Objective:To assess the utility of whole-genome sequencing (WGS) in predicting Mycobacterium tuberculosis resistance to fluoroquinolones (FQs) and to establish a quantitative relationship between resistant gene mutations and resistance levels. Methods:A total of 296 drug-resistant tuberculosis surveillance strains with various resistance profiles, preserved by the National Tuberculosis Reference Laboratory of the Tuberculosis Prevention and Control Center at the Chinese Center for Disease Control and Prevention between 2013 and 2020, were included as study subjects. The Sensititre? MYCOTBI microplate method and WGS were used to assess the phenotypic and genotypic drug sensitivity of Mycobacterium tuberculosis to ofloxacin and moxifloxacin. Sensitivity, specificity, and concordance (Kappa value) of WGS in predicting fluoroquinolone sensitivity were calculated using phenotypic drug susceptibility testing (DST) results as the gold standard. A summary analysis was conducted on the distribution of drug resistance mutation sites and resistance levels. The paired χ 2 test was used to compare the detection rates between the two methods, with P<0.05 indicating statistical significance. Results:Among the 296 Mycobacterium tuberculosis strains with different resistance profiles, 196 were rifampicin-resistant, 50 were resistant to other drugs, and 50 were fully sensitive. WGS identified 81 strains carrying FQs resistance-related mutations, primarily at gyrA codons 94, 90, and 91. Sensitivity, specificity, and consistency (Kappa value) of WGS in predicting ofloxacin resistance were 86.5%, 98.1%, and 0.87, respectively. For moxifloxacin resistance prediction, these values were 80.0%, 99.5%, and 0.83, respectively. There was no statistically significant difference between the phenotypic DST and WGS detection rates for ofloxacin resistance (30.1% vs 27.4%, χ 2=3.06, P=0.08). However, the phenotypic DST detection rate for moxifloxacin resistance (33.8%, 100/296) was significantly higher than that of WGS (27.4%, 81/296) (χ 2=15.43, P<0.01). Analysis of the distribution of resistance mutation sites and resistance levels showed that different mutation sites corresponded to different minimum inhibitory concentrations (MICs). Multiple mutation combinations, including gyrA_D94G, gyrA_D94Y, and gyrA_D94N were mainly associated with high-level resistance, while gyrA_D94A, gyrA_A90V, and gyrA_S91P were primarily linked to low-level resistance. Conclusion:WGS demonstrates favorable sensitivity, specificity, and consistency in predicting FQs resistance and can partially predict resistance levels.

The pharmacy of Y anqing Winter （Paralympic）Olympic Village Polyclinic of 2022 Beijing Winter （Paralympic） Olympic Games will be comprehensively managed from five aspects of man ，machine，material，method and environment by adopting the 4M1E management method of quality management tool . In terms of man ，constantly enhanced training and daily health monitoring have been insisted ；in terms of machine ，drug information entry and drug information management have been improved and maintained ；in terms of drugs ，on the basis of drug management ，the management of drugs banned by the World Anti -Doping Agency，class Ⅱ psychotropic drugs and narcotic drugs have been strengthened ；in terms of regulations and environment ，strict pharmacy workflow and management system have been established ，and epidemic prevention closed -loop management requirements have been strictly implemented . From the pre -opening of Yanqing Winter （Paralympic）Olympic Village on January 23，2022 to the official closing of the Village on March 16，2022，the clinic pharmacy dispensed a total of 156 prescriptions（105 prescriptions during the Winter Olympic Games ，51 prescriptions during the Winter Paralympic Games ），including 18 prescriptions from the National （Regional） Olympic Committee and the National （Regional） Paralympic Games team doctors ，2 prescriptions containing stimulants that do not need to be exempted from treatment，1 prescription for narcotics ，and 37 prescriptions for athletes（23.7%）. Ibuprofen tablets ，Diclofenac diethylamine emulsion，Loratadine tablets and other drugs are widely used . The application of 4M1E management method enables the clinic pharmacy to provide perfect and high -quality pharmaceutical services for large -scale events while doing well in epidemic prevention and control .

Objective:To investigate the effects of quaternary ammonium salt of chitosan combined with closed irrigation on ischemia-reperfusion injury (I/R) in rat skin flap.Methods:Fourty-five adult male SD rats were randomly divided into 3 groups: sham-operated group, I/R group and chitosan group. Before operation, collection of arterial blood for testing PCT and hs-CRP were used as basic values. Right low abdominal island flap fed by the superficial epigastric artery was created in rats of all three groups. After operation, we flushed and dressed the wound with 10% saline of sham-operated group and I/R group. Quaternary ammonium salt of chitosan combined with closed irrigation were used in chitosan group. Flaps were assessed histologically at 3 d, 5 d and 7 d of reperfusion. The tissue was collected to make the SOD, MDA and PCT, hs-CRP testing, pathological section observation and to calculate the flap survival rate.Results:Marked neutrophil infiltration and edema were observed in I/R group, while less neutrophil infiltration was observed in chitosan group. Compared with the sham-operated group, the PCT and hs-CRP levels of the I/R group after modeling for 3 and 5 days were significantly increased. There were statistically significant differences ( P<0.05); Compared with the sham-operated group, the PCT and hs-CRP levels of the Chitosan group after modeling for 3 days, 5 days were significantly increased. There were statistically significant differences ( P<0.05); Compared with the I/R group, the PCT and hs-CRP levels of the Chitosan group after modeling for 3 and 5 days were significantly induced. There were statistically significant differences ( P<0.05); after modeling for 7 days, the values between all groups were no statistically significant differences ( P>0.01). Compared with the sham-operated group, the SOD level of the I/R group after modeling for 3, 5 and 7 days was significantly induced. MDA level was significantly increaesd; compared with the sham-operated group, the SOD level of the Chitosan group after modeling for 3, 5 and 7 days were significantly induced. MDA level was significantly increaesd; Compared with the I/R group, the SOD level of the chitosan group after modeling for 3, 5 and 7 days were significantly increased. MDA level was significantly induced. They all had statistically significant differences ( P<0.05). In the chitosan group, the survival of flaps was (85.34±10.02)%, which was significantly greater than the I/R group, in which only (41.11±14.17)% of the flap survived ( P<0.05). Conclusions:Quaternary ammonium salt of chitosan combined with closed irrigation can significantly improve flap survival, reduce I/R in transplanted rats＇ skin flaps.

Objective To translate English version of sensory-motor profile awake ( SMP-a) into Chinese version (the Chinese Version of SMP-a),and analyze the reliability and validity of the scale before and after craniotomy under awakening anesthesia. Methods Eighty-one patients whose tumors were located near or already in sensory-motor functional area were included in this study. Before and after awake cranioto-my,the Chinese version of SMP-a was used to accurately assess the sensory-motor function of each patient. Finally, the reliability and validity of the scale were analyzed by SPSS statistical software. Results Cronbach's α coefficient in the Chinese version of SMP-a was 0. 971,and Cronbach's α coefficient in the four subscales of face,hand,leg and sensation was 0. 965,0. 989,0. 981 and 0. 970,respectively. The test-retest reliability of the Chinese version of sensorimotor assessment scale was 0. 910,0. 904,0. 884,0. 898 and 0. 695 (total,face,hands,legs and sensory score respectively). The raters' consistency reliability was above 0. 949,0. 960,0. 934,0. 887 and 0. 660,respectively. The Pearson correlation coefficients of sensorimotor function score with SF-36 physiological function factors and KPS score were 0. 868 and 0. 790,respectively. Conclusion Before or after operation,the Chinese version of SMP-a has preferable reliability,internal con-sistency reliability and structural validity. It is feasible in awakening anesthesia craniotomy,and the degree of damage can be determined by repeated measurement of the sensorimotor sites that may be impaired by the patient.

Objective To investigate the effect of superomedial pedicle reduction mammaplasty in the treatment of severe gynecomastia.Methods 24 males with severe gynecomastia were involved in this study,who underwent superomedial pedicle reduction mammaplasty.Results All the patients were followed up for one to two years,and there were no complications such as skin necrosis,nippleareolar complex (N A C) hypaesthesia,hematoma or infection occurred.Minimal wound dehiscence occurred in two cases,and the rest cases were healed well.Conclusions Superomedial pedicle reduction mammaplasty in the treatment of severe gynecomastia could achieve satisfactory results,and good breast shapes are produced.

Objective: To study the expression of H O X A 1 0 gene in endometrial carcinoma and its effect on the apoptosis, migration and invasion of Ishikawa cells. Methods: Twenty-one cases of endometrial carcinoma tissue samples and 25 cases of normal endometrial tissue samples from patients treated at the Department of Obstetrics and Gynecology, Nanjing Drum Tower Hospital from 2012 to 2013 were collected for this study. The mRNA and protein expressions of H O X A 1 0 in endometrial carcinoma and normal endometrial tissues were separately tested by Realtime-qPCR (qRT-PCR) and Western blotting. Ishikawa cells were infected with adenovirus-flagHOXA10 at different multiplicity (5, 10, 20 MOI), and infected by adenovirus-flag-lacz (20 MOI) as control; And the cell apoptosis was tested by Flow Cytometry. Ishikawa cells were transfected with 50 nmol/L si-HOXA10 plasmids and 50 nmol/L si-NC plasmids, as down-regulation group and down-regulation control group, respectively. Ishikawa cells were infected with 20 MOI adenovirus-flagHOXA10 and 20 MOI adenovirus-flag-lacz, as up-regulation group and up-regulation control group, respectively. The ability of migration and invasion was detected by transwell assay. Results: The results of qRT-PCR and Western blotting showed that the expressions of H O X A 1 0 mRNA and protein in endometrial carcinoma samples were both significantly lower than normal samples [mRNA: (0.56± 0.14)vs (1.36±0.33), P<0.01; protein: (1.01±0.25) vs (2.10±0.71), P<0.001]. After the up-regulation of H O X A 1 0 gene in Ishikawa cell line, the cell apoptosis rate in ad-flag-HOXA10 groups (5, 10, 20 MOI) was significantly raised, and most of which was in the early apoptosis [(50.92±8.79)%, (55.17±4.07)%, (76.10±3.65)% vs (7.74 ± 0.15)%, all P <0.01]. The number of migrated cells was markedly up-regulated in si-HOXA10 group [(248±25) vs (135±15), P <0.01] but markedly down-regulated in ad-flag-HOXA10 group [(50±6) vs (100±13), P <0.01]. The number of invasive cells was markedly up-regulated in si-HOXA10 group [(131±18) vs (66±9), P <0.01] but markedly down-regulated in ad-flag-HOXA10 group [(34±8) vs (60±4), P <0.01]. Conclusions: Both mRNAand protein expressions of H O X A 1 0 were down-regulated in endometrial carcinoma samples than in normal endometrium. Up-regulation of H O X A 1 0 gene in Ishi kawa cell line can promote cell apoptosis and inhibit cell migration and invasion.

Objective To discuss the reasons of iatrogenic nipple-areola necrosis and to investigate clinical effect of nipple-areola reconstruction by three-bladed flap combined with free skin graft.Methods Between May 2012 and May 2015,13 patients with nipple-areola necrosis caused by iatrogenic reasons were treated with our new method.We designed a skate flap on the new nipple site.After incising skin,three-bladed flap was cross-stitched to form a new nipple.New areola was reconstructed by free skin graft,which was obtained from perineum or contralateral areola,and margin of the donor area was sutured after proper free from surrounding tissues.Being deducted subcutaneous tissue,the full-thickness skin graft was sutured around the nipple just like letter O.After operation,the areola area was pressurized bandaging,and the nipple was drawn by silk at least one month.Results The incisions were all primary healing.Over a mean period of 8 months (range 6-18),all patients were satisfied with the results.No significant asymmetry occured.The average nipple retraction rate was 10.03％.In 13 patients,there was no infection,bleeding or hematoma and skin flap necrosis.Conclusions Nipple-areola necrosis occurs when the blood supply is not protected carefully during a breast plastic surgery.Therefore,we take full advantage of the scar tissue which has hyperpigmentation.Reconstructive nipple-areola complex has good shape,and there is no need to tattoo after operation.

Early diagnosis and intervention is an important way to delay the progress of Alzheimer′s disease (AD).Compared with cerebrospinal fluid, blood sampling is not invasive and easy to be obtained in clinic practice.In this study, the serum samples of 9 controls, 10 AD and 12 mild cognitive dysfunction (MCI) patients were analyzed and compared through one by one analysis to screen potential markers for AD diagnosis.The experimental results showed that VGFYESDVMGR of α-2-macroglobulin peptide was closely related to the late stage of AD disease, and the large amount degradation of apolipoprotein C-Ⅲ, histone H1.2 and histone H1.4 was significantly related to early stages of AD progression.The characteristics of serum peptidome were different for the early and late AD, and these four proteins may be used as potential biomarkers of AD disease.In addition, the obvious ladder sequence characteristic was observed for apolipoprotein C-Ⅲ and histone H1, which could partly explain why the peptides distribution in different samples was somewhat contingent.On the contrary, the distribution at protein level was more stable.Finally, it was confirmed that the peptides of proteins such as fibrinogen α-chain, thymosin β-4 and patchy proteins were the dominant peptides in all serum samples.Overall, this study showed that the method of using serum peptidomics to diagnose AD was possible.The results may provide evidence and references for the large-scale clinical validation of AD.

A novel probe (DNSBN) towards biothiols on the basis of 4-hydroxynaphthalimide as fluorophores and 2, 4-dinitrobenzenesulfonyloxy group as specific recognition site was designed and synthesized.The result of absorption and fluorescence spectral analyses indicated that the probe had high sensitivity and selectivity towards cysteine (Cys), homocysteine (Hcy) and glutathione (GSH), and the detection was not affected by other 17 kinds of natural amino acids.Meanwhile, it was confirmed that DNSBN was a ratiometric probe through the fluorescence titration experiment, and the fluorescent intensity at 555 nm had a high linear relationship with biothiols concentration in the range of 0-20 μmol/L.The detection limits (3σ) of Cys, Hcy and GSH were 25.9, 92.0 and 77.9 nmol/L, respectively.The absorption, emission and mass spectra indicated that biothiols could be engaged in nucleophilic substitution reaction with 2,4-dinitrobenzenesulfonate, which induced the sulfonic esters decomposed.With the departure of receptor unit, the d-PeT progress (donor-excited photoinduced electron transfer) was blocked with an obvious colorimetric and fluorescence change.Finally, HeLa cell imaging experiments verified that DNSBN had good biocompatibility and could be used to detect exogenous biothiols.

Objective To evaluate the efficacy of medial femoral tubed skin flap combined with penile elongation for repairing long-segment penile defects.Methods From June 2009 to January 2015,13 cases of long-segment penile defects were treated by medial femoral tubed skin flap with pe nile elongation.Results All cases were followed up for 0.5 to 5.0 years.All flaps survived and all patients had the minor scar in donor areas,with good appearance and feeling of penis.The length of pe nis in flaccid state was 6.3±1.6 cm.The length of penis in erection state was 8.5±1.2 cm;The circumference of penis was 5.3±1.9 cm.1.0 cm × 0.7 cm of necrosis at distal parts occurred in one case early after operation,and healed after dressing changes.Two cases had tumid appearance which influenced penile appearance and sexual intercourse.It was improved by local liposuction.No feeling and e rectile dysfunction developed during the following-up.Except 5 cases who did not married,the other married patients and their partners were satisfied or quite satisfied with sexual intercourse activity.Conclusions Medial femoral tubed skin flap combined with penile elongation is a good method for the treatment of long-segment penile defects.The prosthetic penis could have suitable length,good ap pearance,good feeling and erectile function.