BackgroundThe proximal femoral nail anti-rotation Asia (PFNA 2) is an implant designed for unstable osteoporotic intertrochanteric fractures in Asians as the PFNA was designed for Caucasians and had various complications when applied to the Asian population due to the femoral geometrical mismatch. This study observes the functional outcomes and complications associated with PFNA 2 in unstable intertrochanteric fractures in the elderly Indian population.MethodsSixty-one above 60 years old patients with an unstable intertrochanteric fracture who were operated with PFNA 2 were included in this prospective observational study. They were followed up for one year. The functional and radiographic evaluations were done at 6, 12, 20 weeks, and the functional outcome was evaluated at the end of one year. Association of age, American Society of Anaesthesiologists (ASA) grade, AO Foundation classification, osteoporosis to the functional outcome of modified Harris hip score (MHHS) was evaluated.ResultsType A2 fractures demonstrated a statistically higher-good reduction than Type A3 . The difference in mean surgical duration in Type A3 (45.47 minutes) and Type A2 (40.30 minutes) was statistically significant . Mean blood loss was 110.66 ml (SD = 48.40 ml). MHHS at 6, 12, 20 weeks, and one year were 40.37, 63.93, 79.03, and 82.34, respectively. At the end of the year, 46 (82.1%) patients achieved good scores, eight (14.3%) achieved fair scores, and two (3.5%) achieved poor scores. There was one case of nonunion and medial migration of the helical blade. The mortality rate was 6.55% at the end of one year.ConclusionA good reduction was associated with a better functional outcome. PFNA 2 is an efficient implant in managing unstable intertrochanteric fractures in elderly Indian patients with good outcomes, low morbidity rates, and mortality. Implant mismatch was not a problem in the Indian population. However, large multi-centric studies with a larger sample size are required. Moreover, achieving a good reduction cannot be over-emphasized in unstable intertrochanteric fractures, especially in the elderly, to achieve a good functional outcome. Intertrochanteric hip fracture is a major public health problem in the geriatric age group. Definitive management is desired, considering age and associated co-morbidities. Indian population is at higher risk of osteoporosis and associated hip fractures . A studyThe proximal femoral nail anti-rotation (PFNA), a modification of proximal femoral nail (PFN), was introduced in 2003, which features a helical blade. Biomechanical cadaveric studies demonstrated that PFNA fixation using a helical blade was better compared to the sliding hip screw. PFNA, characterized by rotational along with angular stability has biomechanically improved purchase in the osteoporotic bone due to the bony impaction it achieves in the femoral head and neck ,11. PFNAThis study was undertaken to analyze the results of unstable intertrochanteric fracture of femur fixed with PFNA 2, its functional and radiological outcomes in the elderly Indian population. No previous studies have established the role of various factors affecting the outcome of unstable intertrochanteric fractures fixed with PFNA2 in the elderly Indian population with a prospective study.The study was a prospective two-year study after approval from the scientific and ethical committee of the institution. Written informed consent was taken from all patients. Patients > 60 years with unilateral unstable inter-trochanteric fracture femur , ambulatory prior to fracture, mentally sound and asymptomatic contra-lateral lower limb were included. Fractures with subtrochanteric extension, inflammatory arthritis, revision surgery, >2 weeks old fracture, active infection were excludedPatient's demographic data, American Society of Anaesthesiologists (ASA) grading , mode ofStatistical analysis was done using SPSS 21 . Descriptive statistics were computed. Categorical variables were expressed as proportions and the annotations were assessed using Pearson's chi-square test for large frequencies and Fischer's exact test for small frequencies. T-tests were used for the significance of difference and the level of significance was assessed with P value (significant when P < 0.05). The MHHS was analyzed to find its association with age, AO fracture classification, reduction quality, Singh's osteoporotic index, and ASA grade as fixed variables and age as a covariate using the Analysis of Covariance (ANCOVA) test.The total number of patients in our study were 61 which included 41 females (67.2 %) and 20 males (32.7 %). The mean age was 75.39 years (SD = 8.4years). 46 (75.4 %) fractures were of type 31A2 and 15 were of type 31A3 (AO Classification). 93% of patients were grade ≥3 as per Singh's index for osteoporosis, four patients had severe osteoporosis, i.e., grade <3. The mean duration from admission to surgery was 1.59 days (1-8 days). The ASA grading was grade 3 in 57.37% of patients. In 68.9% patients good reduction was achieved . Type A2 fractures had a statistically higher number of good reductions (P < 0.05). The mean duration of surgery was 41.57 minutes (SD = 6.24 minutes). The difference between the mean surgical duration of Type A3 (45.47 minutes) and Type A2 fractures (40.30 minutes) was statistically significant (P < 0.05). Mean TAD was 21.72 mm. Mean TAD for type A2 fracture and type A3 fractures was 21.53 mm and 22.32 mm, respectively. No significant difference was observed between type 31A2 and 31A3 fractures. The type of fracture did not influence the TAD (P > 0.05). 86.8% of the patients had a TAD < 25 mm. Forty-nine out of 61 patients (80.3%) had the Helical blade placed in the center-center zone in the head and neck of the femur. Ten patients had a central-posterior position and one each had an inferior-central and central-anterior position. The mean blood loss was 110.66 ml (SD = 48.40 ml). No blood transfusion was necessary. No early complications related to the implant and no infections were observed. Contralateral femur NSA and post-operated hip NSA were measured, and the mean values were 134.41° (SD = 1.53) and 134.54° (SD = 1.66), respectively. We did not come across implant mismatch in our group of patients. See Table The MHHS at 6, 12, 20 weeks, and 1 year were 40.37, 63.93, 79.03, and 82.34, respectively. At the end of one year, 46 (82.1%) patients had a good score, 8 (14.3%) had a fair score and only 2 (3.5%) had poor scores  patients had a good MHH score. This is comparable to the study by Chiaoling et. al. with good to excellent results in 78.2% patients and by Ming et. al. who found good to excellent outcome in 81.9% patients ,26. A siFracture of the femoral shaft or implant breakage was not observed which is attributed to the improved design of the distal end of the PFNA 2 with a flexible tip ,17. No pThe limitation of our study is the absence of a control group to compare the functional outcome of unstable fractures fixed with other implants. A smaller sample size and no subjective method of measuring osteoporosis may be other drawbacks. We recommend such prospective studies with a larger sample size to gauge the functional outcome and complications associated with this implant using a better method to measure grade of osteoporosis. The message must go out to the young orthopedic surgeons that achieving a good reduction cannot be overemphasized even with a very advanced implant design.An intra-medullary device requiring minimally invasive procedure is the implant of choice in an unstable inter-trochanteric fracture in an osteoporotic elderly patient. It should allow a shorter duration of surgery with minimal blood loss. PFNA 2 fulfils these requirements. We recommend a good fracture reduction with the optimum position of the helical blade  to achieve union and prevent cut out. TAD of < 25 mm is another important factor to prevent the cut-out of the helical blade. Osteoporosis, ASA grade, and type of fracture were not found to be associated with the functional outcome. A prospective study with functional outcomes of PFNA2 in unstable intertrochanteric fractures is lacking and this study should encourage orthopedic surgeons from India to further report larger groups in prospective studies which may help us further understand the implications and outcome of this implant in unstable fractures in the Indian population in long term.


The erratum notes a correction to Fig. 5 for the published article. J. Biomed. Opt.25(3), 035002 (2019) doi: 10.1117/1.JBO.25.3.035002] was originally published online on 6 March 2020 with an error in Figure 5, corresponding to percentage errors for the optical properties.This article [The article included a version of the figure that listed the wrong values. The text of Section 3.1 correctly states, “Average errors were 9.4 and 6.7 for Original figure:Corrected figure:The caption for the figure is unchanged. The results reported were not affected by the error.This article was corrected online on 27 March 2020.


Proteus mirabilis isolated from chronic wound infections. According to the findings, the most frequent factors of infection are Staphylococcus aureus (46.1%), Pseudomonas aeruginosa (35.0%), and Proteus mirabilis (10.6%). Minimal inhibitory concentration and minimal bactericidal concentration values were assigned using the microbroth dilution test according to the Clinical and Laboratory Standards Institute. Biofilm of Proteus mirabilis isolates was formed in 96-well polystyrene plates and treated with Manuka honey (concentrations from 1.88% to 30.0%) and ethanol extract of propolis (1.0% to 40.0%). After 24 h, the biofilm viability was expressed by formazan absorbance (λ = 470 nm). Manuka honey reduced the biofilm viability in all, and ethanol extract of propolis in most, of the concentrations tested. Ethanol extract of propolis at the concentrations of 20.0% and 40.0%, reduced biofilm viability stronger than ethanol itself. With these results comes the conclusion that these substances can reduce biofilm formation.Chronic wound infections are difficult to manage because of the biofilm formation in the wound environment. New measures for eliminating infections are necessary to increase the chance of wound healing. Apitherapy may be the new solution. The aim of this study was to assess the prevalence of wound infection factors and to examine the impact of Manuka honey and ethanol extract of propolis on biofilm formation of  The definition of a chronic wound is not clearly outlined in the literature or clinical practice. Chronic wounds do not heal in a standard time because the healing process does not progress. According to literature reports, a wound is presumed chronic after two to eight weeks of a prolonged healing process ,2,3,4.In this paper, we focused on chronic wounds related to diabetes (diabetic foot ulcers) and pressure lesions. During their lifetime, 12.0 to 25.0% of diabetic patients will develop ulcerations. An infection will occur in up to 10.0% of cases .6 colony-forming units (CFU) per gram of tissue sample) in the microbiological examination is not enough without manifested symptoms [The occurrence of planktonic bacteria or even biofilm in the wound is not sufficient to diagnose the infection. Exhibiting a cut-off value  chronic wounds, constricting and prolonging the wound healing process. It can be infection causative, although its presence is not equivalent to infection. Biofilm prolongs granulation and raises bacteria resistance to antiseptics and antimicrobials . Topical antibiotics are not recommended in infected chronic wound therapy. Systemic use of antibiotics is justified only for antibiotics with high bioavailability. Some antiseptics are toxic for bacteria and also for eukaryote cells, disturbing the process of wound healing . This isLeptospermum scopiarum flowers in New Zealand. The antibacterial activity of MH depends on several factors. Some factors, such as carbohydrates content, high osmolarity, and low pH level (about 3.9) are common for every type of honey, but others, such as methylglyoxal (MGO) or leptosine, are specific only for MH [Manuka honey is produced by bees from y for MH ,15,16,17y for MH ,20,21,22S. aureus growth inhibition zone in agar plates [Unique Manuka factor (UMF) represents the antibacterial activity of honey in phenol concentration causing the same r plates . UMF is r plates ,24. Propolis or bee glue is a bee-derived product made of raisin and beeswax. It is used to seal the beehive and protect it from bacteria and fungi. Propolis consists of 50% raisin (triterpenes), 30% wax, 10% essential oils, 5% pollen, and other organic substances. Flavonoids such as pinocembrin, chrisin, and pinobanksin are the main factors known for their antioxidant, antivirus, and antibacterial activity ,26. Pino® is a medical-grade MH used for acute, chronic, or surgical wounds and superficial burns for autolytic debridement promotion or malodor reduction. Although Medihoney® is promoted as a dressing for infected wounds, the producer does not claim that it has proven antimicrobial activity.There are a few products on the market available, for example, for wound dressings that contain bee-derived products. MedihoneyTo examine the epidemiology of infected chronic wounds at University Hospital No. 1 in Bydgoszcz, Poland, from 2016 to 2018, we evolved epidemiology data. All positive results of chronic wound examinations (contamination excluded), i.e., a total of 1142, were included in the examination. We did not include anaerobes in the present paper.n = 31) were obtained from the clinical isolates collection of the Clinical Microbiology Department of Dr. Antoni Jurasz University Hospital No. 1 in Bydgoszcz. The strains were chosen randomly from chronic wound infection isolates collected through the years 2016–2018. Strains were stored in brain-heart infusion broth  with 20.0% glycerol , at −70 °C. Isolates (®): Escherichia coli ATCC® 25922™ (ECO 25922™), E. coli ATCC® 35218™ (ECO 35218™), Enterococcus faecium ATCC® 29212™ (EFA 29212™), S. aureus ATCC® 25923™ (SAU 25923™), S. aureus ATCC® 29213™ (SAU 29213™), and P. aeruginosa ATCC® 27853™ (PAE 27853™). Strain selection was based on the PN-EN 1040:2006E standard quantitative dilute method for bactericidal activity of disinfectants and antiseptics . The folManuka honey was obtained from Manuka Health, New Zealand and stored at room temperature in the dark. The producer ensures that the honey MGO index is 400+ (at least 400 mg of MGO in 1 kg of honey). Honey sterility assay was conducted using a Tryptic Soy Agar plate  with the addition of MH preparation and incubation at 37 °C for 5 days and checking for any trace of microbe growth. The MH concentrations for the investigation were prepared by a series of dilutions in Tryptic Soy Broth  to the final concentrations of 1.87%, 3.75%, 7.50%, 15.0%, 20.0%, and 30.0%.−1 gallic acid equivalents (GAE) and 701.4 mg·kg−1 GAE.The polyphenol content of MH was examined using the Folin–Ciocalteu colorimetric method modified by Keutgen and Pawelzik  (λ = 736−1.The antioxidant activity of MH was tested with the ferric reducing antioxidant power (FRAP) assay. The test was repeated twice. The obtained values were 0.33 and 0.32 mmol·kg−1), 2.50% (3.7 mg·mL−1), 5.0% (7.4 mg·mL−1), 10.0% (14.7 mg·mL−1), 20.0% (29.4 mg·mL−1), and 40.0% (58.8 mg·mL−1).New Zealand propolis, in the dose of 80.0 g, was mixed with 100 mL of ethanol and incubated at room temperature for 14 days. Next, the solution was stored at −20 °C for 30 days for the insoluble compound sedimentation. Only supernatant, ethanol extract of propolis (EEP), was used in the research. The extract was filtered through a sterile syringe . The mean weight of active substances in 1 mL of EEP was 0.147 g. A series of concentrations of EEP were prepared by the dilution of EEP in TSB to final concentrations of 1.0%  values were impossible to obtain. For this reason, the assessment of minimal bactericidal concentration (MBC) was chosen.The assessment of MBC for planktonic cells was based on the Clinical and Laboratory Standard Institute (CLSI) recommendation standard. MBC and biofilm bactericidal concentration (BBC) for a 24-h mature biofilm (0.5 McFarland standard inoculum undiluted and diluted 1:100) were assigned for reference strains . The wel−1 inoculum, were assigned for the P. mirabilis clinical strains. For each strain, the suspension control was performed by seeding aliquots of the positive control well on CLED agar and CFU·mL−1 counting after 24-h incubation.Minimal biofilm inhibitory concentration (MBIC), biofilm prevention concentration (BPC), and minimal biofilm eradication concentration (MBEC) , for appMBIC and BPC were assigned by simultaneous bacterial inoculation and substances exposure, and MBEC was tested on the 24-h mature biofilm. After the final incubation, MBIC was read by the absence of opacity, while BPC and MBEC were read as the lowest concentration preventing growth after seeding well content on the CLED agar plate.v/v) proportion. Inocula in TSB (0.5 McFarland optic density) were added directly to the wells of polystyrene plate or were primarily diluted in sterile TSB (1:100). Negative control wells proved that the suspension media were sterile. Plates were covered with lids and incubated in a humid atmosphere at 37 °C. After 24 h, the suspension medium with planktonic cells was removed and wells were washed thrice with PBS  (pH 7.2).Bacteria inocula  was simultaneously tested. For each strain, the impact of all concentrations was determined in triplicate. To examine the impact of MH and EEP on the biofilm, serial dilutions of substances in liquid medium were added to wells containing the washed biofilm. Plates were covered with lids and incubated in a humid chamber at 37 °C, for 24 h. To compare the impact of EEP on the mature biofilm formation with ethanol, ethanol solution in PBS (1.25:1 w/v) 2,3,5-triphenyl-2H-tetrazolium chloride  in TSB (1:9 v/v), and incubated in humid conditions at 37 °C. After 4 h, the supernatant was removed, wells were rinsed with water, and filled with methanol (Avantor). The absorbance of formazan was read at 470 nm .To assess the metabolic activity of biofilm cells, polystyrene plates (containing biofilm treated with the solutions tested) were rinsed with PBS, then filled with 1%  means positive control absorbance;A (x) means absorbance after treatment with honey or EEP in defined concentration.The grade of metabolic activity reduction of strains was calculated in Excel 2007 based on the equation:p < 0.05.Statistical analysis was performed using Statistica 13.1. The assessment of independent variables was conducted using Wilcoxon’s test. The null hypothesis was stated as, “Mean grades of metabolic activity after MH, ethanol extract of propolis, and ethanol treatment in all examined concentrations are not statistically different.” The statistical significance (P) was determined at level 0.05. Independent variables were considered to be statistically significant at P. mirabilis is third among the most frequently isolated species (10.6%). Although it is not as an emergent pathogen as multidrug-resistant Klebsiella pneumoniae or Acinetobacter baumannii, its ability to form biofilm in wound environment allows P. mirabilis to survive and be infection causative. The most frequent factors isolated from chronic wounds are S. aureus (isolated from 46.1% of cases) and P. aeruginosa (35.0%). S. aureus (51.2%), the second most frequent is P. aeruginosa (28.4%), and the third most frequent is P. mirabilis (4.2%). Among all cases of chronic wound infections, there are 285 (25.0%) monocultures and 857 (75.0%) mixed cultures. The most frequent factor of infection in monocultures is S. aureus coexisting with P. aeruginosa (12.5%), and P. aeruginosa coexisting with P. mirabilis (5.9%). Moderately frequent tandems are S. aureus coexisting with Enterobacter cloacae, P. mirabilis, Streptococcus agalactiae, or Streptococcus dysgalactiae. These pairs occur in 3 to 5% of cases. Infection is the result of mixed cultures in 857 (75.0%) cases. The most frequent tandems of microorganisms are Enterococcus spp. . Coagulase-negative Staphylococcus spp. and Corynebacterium spp. strains found in insignificant quantities are treated as skin microflora contamination and occurred in 14.7% of all cases.Generally, more than two microorganisms are isolated in 498 cases, which is 58.1% of all chronic wound infections. The bacteria most frequently isolated as accompanying microbiota are A comparison of MBC and BBC values for six reference strains after MH and EEP treatments is presented in v:v) dilution, the BBC values of MH are lower than EEP in five out of six strains. As for EEP, the MBC values are always lower than BBC values. There is no regularity for MH.Lower values of MBC for EEP as compared with MH indicated that EEP suppressed the growth of planktonic cells more effectively. For 24-h mature biofilm cultured from 0.5 McFarland standard, the BBC values of MH are lower than EEP in four out of six strains, while for 1:100  and most strains obtain the value 5.0% . BPC shoThe noticeable tendency of increasing the obtained values arranged in n = 31), the Wilcoxon signed rank tests were taken. The results of the statistical analysis of metabolic activity are presented in Statistical analysis of the obtained data was calculated as described above. Because of the absorbance adjustment for propolis, the percentage values are above 100%. Values below zero prove the acquisition of strain metabolic activity as compared with the positive control. To exhibit the causality between MH or EEP impact and metabolic activity of the mature biofilm derived from clinical strains  and 40.0% concentration of EEP (116.9%). In the concentration range from 40.0% to 5.0%, EEP causes a higher reduction of metabolic activity than MH in the concentration range from 30.0% to 7.50%. EEP concentrations lower than 5.0% result in a lower mean metabolic activity reduction than MH. After the treatment with the lowest concentrations (1.88% for MH and 1.0% for EEP), MH causes a mean metabolic activity reduction of 18.0%, while EEP causes a mean metabolic activity increase of 19.0%. With a decrease in the MH or EEP concentration, an increase in absorbance and a lower level of metabolic activity reduction is visible, except for the absorbance after 3.75% MH treatment. A clear increase in the level of metabolic activity reduction is present as compared with the concentration ranges 7.5%, 15.0%, and 20.0%. Probability values below 0.05 and even 0.01 indicate the presence of statistically significant differences between the absorbances obtained for each MH concentration and the positive control (0.0%).The probability values, obtained for the EEP concentration range from 2.5% to 40.0%, are below 0.05. This indicates that there is a statistically significant difference between the absorbance obtained after EEP treatment (in this range of concentrations) and absorbance obtained with the positive control (0.0%). Only with a concentration of 1.0%, the probability value was above 0.05.To compare the EEP and ethanol (EtOH) impact on the absorbance depicting the metabolic activity of clinical strains, the Wilcoxon signed rank test was taken. The results of the statistical analysis of data are presented in Only after EEP treatment with concentrations of 40.0% and 20.0%, the level of metabolic activity reduction is higher as compared with EtOH. The mean grades of metabolic activity reduction for EEP are 116.9% and 105.0%, respectively, and 90.7% or 93.1% for EtOH. As for the concentration range from 10.0% to 1.00%, the mean grade of metabolic activity reduction for EEP is consistently lower than for EtOH. The mean grades of metabolic activity reduction values for 10.0% to 1.0% EEP are 73.2%, 76.4%, 32.1%, and −19.0%, respectively, while for EtOH, the values are 91.1%, 88.3%, 87.3%, and 71.3%.Statistically significant differences between absorbance values were proven for each tallied pair of EEP and EtOH concentrations .S. aureus, S. epidermidis, and other coagulase-negative staphylococci, as well as Streptococcus spp., P. aeruginosa, and other Gram-negative rods [Bacteriology of chronic wound infections is being constantly monitored by scientist and clinicians all over the world, and the vast majority of articles have reported that the most prevalent bacteria isolated from chronic wounds were ive rods ,30,31,32S. aureus (46.1%), P. aeruginosa (35.0%), and P. mirabilis (10.6%). In most cases (75.0%), chronic wound infections had a mixed etiology. Most infections were caused by tandems of S. aureus and P. aeruginosa (12.5%) or P. aeruginosa and P. mirabilis (5.9%). Mixed etiology of the infection hindered antibiotics penetration to the wound environment, and defective flow through the venous system altogether obstruct the chronic wound healing and decrease the quality of life of patients. Apitherapy products incorporated into dressings may fasten the healing process and attenuate the infection.In the bacterial profile described in this paper, the most frequently isolated bacteria were S. aureus, P. aeruginosa, and S. pyogenes bacterial cells in binding to the human proteins, i.e., fibronectin, fibrinogen, and collagen. Manuka honey treatment also caused a reduction in binding to human keratinocytes by all these species and bacterial invasion of S. pyogenes spp. A publication by Lu J. et al. [® on the S. aureus biofilm using a CV assay. Their results presented a significant decrease of the biofilm biomass in all the strains tested after treatment with honey concentrations ranging from 16.0% to 32.0%. Generally, the metabolic activity of strains decreased proportionally to the biomass. Results of the present experiment confirm that Manuka honey affects the metabolic activity of Proteus mirabilis strains. Emineke S. et al. [P. mirabilis and E. coli was most effective after 24-h incubation; the most abundant biofilm reduction was shown after 24 h as compared with 4 h of honey treatment, and early biofilm formation was inhibited even at a concentration as low as 3.3% if the exposure was long enough (72 h in this case).Several research groups have attempted to verify the antimicrobial effect of Manuka honey and propolis ,33,34,35. et al.  reportedP. mirabilis clinical strains were slightly higher than MIC or MBC values obtained in other papers [In this research, following our expectations, Manuka honey MBIC and MBEC values of r papers ,39,40. Hr papers  and Sherr papers   obtained for . mutans ,50, oralfaecalis . Kouidhifaecalis  obtained in 2007 , it was S. aureus biofilm cultured for 24 h. Propolis derived from two sources decreased the metabolic activity of the biofilm, but with propolis from the third source, a significant difference was noticeable only after treatment with the concentration of 2 mg·mL−1. In this research, the lowest concentration of EEP that caused a significant difference was 3.7 mg·mL−1 (2.50%).In 2018, de Oliveira Dembogurski D.S. et al.  conducteCases of treatment with ethanol extract of propolis have been described in the scientific literature since 1969. Ethanol extract of propolis has been used with chronic wounds, ulcers, surgical site infections, and burns, but also in cases of vaginitis or dental sockets ,55,56,57E. faecalis [Oral uptake of propolis alone and in combination with silver nanoparticles increased fibroblast proliferation, collagen deposition, and provided an anti-inflammatory effect on experimental wounds in rats . There wfaecalis . Chitosafaecalis . Anotherfaecalis . Other mfaecalis  and polyfaecalis .P. mirabilis biofilm in all examined concentrations (1.88% to 30.0%). Ethanol extract of propolis suppresses the metabolic activity of P. mirabilis more tenuously than ethanol in concentrations beneath 20.0% (29.4 mg·mL−1). Antimicrobial activities of Manuka honey and propolis depend on their concentration, MGO content, polyphenols content, antioxidant activity, high osmolarity, and several other factors. However, their antimicrobial activity is incontestable. Manuka honey suppresses the metabolic activity of Further investigation should focus on the selection of the best material for wound dressing enriched with propolis or Manuka honey. The material used for dressing preparation should be a fine carrier for antibacterial substances and air permeable, but should also provide high humidity for the wound environment and be easy to remove.


To assess the clinical value of early clearance of HE4 and CA125 for platinum sensitivity and prognosis in patients with ovarian cancer.HE4 and CA125 value including clinical data of 89 patients with ovarian cancer were collected. The clearance of HE4 and CA125 were assessed base on the platinum sensitivity, two-year PFS, PFS and OS.p< 0.0001, p< 0.0001) as well as CA125 after 1st cycle chemotherapy .Sixteen patients were classified as platinum resistant and 73 as platinum sensitive according to the response to platinum-base chemotherapy. When HE4 clearance after 3rd cycle chemotherapy or CA125 clearance after 1st cycle chemotherapy, it gave the highest AUC of 0.788, with 100% of sensitivity and 57.5% of specificity respectively between platinum resistant and platinum sensitive group. In addition, 59 patients were classified as two-year PFS group and 30 as not achieved two-year PFS group according to obtaining two-year PFS or not. It gave the highest AUC of 0.730, with 83.3% of sensitivity and 62.7% of specificity respectively when HE4 clearance after 3rd cycle chemotherapy or CA125 clearance after 1st cycle. The prolonged PFS and OS were significantly associated by the clearance of HE4 after 3rd cycle chemotherapy (Our data suggested that the early clearance of HE4 and CA125 could predict platinum response and prognosis in patients with ovarian cancer. Monitoring the HE4 and CA125 during first-line chemotherapy might be helpful in predicting platinum sensitivity and risk to progress and relapse. Epithelial ovarian cancer (EOC) is the leading cause of mortality among gynecological cancer . Due to Since patients with EOC have no measurable target lesion after initial surgery, it is difficult to assess the treatment response with gynecological examination and image. Effort has been made to find out reliable biomarker for monitoring therapeutic response and detecting relapse in EOC. Cancer antigen 125 (CA125) is the most common serum biomarker for judging treatment response and monitoring recurrent in EOC. Our previous study demonstrated that the median PFS and OS of patients with serum CA125 who had a logarithmic decrease or a decrease to normal within 1 month after treatment were better than those of with a non-logarithmic decrease or a decrease to normal that took longer than 1 month , 17, 22.Human epididymis protein 4 (HE4), has been proved to be a reliable biomarker for detecting ovarian cancer with a sensitivity of 76%  and a specificity of 94%   and apprThe retrospective study was conducted in the Guangxi Medical University Cancer Hospital of china from July 2012 to December 2018. Patients diagnosed with epithelial ovarian cancer by histopathology with full serum HE4 and CA125 value and clinical record were available for review. Inclusion criteria were: 1) Initially treated with staging surgery or optimal cytoreductive surgery, including total hysterectomy, bilateral oophorectomy and salpingectomy, peritoneal washing, omentectomy, pelvic/para-aortic nodal dissection, and multiple peritoneal biopsies . 2) Patients with stage IC and higher stage received platinum-based combined chemotherapy for 6–8 cycles after surgery. 3) Good nutritional status without other cancer. Exclusion criteria were:1) Death due to non-oncological reasons. 2) Lacking of follow-up after treatment.Responds to treatment and progression were evaluated according to the guidelines of the Gynecology Cancer Intergroup (GCIG)  and the The study was compliant with the Declaration of Helsinki, and approved by the Ethics Committee of Guangxi Medical University Cancer Hospital. Informed consents of all treatments and examinations have been obtained from patients or their families.Serum HE4 and CA125 concentration of each patient were measured at the time of pretreatment, each post-chemotherapy and recurrence. The tests were performed using an electro-chemiluminescence immunoassay  at the departmental laboratory of Guangxi Medical University Cancer Hospital. The normal value range of HE4 is less than or equal to 70 pmol/L as suggested by Moore et al. , and thep < 0.05 was defined as a statistically significant.All the statistical analyses were conducted by SPSS 25.0 Software. T-test was used for the comparison between groups. The area under the curve (AUC) was used to calculate with a receiver operating characteristics (ROC) in predicting platinum sensitivity and two-year survival. The Kaplan-Meier survival curve and long rank test were used to assess the influence of HE4 and CA125 on PFS and OS. Cox regression models was used to conduct the univariate and multivariate analyses. A two-tailed probability of p=0.008) and tumor grade (p=0.000). While the mean CA125 level of pretreatment was only significant increased with FIGO stage (p=0.008) and histology types (p=0.040). However, there were no significant differences in menopausal, platinum response and two-year PFS with pretreatment HE4 or CA125.A total of 89 patients with EOC were included in the study. All patients were followed up to December 31, 2019. At the end of follow-up period, 36 patients were progress, 19 patients were dead and 70 patients were still alive. The median follow-up time was 35 months. Patients’ clinicopathological characteristics and the mean pretreatment level of HE4 and CA125 were presented in Table p value of 0.000. The maximum AUC was 0.731 for CA125 after 1st cycle chemotherapy, reporting 75.0% of sensitivity, 71.2% of specificity, and a p value of 0.004. When the two biomarkers were combined, the result showed that when HE4 clearance after 3rd cycle chemotherapy or CA125 clearance after 1st cycle chemotherapy, it gave the highest AUC of 0.788, with 100% of sensitivity and 57.5% of specificity respectively. When HE4 clearance after 3rd cycle chemotherapy and CA125 clearance after 1st cycle chemotherapy were used at the same time, the AUC, sensitivity, and specificity were 0.723, 50, and 94.5% respectively. The AUC, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), accuracy and p-value base on each parameter were shown in Table In the analyzed patients, 16 were defined as platinum resistant and 73 as platinum sensitive. The capability of HE4 and CA125 clearance after 1st, 3rd, 6th cycle chemotherapy to predict platinum sensitivity were assessed by ROC and AUC. The clearance was defined as the level of HE4/CA125 reduced to normal value or had a reduction rate of 90% at least. The early clearance was defined as the clearance of HE4/CA125 before the 4th cycle of adjuvant chemotherapy. According to the definition, we found in platinum sensitive group that the clearance of HE4 in 55 of 73 (75.3%) cases after 1st cycle chemotherapy, in 59 of 73 (80.8%) cases after 3rd cycle chemotherapy, and in 61 of 73 (83.6%) cases after 6th cycle chemotherapy. The HE4 non-clearance patients of platinum resistant were found in 10 of 16 (62.5%) cases after 1st cycle chemotherapy, in 12 of 16 (75.0%) cases after 3rd cycle chemotherapy, and in 10 of 16 (62.5%) cases after 6th cycle chemotherapy. The maximum AUC was 0.779 for HE4 after 3rd cycle chemotherapy, with 75.0% of sensitivity, 80.8% of specificity, and a p=0.001, 0.011), and the AUC of 0.707, 0.666 respectively (Table p=0.023), with the AUC of 0.648, and a corresponding 56.7% of sensitivity and 72.9% of specificity . When HE4 and CA125 were combined, it was shown that the AUC reached 0.730 when the HE4 value after 3rd cycle chemotherapy or the CA125 after 1st cycle chemotherapy that declined above 90% or normalized (p=0.000). However, when both of the HE4 after 3rd cycle chemotherapy and the CA125 after 1st cycle chemotherapy declined above 90% or normalization, the AUC was 0.625 with p=0.056.In this part Table , 59 patily Table . The earp< 0.0001, p< 0.0001) and CA125 after 1st cycle chemotherapy . The pretreatment levels of HE4 had an impact on the PFS (p=0.014). However, there were no statistical significance between pretreatment levels of CA125 with PFS (p=0.694) and also no statistical significance between pretreatment HE4 and CA125 with OS .The Kaplan-Meier survival curve and a log rank test were conducted to analyze the relationship between patient’s PFS/OS and the value of HE4 and CA125  in predicting platinum sensitivity after the third cycle chemotherapy compared to the maximum AUC=0.731 of CA125 (p=0.004) after 1st cycle chemotherapy respectively. When the two biomarkers were combined, the result showed that when HE4 clearance after 3rd cycle chemotherapy or CA125 clearance after 1st cycle chemotherapy, the AUC, sensitivity and specificity were 0.788, 100 and 57.5% respectively. It means that 100% patients with platinum resistant could be identified through the both non-clearance of HE4 after 3rd cycle chemotherapy and CA125 after 1st cycle chemotherapy. When HE4 after 3rd cycle chemotherapy and CA125 after 1st cycle chemotherapy were both clearance, 94.5% individual with platinum sensitive could be recognized. Previous publications on the application of HE4 and CA125 as a method of predicting chemotherapy response have mainly focused on the pretreatment biomarker value [A prospective study of Roberto Angioli suggested that HE4 reduction with above 47% at the third cycle of chemotherapy were more likely to be platinum sensitive. On the contrary, CA125 value did not correlate with platinum response [er value . Anita eer value . Few stuer value . They foer value . Our stup=0.001, and AUC=0.666, p=0.011). CA125 level normalization or reduction above 90% after 1st cycle chemotherapy correlated with two-year PFS, with a lower AUC of 0.648, p=0.023. When combination of the HE4 after 3rd cycle chemotherapy and CA125 after 1st cycle chemotherapy, it was shown that the AUC reached to 0.730 when one of them declined above 90% or normalized (p=0.000), reporting an 83.3% of sensitivity and a 62.7% of specificity. Approximately 83.3% of patient who relapsed or progressed in 2 years could be identified during first-line chemotherapy with a 62.7% of specificity based on the combination of HE4 and CA125 non-early clearance. Using biomarkers to identify high-risk patients was a simple and non-invasive way and may complement the current definition of high-risk patients.Olivier Colomban et al. used a Kelim model , which cThe result of Kaplan-Meier survival curve and long rank test also demonstrated that both the clearance of HE4 after 3rd cycle chemotherapy and CA125 after 1st cycle chemotherapy were significantly correlation with the PFS and OS. Prolonged PFS was significantly impacted by the pretreatment HE4 value but not pretreatment CA125. As respected to OS, there were no significant correlation between pretreatment value of HE4 and CA125 in our results. Similar results were obtained in other research which analyzed the influence of the pretreatment HE4 on PFS , 15, 24.This study was a retrospective character, single-center setting, relatively small number of patients, and lack of external validation, which may bias the results. A bigger prospective cohort or including more clinical covariate such as ECOG score, oncogene expression might be helpful to validate the result and provide more information. Therefore, further studies are needed to confirm these results.In conclusions, our findings revealed that the early clearance of HE4 and CA125 during first-line platinum chemotherapy were significantly associated with the platinum sensitivity and prognosis. Monitoring the dynamic value of both HE4 and CA125 during treatment might be helpful in future clinical practice.


ObjectiveThe purpose of this study was to compare two different sizes of an endotracheal tube (ETT), with inner diameters (ID) of 6.5 mm and 7.5 mm, for the frequency of postoperative sore throat in patients undergoing elective breast cancer surgery.MethodologyThis study was a randomized controlled trial conducted in the Shaukat Khanum Memorial Cancer Hospital and Research Center from December 3, 2016, to February 3, 2017. This study included 110 patients, 55 from each group, who were selected from the elective surgery list fulfilling the inclusion criteria. Group A patients were intubated with 6.5-mm ID ETTs and group B patients with 7.5-mm ID ETTs. All patients were carefully extubated in the operating room when fulfilling extubation criteria. Any concerns of sore throat were recorded 24 hours postoperatively. All relevant data were recorded on a pro forma.Results2, with a minimum and maximum of 19 kg/m2 and 38 kg/m2, respectively. The mean four-point scale was 2.98 ± 1.3, with a minimum and maximum of 1.0 and 4.0, respectively. Of the 110 patients, 47 patients reported a sore throat at 24 hours after surgery, whereas 63 patients did not report a sore throat. In this study, 14 patients in group A were diagnosed with a postoperative sore throat, whereas 33 patients in group B were diagnosed with a sore throat at 24 hours. A chi-square test was significant, and a poststratification chi-square test was applied to compare sore throat at 24 hours postoperative between the groups with respect to age and BMI.In this study, the mean age was 46.6 ± 13.2 years (range: 18-65 years). The mean body mass index (BMI) was 29.50 ± 07.12 kg/mConclusionsETT sizes produced a significant difference in the frequency of postoperative sore throat in patients undergoing breast surgery. Physicians should consider this impact on clinical practice to optimize patient outcomes. Additional studies with a larger sample size are warranted to further explore this impact. A common complication of anesthesia following endotracheal tube (ETT) placement is a sore throat, which can lead to dissatisfaction after surgery and delay a patient's return to normal activities. The incidence is highest after tracheal intubation and varies from 14.4% to 50% . Safe anAlthough a postoperative sore throat is a minor problem, controlling it imposes a significant impact on a patient's comfort during the postoperative period . The appHypothesisThere is a difference in the frequency of postoperative sore throat in patients undergoing breast surgery with the 6.5-mm ETT compared to the larger 7.5-mm ETT.This randomized controlled trial was conducted at the Shaukat Khanum Memorial Cancer Hospital and Research Center for a period of six months. A sample size of 110 cases (55 in each group) was calculated with an 80% power of test and a 5% level of significance. A nonprobability consecutive sampling technique was used. American Society of Anesthesiology (ASA) class I and II patients undergoing breast cancer in the age group of >18 to 65 years were included. Exclusion criteria were the use of succinylcholine, anesthesia with rapid sequence induction , patients having ongoing respiratory tract infection assessed by history and examination, and patients with a history of deep vein thrombosis.After approval from the Shaukat Khanum Memorial Cancer Hospital and Research Center ethical committee and obtaining informed consent from the patients, a total of 110 patients selected from the elective surgery list fulfilling the inclusion criteria using a lottery method were included in the study. Standard monitoring was applied to all the patients preoperatively. General anesthesia was induced with intravenous 1.5 mg/kg propofol and 0.5/kg atracurium for muscle relaxation. After intravenous muscle relaxant, direct laryngoscopy was performed by an anesthetist (with more than six months' experience) using Macintosh laryngoscope blade no. 3. Group A patients were intubated with 6.5-mm ETT and group B patients with 7.5-mm ETT. After intubation, anesthesia was maintained by routine inhalation agent isoflurane or sevoflurane. After completion of the surgical procedure, a residual neuromuscular block was antagonized with 2.5 mg of neo-pyrolite intravenously. All patients were carefully extubated in the operating room when filling extubation criteria. Any concerns of sore throat were recorded at 24 hours postoperatively. All relevant data were noted on the prescribed pro forma. Sore throat was diagnosed by asking the patients. All the patients were females, and therefore a standard ETT of 7.5 mm was used for anesthesia.Mean and standard deviation for age, body mass index (BMI), and four-point scale scores were calculated. Frequency and percentage for sore throat at 24 hours postoperatively were calculated. The two groups were compared for frequency using a chi-square test with p < 0.05 as the level of significance. Data were stratified for age and BMI to address the effect modifier. Poststratification chi-square test was applied to check the significance with p < 0.05 enabled as significant. All variables were analyzed using IBM SPSS Statistics for Windows, Version 20.0. .2 was 29.50 ± 07.12 kg/m2, with a minimum and maximum of 19 kg/m2 and 38 kg/m2, respectively. The mean four-point scale was 2.98 ± 1.3  are required for future studies to adequately confirm our findings.ETT sizes produced a significant difference in the frequency of postoperative sore throat in patients undergoing breast surgery. Physicians should consider this impact on clinical practice to optimize patient outcomes and contribute to improved patient quality of life. This study will help the patients especially cancer patients who are immunocompromised and fragile and for whom extra care must be made to avoid any complications. Due to such a high incidence of sore throat, we need further larger sample size trials in diverse populations that can help to identify factors preventing this side effect and need of other alternative methods to avoid it.


HOXD11) plays an important role in a variety of cancers, but its precise role in gliomas remains unclear. This study aimed to explore the relationship between HOXD11 and gliomas by combining bioinformatics methods with basic experimental validation.Homeobox D11 , CGGA (749 glioma samples), GEPIA, GEO (GSE4290 and GSE15824). Nine cases of glioma tissue and five cases of normal control brain tissue were collected from the clinical department of Henan Provincial People’s Hospital for further verification. A series of bioinformatic analysis methods were used to confirm the relationship between HOXD11 was significantly related to age, World Health Organization (WHO) grade, chemotherapy status, histological type, and even 1p19q codeletion data and isocitrate dehydrogenase (IDH) mutation. HOXD11, as an independent risk factor, reduces the overall survival of glioma patients and has diagnostic value for the prognosis of glioma. Gene Set Enrichment Analysis (GSEA) showed that HOXD11 was significantly enriched in cell signaling pathway such as cell cycle, DNA replication and so on. Finally, we confirmed that the knockout of HOXD11 can inhibit the proliferation and invasion of U251 glioma cells, and change the biological behavior of tumor cells by preventing the progression of cell cycle.The high expression of HOXD11 may be used as a candidate biomarker for the clinical application of targeted drug and prognostic assessment treatment of glioma. In addition, This study will help to explore the pathological mechanism of glioma. Gliomas are one of the most common primary intracranial neoplasms, accounting for 81% of intracranial neoplasms . The treIn recent years, many studies have reported that epigenetics plays a crucial role in the development and regulation of cancer . EpigeneTCGA is a widely used public database storing the genetic and clinical information of a variety of human malignant tumors, and it has made great contributions to the research on human malignant tumors . In addiHOXD11 belongs to the HOX gene family, which encodes transcription factors that regulate various physiological processes. In recent years, many articles have reported that HOXD11 plays a vital regulatory role in tumors like laryngeal squamous cell carcinoma, ovarian cancers, head and neck cancer, and others  was used to reveal the mechanism of action of HOXD11 in gliomas. Finally, a variety of experimental methods have been used to confirm that HOXD11 can promote the malignant behavior of glioma cells as an oncogene by participating in the regulation of cell cycle signaling pathways. Thus, we believe this study will provide a new diagnostic biomarker for the diagnosis and therapeutic target for gliomas.d others . HoweverHOXD11 in various tumors and tissues, including 163 glioblastoma, 518 low grade glioma and 207 normal brain tissue samples, were obtained from the GEPIA database. To further determine the expression level changes of HOXD11 in glioma, we obtained gene expression data in the form of FPKM (Fragments Per Kilobase per Million) from a glioma tissue microarray (GSE4290) and a glioma cell microarray (GSE15824) from GEO database https://www.ncbi.nlm.nih.gov/geo/ for further analysis. GSE4290 contains 23 non-tumor brain tissue samples and 77 glioma tissue samples (GSE15824 contains three human-derived astrocyte (HA) cell line samples and two LN319, two LN229, two BS149 and two LN018 glioblastoma cell line samples  . This ad samples . GSE1582 samples .http://www.cgga.org.cn/) is a public database focused on glioma research, which contains many types of data and corresponding clinical information  and obtained clinical information of the corresponding patients.In order to verify the effect of Nine glioma tissue samples and five non-tumor brain tissue samples were collected and stored in a liquid nitrogen environment during surgery and then transferred to a -−80 °C freezer until use. Clinical samples were obtained after receiving written informed consent from patients in the clinical department of Henan Provincial People’s Hospital. This study has been approved by the Institutional Review Board of Zhengzhou University.HOXD11. Functional enrichment analysis was performed using the GSEA 4.0.2 jar software. The number of permutations was set to 1000 times, and the gene set database was set to the Kyoto Encyclopedia of Genes and Genomes (KEGG) cell signaling pathway.GSEA is widely used as a bioinformatics analysis tool for gene function annotation and analysis. The RNA-seq data obtained from the CGGA database was batch-corrected and normalized by limma packages and then classified into “group H” high expression group or “group L” low expression group based on the median expression level of Human glioma cell lines  and human-derived astrocyte (HA) were purchased from the Cell Bank of the Chinese Academy of Sciences . All cells were grown in incubators at 37 °C and 5% carbon dioxide and cultured using DMEM medium  plus 10% FBS .HOXD11 in tissues by RT-qPCR. Three glioma cell lines  and one human astrocyte were used to detect the expression level of HOXD11 in glioma cells. First, total RNA was extracted from glioma and normal brain tissue samples using Tri-Reagent . The quality and quantity assessments of total RNA were determined using NanoDrop One spectrophotometer , evaluating 260/280 nm absorbance values. Thereafter, the cDNA was reverse transcribed from the total RNA using the Transcriptor First Strand cDNA Synthesis kit . RT-qPCR was performed following the guidelines for FastStart Universal SYBR Green Master (ROX) . Results were quantified using QuantStudio software  following the manufacturer’s instructions. GADPH was used as an internal reference. The primer sequences used for HOXD11 were 5′-GAACGACTTTGACGAGTGCG-3′(F) and 5′-ACGGTTGGGAAAGGAACGAA-3′(R). The expression level of HOXD11 was measured by the “2–ΔΔCT” method. Statistical differences between the two groups were analyzed by unpaired t-test, and results were considered statistically significant when the p value was < 0.05.9 glioma tissue and 5 peritumoral tissue brain tissue samples were used to detect the expression level of HOXD11 was purchased from Genepharma . The siRNA sequence was sense (5′-GACUUCAACUCUCUCGGAUTT-3′) and antisense(5′-AUCCGAGAGUUGAAGUCTT-3′). The negative control (NC) RNAi sequence was sense(5′-UUCUCCGAACGUGUCACGUTT-3′) and antisense, (5′-ACGUGACACGUUCGGAGAATT-3′). Cells were transfected using Lipofectamine 2000 reagent . Detection of transfection efficiency by RT-qPCR at 48 h after transfection.The small interfere RNA (siRNA) that specifically targeting HOXD11 on the proliferation ability of U251 cells, MTT experiments were conducted. The transfected cells were transferred into 96-well plates (1 × 103 cells/well). Cell proliferation ability was measured at 12 h, 24 h, 48 h and 96 h after transfection. The specific method was to add 20 ul of MTT (5mg/ml) to each well for culture. The liquid was discarded after incubation for 4 h and 150 ul DMSO was added to each well. After 15 min of shaking, the optical density (OD) of 490 nm was detected using microplate reader.To detect the effect of In colony formation assay, U251 cells were planted in six-well plates at a density of about 300 cells/well. After 10 days of culture, the cells were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. Colonies larger than one mm in diameter were counted using ImageJ Software (v1.8.0) after photographing.The exponential growth cells were seeded on the 6-well plate with a fusion degree of 90%. A straight linear scratch was made on the center of the plate with a pipette (200 uL). PBS was washed gently, and serum-free medium was replaced for culture. After that, the scratch width was observed and photographed under the microscope for 24 h and 48 h respectively, then was measured by ImageJ Software (v1.8.0).Firstly, treated cells were fixed with 4% paraformaldehyde for 30 min, and then 0.3% Triton X-100 was used to penetrate for 20 min at room temperature. After blocking with 5% BSA, primary antibody Ki67  was added and placed in a 4 °C refrigerator overnight. After that, washed 3 times with PBS for 5 min each time. Secondary antibody was added and incubated for one hour at room temperature in the dark. After washing with PBS, DAPI was added for nucleation. The photographs were finally observed using a fluorescence microscope. After that, the total number of cells and the number of KI67 positive cells were counted by ImageJ Software (v1.8.0).U251 cells were digested and centrifuged to collect the cells after transfection. Precooled 75% ethanol was added, fixed overnight at −20 °C Each tube was added with 500 uL propidium iodide (PI) dye solution, stained for 30 min in dark, and filtered through a filter. Cell cycle distribution was analyzed by flow cytometry .HOXD11-NC group and the HOXD11-siRNA group is represented by the gray value of the corresponding protein bands.Total protein of transfected U251 cells was extracted using RIPA buffer with protease inhibitor. Separated by 10% SDS-PAGE gel electrophoresis, transferred to polypropylene fluoride membrane (PVDF) treated with methanol for 2 h, transferred to a shaker, sealed with TBS-T blocking solution containing 5% skimmed milk powder for 1 h. Then, the primary antibodies GAPDH , CCNB1 , CDK1  and Cdc25C  were added to incubate overnight at 4 °C. Washed by TBS-T 5 times for 5 min each time, secondary antibody (1:5000) was added to incubate for 1 h at room temperature, Washed by TBS-T 5 times for 5 min each time, and finally ECL color developing solution was added for color development. The developed bands were subjected to gray level analysis by ImageJ Software (v1.8.0), The difference in protein expression levels between the t-test was used for comparison between the two groups. The expression levels of HOXD11 were examined in glioma and non-tumor brain tissue samples by the Wilcox test method. The relationship between the Overall Survival rate of patients and the expression level of HOXD11 was analyzed by the Kaplan–Meier method and Cox regression analysis, and the survival curve was drawn accordingly. The relationship between clinically relevant patient information and the expression level of HOXD11 was analyzed using the Wilcox and Kruskal tests.R software (v.3.6.1 version) and GraphPad Prism (v8.0.2) was used to process data and perform statistical analysis. Unpaired HOXD11 was abnormally highly expressed in a variety of tumors, such as ESCA, GBM, HNSC and HUSC, especially in GBM, as shown in HOXD11 in gliomas relative to normal controls, data from more sources were analyzed. By analyzing the data of glioma tissues and cell lines microarrays obtained from GEO database, it was found that HOXD11 was highly expressed in both glioma tissues and cell lines, as shown in HOXD11 was highly expressed in glioma tissues and cell lines compared with normal brain tissues and human astrocytes, as shown in Through GEPIA database, we found that HOXD11 and the clinical and prognosis of glioma patients, we conducted further studies using glioma sequencing data and relevant clinical information from CGGA database. A total of 749 glioma samples with complete clinical information were obtained from the CGGA database, which contains a variety of clinical information such as Primary-Recurrent-Secondary (PRS) type, age (age at diagnosis), gender, histological type, and chemotherapy status, as well as 1p19q codeletion status and isocitrate IDH mutation status. More detailed information of clinical characteristics is shown in To clarify the relationship between high expression of HOXD11. The expression level of HOXD11 was significantly correlated with PRS type, WHO grade, chemotherapy status, age, 1p19q codeletion data, IDH mutation status and histological type (HOXD11 increases as the WHO grade and age of the glioma increases (p < 0.001). As for the IDH mutation status, there was a higher level of gene expression in wildtype than in the mutant (p < 0.001). Moreover, for 1p19q codeletion status, there were lower gene expression levels in patients with a codeletion of 1p19q than in patients with a non-codeletion of 1p19q (p < 0.001).The Wilcox and Kruskal tests were used to analyze the relationship between clinically relevant patient information and the expression level of cal type –2G. The HOXD11 in patients with gliomas was explored by Kaplan–Meier survival analysis. The 749 patients with gliomas included in the study were divided into two groups: “group H” and “group L,” based on the median expression level of HOXD11. As shown in HOXD11 (group H) compared to those with low levels  in CGGA-sequence, which indicated poor prognosis.The relationship between overall survival and the expression level of HOXD11 expression level can indeed reduce the overall survival time of glioma patients  for survival analysis. The results showed that the prognosis of the group with high HOXD11 expression was significantly worse than that of the group with low HOXD11 expression among the three molecular subtypes, as shown in HOXD11 may lead to poor prognosis of patients with glioma.To improve the credibility of the data results, we further obtained 666 glioma tissues in the TCGA database. The Kaplan–Meier method was used to verify that the increase of patients . To clarHOXD11 could be used as an independent prognostic factor, we performed univariate and multivariate analyses for the date from CGGA-sequence. Univariate analysis was performed by applying Cox regression model . The PRS type, histology, WHO grade, age and other factors showed similar results. Next, multivariate analysis was performed using the Cox regression model , PRS type , and high WHO grade  were significantly associated with poor prognosis. Together, the univariate and multivariate analyses results suggest that high expression levels of HOXD11 may be an independent risk factor for poor prognosis.To determine whether on model . The reson model . High exHOXD11 for glioma prognosis, we plotted the receiver operating characteristic (ROC) curve based on the sequencing data of glioma patients from CGGA and TCGA databases. As shown in HOXD11 had certain evaluation value for one-year, three-year and five-year survival rates. These results indicate that HOXD11 expression level could be related to prognosis in patients with gliomas; moreover, high expression of HOXD11 may be used for clinical prognosis evaluation of patients with glioma.To clarify the diagnostic value of highly expressed HOXD11 could be an independent prognostic factor for glioma and had certain diagnostic value. To verify the effect of HOXD11 on malignant biological behavior of glioma cell lines, we conducted a series of phenotypic experiments. First, we used siRNA to interfere with the expression level of HOXD11 in glioma cell U251. Meanwhile, the transfection efficiency was detected by RT-qPCR  was significantly reduced compared with the control group(HOXD11-NC)  . Meanwhiormation . Meanwhiormation , 5D. In wn group . TherefoHOXD11 plays a role in the pathological process of glioma, we performed GSEA analysis to predict the cell signaling pathways that HOXD11 may participate in. As shown in HOXD11 may be involved in a variety of cancer-related cell signaling pathways such as cell cycle, DNA replication, ECM receptor interaction, and focal adhesion. These results reveal that HOXD11 may be involved in these cancer-related cell signaling pathway as an oncogene.To clarify how HOXD11 may participate in the pathological process of glioma through a variety of cancer-related cell signaling pathways.Through GSEA analysis, we predicted that highly expressed HOXD11 on the cell cycle of glioma cell U251, we performed flow cytometry. The results showed that in the HOXD11 knockdown group, U251 cells were significantly blocked in G2 phase compared with the control group  as a homeobox gene , and knockdown of HOXD11 was able to reduce invasion in HNSCC (HOXD11 may enhance cell growth, clonality, and metastatic potential in Ewing sarcoma (HOXD11 plays a crucial role in cancer progression, the role of HOXD11 in gliomas has not yet been elucidated.Numerous studies have shown that iomarker . Studiesbox gene . In addiigration . HOXD11  factors . Moreoveblastoma . Some stn levels . Similarin HNSCC . HOXD11  sarcoma . AlthougHOXD11, as a novel carcinogenic gene in gliomas, expression levels were significantly higher in gliomas than that in non-tumor brain tissues. This result is completely credible and in line with the rigor of scientific research. Firstly, there are more than a thousand of glioma tissue samples in this result, and the data types contain data from multiple races groups, such as White, Asian and Black or African American. Secondly, the data used for analysis comes from a variety of analysis technologies including RNA sequencing, gene chip technology, RT-qPCR and other technologies, which can complement each other and ensure the authenticity and accuracy of the data. Third, the Kaplan–Meier method and Cox-regression analysis showed that HOXD11 could lead to poor prognosis as an independent risk factor. Finally, it is worth emphasizing that the knockdown of HOXD11 can inhibit the proliferation and migration of U251 glioma cells. Therefore, we have sufficient reason to confirm that HOXD11 can be a valuable biomarker for targeted treatment of glioma patients.We found that HOXD11 leading to poor prognosis in glioma patients, GSEA was used to indirectly reveal the signal pathway activated by HOXD11, which indicated that HOXD11 is significantly enriched in cancer-related cell signaling pathways, such as cell cycle, DNA replication, ECM receptor interaction, and focal adhesion. As one of the most important cell signaling pathways, the cell cycle plays an extremely crucial role in the development of various diseases, especially the progression of cancer. Studies have shown that mutations in genes that regulate the cell cycle may be associated with glioma progression, recurrence, and poor prognosis in patients with gliomas (HOXD11 may play a role in the aforementioned signaling pathway and thus affect the activity, adhesion, proliferation, and migration of glioma cells.In order to reveal the mechanism of  gliomas . Moreove gliomas . Focal a gliomas . It has  gliomas . In glio gliomas . In combHOXD11 leads to poor prognosis in glioma patients was indirectly revealed by functional enrichment analysis via GSEA. We only confirmed that HOXD11 knockdown could block U251 cells to G2 phase by flow cytometry and western blotting. Therefore, other specific mechanisms of HOXD11 affecting glioma need to be further verified by other research. This study only provides a clue to lead more researchers to pay attention to the role of HOXD11 in glioma.Although we have incorporated both the data sources of diverse for analysis, which contains Chinese, African American, and Caucasian individuals, so that the differences observed may not be narrowly limited to one ethnicity. Nonetheless, this research still has certain shortcomings to be improved. For example: based on the WHO glioma classification in 2016, more attention has been paid to the molecular typing of glioma. However, most of our data were obtained from public databases which may not necessarily contain information on the molecular typing of gliomas. Therefore, if conditions permit, we may develop more detailed research programs related to the molecular typing of gliomas. As another limitation, the mechanism whereby high expression of HOXD11 was higher in glioma samples at the cell and tissue level, and may be related to the poor overall survival of patients with glioma. In addition, HOXD11 may be involved in cancer-related cell signaling pathways, such as cell cycle, DNA replication, focal adhesion, and ECM receptor interaction, and thereby affect the malignant progression of glioma by participating in the regulation of the cell cycle. Therefore, HOXD11 could be used as a candidate biomarker for molecular therapy and prognostic evaluation of patients with glioma.This study found that compared with normal controls, the expression level of 10.7717/peerj.10820/supp-1Supplemental Information 1Click here for additional data file.10.7717/peerj.10820/supp-2Supplemental Information 2Click here for additional data file.10.7717/peerj.10820/supp-3Supplemental Information 3Click here for additional data file.10.7717/peerj.10820/supp-4Supplemental Information 4Click here for additional data file.10.7717/peerj.10820/supp-5Supplemental Information 5Click here for additional data file.10.7717/peerj.10820/supp-6Supplemental Information 6Click here for additional data file.10.7717/peerj.10820/supp-7Supplemental Information 7Click here for additional data file.10.7717/peerj.10820/supp-8Supplemental Information 8Click here for additional data file.10.7717/peerj.10820/supp-9Supplemental Information 9Click here for additional data file.10.7717/peerj.10820/supp-10Supplemental Information 10Data analysis of colony formation assay after 10 days of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-11Supplemental Information 11The clones formed from sample No. 1 in the HOXD11 gene silencing group after 10 days of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-12Supplemental Information 12The clones formed from sample No. 2 in the HOXD11 gene silencing group after 10 days of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-13Supplemental Information 13The clones formed from sample No. 3 in the HOXD11 gene silencing group after 10 days of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-14Supplemental Information 14The clones formed from sample No. 4 in the negative control group after 10 days of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-15Supplemental Information 15The clones formed from sample No. 5 in the negative control group after 10 days of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-16Supplemental Information 16The clones formed from sample No. 6 in the negative control group after 10 days of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-17Supplemental Information 17The scratch of sample No. 1 in the negative control group after 0 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-18Supplemental Information 18The scratch of sample No. 2 in the negative control group after 0 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-19Supplemental Information 19The scratch of sample No. 3 in the negative control group after 0 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-20Supplemental Information 20The scratch of sample No. 1 in the negative control group after 24 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-21Supplemental Information 21The scratch of sample No. 2 in the negative control group after 24 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-22Supplemental Information 22The scratch of sample No. 3 in the negative control group after 24 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-23Supplemental Information 23The scratch of sample No. 1 in the negative control group after 48 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-24Supplemental Information 24The scratch of sample No. 2 in the negative control group after 48 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-25Supplemental Information 25The scratch of sample No. 4 in the HOXD11 gene silencing group after 0 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-26Supplemental Information 26The scratch of sample No. 3 in the negative control group after 48 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-27Supplemental Information 27The scratch of sample No. 5 in the HOXD11 gene silencing group after 0 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-28Supplemental Information 28The scratch of sample No. 6 in the HOXD11 gene silencing group after 0 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-29Supplemental Information 29The scratch of sample No. 4 in the HOXD11 gene silencing group after 24 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-30Supplemental Information 30The scratch of sample No. 5 in the HOXD11 gene silencing group after 24 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-31Supplemental Information 31The scratch of sample No. 4 in the HOXD11 gene silencing group after 48 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-32Supplemental Information 32The scratch of sample No. 6 in the HOXD11 gene silencing group after 24 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-33Supplemental Information 33The scratch of sample No. 5 in the HOXD11 gene silencing group after 48 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-34Supplemental Information 34The scratch of sample No. 6 in the HOXD11 gene silencing group after 48 h of cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-35Supplemental Information 35Data analysis of cell cycle distribution ratio by flow cytometry after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-36Supplemental Information 36The cell cycle distribution ratio of sample No. 1 in the negative control group by flow cytometry after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-37Supplemental Information 37The cell cycle distribution ratio of sample No. 4 in the HOXD11 gene silencing group by flow cytometry after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-38Supplemental Information 38The cell cycle distribution ratio of sample No. 5 in the HOXD11 gene silencing group by flow cytometry after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-39Supplemental Information 39The cell cycle distribution ratio of sample No. 2 in the negative control group by flow cytometry after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-40Supplemental Information 40The cell cycle distribution ratio of sample No. 3 in the negative control group by flow cytometry after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-41Supplemental Information 41The cell cycle distribution ratio of sample No. 6 in the HOXD11 gene silencing group by flow cytometry after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-42Supplemental Information 42Data analysis of the expression of key proteins in the G2 phase of the cell cycle by western blotting after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-43Supplemental Information 43The expression of the key protein Cdc25c in the G2 phase of the cell cycle by western blotting after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-44Supplemental Information 44The expression of the internal reference protein GAPDH by western blotting after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-45Supplemental Information 45The expression of the key protein CDK1 in the G2 phase of the cell cycle by western blotting after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-46Supplemental Information 46The expression of the key protein CyclinB1 in the G2 phase of the cell cycle by western blotting after cell transfection.Click here for additional data file.10.7717/peerj.10820/supp-47Supplemental Information 47Click here for additional data file.10.7717/peerj.10820/supp-48Supplemental Information 48Click here for additional data file.


Hemophilia A is an X-linked recessive bleeding disorder caused by mutations in FVIII gene with an incidence of 1 in 5,000 to 10,000 live born males. The Inv22 mutation is a major cause of the disease worldwide, accounting for up to 40%–50% of severe FVIII mutations. The aim of the present study was to screen Inv22 of the FVIII gene in Palestinian patients with severe HA and reveal its role as a predisposing factor for the development of inhibitors.  A cohort of 77 HA individuals including 5 carrier females from 52 unrelated families registered at governmental hemophilia centers in the West Bank area of Palestine was investigated. The demographic data and the clinical history were retrieved from medical files. Molecular analysis of Inv22 mutation in severe HA (30 cases) from Palestine was performed using the subcycling polymerase reaction (S-PCR). FVIII coagulant activities were carried out on an aPTT-based 1-stage clotting assay. FVIII inhibitors were quantified using the Nijmegen modification of the Bethesda assay. p=0.53) between inhibitor development and the Inv22 mutation.  Overall, 41.7% (30/72) of the studied cases were classified as having severe HA, 22.2% (16/72) had moderate HA, and 36.1% (26/72) had mild HA. Five randomly selected carrier mothers were screened for the Inv22 mutation to confirm its transmission to their sons. The Inv22 mutation was detected in 11 severe HA patients (36.6%). Among the severe HA patients with positive Inv22, 45.5% (5/11) had developed inhibitors. The current study showed that there was no association ( Findings on Inv22 are in agreement with worldwide reports, being a major genetic mutation in severe HA. The S-PCR is a simple, rapid, and cost-effective method for the diagnosis of Inv22 in severe HA patients. Although the Inv22 mutation was associated with 36.6% of severe HA phenotype cases, it was not a major predisposing factor for inhibitor formation. In Palestine, thus far, no molecular genotyping services for HA are available. Screening for the molecular defects has become a crucial tool in hemophilia care with respect to prediction of the clinical course and safe genetic counseling of relatives. Therefore, the aim of the present study was to screen Inv22 of the FVIII gene in Palestinian patients with severe HA and reveal its role as a predisposing factor for the development of inhibitors.Hemophilia A (HA) is an X-linked recessive bleeding disorder and the second most common coagulation disorder with an incidence of 1 in 5,000 to 10,000 live born males . More thhttp://bcshguidelines.org) was followed for the patients' selection criteria. Based on their conformation to the analogous FVIII level, 41.7% (30/72) of the studied cases were classified as having severe HA, 22.2% (16/72) had moderate HA, and 36.1% (26/72) had mild HA. All patients were tested for factor VIII activities that were carried out on a 1-stage aPTT-based clotting assay via the Stago blood coagulation analyzer  according to the manufacturer's instructions. 10 ml of peripheral blood samples was collected in two tubes: tripotassium ethylene-diamine-tetraacetate (EDTA K3) and 3.2% trisodium citrate tubes . Blood samples were taken at least 1 week following FVIII infusion. EDTA K3 tubes were stored at 4°C and were used later on for genomic DNA extraction. The 3.2% trisodium citrate tubes were processed immediately and centrifuged at 5000 rpm for 10 minutes, and the plasma was aspirated and stored at −20°C and used later on for FVIII activity measurements assay and the presence of inhibitors.This was a multicenter cohort study. HA patients attending governmental hemophilia centers from three major governmental hospitals in Nablus, Jenin, and Tulkarm in West Bank region were included in the study. A total of 72 male patients with severe hemophilia A and 5 females (mothers) from 52 unrelated families were included. HA diagnosis was determined according to each patient's personal and family history of bleeding, a prolonged activated partial thromboplastin time (aPTT), and a reduced FVIII level. Clinical data such as age, gender, age at first exposure to FVIII, family history of inhibitors, and number of exposure days (EDs) were obtained from the patients using a specially designed questionnaire. Approval for the study was obtained from the Board or Ethics Committee at An-Najah National University, Nablus, Palestine. All of the patients or their guardians for minors provided informed consent for the molecular studies. British Committee for Standards in Hematology Guidelines  developed inhibitors: 14 patients had severe and 3 patients had moderate disease. Inhibitor titrations were carried out using the Nijmegen modification of the Bethesda assay. The majority of patients manifested high titers and 3 patients showed low titers. The patients showed a mean antibody titer of 93.6 BU/mL (range: 7–795 BU/mL).μL of buffy coat using the Qiagen DNA Blood Mini kit , following manufacturer's recommendation. DNA concentration and purity were determined in a Nanodrop 2000 spectrophotometer .DNA was extracted from 200 http://www.genome.ucsc.edu). The primers were used to differentiate between normal, inversion patients and carrier females. Primers P & Q are specific to flanking sequences of int22h1 located at −1212 bp before and at +1334 bp after the homolog, and produce 12 kb segment in DNA of wild type and carrier females. Primers A & B are specific to the flanking sequences of int22h2 and int22h3 located at −167 bp before and at +118 bp after the homologs and always produce 10 kb segment, because at least one copy of Int22h2 and Int22h3 remains intact and could serve as a positive control. Inv22 can occur by homologous recombination between Int22h1 and either Int22h2 or Int22h3; thus, each P & B and A & Q produces 11 kb segments in DNA of male patients and carrier females. S-PCR reaction was performed using Expand Long Template PCR kit  as previously described with minor modification of the original protocol [μl containing 2.5 μl buffer, 0.75 mM MgCl2, 1.75 U enzyme mix, 200 μ mol 7-deaza-dGTP , 300 μ mol GTP and 500 μ mol of each of the other dNTPs , 0.2 μ mol of each forward and reverse primer , 7.5% dimethyl sulphoxide (DMSO), and 50–100 ng of human genomic DNA template. Amplified products were resolved in 1% agarose  in 1x TBA buffer  and visualized using ultraviolet transilluminator documentation system .The primer sequences used for the Inv22 PCR reactions  are shown in In this study, 72 male patients with hemophilia A and 5 carrier mothers from 52 unrelated families aged 1 month to 67 years  were investigated. All patients tested for Inv22 were varying with respect to family history, treatment regimen, first bleeding site, and age at first bleeding. Family history of bleeding was present in 96.7%. Age at first bleeding was from less than 1 month to 2 years. First bleeding was reported in 83.3% at circumcision, 6.7% after trauma, and 10% due to muscle hematoma. According to treatment regimen, all individuals were treated with FVIII concentrate (plasma derived), and 76.7% of them received on-demand treatment, while the rest 23.3% were on prophylactic treatment .The Inv22 mutation was detected in 11 patients (36.6%), and all of them had severe HA.p=0.53) between inhibitor development and the Inv22 mutation.Among the severe HA patients with positive Inv22, 45.5% (5/11) developed inhibitors. Mutational analysis using S-PCR technique was carried out for all 30 HA severe cases. Analysis also included 5 randomly selected mothers from these patients, for evaluation of carrier status of Inv22, and 15 wild type healthy male controls. Affected males showed positive amplification for three bands, AB, PB, and AQ, and absence of PQ amplified products (12 kb segment). Analysis showed that, out of the 30 tested severe hemophilia A cases, 11 (36.6%) were positive for Inv22. A representative gel photograph for S-PCR analysis is shown in p=0.53) between inhibitor development and the Inv22 mutation.To the best of our knowledge, this is the first study to describe the health status of HA patients in the West Bank area of Palestine and to assess the prevalence of Inv22 mutation in X chromosome among severe HA patients as well as suspected carriers, being a major causative of this disease worldwide. Based on clinical data of early symptoms presentation and family history, one can easily predict the severity of the disease in the area. Since family history is strongly correlated with severity of disease, early screening diagnosis for families at risk, performing aPTT before circumcision will be helpful and cost effective. Early diagnosis and adequate and timely prophylaxis treatment can prevent complications like hemarthrosis and life-threatening bleedings. They are also expected to improve the quality of life and decrease the morbidity and mortality rates . The aimAccording to treatment regimen in Palestinian Ministry of Health (MOH), all HA patients are treated with FVIII concentrate (plasma derived). In this study, 76.7% of HA patients received on-demand treatment. Ghosh et al.  reportedSince hemophilia is an X-linked disorder, it was reasonable to carry out pedigree analysis involving expected carrier female cases reported in this study. Data on S-PCR analysis, involving 2 expected carrier mothers of severe HA cases and presented in We report here the first molecular analysis of Inv22 mutation as a major genetic mutation responsible for severe HA patients in Palestine. Our results are consistent with the reports in neighboring countries and countries all over the world. The S-PCR is a simple, rapid, and cost effective method for diagnosis of Inv22 in severe HA patients. Although the Inv22 mutation was associated with 36.6% of severe HA phenotype cases, it was not a major predisposing factor for inhibitor formation. Further studies are recommended with large number of patients as well as screening for other mutations such as Inv1 and full genome sequencing for novel mutations among negative Inv22 patients.


Obesity is an epidemic with severe consequences on the professional development of patients. Bariatric surgery has proven to be a safe treatment with effective results in weight control. The aim of this study is to assess the implications of professional occupation in relation to the development of obesity and weight changes after bariatric surgery. We analyzed 500 obese patients  who underwent one anastomosis gastric bypass surgery at the Centre of Excellence for the Study and Treatment of Obesity and Diabetes (2014–2019), assessing the influence of professional occupation on body composition and evolution of weight loss up to two years after surgery. Preoperative obesity type III and IV was higher in men than in women . Prevalent clinical history in women was depression (46.7%), varicose veins (35.6%), and thyroid disease (9.7%), while in men it was respiratory failure (98.2%), high blood pressure (56.8%), hepatic steatosis (82%). Postoperative weight loss was effective in every professional field, reaching normal weight values from 12 months after surgery. Obesity is a worldwide epidemic that represents one of the most significant public health concerns globally ,2. The tDirect costs caused by obesity represent about 9% of total healthcare expenses in developed countries, where overweight and obesity are more frequent ,4. FurthSome articles describing obesity in relation to professional field, display how it is more frequent in sedentary jobs , which cBariatric surgery has proven to be an effective long-term treatment in the weight control and comorbidities associated with obesity ,12,13. MThere are different surgical techniques of restrictive and malabsorptive type; the criteria of choice depend on each patient and the surgical team. The International Federation for the Surgery of Obesity and Metabolic Disorders (IFSO) has described bariatric surgeries worldwide in order to establish a profile of surgery patients in the future . Among tThe assessment of body composition and anthropometric changes has been evaluated for the different types of surgical techniques, and malabsorptive type surgeries obtain better long-term results . However2, or BMI > 30–35 kg/m2 with the existence of associated comorbidities and poor metabolic control, of legal age and willing to participate in the study by giving their written consent [Cross-sectional descriptive study conducted on 500 obese patients undergoing one anastomosis gastric bypass (OAGB) bariatric surgery at the Centre of Excellence for the Study and Treatment of Obesity and Diabetes, Valladolid (Spain) from January 2014 to July 2019. The inclusion criteria of the patients in the study followed the recommendations approved by the International Federation for the Surgery of Obesity and Metabolic Disorders (IFSO): BMI > 40 kg/mThe preoperative protocol included both the selection criteria for bariatric surgery and the nutritional, psychological, blood, radiological, and cardiorespiratory function assessment, as well as other complementary tests in relation to existing comorbidities depending on the type of patient. This protocol was conducted by a multidisciplinary team following the recommendations in management and collection of clinical parameters described by the IFSO .The clinical assessment of patients was conducted from the first preoperative consultation in which compliance with the selection criteria for surgery prospective patients was evaluated. If a patient met the criteria, he or she was included in the individualized preoperative protocol based on preoperative nutritional monitoring to enhance the commitment to postoperative dietary changes as well as to increase the safety of the surgical procedure.The initial assessment included the collection of clinical background and socio-demographic data to complete the initial medical history of each patient according to their personal characteristics and specificities. In the consultation prior to surgery, the following were evaluated: age, gender, height, clinical and surgical history, professional occupation  , body coBody Mass Index (BMI):Weight loss expression criteria:Percentage of Excess of Body Mass Index Loss (EBMIL):Percentage of Excess Weight Loss (EWL):*The ideal body weight was determined according to Metropolitan Life Insurance Company’s formula.OAGB bariatric surgery has been described in previous publications ,24. It wThe study was conducted in accordance with the recommendations of the 1964 Declaration of Helsinki . It has been approved by the Research Commission of the Nursing School of Valladolid . The informed consent was received from all the participants in the study and the data treatment was anonymous and confidential. There is no conflict of interest and no funding from public or private institutions.We created an Excel database (Microsoft Office 2016) to gather data. The data were recorded by one person and, after review, they were analyzed using the statistical program IBM SPSS Statistics v. 25.0.t test or Mann-Whitney U test were applied, depending on the conditions of application, for two groups; and the ANOVA or Kruskal-Wallis H test, also depending on the conditions of application, for more than two groups. The significance level for all tests was set at p ≤ 0.05.Quantitative variables were described as mean ± standard deviation and their normalization was established with the Kolmogorov-Smirnov test. Qualitative variables were described via absolute and relative frequencies (percentages). To study the association between qualitative variables, the Chi-square test was used with Fisher’s exact test or likelihood ratio, depending on the conditions of application. To study the differences between mean values, the Student The study sample are 500 patients, average age of 40.8 ± 11.6 years, 389 women (77.8%) and 111 men (22.2%). There were no significant age differences between women (40.7 ± 11.7 years) and men (41.5 ± 11.2 years).p < 0.05).Patient distribution based on the degree of overweight and obesity  is detaip < 0.05), thyroid disease (9.7% vs. 3.6%. p < 0.05), depression (46.7% vs. 18.9%. p < 0.05), and were more under psychiatric treatment (23.8% vs. 9.9% p < 0.05).In the anamnesis, clinical history was assessed, and we found respiratory failure, arthropathies, obstructive sleep apnea syndrome, and hepatic steatosis to be the most prevalent conditions. The clinical history described in p < 0.05), treatment with oral antihypertensive drugs (36.9% vs. 16.2%. p < 0.05), obstructive sleep apnea syndrome (86.5% vs. 77.7%. p < 0.05), respiratory failure (98.2% vs. 93.6%. p < 0.05), hepatic steatosis (82% vs. 67.7%. p < 0.05), and the presence of hiatal hernia (56.8% vs. 46.7%. p < 0.05) prevailed in men. In contrast, high blood pressure (56.8% vs. 24.4%. p < 0.05). 11.5% of women had previously carried a gastric balloon compared to 3.6% of the men, and 7.2% had undergone liposuction compared to 0.9% of the men (p < 0.05). Regarding the surgical history of the patients, trauma surgery (14.4%) was the most common, followed by appendectomy (10.8%). Women had a higher frequency of previous surgeries than men in the case of cholecystectomy (7.7% vs. 0.9%. p < 0.05), sales and services (22.3% vs. 15.3%. p < 0.05), and healthcare (8.4% vs. 2.7%. p < 0.05). In contrast, men’s professional occupations are higher in the trade, equipment and transport operators (21.6% vs. 1.7%. p < 0.05), applied and related natural sciences (17.1% vs. 2.5%. p < 0.05), primary and extractive exploitation (4.5% vs. 0.2%. p < 0.05), and in processing, manufacturing, and assembly of goods (2.7% vs. 0.5%. p < 0.05) The professional occupation of patients was distributed in nine major fields , adding p < 0.05. Regardless of preoperative weight, the trend in all patients in all professional fields covered was a significant decrease in weight and BMI.Postoperative changes in weight loss were analyzed according to the type of professional occupation . The higThere are variabilities among professional fields regarding the point of lowest average weight during the first 24 postoperative months. In the finance and administration field, this was reached at 18 months (66.2 ± 14.1); at 12 months in natural, applied, and related sciences (78.4 ± 15.3 kg), healthcare (66.4 ± 11.9 kg), social sciences, education, public administration, and religion (66.7 ± 11.7 kg), primary and extractive exploitation (65 ± 1.4 kg), trades, equipment operators and transport fields (76.3 ± 13.2 kg); and at 24 months in the arts, culture, recreation and sport (65.4 ± 10.1 kg), sales and services (64.8 ± 9.1 kg), processing, manufacturing and assembly of goods fields (70.5 ± 8.1 kg), and students (74.3 ± 11.9 kg).Weight loss expressed in relative terms like % EWL and % EBMIL shows the highest weight loss at 24 months in the processing, manufacturing, and assembly of goods field . In contrast, the primary and extractive exploitation field had the lowest weight loss in relative terms at 18 months: % EWL (78.5 ± 15.5%) and % EBMIL (93.8 ± 18.3%).p < 0.05.In the professional fields where there were more women than men , men were found to have a higher average initial weight in every field: 136.6 ± 19.9 kg vs. 109.5 ± 18.1 kg; 129.3 ± 24.1 kg vs. 108.19 ± 16.87 kg; 142 ± 9 kg vs. 111.7 ± 20.7 kg respectively. Nevertheless, the trend in these three professional areas was similar, with significant weight loss, expressed in % EWL and % EBMIL, in all the controls carried out since the preoperative and in the following ones, as illustrated in Bariatric OAGB surgery has enabled effective weight control in the patients studied, illustrating which professional fields have a greater impact on weight control. Our study revealed some of the main professional areas associated with the obesity of their workers, highlighting a greater presence of women in areas such as art and culture, sales and services, and the healthcare system. The profile of obese patients in different professional fields is highly variable and responds to different causes ,27. TherPsychiatric disorders and psychological problems are clinical determining factors in the development of obesity and are correlated with emotional state and professional performance ,8. AlmosThe evolution of weight loss follows a similar trend to that observed in several reports of patients undergoing malabsorptive surgery . In our Patients with a greater degree of obesity are correlated with a higher rate of unproductive work, especially in the trades, equipment operators, and transport sectors . In our Bariatric surgery has proven to be an effective long-term treatment in the control of obesity and associated comorbidities ,30. PrevAmong the limitations of this study is the lack of randomness of the selected sample, since all patients belong to the same care center where the OAGB was performed. Although the objective of the study was to analyze the results in the first 24 months after surgery, it is necessary to continue with the follow-up process of the patients analyzed in the current study and to assess the long-term impact of the results on weight control and associated comorbidities by comparing the different professional fields.This study describes the profile of the patient undergoing bariatric surgery, its socio-demographic aspects, and the evolution in weight control at 24 months after OAGB surgery. Patients’ professional occupation was a key factor in obesity and its outcome after surgery. However, it is necessary to assess the evolution of each patient individually, considering aspects such as physical activity required for each job, and dietary and lifestyle changes. In all the professional fields analyzed, average normal weight values were reached (BMI 18.5–24.9 kg/m2) from 12 months after surgery. Women predominated in professional fields such as art and culture, sales and services, and healthcare, and they presented a greater weight loss than men.