BACKGROUND:Irisin,a myokine isolated from the transmembrane protein FNDC5 by muscle cells during exercise,has the function of inducing the browning of white adipose tissue,but its effect on lipotoxicity-induced osteogenic differentiation and the mechanism is unclear. OBJECTIVE:To investigate the effect of irisin on the osteogenic ability of palmitic acid-induced bone marrow mesenchymal stem cells and the mechanism of action. METHODS:CCK-8 assay was used to detect the effect of different concentrations of palmitic acid on the proliferation of mouse bone marrow mesenchymal stem cells and the effect of irisin on the proliferation of mouse bone marrow mesenchymal stem cells in the presence of palmitic acid.After pretreatment with irisin and palmitic acid for 24 hours,osteogenic differentiation of mouse bone marrow mesenchymal stem cells was induced by alkaline phosphatase staining as well as qRT-PCR was performed to detect the expression of osteogenesis-related genes on day 7 of osteogenic induction culture.The expression of proteins related to the AMPK/BMP2/SMAD signaling pathway was detected by western blot assay.Alizarin red staining was conducted on day 21 to detect osteogenic differences. RESULTS AND CONCLUSION:(1)The CCK-8 assay results suggested that the amplification of bone marrow mesenchymal stem cells was inversely proportional to the concentration of palmitic acid,but at 0.02 mmol/L concentration,palmitic acid had no significant effect on the amplification of bone marrow mesenchymal stem cells,and irisin did not affect the proliferation of bone marrow mesenchymal stem cells when its mass concentration was in the range of 0.1-20 μg/L.(2)Alkaline phosphatase staining and alizarin red staining showed that palmitic acid inhibited the osteogenic differentiation ability of bone marrow mesenchymal stem cells.Irisin improved palmitic acid-induced osteogenic inhibition of bone marrow mesenchymal stem cells.qRT-PCR results showed that palmitic acid could cause the downregulation of osteogenic-related genes,and irisin could inhibit this trend.(3)Western blot assay results showed that compared with the palmitic acid intervention group,irisin treatment enhanced AMPK/BMP2/SMAD signal transduction in bone marrow mesenchymal stem cells.It is found that irisin can improve the osteogenic differentiation ability of bone marrow mesenchymal stem cells pretreated with palmitic acid,and proposed that the specific mechanism might be mediated by AMPK/BMP/SMAD signaling pathway.

BACKGROUND:At present,the treatment methods for necrotizing fasciitis mostly use negative pressure sealing suction after thorough debridement.This method requires repeated debridement to completely remove necrotic infected tissue,causing serious physical and economic burdens to patients. OBJECTIVE:To introduce a rare clinical case of calf compartment syndrome caused by diabetic foot necrotizing fasciitis,and summarize the clinical experience of using antibiotic-loaded bone cement for treatment and comprehensive management. METHODS:A total of 6 patients with calf compartment syndrome caused by diabetic necrotizing fasciitis admitted to Wuxi 9th Affiliated Hospital of Soochow University from August 2017 to August 2020 were selected,including 5 males and 1 female with an average age of 54 years.During the perioperative period,the patients'general condition was evaluated and systemic nutritional support treatment was given.In the first stage,all patients received complete debridement to control infection,antibiotic-loaded bone cement packing,and negative pressure sealed drainage.In the second stage,bone cement was removed and wound repair was performed.The wound healing,as well as the occurrence of redness,swelling,and exudation was observed during the follow-up. RESULTS AND CONCLUSION:(1)The wounds of four patients were fresh after twice antibiotic-loaded bone cement packing,and the membrane formation was good,and one patient was good after three times of antibiotic-loaded bone cement packing,and the wounds of all five patients healed well after the second stage of skin grafting.Due to the difficulty in maintaining intraoperative blood pressure and infection in all four compartments of the lower leg,a patient underwent emergency knee amputation.Meanwhile,the stump wound was placed with antibiotic-loaded bone cement.The wound was closed directly after the secondary bone cement was removed,and the wound healed in the first stage.(2)The six patients were followed up for 6-24 months after discharge.At the last follow-up,all six patients had good wound healing and no symptoms such as redness,swelling,and exudation.The quality of life of the patients was significantly improved,and all of them were satisfied with the curative effect.(3)The occurrence of calf compartment syndrome should be vigilant when diabetic foot necrotizing fasciitis is highly suspected.Early diagnosis and timely incision decompression are of great importance.Besides,the application of antibiotic-loaded bone cement in the treatment of calf compartment syndrome caused by diabetic necrotizing fasciitis has a good short-term effect.

Objective:To compare the palmar minimally invasive approach by a single incision versus palmar traditional approach for implantation of a locking plate in the treatment of distal radius fractures.Methods:A retrospective analysis was conducted of the 116 patients with distal radius fracture who had been treated with locking plates implanted through a palmar approach at Department of Traumatic Orthopedics, Wuxi Ninth People's Hospital from January 2019 to December 2022. There were 51 males and 65 females aged (57.0±10.6) years. According to the difference in palmar approach, they were divided into a traditional group of 65 cases treated with a traditional palmar approach and a minimally invasive group of 51 cases treated with a minimally invasive palmar approach through a single 1.5 cm incision. The surgical time, intraoperative bleeding, incision length, incision healing, fracture healing time, pain visual analogue scale (VAS) on the 1st and 7th days after surgery, range of motion and grip strength of the affected wrist, and imaging results at the last follow-up were recorded and compared between the 2 groups. The patient rated wrist evaluation (PRWE) and Gartland-Werley wrist evaluation were used to assess functional recovery of the wrist affected at the last follow-up.Results:There was no statistically significant difference in the baseline data between the 2 groups, indicating comparability ( P>0.05). All patients were followed-up for (13.6±2.1) months. All fractures healed. The intraoperative bleeding [(27.3±3.6) mL] in the minimally invasive group was significantly less than that in the traditional group [(41.2±6.8) mL], the incision length [(1.6±0.1) cm] in the former was significantly shorter than that in latter [(6.6 ± 0.6) cm], the postoperative VAS for pain on the 7th day after surgery [(1.8±0.8) points] and incidence of complications [2.0% (1/51)] in the former were significantly lower than those in the latter [(3.0±0.6) points, 12.3% (8/65)] ( P<0.05). However, the surgical time [(70.4±6.9) min] in the former was significantly longer than that in the latter [(67.2±8.0) min] ( P<0.05). There was no statistically significant difference between the 2 groups in terms of incision healing, fracture healing time, VAS for pain on the first postoperative day, wrist range of movement at the last follow-up, grip strength, PRWE score, Gartland-Werley functional recovery, fracture alignment, palm angle, ulnar deviation angle, and radial height at the last follow-up ( P>0.05). Conclusions:Both the palmar minimally invasive approach by a single incision and the traditional palmar approach are effective for implantation of a locking plate in the treatment of distal radius fractures. However, the minimally invasive palmar approach is superior to the traditional palmar approach in terms of incision aesthetics, intraoperative bleeding, surgical trauma response, surgical time and surgical risk.

Objective:To investigate the effect of sodium tanshinone ⅡA sulfonate (STS) on pyroptosis of human umbilical vein endothelial cells (HUVECs) induced by H 2O 2 and its possible mechanism. Methods:From November 2021 to September 2022, HUVECs were used as the research subjects at Wuxi Ninth People’s Hospital. The experiment was divided into four groups: the blank control group (normal condition), blank + STS group, H 2O 2 group and H 2O 2 + STS group. When the cells reached 80% fusion, 500.00 μmol/L of H 2O 2 was added to H 2O 2 group and H 2O 2 + STS group for 3 hours, and then the medium containing 500.00 μmol/L H 2O 2 was removed. After that, the blank+ STS group and the H 2O 2+ STS group were each supplemented with 5.00 μg/ml of STS and co-cultured with HUVECs for 24 hours. CCK-8 was used to assess the impact of STS at various concentrations (0.00, 0.05, 0.50, 5.00, 50.00, 500.00 μg/ml) on the proliferation of HUVECs. DNA damage-positive cells were detected with TUNEL staining. The expression of NOD-like receptor protein 3 (NLRP3) was detected using real-time PCR (RT-PCR) to investigate the optimal concentration of pyroptosis induced by H 2O 2. A detection kit was used to measure the expression of reactive oxygen species (ROS) induced by H 2O 2. The effect of STS on the migration and tube formation of HUVECs during pyroptosis was examined using a cell scratch test and a matrix gel tube formation test. The expressions of NLRP3, caspase-1, interleukin-18, and interleukin-1β were detected using RT-PCR and Western blotting. Repeated measures ANOVA was used to compare the concentrations at different time points, t-tests were used to compare data between two groups, and one-way ANOVA was used to compare data between multiple groups. P<0.05 was considered statistically significant. Results:STS below 50.00 μg/ml had no effect on the proliferation of HUVECs, while 500.00 μmol/L H 2O 2 had the most significant effect on inducing pyroptosis in HUVECs. TUNEL staining showed that compared with the control group, the number of TUNEL-positive cells in H 2O 2 group was significantly increased, and the difference was statistically significant ( P<0.01). However, there was no significant difference in the number of TUNEL-positive cells in the H 2O 2+ STS group ( P>0.05). The results of ROS detection showed that compared with the H 2O 2 group, intracellular ROS levels in the H 2O 2+ STS group was significantly decreased, and the difference was statistically significant ( P<0.01). Cell scratch and tube formation in vitro experiments showed that compared with the control group, cell mobility and tube formation ability were significantly decreased in the H 2O 2 group (all P<0.01), and there was no statistical significance in the H 2O 2+ STS group (all P>0.05). RT-PCR and Western blotting results showed that, compared with the H 2O 2 group, the expression of pyroptosis-related factors in the H 2O 2+ STS group was significantly decreased (all P<0.05). Conclusion:STS can inhibit the excessive production of ROS, promote the cell migration and tubular formation of HUVECs after pyroptosis induction, and alleviate H 2O 2-induced pyroptosis of HUVECs, thereby promoting angiogenesis.

Replantation of traumatic amputation in children has its own characteristics. This consensus primarily focuses on the issues related to the treatment of traumatically amputated limb injuries in children. Organised along a timeline, the consensus summarises domestic and international clinical experiences in emergency care and injury assessment of traumatic limb amputation limbs, indications and contraindications for replantation surgery, principles and procedures of replantation surgery, postoperative medication and management, as well as rehabilitation in children. The aim of this consensus is to propose standardise the treatment protocols for limb replantation for children therefore to serve as a reference for clinical practitioners in medical practices, and further improve the treatment and care for the traumatic limb amputations in children.

Objective:To investigate the effect of sodium tanshinone ⅡA sulfonate (STS) on pyroptosis of human umbilical vein endothelial cells (HUVECs) induced by H 2O 2 and its possible mechanism. Methods:From November 2021 to September 2022, HUVECs were used as the research subjects at Wuxi Ninth People’s Hospital. The experiment was divided into four groups: the blank control group (normal condition), blank + STS group, H 2O 2 group and H 2O 2 + STS group. When the cells reached 80% fusion, 500.00 μmol/L of H 2O 2 was added to H 2O 2 group and H 2O 2 + STS group for 3 hours, and then the medium containing 500.00 μmol/L H 2O 2 was removed. After that, the blank+ STS group and the H 2O 2+ STS group were each supplemented with 5.00 μg/ml of STS and co-cultured with HUVECs for 24 hours. CCK-8 was used to assess the impact of STS at various concentrations (0.00, 0.05, 0.50, 5.00, 50.00, 500.00 μg/ml) on the proliferation of HUVECs. DNA damage-positive cells were detected with TUNEL staining. The expression of NOD-like receptor protein 3 (NLRP3) was detected using real-time PCR (RT-PCR) to investigate the optimal concentration of pyroptosis induced by H 2O 2. A detection kit was used to measure the expression of reactive oxygen species (ROS) induced by H 2O 2. The effect of STS on the migration and tube formation of HUVECs during pyroptosis was examined using a cell scratch test and a matrix gel tube formation test. The expressions of NLRP3, caspase-1, interleukin-18, and interleukin-1β were detected using RT-PCR and Western blotting. Repeated measures ANOVA was used to compare the concentrations at different time points, t-tests were used to compare data between two groups, and one-way ANOVA was used to compare data between multiple groups. P<0.05 was considered statistically significant. Results:STS below 50.00 μg/ml had no effect on the proliferation of HUVECs, while 500.00 μmol/L H 2O 2 had the most significant effect on inducing pyroptosis in HUVECs. TUNEL staining showed that compared with the control group, the number of TUNEL-positive cells in H 2O 2 group was significantly increased, and the difference was statistically significant ( P<0.01). However, there was no significant difference in the number of TUNEL-positive cells in the H 2O 2+ STS group ( P>0.05). The results of ROS detection showed that compared with the H 2O 2 group, intracellular ROS levels in the H 2O 2+ STS group was significantly decreased, and the difference was statistically significant ( P<0.01). Cell scratch and tube formation in vitro experiments showed that compared with the control group, cell mobility and tube formation ability were significantly decreased in the H 2O 2 group (all P<0.01), and there was no statistical significance in the H 2O 2+ STS group (all P>0.05). RT-PCR and Western blotting results showed that, compared with the H 2O 2 group, the expression of pyroptosis-related factors in the H 2O 2+ STS group was significantly decreased (all P<0.05). Conclusion:STS can inhibit the excessive production of ROS, promote the cell migration and tubular formation of HUVECs after pyroptosis induction, and alleviate H 2O 2-induced pyroptosis of HUVECs, thereby promoting angiogenesis.

Objective To investigate the effect of lifestyle on high-altitude de-adaptation(HADA)through questionnaire during the whole process of entering and existing the plateau in order to provide scientific basis for prevention and treatment of the disease.Methods A case-control trial was conducted on 1 751 participants from a certain unit who entered and existed the plateau together during 2021 and 2022.In 1 to 2 weeks after they returning from the plateau,they were surveyed,and finally,1 544 valid questionnaires were obtained.According to the score of the plateau deacclimation scale ≥6 or not,the subjects were divided into plateau deacclimation group(n=192)and control group(n=1 352).They were further surveyed for their lifestyles and general conditions.Rank sum test was used to analyze the differences of lifestyles between the 2 groups,and unconditional logistic regression analysis was employed to identify the independent risk factors of HADA.Results Hair loss(19.95％),drowsiness(16.58％)and tiredness(12.31％)were the most common symptoms of HADA.High salt diet before entering the plateau,smoking at the plateau,altitude sickness(OR=1.893,95％CI:1.142～3.137,P=0.013),leaving the plateau by plane(OR=1.688,95％CI:1.082～2.634,P=0.021),and drinking much tea,excessive exercise intensity and insufficient sleep after leaving the plateau were independent risk factors for HADA.Conclusion Low salt diet before entering the plateau,active prevention for altitude sickness,smoking cessation at the plateau,taking a slower means of transportation to enter and leave the plateau,drinking less tea,moderate exercise intensity and keeping enough sleep after leaving the plateau can effectively reduce the risk of HADA.

Objective:To investigate the efficacy of 3D-printed quantitative bone implants assisting second-stage Masquelet technique for the treatment of long-segment bone defect following Gustilo type IIIB and IIIC tibial fractures.Methods:A retrospective case series analysis was made on 26 patients with long-segment bone defect following Gustilo type IIIB and IIIC tibial fractures treated in Wuxi Ninth People′s Hospital from July 2015 to December 2020, including 20 males and 6 females; aged 19-63 years [(46.5±4.5)years]. Gustilo classification was type IIIB in 23 patients and type IIIC in 3. In the first stage, all patients had thoroughly emergent debridement, removal of all free bone pieces, restoration of the length and force line plus externally fixion, and vacuum sealing drainage (VSD) of the residual wound. After 2-7 days, the external fixation was removed and replaced by internal fixation, with the bone cement filling in the defect area and the free flap covering the wound. The length of tibial bone defect was 5-14 cm [(6.3±0.4)cm], and the tibial defect volume was 12.2-73.1 cm 3 [(33.6±9.2)cm 3]. In the second stage (6-19 weeks after injury), the bone cement was removed, followed by autologous bone grafting. Prior to bone grafting, digital technology was used to accurately calculate the bone defect volume, and an equal volume of bone harvesting area was designe to produce the 3D printed osteotomy template. Bone grafting was conducted after bone removal according to the osteotomy template during operation. The success rate of one-time iliac bone extraction, bone harvesting time, and bleeding volume were recorded. Pain in the bone extraction area was evaluated by visual analogue score (VAS) at 1 day and 1 month after operation and at the last follow-up. Wound healing, complications, and bone healing were observed. Life quality was evaluated by health survey brief form (SF-36) including scores of physical component summary (PCS) and mental component summary (MCS) before bone grafting and at the last follow-up. Results:All the patients were followed up for 13-53 months [(32.3±12.5)months]. One-time iliac bone extraction was successful in all the patients. Bone harvesting time was 15-30 minutes [(21.0±2.5)minutes]. The bleeding volume was 50-120 ml [(62.3±29.0)ml]. The VAS was 1-4 points [(1.2±0.9)points] at 1 day after operation, higher than these (0.0±0.0)points at 1 month after operation and at the last follow-up (all P<0.01). Totally, 25 patients obtained wound healing after operation, except for 1 patient with superficial wound infection after bone grafting that was healed by dressing change. There was 1 patient with bone infection after 3 months of bone grafting that was healed by repeated surgery with Masquelet technique in the first and second stage. Besides, 2 patients had symptoms of cutaneous nerve injury in the iliac donor area. The time of bone healing was 4-7 months [(5.8±0.8)months]. The scores of PCS and MCS in SF-36 at the last follow-up were (73.6±12.8)points and (83.6±13.2)points, significantly higher than those before bone grafting [(46.8±0.5)points, (60.7±2.0)points] (all P<0.01). Conclusion:Second-stage Masquelet technique with 3D printed quantitative bone implants for the treatment of long-segment bone defect following Gustilo type IIIB and IIIC tibial fractures is associated with shortened bone harvesting time, attenuated pain, reduced complications, accelerated bone healing and improved function.

Objective:To develop an in vitro neuroinflammation model by establishing a microglia-neuron co-culture system. Methods:Mouse microglia (BV-2), motor neurons (NSC34) and hippocampal neurons (HT-22) were selected.This experiment was performed in two parts.Experiment Ⅰ BV-2 microglia were stimulated with different concentrations of lipopolysaccharide (LPS, 10, 100, 500 and 1 000 ng/ml). Microglia culture supernatant(Conditioned Medium) was extracted and two types of neurons were cultured separately.The concentration of LPS that resulted in a significant 50% decrease in neuronal viability was selected using the CCK-8 method for establishment of the Transwell co-culture system.Experiment Ⅱ Microglia were cultured in the upper chamber of Transwell, and neurons were seeded in the lower chamber.Microglia were divided into 2 groups ( n=12 each) using the random number table method: control group and LPS group.In control group and LPS group, microglia were cultured for 6 h with cell culture medium and LPS, respectively, then the medium was replaced with fresh medium, microglia were continuously incubated for 12 h, and then the cells in the upper and lower chambers were combined.The cells were incubated using the BV-2-NSC34 Transwell co-culture system for 12 h and using the BV-2-HT-22 Transwell co-culture system for 24 h. The concentrations of interleukin-1beta (IL-1β) and IL-18 in neuronal culture supernatant were measured by enzyme-linked immunosorbent assay, the apoptotic rate of neurons was determined by flow cytometry, the expression of Bcl-2 and Bax mRNA in neurons was detected by quantitative real-time polymerase chain reaction, and the expression of cleaved caspase-3, Bcl-2 and Bax in neurons was detected by Western blot. Results:Experiment Ⅰ LPS concentration for stimulation was 10 ng/ml in BV-2-NSC34 Transwell co-culture system and 1, 000 ng/ml in BV-2-HT-22 Transwell co-culture system.Experiment Ⅱ Compared with control group, the concentrations of IL-1β and IL-18 and apoptotic rate of neurons were significantly increased, Bax protein and mRNA expression was up-regulated, Bcl-2 protein and mRNA expression was down-regulated, and cleaved caspase-3 expression was up-regulated in LPS group ( P<0.05 or 0.01). Conclusions:The microglia-neuron co-culture system is successfully established by the conditioned medium technique and Transwell co-culture system, which provides an experimental protocol for establishment of neuroinflammation models associated with postoperative cognitive dysfunction.

Objective:To investigate the long-term death of patients with ischemic stroke and its influencing factors.Methods:Based on the data of patients with ischemic stroke in the multi-center oral fibrinogen-lowering drug secondary prevention database, the follow-up patient information and the cause of death were registered through the epidemiological investigation method, and then compared with the baseline data of patients in the original database.Results:A total of 278 patients completed the follow-up, and 166 were in lumbrokinase group and 112 were in control group. There were 124 deaths (44.6%) within 10 years, of which 92 (74.2%) were vascular deaths. In the lumbrokinase group, 74 patients (44.6%) died of all causes and 55 (33.1%) died of vascular diseases; in the control group, 50 (44.6%) died of all causes and 37 (33.0%) died of vascular diseases. Cox proportional risk model analysis showed that lumbrokinase treatment had no significant effect on the 10-year survival rate of patients with ischemic stroke. The analysis of death influencing factors showed that the baseline international normalized ratio (INR) was significantly associated with the 10-year non-vascular death risk of patients (hazard ratio [ HR] 1.98, 95% confidence interval [ CI] 1.21-3.25; P=0.006). The greater the decrease of tissue plasminogen activator (tPA) within half a year, the lower the 10-year all-cause mortality risk ( HR 0.94, 95% CI 0.90-0.99; P=0.011); the greater the decrease in INR within one year , the lower the 10-year vascular death risk ( HR 0.41, 95% CI 0.17-0.96; P=0.040); the greater the decrease of D-dimer within one year , the higher the risk of the 10-year vascular death ( HR 1.37, 95% CI 1.02-1.83; P=0.034). The greater the decrease of INR in patients with ischemic stroke within one year, the higher the 10-year non-vascular death risk ( HR 2.15, 95% CI 1.29-3.59; P=0.004). Conclusions:The 10-year mortality rate of patients with ischemic stroke is higher, and about 3/4 are vascular deaths. The fibrinogen-lowering treatment in the acute stage has no significant effect on the 10-year all-cause mortality of patients with ischemic stroke. The greater the decrease of tPA in half a year, the lower the all-cause mortality; the greater the decrease of D-dimer level at baseline and within 1 year, the higher the 10-year vascular death; the greater the decrease of INR at baseline and within 1 year, the higher the 10-year non-vascular death risk.