Chronic refractory wound (CRW) is one of the most challengeable issues in clinic due to complex pathogenesis, long course of disease and poor prognosis. Experts need to conduct systematic summary for the diagnosis and treatment of CRW due to complex pathogenesis and poor prognosis, and standard guidelines for the diagnosis and treatment of CRW should be created. The Guideline forthe diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients ( version 2023) was created by the expert group organized by the Chinese Association of Orthopedic Surgeons, Chinese Orthopedic Association, Chinese Society of Traumatology, and Trauma Orthopedics and Multiple Traumatology Group of Emergency Resuscitation Committee of Chinese Medical Doctor Association after the clinical problems were chosen based on demand-driven principles and principles of evidence-based medicine. The guideline systematically elaborated CRW from aspects of the epidemiology, diagnosis, treatment, postoperative management, complication prevention and comorbidity management, and rehabilitation and health education, and 9 recommendations were finally proposed to provide a reliable clinical reference for the diagnosis and treatment of CRW.

Objective:To investigate changes of natural killer (NK) cell subsets and interleukin-18 (IL-18) level in peripheral blood of patients with alopecia areata, and to assess the regulatory effect of IL-18 on NK cell activity.Methods:A total of 67 patients with alopecia areata (alopecia areata group) and 25 healthy volunteers (control group) were collected from Shanxi Provincial People′s Hospital between December 2019 and January 2021. Peripheral blood mononuclear cells (PBMCs) and plasma were isolated. The percentage of NK cell subsets was investigated by flow cytometry, and plasma IL-18 level was measured by enzyme-linked immunosorbent assay. PBMCs were stimulated with recombinant human IL-18, and co-culture systems of PBMCs with 721.221 cells, K562 cells and P815-Ab cells were established separately. NK cell function was assessed by determining the percentage of CD107a-expressing NK cells and fluorescence intensity of CD16 + NK cells. Comparisons between groups were performed using t test or paired t test. Results:Compared with the control group, the alopecia areata group showed significantly decreased percentage of CD56 +CD16 - NK cells (8.12% ± 3.14% vs. 10.78% ± 4.08%, t = 3.33, P = 0.001) , but significantly increased percentage of CD56 +CD16 + NK cells (46.08% ± 15.21% vs. 32.14% ± 10.45%, t = 4.22, P < 0.001) , and there was no significant difference in the percentage of CD56 -CD16 + NK cells between the alopecia areata group and control group (28.81% ± 8.65% vs. 27.09% ± 7.62%, t = 0.88, P = 0.383) . The plasma IL-18 level was significantly higher in the alopecia areata group than in the control group (112.0 ± 23.72 pg/ml vs. 99.34 ± 15.15 pg/ml, t = 2.48, P = 0.015) . After co-culture with 721.221 cells, K562 cells and P815-Ab cells, the percentage of CD107a-expressing NK cells was significantly higher in NK cells from the alopecia areata group (9.53% ± 1.70%, 5.15% ± 1.35%, 6.50% ± 1.64%, respectively) than in those from the control group (5.00% ± 1.17%, 4.40% ± 1.09%, 5.13% ± 1.36%, respectively, all P < 0.05) . After the stimulation with P815-Ab cells, the alopecia areata group showed significantly decreased fluorescence intensity of CD16 + NK cells (151.10% ± 59.30%) compared with the control group (221.90% ± 93.56%, t = 4.31, P < 0.001) . After IL-18 stimulation, the percentage of CD107a-expressing NK cells significantly increased in the co-culture system of NK cells with 721.221 cells compared with the unstimulated co-culture system (14.47% ± 2.67% vs. 9.93% ± 1.94%, t = 6.00, P < 0.001) , while there was no significant difference between the IL-8-stimulated co-culture system of NK cells with K562 cells or P815-Ab cells and the unstimulated co-culture systems (both P > 0.05) . Conclusion:IL-18 could enhance NK cell activity in patients with alopecia areata, likely by promoting natural cytotoxicity receptor-mediated cytotoxicity.

Objective:To investigate changes in the peripheral interleukin-35 (IL-35) level in patients with alopecia areata, and to assess its modulatory effect on regulatory T (Treg) cell activities.Methods:Totally, 81 patients with alopecia areata (alopecia areata group) and 27 healthy volunteers (control group) were enrolled from Shanxi Provincial People′s Hospital between December 2019 and January 2021. Sera and peripheral blood mononuclear cells (PBMCs) were isolated. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum IL-35 level, real-time fluorescence-based quantitative PCR to determine the mRNA expression of IL-35 subunits EBI3 and IL-12p35, and flow cytometry to determine the proportion of CD4 + CD25 + CD127 dim/- Treg cells. Sorted Treg cells were stimulated by recombinant human IL-35, ELISA was performed to detect levels of perforin and granzyme B in the culture supernatant, and real-time fluorescence-based quantitative PCR to determine the mRNA expression of EBI3, IL-12p35, and immune checkpoint molecules, such as programmed death protein 1 (PD-1) , T cell immunoglobulin and mucin protein-3 (Tim-3) , cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and lymphocyte activation gene-3 (LAG-3) in Treg cells. IL-35-stimulated or unstimulated Treg cells were co-cultured with autologous PBMCs, and cell counting kit-8 (CCK8) assay was conducted to assess cellular proliferative activity. Measurement data were compared between 2 groups by using t test, comparisons among multiple groups were carried out by using one-way analysis of variance, correlation analysis was carried out by using Pearson correlation analysis, and enumeration data were compared by using chi-square test. Results:Compared with the control group, the alopecia areata group showed significantly decreased IL-35 levels (90.10 ± 11.98 ng/L vs. 100.74 ± 28.71 ng/L, t= 2.71, P= 0.008) , mRNA expression of EBI3 and IL-12p35 in PBMCs (EBI3: 1.06 ± 0.15 vs. 1.25 ± 0.11, t= 6.09, P < 0.001; IL-12p35: 1.00 ± 0.15 vs. 1.38 ± 0.22, t= 10.16, P < 0.001) , and proportions of Treg cells (5.91% ± 1.17% vs. 6.85% ± 1.23%, t= 3.54, P= 0.001) . In the alopecia areata group, the proportion of Treg cells was positively correlated with the serum IL-35 level ( r= 0.25, P= 0.026) , and the mRNA expression of EBI3 and IL-12p35 in PBMCs ( r= 0.31, 0.24, P= 0.004, 0.032, respectively) . Compared with the control group, the unstimulated Treg cells from the alopecia areata group showed significantly decreased supernatant levels of perforin and granzyme B, mRNA expression of EBI3, IL-12p35 and immune checkpoint molecules ( P < 0.05 or 0.001) , as well as weakened inhibitory effect on the proliferative activity of PBMCs ( P= 0.013) . There was no significant difference in the level of perforin or granzyme B between the recombinant human IL-35-stimulated and unstimulated Treg cells from the patients with alopecia areata (both P > 0.05) . However, the mRNA expression of EBI3, IL-12p35 and immune checkpoint molecules was significantly higher in the IL-35-stimulated Treg cells than in the unstimulated Treg cells in the alopecia areata group ( P < 0.05 or 0.001) , and the inhibitory effect on the proliferative activity of PBMCs was also significantly enhanced in the IL-35-stimulated Treg cells compared with the unstimulated Treg cells ( P= 0.037) . Conclusion:The peripheral IL-35 level was significantly decreased in the patients with alopecia areata, which was closely associated with reduced activities of Treg cells, and IL-35 may be involved in the occurrence of alopecia areata.

Objective:To explore the short-term therapeutic outcomes of femoral neck fractures fixated by Femoral Neck System (FNS).Methods:A retrospective analysis was conducted of the 16 patients with femoral neck fracture who had been admitted to the Department of Traumatology & Orthopedics, The Second Affiliated Hospital to Anhui Medical University from January to December 2019. They were 6 males and 10 females, aged from 24 to 69 years (average, 47.5 years). According to the Garden Classification, there were 3 cases of type Ⅱ, 7 cases of type Ⅲ, and 6 cases of type Ⅳ. All the 16 patients were fixated with FNS. Recorded were operation time, frequency of intraoperative fluoroscopy, intraoperative blood loss, quality of postoperative fracture reduction, hospital stay, fracture healing time, and efficacy and complications at the last follow-up.Results:The 16 patients were followed up for 5 to 16 months after operation (average, 9.3 months). According to the Garden indexes, fracture reduction was evaluated as satisfactory in all the 16 patients. The operation time in this group ranged from 25 to 115 min with an average of 41.5 min, the frequency of intraoperative fluoroscopy from 14 to 47 times with an average of 26 times, the intraoperative blood loss from 35 to 210 mL with an average of 76 mL, the hospital stay from 3 to 9 days with an average of 4.6 days, and the fracture healing time from 3 to 6 months with an average of 4.5 months. By the Harris hip scores at the last follow-up, the function of the affected hip was rated as excellent in 10 cases, good in 5 and fair in one. Follow-ups revealed no complications like infection or implant loosening.Conclusions:The FNS is a new type of simple, reliable, safe and minimally invasive internal fixation for the treatment of femoral neck fractures. The plate, bolt, anti-rotation screws and locking screws in FNS can be inserted through a single minimally invasive incision, leading to reliable fixation, limited irritation to the lateral muscular soft tissues, and fine short-term outcomes.

Objective:To compare of the efficacy of Steinman pin-assisted and manual reduction for the treatment of distal femoral fracture with anterograde intramedullary nail.Methods:From January 2014 to August 2018, data of 54 patients with distal femoral fracture were retrospectively analyzed. According to the fracture reduction methods, patients were divided into two groups: bare-handed reduction group and Steinman pin-assisted reduction group (referred to Steinman pin group). There were 16 males and 10 females in bare-handed reduction group, with age of 37.5±9.2 years (range, 21-59 years). According to AO/OTA classification, 16 cases were type 32-A, 7 type 32-B, 3 type 32-C. There were 19 males and 9 females in Steinman pin reduction group, with age of 36.4±9.8 years (range, 18-55 years). According to AO/OTA classification, 19 cases were type 32-A, 7 type 32-B, 2 type 32-C. The reduction time, fluoroscopy times, intraoperative blood loss, fracture healing time, and knee joint function score of American hospital for special surgery (HSS) were compared between the two groups.Results:The amount of blood loss during operation was 142.78±29.76 ml in the bare-handed group, and 94.81±17.71 ml in the Steinman pin group. The reduction time of fracture was 14.19±2.50 min in the bare-handed group and 5.02±1.69 min in the Steinman pin group. The times of fluoroscopy during reduction was 12.56 ±2.01 in the bare-handed group and 5.01±1.51 in the Steinman pin group. There were significant differences in the above indexes ( t=12.19, 4.02, 5.47; all P < 0.05). All 54 patients were followed up for 12-51 months, with an average of 23.4 months. All the fractures healed, and there was no delayed union or nonunion. The healing time was 7.01±1.15 months in the bare-handed group and 5.99±0.97 months in the Steinman pin group. There were no significant difference. The HSS score of knee joint function was 23.7±4.1 before operation, 61.3±4.5 at 1 month after operation, 70.2±4.2 at 2 months after operation, 78.9±5.9 at 3 months after operation, 87.9±4.6 at 6 months after operation, and 93.1±5.8 at 12 months after operation, in the bare-handed group. Meanwhile, in the Steinman pin group, 22.5±3.8 before operation, 62.2±5.1 at 1 month after operation, 69.1±4.7 at 2 months after operation, 79.2±4.3 at 3 months after operation, 88.6±5.3 at 6 months after operation, and 92.3±6.1 at 12 months after operation. There were no significant difference between the two groups ( t=1.113, 0.689, 0.908, 0.212, 0.519, 0.494, P > 0.05). There were significant differences between the two groups at each time point before and after operation ( F=716.42, 815.52, P < 0.001). There were no complications such as injection point infection, vascular and nerve injury, failure of internal fixation and so on. Conclusion:Both groups had good functional recovery after operation. However, compared with bare-handed reduction, Steinman pin groupreduction has less intraoperative blood loss, shorter reduction time and less fluoroscopy times, which is a safer reduction method.

Objective:To compare four reconstruction algorithms of 18F-fluorodeoxyglucose (FDG) PET/CT on standardized uptake value (SUV) of pulmonary nodules. Methods:A total of 46 patients (27 males, 19 females; median age: 66 (range: 44-82) years) with solid pulmonary nodules from February 2018 to July 2019 in the First Hospital of Shanxi Medical University who performed 18F-FDG PET/CT imaging were enrolled. All PET/CT images were retrospectively reconstructed by using four algorithms reconstructions including ordered subset expectation maximization (OSEM), OSEM+ time of flight (TOF), OSEM+ TOF+ point spread function (PSF) and block sequential regularized expectation maximization (BSREM) (G1-G4). Nodule and background parameters were analyzed semi-quantitatively and visually. The maximum of SUV(SUV max), mean of SUV(SUV mean) and peak of SUV (SUV peak) were collected by the region of interest (ROI). Nodules were divided into small nodule group (diameter ≤10 mm) and large nodule group (10 mm < diameter ≤30 mm). Kruskal-Wallis rank sum test and Bonferroni method were performed to compare the differences of SUVs between G1-G4, and Spearman correlation analysis was used to analyze the correlation between the change rate of SUV (%ΔSUV) and the diameter of nodules. The receiver operating characteristic (ROC) curve analysis was used to analyze the diagnostic efficacy of SUV for the differential diagnosis of pulmonary nodules and to get the optimal threshold. Results:There were 114 pulmonary nodules (large nodules, n=55; small nodules, n=59). In visual analysis, the visual detection rates of small nodules in G4 were 55.93%(33/59), 44.07%(26/59), 20.34%(12/59) higher than those in G1-G3. Of 114 pulmonary nodules in 46 patients, there were differences in SUV max and SUV mean between G1-G4 (median SUV max : 2.65-5.29, median SUV mean: 2.05-2.99; H values: 20.628 and 17.749, respectively, both P<0.001), G4 had significant increases compared to G1 in SUV max (median 5.29 and 2.65, P<0.001) and SUV mean (median 2.99 and 2.05, P<0.001). The %ΔSUV max (median: 4.45%-52.96%) and %ΔSUV mean (median: 1.69%-47.56%) were negatively correlated with the diameter of nodules (9.75(6.20, 16.58) mm; r s values: -0.371 to -0.354, -0.371 to -0.320, all P<0.001). In 59 small nodules, G1 significantly increased the SUV max of G4 (median 4.05 and 2.14, H=18.327, P<0.001), while G4 significantly increased the SUV mean of G1 and G3 (median 2.31, 1.26 and 1.53, H=16.808, P<0.05). There was no significant difference in SUVs between G1-G4 in 55 large nodules ( H values: 0.812-7.290, all P>0.05). The optimal threshold values of SUV max in G1-G4 were 4.335, 5.185, 5.410, 5.745 and the area of under curves (AUCs) were 0.747, 0.699, 0.756, 0.778 respectively. The AUC of SUV mean and SUV peak also showed a similar trend. Conclusion:Among the four reconstruction algorithms, BRERM can not only enhance the image quality, but also significantly improve the SUV max and SUV mean of lung nodules diameter below 10 mm, and thus its diagnostic threshold of SUV should be appropriately increased.

Objective To propose a concept of irreducible tibial shaft fractures and to discuss their imaging characteristics and clinical significance.Methods A retrospective study was performed in 21 patients with tibial shaft fracture who had received intraoperative intramedullary nailing after limited open reduction at Department of Orthopaedics,The Second Affiliated Hospital to Anhui Medical University from November 2013 to June 2018.They were 14 males and 7 females,aged from 21 to 66 years (average,34.9 years).There were 15 left and 6 right sides.Firstly,closed reduction was performed followed by traction,folding and rotation,but repeated attempts failed to achieve smooth reduction or insertion of guide wire.Next,local limited open reduction had to be performed for intramedullary nailing.The X-ray and CT images of the tibial fractures were collected to analyze their imaging characteristics.The imaging manifestations were characterized into 4 types:single-segment type with intact fibula,multiple-segment type,interlocking type where the distal and proximal ends interlock commonly seen in short spiral and short oblique fractures,and incarceration type where the fracture interspace is blocked by a bone fragment.The therapeutic efficacy was evaluated at the final follow-up by knee scores of The Hospital for Special Surgery (HSS) and Kofoed ankle scores.Results Of the 21 patients,2 were single-segment type,4 multiple-segment type,13 interlocking type and 2 incarceration type.They were followed up for 7 to 50 months (average,22.7 months).The fractures united after 5 to 16 months (average,7.3 months).Postoperative knee pain was observed in 3 cases and delayed fracture union in 2.Osteomyelitis,superficial wound infection,implant breakage or malunion occurred in none of the patients.The therapeutic efficacy evaluated at the final follow-up by HSS knee scores and Kofoed ankle scores revealed 15 excellent,4 good and 2 fair cases,yielding an excellent to good rate of 90.5％.Conclusion The concept of irreducible tibial shaft fractures may lead to preoperative awareness on the part of the surgeons so that ineffective repeated reductions can be spared and the damage to the blood supply to the fracture ends and the operation time can be reduced.

Objective To investigate antimicrobial resistance of Escherichia coli (E.coli)and Klebsiella pneumoniae (K.pneumoniae),antimicrobial use density(AUD),as well as relation between antimicrobial resistance and AUD in a ter-tiary first-class hospital.Methods Antimicrobial resistance rates of clinically-isolated E.coli and K.pneumoniae,AUD of carbapenems and quinolones,as well as relation between resistance and AUD in 2013-2015 were statistically analyzed. Results Correlation analysis of antimicrobial resistance of bacteria and AUD showed that the decrease in resistance rate of E.coli to levofloxacin was related to the decrease in the use density of quinolones(r=0.61,P=0.03);increase in resist-ance rate of K.pneumoniae to imipenem was related to the increase in the use density of carbapenems(r=0.78,P<0.01). Conclusion Antimicrobial use is one of the causes of bacterial resistance,management on antimicrobial use needs to be strengthened to reduce the threat of bacterial resistance to human health.

Abstract A chiral separation and residue determination method for cis-epoxiconazole enantiomers in apple, grape and tea samples was developed and validated by ultra performance convergence chromatography combined with quadrupole time-of-flight mass spectrometry ( UPC2-QTOF/MS) . The Chrial CCA column was used to separate cis-epoxiconazole enantiomers and the chromatography conditions ( mobile phase modifier and proportion, column temperature, automated backpressure regulator, and auxiliary solvent ) were optimized. Samples were extracted by acetonitrile, and respectively purified by Cleanert TPT or Pesti-Carb solid phase extraction ( SPE ) columns, then analyzed by UPC2-QTOF/MS. The optimum conditions were as follows:mobile phase was CO2/isopropanol (95: 5, V/V), flow-rate was 2. 0 mL/min, automated backpressure regulator (ABPR) was 13. 79 MPa, column temperature was 30℃, with a post-column mauxiliary solvent of methanol/water (1:1, V/V) containing 2 mmol/L ammonium formate. The analyte was quantified by matrix external standard method. The results showed that linear range of this method was 0. 01-1. 00 mg/L, and the correlation coefficients were above 0 . 99 . The recoveries of cis-epoxiconazole enantiomers at three spiked levels (0. 005, 0. 025 and 0. 25 mg/kg) in fruit matrix were 67. 9%-92. 8% with relative standard deviations (RSDs, n=6) less than 10%, and the limit of quantification (LOQ) of enantiomers was 0. 005 mg/kg. The recoveries of cis-epoxiconazole enantiomers at three spiked levels (0. 01, 0. 05 and 0. 5 mg/kg) in black tea were 74 . 1% -84 . 0% with RSDs ( n=6 ) less than 8%, and the LOQ for these two enantiomers was 0. 01 mg/kg. This method is rapid, convenient and reliable, and could meet the requirement of residue analysis.

Objective:To investigate the gene subtypes of cervical human papilloma virus (HPV) infection in pregnant and non-pregnant women .Methods :From February 2012 to May 2013 ,cervical epithelial cells in 684 women of childbearing age (non-pregnant women) and 857 pregnant women in Jinshan Hospital ,Fudan University were collected .Polymerase chain reaction (PCR) and gene-chip technique were used to detect HPV gene subtype .Results :A total of 195 positive results were found in non-pregnant women with the HPV infection rate of 28 .5% .The predominant types of HPV infection were type 43 ,16 and 58 .A total of 102 positive results were found in pregnant women with the HPV infection rate of 11 .9% .The predominant types of HPV infection were type 43 ,16 and 58 .The infection rates of pregnant women in less than 30 years of age ,30 to 40 years old ,more than 40 year old were 10 .9% ,15 .1% and 30% ,respectively .The infection rates of non-pregnant women in less than 30 years ,30 to 40 years old ,more than 40 year old were 25 .3% ,25 .2% and 32 .2% ,respectively .Conclusions :The HPV infection rate of pregnant women was lower than that of non-pregnant women .The main HPV infection subtypes of the pregnant women and the non-pregnant women were the same .HPV43 ,16 and 58 were the predominant subtypes in this hospi-tal .The infection rate of non-pregnant women increased significantly in accordonce with age .