OBJECTIVE To investigate the metabolic changes in MEG-01 megakaryocytes after treatment with linezolid （LZD） from metabolomic perspective and its impact on the the proliferation of cells and generation of platelet precursors. METHODS MEG-01 cells were seeded in proliferation medium and divided into blank control group （untreated）， solvent control group （4‰DMSO）， and 100， 200， 400， 800 μg/mL LZD groups. The proliferation status of cells in each group was observed under the microscope； cell proliferation and viability were assessed. Cells were also seeded in differentiation medium and divided into blank control group （untreated）， solvent control group （4‰DMSO）， and 400 μg/mL LZD group； after 14 days of culture， platelet precursor generation was observed under the microscope； immunofluorescence staining was performed to count the proportion of cells producing pseudopodia， the relative length of pseudopodia was measured， and the expression levels of CD41 and CD42b mRNA were assessed. Cells from the solvent control group and the 400 μg/mL LZD group， cultured in differentiation medium for 14 days， were extracted and subjected to non-targeted metabolomics and targeted energy metabolomics analysis using liquid chromatography-tandem mass spectrometry. The relative content of pyruvate in cells， after being cultured for 14 days with the addition of pyruvate （10 mmol/L） or LZD （400 μg/mL）+pyruvate （10 mmol/L）， was measured and observed， as well as its effects on cell proliferation and platelet precursor generation. RESULTS 400 μg/mL LZD could significantly inhibit the proliferation of MEG-01 cells and the generation of platelet precursors （P＜0.01）. Non-targeted metabolomic analysis of MEG-01 cells after 400 μg/mL LZD treatment revealed significant changes in energy metabolism-related pathways such as mTOR signaling pathway， alanine， aspartate and glutamate metabolism， and central carbon metabolism in cancer. Targeted energy metabolomic analysis further showed that the relative contents of adenosine triphosphate， guanosine triphosphate， and pyruvate in MEG-01 cells were significantly reduced （P＜0.01）， while the relative contents of lactate were significantly increased （P＜0.01）. Compared with the LZD group， the relative content of pyruvate， cell count， the proportion of cells producing pseudopodia and the relative length of pseudopodia were significantly increased in the LZD+pyruvate group （P＜0.01）. CONCLUSIONS LZD may reduce pyruvate production by inhibiting mitochondrial energy metabolism， thereby suppressing megakaryocyte proliferation and platelet precursor production， ultimately leading to the occurrence of thrombocytopenia.

As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate?adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.

Objective To establish an intelligent discipline inspection and supervision platform in university-affiliated hospitals using modern information technology,strengthen the supervision and early warning of important integrity risks,promote the deep integration of manual supervision and intelligent supervision,and achieve full recording,tracing,and early warning of the exercise of power,allowing power to operate in the sunlight.Methods The intelligent discipline inspection and supervision platform information system adopts big data technology,focusing on the management of drugs and medical consumables,and es-tablishes a supervision platform and a standardized supervision system with five major risk analysis and early warning functions from the perspectives of hospitals,departments,and doctors.Results The construction of the intelligent discipline inspection and supervision platform has successfully achieved interoperability with the main core systems of the hospital,covering key busi-ness areas of the hospital.It has formed an integrated and coordinated working mechanism among the discipline inspection depart-ment,the department of ethical conduct,and the department of drug and consumable management,enhancing regulatory synergy.It gradually constructs a new supervision model of"strict management+intelligent management"and continuously optimizes and improves hospital management and service levels.Conclusion The construction of the intelligent discipline inspection and su-pervision platform in hospitals is conducive to enhancing the awareness of integrity risk prevention among Party members,cadres,and employees,realizing dynamic monitoring and early warning management of important integrity risks,promoting institutional reconstruction,process reengineering,and system reshaping,and helping to enhance the construction of Party conduct and integ-rity in university-affiliated hospitals.

Objective:To evaluate the efficacy and safety of standardized dust mite allergen subcutaneous immunotherapy (SCIT) in children with allergic rhinitis (AR) during treatment.Methods:A total of 283 children with AR diagnosed with definite dust mite allergy and completed 2 to 3 years of SCIT who attended the Department of Otorhinolaryngology Head and Neck Surgery, Xiangya Hospital, Central South University, from August 2019 to October 2021 were included, including 205 males and 78 females, with a mean age of 10.8 years. The total nasal symptoms score (TNSS), symptom medication score (SMS), rhinoconjunctivitis quality of life questionnaire (RQLQ) and visual analogue scale (VAS) before and after 2 to 3 years′ treatment were recorded, and the differences before and after treatment were compared. Adverse reactions during SCIT were recorded to evaluate its safety. SPSS 22.0 software was used for statistical analysis.Results:The overall effectiveness rate during SCIT in 283 children with AR was 89.4% (253/283). Compared with baseline, all symptom scores, medication scores and quality of life scores were significantly lower after 2 to 3 years of SCIT (all P<0.05). Further group comparisons showed positive efficacy in patients with different clinical characteristics, including age, gender, smoking status, family history of AR, symptom severity, mono-or poly-allergy, and second immunization, with no statistically significant differences between groups (all P>0.05). A total of 12 735 injections were administered during the SCIT, and a total of 213 (1.67%) injections of local adverse reactions occurred, mainly in the initial treatment phase, and the diameter of the local air mass was mostly 5 to 20 mm; 71 (0.56%) injections of systemic adverse reactions occurred, mainly in the initial treatment phase, and most of them were grade 1 reactions with no serious systemic adverse reaction such as shock. Conclusion:Standardized dust mite SCIT has a good safety profile and definite efficacy in treating AR children with different clinical characteristics. It can significantly improve all symptoms, reduce the use of symptomatic drugs and improve their quality of life.

Objective: To investigate the effects of sodium butyrate on intestinal barrier of the severe scald mice and the related mechanism. Methods: Eighteen C57BL/6 female mice, aged eight to twelve weeks, were divided into sham scald group, pure scald group, and scald+ sodium butyrate group according to random number table, with 6 mice in each group. Back of each mouse in pure scald group and scald+ sodium butyrate group were immersed into 90 ℃ water for 9 s, causing full-thickness scald of 30% total body surface area, while back of each mouse in sham scald group were immersed into 37 ℃ water for 9 s, causing sham injury. All of the mice in 3 groups were intraperitoneally injected with 1 mL sterile lactated Ringer′s solution immediately after injury. Besides, mice in scald+ sodium butyrate group were intraperitoneally injected with 300 mg/kg sodium butyrate at 30 min before injury and immediately after injury, while mice in sham scald group and pure scald group were intraperitoneally injected with the same volume of sterile phosphate buffer solution. At post injury hour (PIH) 24, portal vein of mice in 3 groups was harvested, intestinal permeability was measured by fluorescin isothiocyanate-dextran fluorescence probe tracing method, then lileal tissue of mice in 3 groups was harvested, protein expressions of zonula occludens l (ZO-1), occludin, claudin-1, claudin-2, nucleotide-binding oligomerization domain-containing protein-like receptor family pyrin domain containing 3 (NLRP3), interleukin-1β (IL-1β), and IL-18 were detected by Western blotting, and distribution of ZO-1 in intestinal mucosa was observed by indirect immunofluorescence. Data were processed with one-way analysis of variance, least-significant difference test, and Bonferroni correction. Results: (1) At PIH 24, the intestinal permeability of mice in sham scald group, pure scald group, and scald+ sodium butyrate group was 0.88±0.19, 2.62±0.48, 1.23±0.16, respectively. Compared with that in sham scald group, the intestinal permeability of mice in pure scald group was significantly elevated (P<0.01), while the intestinal permeability of mice in scald+ sodium butyrate group showed no obvious change (P>0.05). Compared with that in pure scald group, the intestinal permeability of mice in scald+ sodium butyrate group was significantly decreased (P<0.01). (2) At PIH 24, compared with those in sham scald group, the protein expressions of ZO-1, occludin, and claudin-1 of mice in pure scald group and scald+ sodium butyrate group were significantly decreased (P<0.05), while the protein expression of claudin-2 was significantly increased (P<0.05). At PIH 24, compared with those of pure scald group, the protein expressions of ZO-1 and occludin of mice in scald+ sodium butyrate group were significantly elevated (P<0.05), while the protein expression of claudin-2 was significantly decreased (P<0.05), the protein expression of claudin-1 showed no significant difference (P>0.05). (3) At PIH 24, compared with those in sham scald group, the protein expressions of NLRP3, IL-1β, and IL-18 of mice in pure scald group and scald+ sodium butyrate group were significantly increased (P<0.05). Compared with those of pure scald group, the protein expressions of NLRP3, IL-1β, and IL-18 of mice in scald+ sodium butyrate group were significantly decreased (P<0.05). (4) At PIH 24, ZO-1 in intestinal mucosa of mice in sham scald group was distributed smoothly, continuously and homogeneously along the membrane. ZO-1 in intestinal mucosa of mice in pure scald group was distributed unsmoothly with breaks. The distribution of ZO-1 in intestinal mucosa of mice in scald+ sodium butyrate group was ameliorated compared with that in pure scald group. Conclusions Sodium butyrate can inhibit the activation of NLRP3 inflammasome and decrease the production of IL-1β and IL-18 in intestinal mucosa of severe scald mice, which protects the intestinal barrier function by alleviating the alteration of tight junction protein expression and localization.

Objective To investigate the efficacy of total glucosides of paeony ( TGP ) in the treatment of hematuria and proteinuria purpura nephritis ( HSPN ) and serum interleukin?6 ( IL?6 ), interleukin?1 beta (IL?1 beta) and interleukin?18 (IL?18).Methods From June 2017 to December 2018, 64 cases of children with primary hematuria and proteinuria purpura nephritis admitted to the department of pediatrics affiliated hospital of Xuzhou medical university were selected as study subjects,and were divided into control group (33 cases) and observation group (31 cases) according to random number method.The children in the control group were given oral administration of poniasone acetate,and the observation group was treated with TGP on the basis of the control group.Both groups received continuous treatment for 4 weeks.Two groups′ before and after treatment expression level of serum interleukin?6, interleukin?1β and interleukin?18,and level of clinic indicator such as Urinary red biood cell (URBC),24 hour urinary protein quantity, urinary immune globulin G ( IGU ), micro?albuminuria ( urinary microalbumin, MAU ), α?Microglobulin (α1?MG) and urinary transferrin ( TFRU) is made a comparison.Results After 4weeks of treatment,the total clinical effective rate of the observation group (93.5%(29／31)) was significantly higher than that of the control group (72.7%(24／33)),and the difference between the two groups was statistically significant (χ2＝4.868,P＜0.05).Before treatment,difference between serum interleukin?6,interleukin?1β and interleukin?18 from two groups and each clinic indicators level has no statistic significance (all P＞0.05) .After treatment,in the observation group,Serum IL?6 ( (16.68±6.83) ng／L and (32.24±6.99) ng／L,t＝12.373),IL?1β((63.83 ±8.97)ng／L and (85.59±9.42) ng／L,t＝9.758),IL?18((64.52±5.46) ng／L and (88.50±5.54) ng／Ll,t＝18.899),24 h urinary protein quantification ( 0.3 (0.21,0.36) g／24 h and 1.0 (0.65,1.23) g／24 h,Z＝－4.861),URBC (15.30 (3.80,36.80)×106／L and 168.9 (58.4, 324.0)×106／L,Z＝－4.840),were lower than before treatment ( all P＜0.05); After treatment, in the control group,Serum IL?6 ((23.62±5.95) pg／ml and (33.44±4.68) pg／ml,t＝9.149),IL?1β ((68.67 ±6.31) pg／ml and (86.59±8.71) pg／ml,t＝10.617),IL?18((71.25±9.69) pg／ml and (89.87±6.68) pg／ml,t＝11.506),24 h urinary protein quantification (0.42 (0.33,0.56) g／24 h and 0.94 (0.74,1.25) g／24 h,Z＝－5.013),URBC ( 57.00 ( 39.25,77.50)×106／L and 145.60 ( 58.20,360.85)×106／L,Z＝－4.762),were lower than before treatment ( all P＜0.05).The difference between the two groups was statistically significant (P＜0.05).Conclusion The adjuvant effect of total glucoside of white peony root on hematuria and proteinuria purpura nephritis is significant, which can reduce the level of hematuria and proteinuria and improve the renal symptoms of children.Its mechanism of action may be to reduce renal inflammation and protect the kidney by down?regulating the expression of serum IL?6,IL?1 and IL?18.

Objective: To explore the limb salvage strategies for patients with high voltage electric burns of extremities on the verge of amputation. Methods: From January 2003 to March 2019, 61 patients with high voltage electric burns of extremities on the verge of amputation were treated in our hospital. All of them were male, aged 15-58 years, including 49 cases of upper limbs and 12 cases of lower limbs. The wound area after thorough debridement ranged from 15 cm×11 cm to 35 cm×20 cm. Emergency surgery for reconstruction of the radial artery with saphenous vein graft under eschar was performed in 5 cases. The arteries of 36 patients (including 7 cases with simultaneous ulnar artery and radial artery reconstruction) were reconstructed with various forms of blood flow-through after debridement, among them, the radial artery of 13 cases, the ulnar artery of 8 cases, the brachial artery of 8 cases, and the femoral artery of 2 cases were reconstructed with saphenous vein graft; the radial artery of 3 cases and the ulnar artery of 7 cases were reconstructed with the descending branch of the lateral circumflex femoral artery graft; the radial artery of 2 cases were reconstructed with greater omentum vascular graft; the reflux vein of 3 cases with wrist and forearm annular electric burns were reconstructed with saphenous vein graft. According to the actual situation of the patients, 12 cases of latissimus dorsi myocutaneous flap, 6 cases of paraumbilical flap, 28 cases of anterolateral thigh flap, 10 cases of abdominal combined axial flap, 5 cases of greater omentum combined with flap and/or skin grafts were used to repair the wounds after debridement and cover the main wounds as much as possible. Some cases were filled with muscle flap in deep defect at the same time. The area of tissue flaps ranged from 10 cm×10 cm to 38 cm×22 cm. For particularly large wounds and annular wounds, the latissimus dorsi myocutaneous flap, the paraumbilical flap, the abdominal combined axial flap, and the greater omentum combined with flap and/or skin grafts were used more often. Donor sites of three patients were closed directly, and those of 58 patients were repaired with thin and medium split-thickness skin or mesh skin grafts. The outcome of limb salvage, flap survival, and follow-up of patients in this group were recorded. Results: All the transplanted tissue flaps survived in 61 patients. Fifty-six patients had successful limb salvage, among them, 31 limbs were healed after primary surgery; 20 limbs with flap infection and tissue necrosis survived after debridement and flap sutured in situ; 5 limbs with flap infection, radial artery thrombosis, and hand blood supply crisis survived after debridement and radial artery reconstruction with saphenous vein graft. Five patients had limb salvage failure, among them, 3 patients with wrist electric burns had embolism on the distal end of the transplanted blood vessels, without condition of re-anastomosis, and the hands gradually necrotized; although the upper limb of one patient was salvaged at first, due to the extensive necrosis and infection at the distal radius and ulna and the existence of hand blood supply under flap, considering prognostic function and economic benefits, amputation was required by the patient; although the foot of one patient was salvaged at first, due to the repeated infection, sinus formation, extensive bone necrosis of foot under flap, dullness of sole and dysfunction in walking for a long time, amputation was required by the patient. During the follow-up of 6 months to 5 years, 56 patients had adequate blood supply in the salvaged limbs, satisfied appearance of flaps, and certain recovery of limb function. Conclusions Timely revascularization, early thorough debridement, and transplantation of large free tissue flap, combined tissue flap, or blood flow-through flap with rich blood supply are the basic factors to get better limb preservation and recovery of certain functions for patients with high voltage electric burns of limbs on the verge of amputation.

Objective: To investigate the significance of intestinal fatty acid binding protein (IFABP) in the evaluation of intestinal barrier dysfunction of mice at the early stage of severe burn injury. Methods: Thirty-six 8-week-old C57BL/6 male mice were collected and divided into normal control group (n=6) and scald group (n=30) according to random number table. Back of each mouse in scald group was placed into hot water of 90 ℃ for 10 s, causing full-thickness scald (hereinafter refer to as burn) of 30% total body surface area, while mice in normal control group were not inflicted with burns. Six mice in normal control group were taken, and 6 mice in scald group at 1, 2, 6, 12, and 24 h post injury were taken respectively. The portal vein blood of each mouse was extracted and the plasma was separated to measure intestinal permeability with fluorescin isothiocyanate-dextran fluorescence probe tracing method and plasma IFABP content by enzyme-linked immunosorbent assay. The distal ileum tissue of mice in normal control group and scald group at each time point post injury was collected to observe the morphology of the intestinal mucosa tissue by hematoxylin-eosin staining. Data were processed with one-way analysis of variance and Student-Newman-Keuls test, and pearson correlation test was used to analyze the correlation between intestinal permeability and plasma IFABP content of burned mice. Results: (1) At 1, 2, 6, 12, and 24 h post injury, the intestinal permeability of mice in scald group was 2.7±0.8, 5.4±2.5, 7.3±4.2, 12.4±6.1, 1.4±0.7, respectively, obviously higher than 1.0±0.4 of normal control group (P<0.05 or P<0.01). The intestinal permeability of mice in scald group showed an increasing trend post injury, reaching the peak at 12 h post injury, and rapidly falling back at 24 h post injury. (2) At 1, 2, 6, 12, and 24 h post injury, the plasma IFABP content of mice in scald group was (64±11), (59±12), (76±18), (111±22), and (66±10) ng/mL, obviously higher than (35±8) ng/mL in normal control group (P<0.05 or P<0.01). The plasma IFABP content of mice in scald group showed an increasing trend post injury, reaching the peak at 12 h post injury, and rapidly decreasing at 24 h post injury. (3) Uniform thickness of mucosa, intact epithelia, regularly arranged villi, and no inflammatory cell infiltration were observed in ileum of mice in normal control group. In ileum of mice in scald group, shortened villi of mucosa with different degrees, edema of lamina propria, and infiltration of neutrophils were observed at 1 and 2 h post injury; obviously damaged and partially exfoliated ileal mucosa, disorderly arranged and broken villi, degenerated and necrotic epithelial cells, dilated central lacteal, and infiltration of lymphocytes and neutrophils were observed at 6 and 12 h post injury; the damage of ileal mucosa was alleviated, and basically intact epithelia, dilated central lacteal, and infiltration of inflammatory cells were observed at 24 h post injury. (4) There was a significantly positive correlation between the intestinal permeability and the plasma IFABP content of burned mice (r=0.841, P<0.05). Conclusions The plasma IFABP can be used as a good biological indicator for the evaluation of intestinal barrier dysfunction of mice at the early stage of severe burn injury.

OBJECTIVE： To establish the method for simultaneous determination of clopidogrel （CLP）， its intermediate metabolite （2-O-CLP）， inactive metabolite （CLPCA） and active metabolite （CLPTM） in human plasma. METHODS： Totally 90 patients diagnosed as stroke were selected from the First Affiliated Hospital of Army Medical University. They were given one CLP tablet （75 mg/tablet） orally on an empty stomach in the morning. Blood samples were collected 2 h after taking the tablet. CLPTM- D was formed by derivation of CLPTM with 2-bromo-3’-methoxyacetophenone and extracted by precipitation of acetonitrile protein together with the other three substances to be measured. LC-MS/MS method was adopted. The determination was performed on Agilent poroshell 120 EC-C18 column with mobile phase consisted of acetonitrile （0.1％ formic acid） and water （0.1％ formic acid） （90 ∶ 10， V/V）. The quantitation analysis was performed using multiple reaction monitoring at the specific ion transitions of m/z 308.1→198.1 （CLPCA）， 322.3→212.0 （CLP）， 338.3→155.0 （2-O-CLP）， 504.4→354.1 （CLPTM-D） and 264.0→154.1 （ticlopidine， internal standard）， respectively. RESULTS： The retention time of CLPCA， CLP， 2-O-CLP， CLPTM-D and internal standard were 2.01， 3.32， 2.83， 2.68， 1.87 min， respectively. The linear range of CLPCA， CLP， 2-O-CLP and CLPTM-D were 100-10 000， 0.2-20， 0.3-30， 0.5-50 ng/mL （all r≥0.999 5）. The intra-day and inter-day RSD were all less than 9.5％ （n＝5）. Accuracy ranged from 93.5％-98.9％ （n＝5）， and extraction recovery was from 85.4％ to 95.9％ （n＝5）. The matrix effect ranged from 2.7％-6.2％ （n＝5）. In stability tests （storing at －80 ℃ for 3 months， 3 freeze-thaw cycles， storing at 4 ℃ for 8 h）， RE of CLP， CLPCA and CLPTM-D were all lower than 10.0％ （n＝5）. CONCLUSIONS： Established LC-MS/MS method has the advantages of high specificity， accuracy and reliability， and can be used to detect the concentration of CLP and its three metabolites in human plasma.

Objective To report the clinical outcomes of latissimus dorsus flap used to reconstruct severe trauma on lower leg.Methods From May 2011 to September 2016,12 patients were treated for severe trauma on lower leg at Orthopaedic Department,General Hospital of Rocket Force.They were 10 men and 2 women,aged from 22 to 54 years (average,37.5 years).According to the Gustilo classification,their open fractures were of type ⅢA in 2 cases and of type ⅢB in 10.The areas of soft tissue defect ranged from 15 cm × 8 cm to 35 cm × 12 cm.Their wounds were covered with latissimus dorsus flaps;the dorsal expansion of the ankle joint was reconstructed after their bone infection was controlled.Two flaps were anastomosed directly to the anterior tibial vessels and one flap to the femoral vessels.The other 9 flaps were anatomosed to the lateral circumflex femoral artery,bridging with the greater saphenous vein (average length:32 cm).Ten flaps were neuroanatomosed with the motor branch of common peroneal nerve and 2 with the motor branch of vastus lateralis.The therapeutic outcomes were evaluated in terms of repair of soft tissue defects,control of infection and range of active dorsal flexion of the ankle at neutral position.Results The 12 patients were followed up for an average of 3.5 years (from 1 to 6 years).All the 12 flaps survived and infections were eradicated.The fractures were healed with muscle strength of grade 3 to grade 4.Their ankle extension was reconstructed.Their limbs were preserved with satisfactory limb function.The bone exposures of 5 cm × 3 cm and 6 cm × 3 cm in area after debridement respectively in 2 cases of Gustilo type ⅢA were covered with latissimus dorsus flaps.The range of active dorsal flexion of the ankle at neutral position increased from preoperative 10.5° to postoperative 19.1° on average.Conclusion In treatment of severe trauma on lower leg,latissimus dorsus flaps can be used to repair skin wound,control infection and reconstruct ankle function simultaneously,with advantages of reduced operative procedures,a high rate of limb preservation,and better function recovery.