Objective:To evaluate Masquelet technique plus flap transfer in repair of tibial infected defects complicated with extensive soft tissue defects in the lower leg.Methods:A retrospective analysis was performed in the 23 patients who had been treated by Masquelet technique plus flap transfer at Institute of Orthopedics and Trauma, 920 Hospital for tibial infected defects complicated with extensive soft tissue defects in the lower leg from March 2016 to June 2019. They were 15 males and 8 females, aged from 18 to 59 years (average, 38.4 years). The duration of disease ranged from 6 to 312 months (average, 23.6 months). All patients underwent surgery by 2 stages:1) debridement, locking compression plate fixation, formation of induced membrane by antibiotic-loaded bone cement, and repair of soft tissue defects with lower leg flaps; 2) removal of bone cement and fixation 6 to 8 weeks after infection control, fixation of broken ends after rinse, followed by grafting of cancellous bone particles in the induced membrane. The area of wound soft tissue defects after debridement ranged from 4.0 cm × 3.5 cm to 18.0 cm × 6.0 cm, and the length of bone defects from 6 to 12 cm (average, 8.4 cm). Locally grafted were pedicled fasciocutaneous flap in 4 cases, sural nerve nutrition skin flap in 9 cases (including 4 anterograde and 5 retrograde ones), saphenous nerve nutrition vascular flap in 7 cases (including 2 anterograde and 5 retrograde ones), retrograde superficial peroneal nerve nutrient vessel flap in one and free flap in 2 cases. The curative efficacy was evaluated according to the Paley fracture healing scores.Results:All the 23 patients were followed up for 9 to 46 months (average, 15.6 months). Flaps healed by the first stage in 18 cases and after skin grafting in 3 cases; skin flap transfer was conducted again in 2 cases. Infection was controlled in 21 cases but recurred in 2 cases at 9 and 14 months respectively after secondary surgery. The time for bone reunion ranged from 4 to 11 months (average, 6.2 months). According to the Paley criteria for fracture healing, 21 cases were excellent, one was good and one poor.Conclusion:In the treatment of tibial infected defects complicated with extensive soft tissue defects, Masquelet technique plus transfer of a variety of lower leg flaps can result in reliable outcomes because it controls infection, promotes formation of complete induced membrane and accelerates the process of bone reconstruction along with repair of soft tissue defects.

Objective To observe the clinical outcomes of the superior gluteal neurocutaneous flap for sacrococcygeal pressure sores. Methods Twelve cases with sacrococcygeal pressure sores were covered by the superior gluteal neurocutaneous flap from May 2005 to Nov. 2009. The sore size ranged from 15 cm ×30 cm to 5 cm × 8 cm, while the flap size ranged from 17 cm × 32 cm to 10 cm× 12 cm. Results All 12 flaps survived totally with the pressure sores healed. The longest follow-up time was four years, the short follow-up time was half a year, the average time was 2.5 years. The superior gluteal neurocutaneous flap was good blood circulation, pressure sores not recur. Conclusion The superior gluteal neurocutaneous flap is a good treatment for sacrococcygeal pressure sores for its reliable blood supply and simple harvesting.

Aim To observe the effect of total flavonoid from Mori folium(TFMF) on renal interstitial fibrosis in type 1 diabetic mice and its possible mechanism.Methods Diabetic mice were induced by intraperitoneal injection of streptozotocin(STZ) dissolved in 0.01 mol·L-1 citrate buffer(pH 4.5) at 150 mg·kg-1 body weight after 12 h of food deprivation.Forty model mice were divided randomly into four groups: model group, and low-(0.25 g·kg-1), moderate-(0.5 g·kg-1), high-dose groups(1 g·kg-1) fed with TFMF once daily.In addition, eight normal mice were used as normal group.After 12 weeks, the fasting blood glucose(FBG), serum creatinine(Cr), blood urea nitrogen(BUN) and microalbuminuria(mAlb) were measured.Masson staining, Sirius red staining and collagen type Ⅳ immunohistochemical staining were used to detect the expression of collagen protein in the cortex, while laminin staining to assess the degree of glomerular and renal tubular basement membrane thickening.The protein expressions related to epithelial-mesenchymal transition and PI3K/Akt/mTOR in the renal cortex of mice were detected by Western blot.Results The moderate and high dose of TFMF could significantly decrease the levels of FBG, Cr, BUN and mAlb in diabetic mice, meanwhile decreasing the expression of α-SMA protein by inhibiting the activation of PI3K/Akt/mTOR signaling pathway, which led to the amelioration of the pathological alterations of renal tissue.Conclusions The moderate and high dose of TFMF can reduce the level of renal interstitial fibrosis in type 1 diabetic mice, and its mechanism may be related to the inhibition of activation of PI3K/Akt/mTOR signaling pathway.

Objective:To investigate the clinical effect of induced membrane technique combined with staged internal fixation for treatment of infected femoral nonunion.Methods:A retrospective case series study was conducted to analyze the clinical data of 21 patients with infected femoral nonunion treated from January 2016 to December 2018 in 920th Hospital of Joint Logistics Support Force of PLA. There were 13 males and 8 females, with the age of 18-57 years [(38.9±6.7)years]. The duration of nonunion was 7-78 months [(27.1±11.4)months]. All patients were treated by induced membrane technique in two stages. At stage I, the original internal fixation was removed and debrided thoroughly, then the antibiotic-loaded bone cement and locking compression plate (LCP) were placed. The length of bone defect following debridement was 5-15 cm[(7.4±1.9)cm]. At stage II, the bone defect was reconstructed with bone grafts and fixed with the intramedullary nail and/or LCP. The wound condition, white blood cell count, C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were recorded after stage I surgery and at the last follow-up to measure infection control. The complications and bone healing time were recorded. The bone healing was evaluated by the Paley criteria and the functional recovery of the affected limb was evaluated by the range of motion of the knee at the last follow-up.Results:All patients were followed up for 23-43 months [(31.9±6.7)months]. The infection recurred in 4 patients after stage I surgery, and the wound healed after repeated debridement. There was no infection recurrence after stage II surgery. The white blood cell count, CRP and ESR were (6.1±1.8)×10 9/L, (10.1±3.1)mg/L, (10.2±3.4)mm/h at the last follow-up, significantly decreased from preoperative (15.0±4.8)×10 9/L, (69.8±14.8)mg/L, (66.2±13.2)mm/h ( P<0.05). The incidence of complications was 43%. Besides infection recurrence in 4 patients after stage I surgery, the donor site at the posterior superior iliac spine in 3 patients showed delayed healing, and the limb shortening occurred in 2 patients with the discrepancy of 3 cm and 4 cm. Bony union was observed in all patients within 6-16 months [(8.8±2.7)months]. The results were excellent in 19 patients and good in 2 patients according to the Paley criteria at the last follow-up. The knee range of motion was significantly improved from preoperative 30.0°(15.0°, 110.0°) to 90.0°(61.5°, 120.0°) at the last follow-up ( P<0.05). Conclusion:For infected femoral nonunion, the induced membrane technique combined with staged internal fixation can effectively control infection, achieve bony union, and promote functional recovery.

Objective To investigate the potential value of exosomes derived from rat ectoderm mesenchymal stem cells(EMSCs-exo)for repairing secondary spinal cord injury.Methods EMSCs-exo were obtained using ultracentrifugation from EMSCs isolated from rat nasal mucosa,identified by transmission electron microscope,nanoparticle tracking analysis(NTA),and Western blotting,and quantified using the BCA method.Neonatal rat microglia purified by differential attachment were induced with 100 μg/L lipopolysaccharide(LPS)and treated with 37.5 or 75 mg/L EMSCs-exo.PC12 cells were exposed to 400 μmol/L H2O2 and treated with EMSCs-exo at 37.5 or 75 mg/L.The protein and mRNA expressions of Arg1 and iNOS in the treated cells were determined with Western blotting and qRT-PCR,and the concentrations of IL-6,IL-10,and IGF-1 in the supernatants were measured with ELISA.The viability and apoptosis of PC12 cells were detected using CCK-8 assay and flow cytometry.Results The isolated rat EMSCs showed high expressions of nestin,CD44,CD105,and vimentin.The obtained EMSCs-exo had a typical cup-shaped structure under transmission electron microscope with an average particle size of 142 nm and positivity for CD63,CD81,and TSG101 but not vimentin.In LPS-treated microglia,EMSCs-exo treatment at 75 mg/L significantly increased Arg1 protein level and lowered iNOS protein expression(P<0.05).EMSCs-exo treatment at 75 mg/L,as compared with the lower concentration at 37.5 mg/L,more strongly increased Arg1 mRNA expression and IGF-1 and IL-10 production and decreased iNOS mRNA expression and IL-6 production in LPS-induced microglia,and more effectively promoted cell survival and decreased apoptosis rate of H2O2-induced PC12 cells(P<0.05).Conclusion EMSCs-exo at 75 mg/L can effectively reduce the proportion of M1 microglia and alleviate neuronal apoptosis under oxidative stress to promote neuronal survival,suggesting its potential in controlling secondary spinal cord injury.

Objective To investigate the potential value of exosomes derived from rat ectoderm mesenchymal stem cells(EMSCs-exo)for repairing secondary spinal cord injury.Methods EMSCs-exo were obtained using ultracentrifugation from EMSCs isolated from rat nasal mucosa,identified by transmission electron microscope,nanoparticle tracking analysis(NTA),and Western blotting,and quantified using the BCA method.Neonatal rat microglia purified by differential attachment were induced with 100 μg/L lipopolysaccharide(LPS)and treated with 37.5 or 75 mg/L EMSCs-exo.PC12 cells were exposed to 400 μmol/L H2O2 and treated with EMSCs-exo at 37.5 or 75 mg/L.The protein and mRNA expressions of Arg1 and iNOS in the treated cells were determined with Western blotting and qRT-PCR,and the concentrations of IL-6,IL-10,and IGF-1 in the supernatants were measured with ELISA.The viability and apoptosis of PC12 cells were detected using CCK-8 assay and flow cytometry.Results The isolated rat EMSCs showed high expressions of nestin,CD44,CD105,and vimentin.The obtained EMSCs-exo had a typical cup-shaped structure under transmission electron microscope with an average particle size of 142 nm and positivity for CD63,CD81,and TSG101 but not vimentin.In LPS-treated microglia,EMSCs-exo treatment at 75 mg/L significantly increased Arg1 protein level and lowered iNOS protein expression(P<0.05).EMSCs-exo treatment at 75 mg/L,as compared with the lower concentration at 37.5 mg/L,more strongly increased Arg1 mRNA expression and IGF-1 and IL-10 production and decreased iNOS mRNA expression and IL-6 production in LPS-induced microglia,and more effectively promoted cell survival and decreased apoptosis rate of H2O2-induced PC12 cells(P<0.05).Conclusion EMSCs-exo at 75 mg/L can effectively reduce the proportion of M1 microglia and alleviate neuronal apoptosis under oxidative stress to promote neuronal survival,suggesting its potential in controlling secondary spinal cord injury.

Objective:To evaluate the efficacy and tolerability of crisaborole 2% ointment in the treatment of childhood atopic dermatitis (AD) at the early stage, and to compare the efficacy of every-other-day (Qod) regimen versus twice-a-week (Biw) regimen against recurrence in the remission stage of AD.Methods:A multicenter, randomized, open-label clinical trial was conducted. Totally, 150 children with mild to moderate AD aged 2 - < 18 years were enrolled from 6 hospitals (including Beijing Children′s Hospital, Capital Medical University, etc), and randomly divided into the Qod group (76 cases) and the Biw group (74 cases). In the acute stage of AD, both groups were treated with topical crisaborole 2% ointment on skin lesions twice a day for 2 - 4 weeks, as well as with emollients throughout the whole body. The improvement of early clinical symptoms was evaluated, and the occurrence of adverse reactions was recorded in the follow up. Once the investigator′s static global assessment (ISGA) scores decreased to 1 point or less, the patient would be enrolled into the remission stage. In the remission stage of AD, patients in the Qod group and Biw group were treated with crisaborole ointment every other day and twice a week respectively; the recurrence rate of AD in the remission stage was evaluated, as well as the severity of skin lesions, itching, life quality, and the occurrence of adverse reactions at weeks 4, 8, and 12. Statistical analysis was carried out with SPSS 23.0 software by using t test for comparisons of normally distributed continuous data between two groups, Mann-Whitney U test for non-normally distributed data, chi-square test for enumeration data, and Kaplan-Meier method for analysis of survival rates. Results:A total of 142 patients were enrolled in the modified intention-to-treat population, including 71 in the Qod group and 71 in the Biw group. In the acute stage of AD, the improvement of itching and skin lesions self-reported by the children or their family members occurred on days 1.9 (1.0, 3.0) and 2.0 (1.0, 4.1) after the application of crisaborole ointment, respectively. At the end of treatment in the acute stage, 89 children (62.7%) achieved ISGA 0/1 and successfully transferred into the remission stage. The follow-up in the remission stage was completed in 83 patients (44 in the Qod group and 39 in the Biw group). In addition, recurrence occurred in 19 (43.2%) and 12 (30.8%) patients in the Qod group and Biw group respectively, and there was no significant difference in the recurrence rate between the two groups ( χ2 = 1.36, P = 0.243) ; the average time to recurrence was 64.25 (95% CI: 53.33 - 75.17) days and 75.78 (95% CI: 65.46 - 86.10) days in the Qod group and Biw group respectively. Among the patients who were in the remission stage and had not yet experienced relapse at weeks 4, 8, and 12, there were no significant differences in the eczema area and severity index (EASI) scores, ISGA scores, pruritus numerical rating scale (NRS) scores, or quality-of-life scores between the two groups (all P > 0.05) at any time points, except for the ISGA scores at week 12 (Biw group: 0 [0, 1] point vs. Qod group: 1 [0, 1] point; Z = -2.31, P = 0.021). A total of 146 patients were enrolled in the safety set. During the study period, 70 adverse events occurred in 65 patients, with an incidence rate of 44.5%, and all were mild or moderate adverse events; 55 (37.7%) patients experienced discomfort at the medication site, which mainly referred to pain (45 cases, 30.8%) and mostly occurred in the tender and skinfold areas. Conclusions:Crisaborole 2% ointment could effectively relieve clinical symptoms in children with mild to moderate AD in the early stage, and intermittent treatment could continuously relieve clinical symptoms in the remission stage. The common adverse reaction was discomfort at the application site in the early stage of AD. There was no significant difference in the impact on AD recurrence in the remission stage between the Qod regimen and Biw regimen.

Objective RNA sequencing(RNA-seq)and bioinformatic analysis were combined and used to explore the anti-inflammatory effects and mechanisms of naringenin(Nar)in lipopolysaccharide(LPS)-stimulated human peri-odontal ligament stem cells(hPDLSCs).Methods Cell counting kit-8,quantitative real-time reverse transcription poly-merase chain reaction(qRT-PCR),and enzyme-linked immunosorbent assay(ELISA)were adopted to detect the effects of Nar on the proliferation and expression of inflammatory factors in LPS-stimulated hPDLSCs,screening for the opti-mal anti-inflammatory concentration of Nar.Differentially expressed genes(DEGs)were screened using|log2FC|≥1 and P≤0.05 as criteria.Volcano plot analysis,Kyoto En-cyclopedia of Genes and Genomes(KEGG)pathway en-richment analysis,the String database,and the MCODE module of Cytoscape were utilized to select core genes and enriched pathways.The effects on the nuclear factor κB(NF-κB)signaling pathway were verified using ELISA,qRT-PCR,and Western blot.Results Appropriate concentrations of Nar could alleviate the expression of inflammatory fac-tors and promote the proliferation of hPDLSCs stimulated by LPS.The best anti-inflammatory effect was achieved with 20 μmol/L Nar.RNA-seq showed significant enrichment of inflammation-related signaling pathways.The anti-inflamma-tory effect of Nar was mediated by inhibiting the NF-κB signaling pathway,similar to the effect of the NF-κB inhibitor BAY 11-7802.Conclusion Nar could exert its anti-inflammatory effects by inhibiting the NF-κB signaling pathway,making it a potential therapeutic option for the adjuvant treatment of periodontitis.

The construction of experimental animal models plays an important supporting role in research into the mechanisms of action of Chinese medicines.There have been increasing reports of the construction and evaluation of animal models of spleen deficiency;however,the construction method have involved different standards and there has been insufficient objectification of the evaluation indexes.In this review,we summarize the construction and evaluation method of animal models of spleen deficiency from the aspects of animal selection,model establishment,macroscopic characterization,behavioral experiments,and objective indexes of spleen deficiency,with a view to providing theoretical guidance for the construction of experimental animal models of spleen deficiency and references for the selection of animal model platforms for spleen deficiency.