multiethnic adults.In China, due to rapid economic growth and lifestyle changes, the prevalence of CVD and morbidities related to being hypertensive, diabetic, dyslipidemic, and overweight have gradually increased. Recent data suggest that the prevalences of coronary heart disease, stroke, and CVD in adults aged 20 or older have been up to 0.63%, 0.83%, and 1.44%, respectively, and the standardized prevalences in the 2006 population were 0.60%, 0.80%, and 1.37%, respectively.In this study, we used data from a nationally representative sample of multiethnic adults to evaluate the current status of CVRFs and to assess ethnic differences of cardiovascular risk in the general population of China.18 Briefly, we selected 6 provinces  from different geographical regions (south or north) in China. A 3-stage cluster sampling method was used to select eligible subjects in each province. First, 2 or 3 cities were sampled based on their economic status and the presence of ethnic minorities; then, several communities and villages were randomly selected from each city.The data used in this study are from the Chinese Physiological Constant and Health Condition (CPCHC) survey, a population-based, cross-sectional survey, which was conducted from 2007 to 2011. This survey used a random, multistage, stratified sampling method to obtain a nationally representative sample of the general Chinese population.A total of 36,215 participants were randomly selected to complete the survey and blood biochemical tests after excluding individuals who suffered from systemic disease and confirmed cardiovascular, renal, gastrointestinal, and pulmonary disease or cancer. A schematic of the screening process is presented in Figure We have performed the calculation of sample size before using these data. According to the existing literatures, we estimated that minimum population prevalence for these risk factors is 5%, and then a minimum sample size should be 7299 if the allowable error is controlled within a range of 0.5%. Actually, the final sample size in this study  fully meets the requirements for minimum sample size. Written informed consent was obtained from each participant prior to data collection. The protocol was approved by the institutional review board of the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences.All of the participants were asked to complete a standard questionnaire, which included demographic characteristics , socioeconomic data , lifestyle risk factors , and their past medical history.2). Waist circumference (WC) was measured with flexible and inelastic tape at the end of a gentle expiration. Blood pressure was measured using an Omron HEM-7000 electronic sphygmomanometer  after the participant had rested for at least 10 minutes.Body weight and height were measured and BMI was calculated as body weight divided by the square of the height , high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C), were measured using the Beckman AU Series Automatic Biochemical Analyzer and Sekisui Medical (Japan) reagents. Uric acid (UA), FBG, creatine (Cr), and blood urea nitrogen (BUN) levels were measured using the same instrument and Beckman AU reagents. The biochemical laboratories participating in the survey followed the common internal quality control program, which was standardized by the Peking Union Medical College Hospital.2. Current cigarette smoking was classified as self-reported responses of “yes” to the question “Do you smoke cigarettes now?”Hypertension was defined as having an average systolic blood pressure (SBP) ≥140 mm Hg and/or diastolic blood pressure (DBP) ≥90 mm Hg, and/or current antihypertensive medication use. Diabetes was defined as FBG ≥ 7.0 mmol/L and/or current antidiabetes medication use. Dyslipidemia was defined as having at least one of the following: TC ≥ 5.2 mmol/L, TG ≥ 1.7 mmol/L, HDL-C < 1.0 mmol/L, LDL-C ≥ 3.4 mmol/L, and/or current cholesterol-lowering medication use. Overweight/obesity was defined as having a BMI ≥ 25 kg/mt test. Variables with a skewed distribution (age and TG) are presented as the median (interquartile range) and were compared by the Wilcoxon rank sum test. Categorical data  are presented as percentages and were compared by the χ2 test. The tests were performed to compare the variables with the relevant groups. Two-tailed values of P < 0.05 were considered statistically significant. The direct standardization method was used for the “age-standardized” prevalence estimates and prevalence estimates of different CVRFs were calculated based on the overall population according to age, in which age-standardized prevalence was determined using the population distribution of China in 2006.7 A multivariable logistic regression model was used to explore the association between demographic characteristics and CVD risk factor clustering. We assessed the goodness-of-fit of the logistic regression model using Hosmer–Lemeshow test. For the selection of the explanatory variables, firstly univariate logistic analysis was applied to examine association between each independent variable and clustered risk factors. Then in multivariable logistic regression analyses, all independent variables that were statistically significant and variables that had been shown to have clinical significance were included as potential confounding variables and the adjusted odds ratios (ORs) and 95% confidence intervals were calculated. The data were adjusted for age and sex, and the ORs and 95% confidence intervals were used to assess the risk of CVD risk factor clustering.The database was constructed with EPI3.02 software by 2 data managers and was corrected to guarantee the accuracy and integration of the data. The data were analyzed using SPSS 16.0 software . Normally distributed continuous variables  are presented as the mean ± standard deviation and were analyzed by P = 0.473). Additionally, men had significantly higher BMI, WC, SBP, DBP, UA, FBG, TG, TC, HDL-C, Cr, and BUN levels compared with women (P < 0.001).As shown in Table P < 0.001). The prevalence of all of these risk factors increased significantly with increasing age (P for trend <0.001), but slightly decreased in participants ≥65 years for dyslipidemia, overweight, and current smoking relative to the total population and regardless of gender (Tables P < 0.01). Notably, there are significant differences in the prevalence of these risk factors among different ethnic groups (P < 0.001). The prevalence of hypertension, diabetes, dyslipidemia, overweight, and current smoking was highest in the Hui (36.8%), Korean (4.9%), Yi (61.3%), Mongolian (44.7%), and Miao (31.3%) populations, respectively. Hui, Yi, and Mongolian individuals (of both sexes) had the highest prevalence of hypertension, dyslipidemia, and overweight, respectively. Miao men and Tujia women had the highest rates of diabetes . Miao men and Yi women had the highest rates of current smoking . In the higher education level group, there was a lower prevalence of these CVD risk factors in women, but a higher prevalence of diabetes, dyslipidemia, and overweight in men (Table Tables r Tables  and 3. EP < 0.001). With increasing age, the prevalence of CVD risk factor clustering increased within the total sample and in women. However, the proportion of men who had ≥1, ≥2, or ≥3 risk factors was higher among subjects of older age until 55 years old, when the prevalence began to be lower  was included and the prevalence of most CVRFs increase with age. However, the prevalence of overweight is still higher in our study, which indicates that over the past decade, more people have become overweight/obesity in China and the prevention and control of CVD should emphasize overweight/obesity. In fact, in 2002, 21.8% of Chinese adults aged 18 years and older were overweight or obese based on the WHO standards, and in 1992, the figure was only 14.6%.19The results from our study showed that 24.3%, 4.3%, 49.3%, 32.0%, and 21.7% adults aged 18 years or over in China have hypertension, diabetes, dyslipidemia, overweight, and current smoking, respectively, which are conventional CVRFs used to predict CVD morbidity and mortality. In 2005, a previous study reported that the prevalence of these CVRFs among Chinese adults aged 35 to 74 years was 26.1%, 5.2%, 53.6%, 28.2%, and 34.4%, respectively.21 the data in this study showed that CVRFs, including hypertension, dyslipidemia, overweight, and current smoking, were more common among people living in rural compared with urban areas. This difference may be explained by the rapid urbanization of rural areas23 and the improvement of the health care system and people's health conditions in urban areas in China during recent years.24 A cohort study in New Zealand obtained similar results and showed that rural Maori had significantly higher levels of type 2 diabetes, diagnosed hypertension, treated dyslipidemia, current smoking, and age-adjusted BMI compared with urban Maori.25 Another study in China demonstrated that from 1990 to 2003, the mortality rate of CVD increased in rural areas, whereas the figures decreased slightly in urban areas.19Although several studies have demonstrated that the prevalence of some CVRFs is higher in urban populations,28 Our study revealed that 70.3%, 40.3%, and 16.7% of Chinese adults had ≥1, ≥2, or ≥3 CVRFs, respectively. The significantly higher prevalence of ≥1, ≥2, or ≥3 risk factors among men, northern and rural residents compared with their women, southern and urban counterparts reflects the fact that most CVRFs investigated in this study were higher among men, northern and rural residents. The different patterns between genders, rural–urban areas, and regions are not only a result of the differences in socioeconomic status or economic development but may also be a result of differences in education levels.30 In this study, we found that the clustering of CVRFs was associated with education levels. Interestingly, in women, there was a decreased risk of clustered CVRFs with higher education levels, but in men, an increased risk of clustered CVRFs was found with higher education levels, which is likely due to differences in occupations, family responsibilities, and health concerns. Although highly educated people tend to have more health awareness, the gender-specific conception of health may be influenced by education levels.31 Highly educated women often pay more attention to their body shape and health and consider thinness to be desirable. In contrast, highly educated men attend more social occasions and eat out more frequently (including drinking and overeating high-fat and high-energy foods), which easily causes diabetes, dyslipidemia, and overweight. In this study, we also found that, in men, the CVRFs increased with increasing age up to the age of 55 years, but decreased thereafter. A possible reason for this observation is as follows: younger men tend to have adverse eating and lifestyle habits . However, older men gradually pay more attention to their own physical health because they have more time and are more worried about health after retirement.A large number of studies have demonstrated that CVD incidence and all-cause mortality increased markedly in the presence of CVRF clustering.33 China is a multiethnic country, and the different ethnic groups have specific dietary habits and lifestyles, which may affect their cardiovascular health status. In this study, the population was ethnically diverse, and the prevalence of CVRFs was significantly different between ethnicities. Compared with Han residents, Hui and Mongolian residents were more likely, and Tujia and Miao residents were less likely, to have ≥1, ≥2, or ≥3 risk factors. A recent study reported that hypertension prevalence in ethnic Hui and Mongolian minorities was significantly higher than in the Han population,34 which is consistent with our findings. Notable differences exist in the CVRF assessment by ethnicity, region, and education that require central and local governments to develop related policies and guidelines to improve the health care system and strengthen interventions.Differences in the prevalence of CVD and associated risk factors have been noted across ethnic groups both within and between countries.A major strength of the present study is that it is a population-based study with a representative sample of the general Chinese adult population, including a plurality of ethnic minorities. Additionally, the larger sample size ensures sufficient power in estimating the prevalence and clustering of major CVRFs, as well as determining the association between clustered risk factors and sex, age, regions, ethnic group, and education.The study also has several limitations. First, the study is based on a cross-sectional survey, and thus cannot determine causality or the temporal relationship between CVRF clustering and the incidence of CVD. However, previous studies have demonstrated the importance of CVRF clustering to the incidence of CVD. Second, the population sizes of some minorities were relatively small because of the limited ethnic group population in the study regions compared to the Han population; however, a 4-stage randomly stratified cluster sampling method was used to make the study population representative. Third, we assessed these major CVRFs according to the commonly used international standards and did not have physician diagnoses for diseases, although a history of self-reported disease was included. Last, there are geographical variations in the distribution of ethnic minorities in China and we also found the significant differences in the prevalence of CVRFs between ethnicities, so the sampling design may cause the presence of some degree of intracluster correlation, which may be a major limitation of the study.In conclusion, our data highlight the high prevalence and clustering of CVRFs in Chinese adults, especially among some minority groups such as the Hui and Mongolian populations. A healthy lifestyle, such as weight control, the avoidance or cessation of smoking, eating a healthy diet, and undertaking regular physical activities should be emphasized not only in the Han population but also in minority ethnic groups. Corresponding public health programs are required to improve this situation in the Chinese population.


The objective of this study was to systematically review the literature for a more comprehensive understanding of the complications of open elbow arthrolysis in patients with post-traumatic elbow stiffness and provide a reference for better prevention and treatment of them.The PubMed, EMBASE, Cochrane Library, and Google Scholar databases were searched for therapeutic studies with a set of inclusion and exclusion criteria. Data were extracted from selected articles, and a statistical analysis was performed to evaluate related factors and management of the complications.Twenty-eight articles published between 1989 and 2013, involving 810 patients, were included. Most of the complications included in the selected articles were nerve complications, heterotopic ossification, elbow instability, infection, pin-related complications and repeat elbow contracture. The total complication rate was 24.3% ± 3.0%, and the reoperation rate was 34.0%. Furthermore, the statistical analysis revealed that preoperative range of motion  and proportion of female  were the independent factors affecting the total complication rate.Various risk factors are related to each of the complications, and we found that patients with less preoperative ROM and a higher proportion of female gender may point to a higher total complication rate. Therefore, to further improve the overall outcomes of this procedure, more and larger prospective studies should be performed to further elucidate the effects of prophylactic interventions targeting the risk factors, thus improving the methods of prevention and treatment of complications. Elbow stiffness, a common condition associated with significant morbidity, is generally defined as elbow range of motion (ROM) <30°–130° . Trauma Both non-operative and operative methods are used to treat elbow stiffness. Generally, surgical release should be considered when non-operative treatment fails to improve the motion after 6 months . Open arIn 2013, Kodde et al.  publisheWe searched the PubMed, EMBASE, Cochrane Library, and Google Scholar databases in August 2014 performed an updated search in October, without publication status restrictions, using the following keywords (elbow and [stiff* or contracture or ankylos*] and [releas* or arthrolysis]) to ensure the inclusion of all possible studies. The search was restricted to articles written in English. We also used terms included under Medical Subject Headings (MeSH) in all searches, when possible. In addition, references regarding elbow stiffness were hand-searched for potential studies. All searches were conducted independently by 2 researchers (JYC and WC), and the differences were checked and resolved by discussion.To be eligible for inclusion, the studies had to: (1) be published clinical trials; (2) meet the diagnostic criteria for post-traumatic elbow stiffness ; (3) report operative treatment and outcomes of post-traumatic elbow stiffness in human adults; and (4) reach an average of at least one-year follow-up. Studies were excluded if they: (1) were cadaveric or biomechanical studies, reviews, expert opinions, case reports (number of cases <5), or conference papers that were not published as full reports; (2) reported on stiffness after a burn or central nervous system injury, results of arthroscopic release, or arthroplasty; and (3) were unable to extract the relevant data from the outcomes. We carefully reviewed the full text to exclude studies that partially involved non-posttraumatic stiffness and those that did not mention complications in their follow-up.Data were extracted and entered into an Excel database by 2 independent authors (JYC and WC). The data included: author details, year, number of cases, population demographics , mean follow-up, and outcome measures. Disagreements were resolved by involving a third reviewer (WW) in the discussion.The outcome measures were functional outcomes and complications .ROM was calculated as mean ± standard deviation (SD). The type-specific complication rates were expressed as percentages with 95% confidence intervals (CIs). We also divided the patients into two groups based on whether they had been treated with hinged external fixation. A Chi-square test or Fisher exact test was applied for comparison of the complication rates of the two groups. Stepwise multiple linear regression analysis was conducted to determine the independent factors , time from injury, duration of follow-up, preoperative ROM, postoperative ROM) affecting the total complication rate. The significance level was set at P < 0.05. The statistical analyses were performed using SPSS 19.0 statistical software .A total of 677 studies were identified in the initial search. After a careful review of the lists, full texts were retrieved for 142 articles. We subsequently excluded 115 articles that did not meet the inclusion criteria. A search of the reference lists of selected articles identified 1 more relevant article; our search was updated in October 2014 with no more relevant articles, leaving a total of 28 articles, involving 810 patients (814 elbows) , for theThe main characteristics of the studies included in the systematic review are summarized in Among the 810 patients, the total number of complications after open elbow arthrolysis was 197 (24.3% ± 3.0%), which included nerve complications, heterotopic ossification (HO), infection, and elbow instability and others. 67 patients (34.3%) needed reoperations because of the persistent symptoms after the index release. The complications are recorded from each of the selected studies in Nerve complications, defined as new-onset or exacerbation of nerve symptoms postoperatively, occurred in 70 patients (8.6% ± 1.9%) and consisted of ulnar, radial, and median nerve symptoms and reflex sympathetic dystrophy. We found that nerve complications involving the ulnar nerve, also known as delayed-onset ulnar neuropathy (DOUN), were the most common (66 patients); thereafter, 13 of 70 patients (18.6%) needed a subsequent surgery to decompress the ulnar nerve with or without transposition.HO occurred in 45 patients (5.6% ± 1.6%) and consisted of radiographic and clinical new-onset or recurrence. Patients who failed to regain a functional arc of motion due to HO postoperatively are considered to have clinical new-onset or recurrence. 13 of 45 patients (28.9%) underwent a second surgery for HO, and all of them had clinical recurrence of HO.Elbow instability occurred in 8 patients (1.0% ± 0.7%), which included valgus and varus instability, as well as elbow subluxation. Hinged external fixators were placed at 3 and 7 weeks post-release in 2 of 8 patients (25.0%), and the remaining patients with mild instability that did not affect daily function did not require surgical treatment.Infection occurred in 13 patients (1.6% ± 0.9%), which included superficial infection and deep infection (pin track infection was excluded). Seven of the 13 patients (53.8%) underwent a reoperation as irrigation and debridement. As a result, the infection was under control in 3 and the outcomes of the remaining 4 patients were not reported in the studies.Among the 86 patients with routine hinged external fixation, pin-related complications occurred in 15 patients (17.4% ± 8.0%), which included 9 patients with pin track infection, 2 with pin breakage, 2 with ulnar nerve irritation, and 2 with pin-site fracture. Two patients (13.3%) needed a second surgery: 1 patient with pin track osteomyelitis underwent debridement and 1 with a pin-site fracture was treated with open reduction and internal fixation.The other complications included 33 repeat elbow contractures for unknown or unreported reasons (excluding elbow contracture associated with HO mentioned in the studies), 4 re-fractures , 3 hematomas, 2 cases of synovitis, 1 tricep avulsion, 1 incomplete wound healing, 1 wound dehiscence and 1 intrarticular bleeding. Twenty-nine patients (63.0%) had subsequent surgeries to deal with the complications: 4 patients with a re-fracture and 25 patients with a repeat elbow contracture. The other 8 with a repeat elbow contracture refused a second arthrolysis.There was a significant difference in the reoperation rate between the patients treated with and without hinged external fixation (P = 0.04). However, difference of total complication rate between the two groups was not significant (P = 0.18) . ResultsIn this systematic review, we have examined the complications following open elbow arthrolysis for post-traumatic elbow stiffness. Twenty-eight studies involving 810 patients were identified for the final inclusion.Nerve complications, especially DOUN, were most common in the examined studies. The associated clinical symptoms are attributable to several factors. First, the procedure can cause iatrogenic injury of the nerve when the dissection is not meticulous or complete ,30. SecoHO is a major complication, and its specific mechanism is still unclear. Since many studies did not report the results of the postoperative radiographic evaluation, the radiographic new-onset or recurrence of HO might have been neglected by the authors, and the actual incidence was much higher than expected. HO of the elbow can develop after direct trauma , central nervous system injury, burn injury, and among those with genetic diseases. Furthermore, surgical dissection, which may lead to soft-tissue insult, is also related to the development of HO . ClinicaElbow instability is a common complication after complete arthrolysis for severe elbow stiffness. With a careful stress test for stability during the surgery as well as the development of the surgical techniques, instability could be recognized and avoided. The main causes of postoperative instability are the initial injuries and iatrogenic violation of the collateral ligaments, which play a vital role in maintaining elbow stability . GeneralIn order to achieve greater stability after extensive release, a hinged external fixator was applied during the surgery in 5 studies and was maintained in place for 4–8 weeks ,12,14,23The occurrence of infection as a complication cannot be underestimated in any joint surgery, including open elbow arthrolysis. It can spiral out of control, and subsequent disability is inevitable. As recorded above, 7 of 13 patients underwent a second surgery and achieving satisfactory outcomes was difficult. In the selected studies, the treatment for infection consisted of intravenous or oral antibiotics as well as irrigation and debridement. Regrettably, the risk factors and more detailed treatments were not described. One study from our institution showed that the severity of elbow stiffness with less active ROM, longer operative time, diabetes and increased previous operation times seemed to be potential risk factors for infection after open elbow arthrolysis . MoreoveWe reviewed all the studies ,28,31,34Based on the results of the multiple linear regression analysis, preoperative ROM  and proportion of female  were the independent factors affecting the total complication rate. Less preoperative ROM, associated with higher severity level of elbow stiffness, was one of the risk factors for many complications including DOUN, HO, elbow instability and infection discussed above. Therefore, it would significantly predict a higher total complication rate. In this regard, surgeons should pay more attention to the patients with less preoperative ROM, and consider the use of prophylactic treatment to reduce complication rate. Meanwhile, the results also showed a higher proportion of female was associated with a higher total complication rate. We assume the different anatomy of the elbow between male and female may primarily account for this finding. This is another point of interest for future research.Although most of these studies did not report the results of subsequent surgeries, to our knowledge, surgeries performed after the index surgery usually do not have satisfactory outcomes. Therefore, more emphasis should be placed on the persistent complications, especially those with a relatively high operation rate, including repeat elbow contracture (75.8%), infection (53.8%) and HO (28.9%). Complication-related reoperation will lead to decreased confidence among patients as well as the surgeons. More importantly, the persistent complications would increase the economic and psychological burden on the patients. This is why use of several prophylactic interventions for preventing complications, especially the persistent ones, is important. In our view, prophylactic interventions tend to exert an increasingly important effect on the safety and outcome of open elbow arthrolysis.This systematic review has some limitations that should be taken into account. First, since the studies identified were level 3 or level 4 on the level of evidence, the general strength of the evidence from these studies was not of the highest quality. Second, as in other meta-analyses or systematic reviews that study specific outcomes, there exists the inherent possibility of publication bias; a lower complication rate or better ROM, for example, may be more likely to be published. Third, though the studies were selected strictly on the basis of inclusion and exclusion criteria, there was a lack of homogeneity in the severity of the stiff elbow, surgical techniques, rehabilitation, and duration of follow-up among the studies and it was difficult to perform a more specific statistical analysis between patients and each of the complication. We must declare, however, that the characteristics of complications were arrived at based on the findings of 28 studies. Moreover, further discussion of the risk factors and prevention and treatment strategy in this study will provide a reference to reduce the complication rate and improve the outcomes of open elbow arthrolysis.In summary, the complications associated with open elbow arthrolysis mainly include nerve complications, HO, elbow instability, infection, pin-related complications and repeat elbow contracture. Various risk factors are related to each of the complication. According to the literature review, a diagnosis of HO, preoperative nerve symptoms and preoperative ROM are risk factors for nerve complications; a high BMI, longer injury time and preoperative ROM are the risk factors for HO; higher severity level of stiffness is a risk factor for elbow instability; severity of stiffness with less ROM, longer operative time, diabetes and increased previous operation times are risk factors for infection. Besides, the statistical analysis reveals that patients with less preoperative ROM and a higher proportion of female gender may point to a high total complication rate. Moreover, the persistent complications with a significant reoperation rate and an unsatisfactory outcome are difficult to handle. Therefore, to further improve the overall outcomes of this procedure, more and larger prospective studies should be performed to further elucidate the effects of prophylactic interventions and to determine which combination of risk factors can be effectively ameliorated by prophylactic interventions like ulnar nerve decompression for nerve symptoms, radiotherapy and NSAIDs for HO, application of suture anchor repair technique and external fixation for instability and the regional antibiotic prophylaxis for infection, thus improving the methods of prevention and treatment of complications, especially the persistent ones.S1 PRISMA Checklist(DOC)Click here for additional data file.


A male patient, aged 77 years, was admitted to hospital with the chief complaint of persistent hyperpyrexia that had presented for four days. The patient also suffered from hypoxemia, and a large white shadow in the left lung was observed on a chest radiograph, indicating inflammation. No therapeutic effect was observed with anti-infection treatment. The patient admitted a history of direct contact with live chickens two weeks prior to hospital admission. The day after admission to the Jingnan District Centre Hospital of Shanghai , the patient was diagnosed with severe H7N9 avian influenza infection by nasopharyngeal swab and blood sampling detection. Although the patient received anti-infective drugs, intubated assisted ventilation and circulation support, the condition of the patient continued to rapidly deteriorate. Oxygen saturation decreased and gastrointestinal bleeding occurred, with the body temperature fluctuating between 39 and 40°C. By day 6 after admission, the patient presented with circulatory failure, with liver and renal failure. On day 7, the blood pressure of the patient was unable to be measured, and the patient was diagnosed with multiple organ dysfunction. Subsequently, clinical death was declared with the patient exhibiting asystole and no spontaneous breathing. Human infections with H7N9 usually occur following recent exposure to poultry, which causes upper respiratory tract disease to progress into pneumonia and subsequently multiple organ failure . A diagnIn the present study, the patient exhibited a rapid deterioration; however, a diagnosis of H7N9 avian influenza was only confirmed after five days of continuous fever. Thus, the treatment process has provided experience for dealing with cases of H7N9, particularly for community health centers.rd 2013, the patient experienced chills and fever, without evident cause or regularity, with a maximum temperature of 39.4°C. The patient did not suffer from a cough, expectoration, sore throat, runny nose, chest tightness, chest pain, pant or whole muscle and joint ache. A chest radiograph, obtained in the initial hospital, revealed a fuzzy shadow in the left lower lung  and a C-reactive protein level of 192 mg/l. A chest computed tomography scan  revealed a ‘frosted glass’ appearance in both lungs, and a high density shadow was observed in the left lower lobe  after presenting with a fever for four days. On April 3wer lung . After iwer lobe . In addiThe patient had >30 years history of paroxysmal atrial fibrillation, and had been treated with amiodarone . In addition, the patient had >20 years history of hypertension, up to a maximum of 150/90 mmHg; thus, amlodipine (5 mg every day) treatment had been used to control the blood pressure, which was determined to be of proper control. The patient denied history of diabetes, coronary heart disease, chronic bronchitis, asthma and chronic kidney disease, viral hepatitis, tuberculosis, typhoid fever and infectious disease.Physical examination revealed a body temperature of 39.1°C, a pulse of 98 bpm, a respiratory rate (RR) of 22 and a blood pressure of 130/70 mmHg. The patient was clear in mentality, had regular respiration, was cooperative on examination and their walking was not affected. The skin and mucosa exhibited no rash or hemorrhagic spots, and the lip mucosa had no cyanosis. Pharyngeal congestion was mild and the double tonsils were not enlarged. Breath sounds of the bilateral lung were coarse, and scattered rales were heard in the left lung, but without wheezing rale and pleural friction sounds. The heart rate of the patient was 98 bpm. Cardiac rhythm was regular, and there were no marked pathological murmurs heard in each valve area. The abdomen was soft, with no muscle tension, tenderness or rebound tenderness. No organomegaly or masses were observed, and the liver and spleen were not palpated under the ribs. There was no edema observed in the lower extremities, and the four limbs exhibited normal muscle force and muscle tension.2 of 38.6 mmHg, a PaO2 of 52.6 mmHg and a SaO2 of 87% (without oxygen). An electrocardiogram (ECG) demonstrated sinus rhythm, atrial premature beats, poor R wave progression in the anterior wall and T wave changes.Hospital emergency blood gas analysis revealed a pH of 7.49, a PaCOth, a diagnosis of severe human infection with H7N9 avian influenza was confirmed  and azithromycin  were applied as anti-infective agents. However, the patient continued to suffer from a fever with a body temperature of up to 39.1°C, experiencing distress on day 2 following admission. With repeated inquiries into the medical history of the patient, it was found that the patient had come into contact with chickens two weeks previously. Combining the epidemiological history and the rapid progression in the pulmonary lesions of the patient , a diagnRNA was extracted from the throat-swab samples using the QIAamp Viral RNA Mini kit , according to the manufacturer’s instructions. Specific RT-qPCR assays were performed to assess the presence of seasonal influenza viruses , H5N1, severe acute respiratory syndrome coronavirus and novel coronavirus. RT-qPCR assays with self-designed specific primer and probe sets were subsequently performed for the detection of H1 to H16 and N1 to N9 subtypes, in order to verify the viral subtypes.th, the patient presented with cyanosis of the lips and limbs. In addition, the fingertip pulse oximeter monitor displayed a SPO2 of 45%, and the patient underwent an emergency tracheal intubation for assisted ventilation , while improving microcirculation and anti-leakage. On April 10th, the patient exhibited a damaged liver performance, and was administered nutritional support treatment , intravenous immunoglobulin to strengthen the immunity, drugs for liver protection , daily fluid therapy , and psychological treatment. The chest radiograph showed progression of the lesions in the right lung  . Severe In addition, attention should be paid to the control of complications, nutritional support, reconstruction and stabilization of the internal environment and immune homeostasis. Early administration of glucocorticoids can inhibit the inflammatory reaction, reduce the release of cytokines and inflammatory mediators, and reduce alveolar exudation. Furthermore, timely trachea intubation and application of respiratory support technology can correct hypoxia, protect major organ functions and prevent multiple organ dysfunction . For patIn conclusion, severe human infection of H7N9 avian influenza often involves multiple systems, with rapid progression and poor prognosis, emphasizing the requirement for multidisciplinary, comprehensive management. Airway management, prevention of infection, mechanical ventilation, water balance support, electrolyte and acid-base balance, nutritional support and the maintenance of important organ function are key to a successful outcome for critically ill patients .


The sequence of cattle genome provided a valuable opportunity to systematically link genetic and metabolic traits of cattle. The objectives of this study were 1) to reconstruct genome-scale cattle-specific metabolic pathways based on the most recent and updated cattle genome build and 2) to identify duplicated metabolic genes in the cattle genome for better understanding of metabolic adaptations in cattle. A bioinformatic pipeline of an organism for amalgamating genomic annotations from multiple sources was updated. Using this, an amalgamated cattle genome database based on UMD_3.1, was created. The amalgamated cattle genome database is composed of a total of 33,292 genes: 19,123 consensus genes between NCBI and Ensembl databases, 8,410 and 5,493 genes only found in NCBI or Ensembl, respectively, and 266 genes from NCBI scaffolds. A metabolic reconstruction of the cattle genome and cattle pathway genome database (PGDB) was also developed using Pathway Tools, followed by an intensive manual curation. The manual curation filled or revised 68 pathway holes, deleted 36 metabolic pathways, and added 23 metabolic pathways. Consequently, the curated cattle PGDB contains 304 metabolic pathways, 2,460 reactions including 2,371 enzymatic reactions, and 4,012 enzymes. Furthermore, this study identified eight duplicated genes in 12 metabolic pathways in the cattle genome compared to human and mouse. Some of these duplicated genes are related with specific hormone biosynthesis and detoxifications. The updated genome-scale metabolic reconstruction is a useful tool for understanding biology and metabolic characteristics in cattle. There has been significant improvements in the quality of cattle genome annotations and the MetaCyc database. The duplicated metabolic genes in the cattle genome compared to human and mouse implies evolutionary changes in the cattle genome and provides a useful information for further research on understanding metabolic adaptations of cattle. B. taurus and B. indicus) produce meat and milk, which contribute 15% of the total protein consumed by humans in the world [One of the most important purposes of livestock is to supply high-quality food  for humans . Ruminanhe world . Therefohe world . In thishttp://168.188.16.73:8080/CATTLE) is the first pathway genome database (PGDB) developed in livestock animals, generated using Pathway Tools and the BioCyc platform [One of the ways to link genetic and metabolic traits of an organism is to perform a genome-scale model via metabolic reconstruction . To recoplatform . An updaMoreover, although metabolic gene deletions in the cattle genome compared with the mouse and human genomes were identified previously , metabolTherefore, the objective of this study was to reconstruct genome-scale cattle-specific metabolic pathways based on the updated reference cattle genome build UMD_3.1 and compare the newly reconstructed cattle metabolic pathways with those in CattleCyc (based on Btau_3.1). This study also identified the cattle-specific gene duplications of metabolic genes compared to the human and mouse genomes.The general scheme of the metabolism-centered approach used for reconstruction of metabolic pathways and identification of duplicated metabolic genes in the cattle genome is represented in A revised amalgamation pipeline was developed on the basis of the previous one as described by Seo and Lewin . BrieflyGenome-scale reconstruction of metabolic pathways from the amalgamated cattle genome database was performed using the PathoLogic algorithm in the Pathway Tools software version 19.0 . PathwayThe initial automated reconstruction identified 316 metabolic pathways in the amalgamated cattle genome based on UMD_3.1. The initial cattle-specific PGDB intrinsically contained some errors such as pathway holes in which the organism-specific enzyme has not yet been identified and false-positive or false-negative pathways. Therefore, an intensive manual curation was performed to revise improper enzymatic reactions and metabolic pathways using various approaches including literature reviews, database searches, and comparison of metabolic pathways with other organisms.A metabolic pathway was deleted in the new cattle-specific PGDB if 1) redundant pathways existed , 2) either the input or the output of the pathway was not present in any mammal (e.g. peptidoglycan biosynthesis III), 3) neither enzyme activity was reported nor homologs were identified in any mammal, and an alternative pathway exists with strong biochemical evidence (e.g. lysine degradation V). When representation of a mammalian pathway in MetaCyc was not adequate, the pathway was modified based on information from the literature, KEGG, and Brenda . The proThe metabolic pathways in the new cattle PGDB was also compared with those in the CattleCyc (version 1.2.1) in the BioCyc databases . With thhttp://168.188.16.73:8080/CATTLE and http://168.188.16.73:8080/TAURUS, respectively. Both PGDB are also deposited into the Figshare data repository (https://dx.doi.org/10.6084/m9.figshare.3082039).The CattleCyc (version 1.2.1) and the new cattle PGDB are freely accessible via http://www.biomart.org) was used. First we obtained a list of human protein coding genes having one-to-one mouse and one-to-many cattle orthologs and a list of mouse protein coding genes having one-to-one human and one-to-many cattle orthologs from the Ensembl release 72. By comparing these list with the metabolic genes in the new cattle-specific PGDB, putative duplicated metabolic genes in the cattle genome were identified.To identify cattle-specific duplicated genes, an orthology table for human, mouse and cattle generated by BioMart  and literature. The putative duplicated genes were ranked by the following criteria: A) supported by the information of nucleotide chain, expressed sequence tag (EST), evidence for expression , and completeness of the sequencing and assembly, B) supported by the evidence of nucleotide chain, EST, and mRNA or protein evidence, but with a possibility of assembly error (i.e. scaffold gaps), C) supported by the evidence of nucleotide chain, but lacking information of either EST or mRNA (or protein) evidence and with a possibility of assembly error, D) supported by the evidence of nucleotide chain, but lacking both EST and mRNA (or protein) evidence and with a possibility of assembly error. The identified cattle-specific duplicated genes were also analyzed for their biological and metabolic functions using the amalgamated functional annotations in the new cattle PGDB.Total numbers of 27,786 and 24,616 genes were contained in the NCBI and Ensembl cattle genome database based on UMD_3.1. Through the matching process, 2,467, 5,742, 10,009, 11,874, 16,317, 16,423 and 1,255 consensus genes between the NCBI and Ensembl cattle genomes were non-exclusively identified based on the criteria of gene coordinate, gene name (or synonym), function name, unigene ID, cross-referenced gene ID, protein accession, or EC numbers, respectively .The sequential one-to-one matching between the NCBI and Ensembl cattle genome annotations, a total of 19,123 consensus gene models were found. Among these, 854 genes were inconsistently annotated for gene type, coding strand, or both between the NCBI and Ensembl genome databases: 821, 21 and 12 genes were differently annotated in gene type, coding strand, or both, respectively. In the consensus gene set 5,295, 5,963, 4,693, 1,398, 1,511, 262 and one genes were sequentially matched to have common gene coordinates, gene name or synonyms, function names, unigene ID, cross-referenced gene ID, protein accessions and EC numbers, respectively . CompareAs a result, the amalgamated cattle genome database, based on the reference cattle genome build UMD_3.1, contained a total of 33,292 genes consisting of 19,123 consensus genes, 8,410 and 5,493 genes only found in NCBI or Ensembl, respectively, and additional 266 genes from NCBI scaffolds genes .The initial cattle PGDB automatically constructed 316 metabolic pathways using the PathoLogic algorithm. The initial automated reconstruction identified 2,460 reactions, including 2,371 enzymatic, 73 spontaneous and 16 transport reactions, and 1,489 compounds . An enzyIn addition, the metabolic pathways in the new cattle PGDB were compared with those in the CattleCyc, a previously developed cattle PGDB based on the genome build Btau_3.1. The CattleCyc is publicly available and can be downloaded from the BioCyc registry. There were 169 and 104 pathways, contained only in the new cattle PGDB or in the CattleCyc, respectively . Among tThe cattle-specific duplicated genes were identified using an orthology table for human, mouse and cattle. The total protein coding genes in human, mouse and cattle genome were 22,691, 22,709 and 19,994, respectively. A total of 315 out of 22,691 human protein coding genes in the Ensembl release 72 had one-to-one mouse and one-to-many cattle orthologs. Among these, 55 genes were the metabolic genes in the cattle metabolic reconstruction, involved in 30 metabolic pathways. Those genes underwent comprehensive analysis of sequence similarity at the nucleotide and protein levels and synteny analysis, followed by intensive manual examination on the basis of our criteria. The total number of apparently duplicated metabolic genes in the cattle genome was eight being involved in 12 core metabolic pathways .HEXB and GSTO1, have been annotated with EST and protein accession so they were assigned in ‘B’ category. Five genes  had at least one of either EST or protein evidence were classified as ‘C’. The BPGM gene was ranked ‘D’ according to our criteria due to lack of expression evidence. Nevertheless, BPGM showed an interesting feature based on synteny analysis among human, mouse and cattle . The reaction of EC 3.1.1.- in this pathway was also replaced by EC 3.1.1.3. In addition, 11 redundant pathways, and 25 bacteria or plant-specific pathways were deleted from the initial reconstruction.The initial reconstruction in this study, however, contained less false-positive and -negative pathways than the one when developing CattleCyc, mainly because the well-curated CattleCyc as well as MetaCyc was also used as a base for the new metabolic reconstruction. Still, the unknown enzyme, enzymatic reactions and enzyme encoded protein in metabolic pathway, called pathway holes, needed to be filled with correct annotations for increasing the quality of the cattle PGDB. For example, in the triacylglycerol degradation pathway, the pathway hole for the reaction of EC 3.1.1.34 was modified to EC 3.1.1.79 and filled with CRYL1 was missing in the previous cattle genome build; however, it was annotated in the current build and thus included in the new cattle PGDB. The primary reason that the 83 metabolic pathways exist in the CattleCyc were not initially included in the new cattle PGDB was the updates of MetaCyc. For example, the ‘pyruvate oxidation’ pathway was included in the CattleCyc. This pathway oxidatively catalyzes pyruvate to acetate and CO2 by pyruvate:quinone oxidoreductase (EC 1.2.2.2) [Comparative metabolic analysis indicated that the new cattle PGDB was better than CattleCyc. This became possible primarily due to the improvement in the cattle genome build and in the MetaCyc database. For example, the ‘glucuronate degradation’ pathway did not exist in the CattleCyc because L-gulonate 3-dehydrogenase (EC 1.1.1.45) encoded by 1.2.2.2) . Upon th1.2.2.2) , and theThese results indicated that a comparative analysis of metabolic pathway is helpful in identifying and evaluating present gaps in our knowledge. A well-curated PGDB like the new cattle PGDB developed in this study will facilitate computational reconstruction of metabolic pathways for other mammalian genomes with greater reliability. In addition, it is suggested that an integrated database like the cattle PGDB needs to be recurrently revised as genome annotations and biological databases are updated, and it is important to follow up the current knowledge when conducting a functional genomics study. Like the previous version , the new cattle PGDB does not include Y-chromosome specific genes and metabolic pathways because the genome assembly UMD_3.1 does not include Y chromosome. Further work will be required to include the male-specific metabolic pathways by integrating genome annotations with bovine Y chromosome. Also, integrating the information of metabolites to metabolic networks may be a necessary step to improve the quality of metabolic networks and their application. This would be done in a future research on constructing an in silico simulation model of bovine metabolism based on the new CattleCyc developed in this study.Gene duplication is one of the most important ways to create new genes during a course of evolution. Gene gain can occur either on a large scale from a whole genome duplication or on small scale when chromosomal sections or individual genes are copied. Gene gain is frequently followed by differential gene loss either by mutation in one gene isoform leading to the creation of a pseudogene or by complete deletion. Gene deletion is believed to closely follow duplication in many cases . In the This study identified a total of eight duplicated genes in 12 metabolic pathways of cattle compared to human and mouse. The duplicated genes can be classified according to their possible mechanism, such as segmental duplication and chromosomal rearrangement. An example for each mechanism is given here.AANAT gene  was due to a segmental duplication in cattle genome. The biological role of vertebrate AANAT is to acetylate serotonin in the synthesis of melatonin. It was reported that melatonin promotes sleep, inhibits reproduction, and promotes weight gain [AANAT is associated with biological timing: daily changes in the activity of this enzyme regulate the daily rhythm in melatonin synthesis, which is essential for optimal temporal coordination of biological functions with day and night and seasonal changes [AANAT is known to be consistently expressed at significant levels only in two tissues, the pineal gland and retina; both of which are photosensitive organs [AANAT was associated with carcass traits [Segmental duplication is one of the possible outcomes of ‘unequal crossing over’, which results from homologous recombination between paralogous sequences . Duplicaght gain . Vertebr changes . Vertebre organs –35. In cs traits , 37.SOD1 gene shows an example of outcomes from chromosomal rearrangement. Chromosomal rearrangement by ectopic pairing and recombination between interspersed repeat sequences presents a mechanism for dramatic reorganization of eukaryotic genomes. The SOD1 gene is located on chr1: 3,113,948–3,122,613 and chr13: 51,930,067–51,930,888 in the cattle genome. The major function of SOD1 is destruction of free superoxide radicals in the body. The superoxide radical (O2-) is generated in biological systems as by-product of the partial reduction of dioxygen during respiration. Cytoplasmic Cu, Zn superoxide dismutase catalyzes the dismutation of two superoxide anions into dioxygen and hydrogen peroxide and thus plays a central role in the cellular defense against oxidative stress [SOD1, is a critical component of the cellular defenses against reactive oxygen species and catalyzes the dismutation reaction of the superoxide radical anion to hydrogen peroxide and oxygen via the cycle reduction and re-oxidation of copper [SOD1 gene had a positive correlation with meat tenderness in semitendinosus muscle.On the other hand, the duplication of the e stress . Cytosolf copper . In a stf copper  reportedBPGM gene can be classified as either chromosomal arrangement or ‘retrotransposition’. Retrotransposition is the integration of reverse transcribed mature RNAs at random sites in a genome. The duplicated gene due to retrotransposition is called as a retrogene, which lacks introns and has poly-A tails [BPGM on chromosome 10 also lacks introns and has a cleavage and polyadenylation specificity factor (CPSF) site and poly-A. The Bisphosphoglycerate mutase, encoded by BPGM, is an erythrocyte-specific trifunctional enzyme. The main activity is a synthase (EC 5.4.2.4), catalyzing the formation of 2, 3-bisphosphoglycerate  from 1, 3-bisphosphoglycerate . The second activity is a mutase  catalyzing the interconversion between 2- and 3-phosphoglycerate. The third activity, as a phosphatase , is to catalyze the hydrolysis of 2,3-BPG to 3- or 2-phosphoglycerate and a phosphate [BPGM has been studied mostly in erythrocytes and placental cells [BPGM was related with growth rate based on QTL association study [Duplication of the -A tails . The cathosphate . Since thosphate , 44, BPGal cells , 45, 46 on study .AANAT was found only in the cattle genome Click here for additional data file.S2 FigThe phylogenetic tree was generated for putative duplicated genes using MEGA 6.0 . The pro(EPS)Click here for additional data file.S3 FigThe phylogenetic tree was generated for putative duplicated genes using MEGA 6.0 . The pro(EPS)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.