Objective:To investigate epithelial cell heterogeneity in ovarian cancer(OC)and its impact on the tumor microenvironment(TME)by constructing OC epithelial subtypes using publicly available single-cell RNA sequencing(scRNA-seq)data and bioinformatics approaches.Methods:Eight cell types and gene expression matrices were identified from the GSE118828 OC scRNA-seq dataset,and a deconvolution approach was used to estimate cellular composition in bulk RNA sequencing(bulk RNA-seq)data.OC samples were stratified into high-epithelial(C1)and low-epithelial(C2)subtypes based on epithelial cell abundance.Differences in overall survival(OS)and immune microenvironment features between patients with C1 and C2 subtypes were analyzed.Differentially expressed genes(DEGs)between C1 and C2 subtypes,as well as between early-stage(stageⅠ～Ⅱ)and late-stage(stageⅢ-Ⅳ)OC groups were identified to screen epithelial-related genes associated with OC progression,and their influence on the TME was analyzed.Protein expression of the epithelial-related genes in OC progression was validated using data from the Human Protein Atlas(HPA)database.Results:Patients with the C1 subtype exhibited significantly better OS compared to those with C2 subtype(P<0.05).The C1 and C2 subtypes demonstrated distinct immune microenvironment infiltration profiles.Infiltration levels of T helper(Th)cells,M1 macrophages,and activated dendritic cells(DCs)were significantly higher in the C1 subtype(all P<0.05).In late-stage OC patients with C1 subtype,the expression of HAS1,DAPL1,and ADH1B was significantly upregulated(all P<0.05).Furthermore,DAPL1 expression was positively correlated with the degree of macrophage and regulatory T cells(Treg)infiltration in OC(P<0.05).HPA database analysis confirmed that DAPL1 protein expression was significantly upregulated in OC tissues.Conclusion:The high-epithelial subtype of ovarian cancer may exert anti-tumor effects through extensive infiltration of T helper(Th)cells,M1 macrophages,and activated dendritic cells within the tumor microenvironment.Conversely,upregulation of the epithelial-related gene DAPL1 may induce the infiltration of macrophages and Treg,thereby promoting OC progression.

Objective To establish a rapid on-site detection method for methamphetamine(MA)based on surface-enhanced Raman spectroscopy(SERS).Methods Laboratory-synthesized colloidal silver nanoparticles(AgNPs)were employed as SERS substrates to rapidly detect methamphetamine in typical crime scene matrices,including beer,cigarette ash,and plastic containers.Results The method requires no sample pretreatment,is simple to operate,and offers high sensitivity with fast detection speed.The limit of detection(LOD)reached 10 ppb(signal-to-noise ratio,S/N＞3).The qualitative results of methamphetamine analysis from forensic case samples were highly consistent with gas chromatography-mass spectrometry(GC-MS)findings.Conclusion Methamphetamine is frequently encountered in illicit settings and criminal activities.This study achieved rapid detection of methamphetamine in complex matrices(beer,cigarette ash,and plastic containers),demonstrating good application prospects in practice and providing important value for law enforcement,public safety,and criminal investigation.

Objective:To evaluate the safety and efficacy of digital technology-assisted debridement and bone and soft tissue reconstruction in the treatment of chronic osteomyelitis of the tibia.Methods:This retrospective study analyzed clinical data from 38 patients (26 males, 12 females; mean age 45.61±18.36 years, range 16-83 years) with chronic post-traumatic osteomyelitis complicated by soft tissue defects in the tibia, treated at the Department of Orthopaedics, Fourth Medical Center of PLA General Hospital between June 2021 and June 2023. There were 18 cases of traffic accidents, 6 cases of high-altitude falls, 6 cases of heavy objects and 6 cases of low-energy falls. Lesion sites: 12 cases in the upper segment of the tibia, 12 cases in the middle segment, and 14 cases in the lower segment. According to the Cierny-Mader classification, there were 24 cases of type III and 14 cases of type IV. Before the operation, the "3D point cloud technology" was used to plan the debridement range of bone and soft tissue. During the operation, the optical navigation system was used to monitor the debridement range in real time. The flap was designed by combining "Reading Tablet Treasure" with CT angiography, and the amount of bone graft was predicted by AI algorithm. The surgical method adopted is the Masquelet technique, namely thorough debridement, bone cement occupation and soft tissue repair in the first stage, and bone reconstruction in the second stage. Comparing the calculated bone defect amount and soft tissue defect area before the operation with the actual measured values after debridement. The cure rate of infection was evaluated by using the McKee bone infection treatment criteria after the operation. The visual analogue scale (VAS) for pain, lower extremity functional scale (LEFS), and self-rating anxiety scale (SAS) were used to evaluate the improvement of the quality of life of postoperative patients.Results:38 patients were followed up with a mean follow-up of 13.53±2.37 months. 37 patients' infections were effectively controlled, and 1 patient had a recurrence of infection, with an effectiveness rate of 97% for the treatment of McKee's infections. The preoperative calculated bone defect amount and soft tissue defect area were 51.05±26.31 cm 3 and 68.42±43.45 cm 2 respectively, and the actual measured values after debridement during the operation were 51.66±26.35 cm 3 and 68.82±43.76 cm 2 respectively. There was no statistically significant difference before and after the operation ( P>0.05). The interval between the first and second stage surgeries was 10.36±1.61 weeks, and all flaps survived after the first stage surgery. Two months after the operation, one case had recurrent osteomyelitis. After palliative treatment, sinus tracts were formed at the infection site. At the 12-month regular dressing change follow-up, there were still purulent secretions in the sinus tracts. There was no recurrence of infection after primary debridement in 37 cases, and the bone grafts healed. The bone healing time was 3.58±0.97 months. The postoperative VAS, LEFS, and SAS scores were 1.00±0.91, 66.68±7.91, and 34.30±4.29, respectively, which were significantly improved compared with the preoperative scores of 7.54±1.52, 21.22±4.29, and 52.70±6.74, respectively, and the difference was statistically significant ( P<0.05). Conclusions:The application of digital technology to precisely design the debridement range of bone and soft tissue, and the real-time holographic visualization monitoring of the debridement range during the operation can achieve precise debridement of bone and soft tissue and personalized and precise repair of soft tissue defects. It is a safe and effective technique for the treatment of chronic osteomyelitis of the tibia.

Objective:To evaluate the safety and efficacy of digital technology-assisted debridement and bone and soft tissue reconstruction in the treatment of chronic osteomyelitis of the tibia.Methods:This retrospective study analyzed clinical data from 38 patients (26 males, 12 females; mean age 45.61±18.36 years, range 16-83 years) with chronic post-traumatic osteomyelitis complicated by soft tissue defects in the tibia, treated at the Department of Orthopaedics, Fourth Medical Center of PLA General Hospital between June 2021 and June 2023. There were 18 cases of traffic accidents, 6 cases of high-altitude falls, 6 cases of heavy objects and 6 cases of low-energy falls. Lesion sites: 12 cases in the upper segment of the tibia, 12 cases in the middle segment, and 14 cases in the lower segment. According to the Cierny-Mader classification, there were 24 cases of type III and 14 cases of type IV. Before the operation, the "3D point cloud technology" was used to plan the debridement range of bone and soft tissue. During the operation, the optical navigation system was used to monitor the debridement range in real time. The flap was designed by combining "Reading Tablet Treasure" with CT angiography, and the amount of bone graft was predicted by AI algorithm. The surgical method adopted is the Masquelet technique, namely thorough debridement, bone cement occupation and soft tissue repair in the first stage, and bone reconstruction in the second stage. Comparing the calculated bone defect amount and soft tissue defect area before the operation with the actual measured values after debridement. The cure rate of infection was evaluated by using the McKee bone infection treatment criteria after the operation. The visual analogue scale (VAS) for pain, lower extremity functional scale (LEFS), and self-rating anxiety scale (SAS) were used to evaluate the improvement of the quality of life of postoperative patients.Results:38 patients were followed up with a mean follow-up of 13.53±2.37 months. 37 patients' infections were effectively controlled, and 1 patient had a recurrence of infection, with an effectiveness rate of 97% for the treatment of McKee's infections. The preoperative calculated bone defect amount and soft tissue defect area were 51.05±26.31 cm 3 and 68.42±43.45 cm 2 respectively, and the actual measured values after debridement during the operation were 51.66±26.35 cm 3 and 68.82±43.76 cm 2 respectively. There was no statistically significant difference before and after the operation ( P>0.05). The interval between the first and second stage surgeries was 10.36±1.61 weeks, and all flaps survived after the first stage surgery. Two months after the operation, one case had recurrent osteomyelitis. After palliative treatment, sinus tracts were formed at the infection site. At the 12-month regular dressing change follow-up, there were still purulent secretions in the sinus tracts. There was no recurrence of infection after primary debridement in 37 cases, and the bone grafts healed. The bone healing time was 3.58±0.97 months. The postoperative VAS, LEFS, and SAS scores were 1.00±0.91, 66.68±7.91, and 34.30±4.29, respectively, which were significantly improved compared with the preoperative scores of 7.54±1.52, 21.22±4.29, and 52.70±6.74, respectively, and the difference was statistically significant ( P<0.05). Conclusions:The application of digital technology to precisely design the debridement range of bone and soft tissue, and the real-time holographic visualization monitoring of the debridement range during the operation can achieve precise debridement of bone and soft tissue and personalized and precise repair of soft tissue defects. It is a safe and effective technique for the treatment of chronic osteomyelitis of the tibia.

Objective:To evaluate the efficacy and safety of the Masquelet technique on the basis of characterization of hematogenous and traumatic chronic osteomyelitis of the tibia.Methods:A retrospective analysis was conducted with chronic tibial osteomyelitis who had been treated by the Masquelet technique at Department of Orthopedic Surgery, The Fourth Medical Center, PLA General Hospital from February 2021 to June 2023. The patients were classified into 2 groups based on the underlying etiology of their conditions: a hematogenous group and a traumatic group. Treatment efficacy was evaluated in terms of infection control rate, bone graft healing time, visual analog scale (VAS) pain score, anxiety measured by the self-rating anxiety scale (SAS), and lower limb function assessed by the lower extremity functional scale (LEFS).Results:A total of 66 patients with chronic tibial osteomyelitis were included in the study. There were 17 cases in the hematogenous group. They were 11 males and 6 females with a median age of 31.0 (15.0, 45.0) years and a median disease duration of 3.0 (1.5, 8.0) months. The median interval between the first and second-stage surgeries was 11.5 (8.0, 13.0) weeks, the median volume of bone defect 25.0 (15.0, 40.0) cm 3, and the median bone graft healing time 4.0 (3.0, 4.0) months. No bacteria were detected in secretion culture in 6 cases and Staphylococcus aureus was found in 6 cases. At 12 months postoperatively, the median VAS score was 1.0 (0.0, 2.0) point, the median SAS score 27.0 (20.0, 32.0) points, and the median LEFS score 78.0 (75.0, 80.0) points. There were 49 cases in the traumatic group. They were 36 males and 13 females with a median age of 52.0 (42.0, 63.0) years and a median disease duration of 6.0 (3.0, 36.0) months. The median interval between the first and second-stage surgeries was 10.0 (8.0, 17.0) weeks, the median volume of bone defect 30.0 (22.0, 53.0) cm 3, and the median bone graft healing time 3.5 (3.0, 4.5) months. No bacteria were detected in secretion culture in 10 cases and Staphylococcus aureus was found in 19 cases. At 12 months postoperatively, the median VAS score was 2.0 (1.0, 3.0) points, the median SAS score 35.0 (28.0, 42.0) points, and the median LEFS score 54.0 (42.0, 60.0) points. According to the McKee criteria for infection treatment, 14 cases achieved complete recovery, 2 ones showed improvement, and 1 case experienced recurrence in the hematogenous group, while 44 cases achieved complete recovery, 4 ones showed improvement, and 1 case experienced recurrence in the traumatic group. Conclusions:Although hematogenous and traumatic cases of chronic osteomyelitis of the tibia differ in terms of age of onset, disease duration, and lesion extent, Staphylococcus aureus is the predominant pathogen in both conditions. Application of the Masquelet technique has proven to be an effective treatment for both conditions with comparable bone graft healing time.

Objective To establish a rapid on-site detection method for methamphetamine(MA)based on surface-enhanced Raman spectroscopy(SERS).Methods Laboratory-synthesized colloidal silver nanoparticles(AgNPs)were employed as SERS substrates to rapidly detect methamphetamine in typical crime scene matrices,including beer,cigarette ash,and plastic containers.Results The method requires no sample pretreatment,is simple to operate,and offers high sensitivity with fast detection speed.The limit of detection(LOD)reached 10 ppb(signal-to-noise ratio,S/N＞3).The qualitative results of methamphetamine analysis from forensic case samples were highly consistent with gas chromatography-mass spectrometry(GC-MS)findings.Conclusion Methamphetamine is frequently encountered in illicit settings and criminal activities.This study achieved rapid detection of methamphetamine in complex matrices(beer,cigarette ash,and plastic containers),demonstrating good application prospects in practice and providing important value for law enforcement,public safety,and criminal investigation.

Objective:To evaluate the efficacy and safety of the Masquelet technique on the basis of characterization of hematogenous and traumatic chronic osteomyelitis of the tibia.Methods:A retrospective analysis was conducted with chronic tibial osteomyelitis who had been treated by the Masquelet technique at Department of Orthopedic Surgery, The Fourth Medical Center, PLA General Hospital from February 2021 to June 2023. The patients were classified into 2 groups based on the underlying etiology of their conditions: a hematogenous group and a traumatic group. Treatment efficacy was evaluated in terms of infection control rate, bone graft healing time, visual analog scale (VAS) pain score, anxiety measured by the self-rating anxiety scale (SAS), and lower limb function assessed by the lower extremity functional scale (LEFS).Results:A total of 66 patients with chronic tibial osteomyelitis were included in the study. There were 17 cases in the hematogenous group. They were 11 males and 6 females with a median age of 31.0 (15.0, 45.0) years and a median disease duration of 3.0 (1.5, 8.0) months. The median interval between the first and second-stage surgeries was 11.5 (8.0, 13.0) weeks, the median volume of bone defect 25.0 (15.0, 40.0) cm 3, and the median bone graft healing time 4.0 (3.0, 4.0) months. No bacteria were detected in secretion culture in 6 cases and Staphylococcus aureus was found in 6 cases. At 12 months postoperatively, the median VAS score was 1.0 (0.0, 2.0) point, the median SAS score 27.0 (20.0, 32.0) points, and the median LEFS score 78.0 (75.0, 80.0) points. There were 49 cases in the traumatic group. They were 36 males and 13 females with a median age of 52.0 (42.0, 63.0) years and a median disease duration of 6.0 (3.0, 36.0) months. The median interval between the first and second-stage surgeries was 10.0 (8.0, 17.0) weeks, the median volume of bone defect 30.0 (22.0, 53.0) cm 3, and the median bone graft healing time 3.5 (3.0, 4.5) months. No bacteria were detected in secretion culture in 10 cases and Staphylococcus aureus was found in 19 cases. At 12 months postoperatively, the median VAS score was 2.0 (1.0, 3.0) points, the median SAS score 35.0 (28.0, 42.0) points, and the median LEFS score 54.0 (42.0, 60.0) points. According to the McKee criteria for infection treatment, 14 cases achieved complete recovery, 2 ones showed improvement, and 1 case experienced recurrence in the hematogenous group, while 44 cases achieved complete recovery, 4 ones showed improvement, and 1 case experienced recurrence in the traumatic group. Conclusions:Although hematogenous and traumatic cases of chronic osteomyelitis of the tibia differ in terms of age of onset, disease duration, and lesion extent, Staphylococcus aureus is the predominant pathogen in both conditions. Application of the Masquelet technique has proven to be an effective treatment for both conditions with comparable bone graft healing time.

Limb preservation techniques of severed limbs play an important role in autologous replantation or allografting. At present, static cold storage (SCS) remains the gold standard for the in vitro preservation of severed limbs, which minimizes oxygen and energy metabolism. However, with a relatively short time window of 4-6 hours, SCS is insufficient for clinical transfer, injury management and other medical resources. To address this issue, researchers have initiated investigations into alternative methods of supplying oxygen and nutrients to severed limbs, leading to the development of mechanical perfusion technology and per-sufflation technology. Mechanical perfusion technology aims to clear metabolic waste and provide essential nutrients by mimicking a physiological state, thereby prolonging the preservation time and improving the outcome of the repair. Per-sufflation technology aims to deliver oxygen directly to the tissue using a vascular system to maintain tissue integrity. With these emerging technologies showing great promise, further exploration and refinement are needed to address their time constraints and complexity in clinical work. Furthermore, for severed limbs that are not suitable for immediate reimplantation, autologous heterotopic parasitism techniques and temporary endovascular shunts offer promising avenues for preservation. Autologous heterotopic parasitism techniques maintain tissue viability by constructing a temporary parasitic environment, while temporary endovascular shunts establish a temporary vascular access at the damaged site to deliver donor blood, thereby maintaining blood flow and promoting repair. Xenogeneic cross-circulation techniques combine recent advances from multiple disciplines and demonstrate remarkable potential; however, they currently face challenges such as ethics, immune rejection, and technical complexity. The combination of these novel techniques has led to a significant advancement in the field of limb preservation and repair. This article aims to provide an overview of the current research status and development of static cold storage, pre-sufflation, mechanical perfusion preservation and temporary ectopic parasitic preservation techniques in limb preservation over recent years, providing information for clinical work, promoting the development of limb reimplantation and transplantation techniques in China and further consolidating China's leadership in the field of amputation reimplantation and allograft transplantation.

Objective The oxidative damage of DNA can be caused by excessive levels of Reactive oxygen species(ROS).Monitoring of DNA oxidative damage enables effective evaluation of ROS damage effects.Although the detection of DNA damage effects based on microbial sensor allows quantitative analysis of oxidative damage,the ROS clearance mechanism existed in bacterial will affect the sensitive of detection.The work of this article is to knockout the key genes of ROS clearance mechanism and construct an antioxidant gene-knock-out microbial sensor.The microbial sensor can realize sensitive monitoring of DNA damage effects and then evaluates the damage effects of cells by ROS.Methods The antioxidant damage genes of bacterial ahpCF,katE and katG were knocked out by λ-Red homologous recombination and antioxidant gene-knockout microbial sensor was constructed.The nalidixic acid sodium salt and UV irradiation were used to characterize the performance for monitoring of DNA damage effects.Results The antioxidant gene-knockout microbial sensors ΔahpC,ΔahpCF/ΔkatEG and ΔahpCF/ΔkatE/ΔkatG were constructed successfully.The results showed that the microbial sensor ΔahpCF/ΔkatE/ΔkatGl had the highest sensitive of damage effects and the limit of detection for nalidixic acid sodium salt was 0.40 μmol/L.In addition,1.80 min of UV irradiation(254 nm)was sufficient to induce a significant fluorescent expression effect in the engineered bacteria.Conclusion In this article,antioxidant gene-knockout microbial sensors had been constructed to realize active and sensitive monitoring of DNA damage effects such as DNA damage reagents and UV irradiation.The sensors could provide an active,effective,and sensitive potential monitoring method for future evaluation of radiation effects in space.

Objective:To compare the efficacy and safety of insulin degludec and insulin glargine U100 in patients with type 2 diabetes mellitus.Methods:This study was a retrospective cohort study. The subjects were patients with type 2 diabetes mellitus who were hospitalized in 13 3A-level general hospitals in Shandong Province from September 2018 to December 2021. According to the type of basal insulin used, the patients were divided into insulin degludec group and insulin glargine U100 group. The basic information and laboratory test results in patients in the 2 groups were collected, the differences of fasting blood glucose level and incidence of hypoglycemia between the 2 groups were compared. The patients with complete blood glucose monitoring data in the 2 groups were selected and their blood glucose fluctuations were compared.Results:A total of 1 152 patients were entered in the study, including 552 patients in the insulin degludec group and 600 patients in the insulin glargine U100 group. The difference in the basic conditions in patients in the 2 groups was not statistically significant (all P>0.05). After treatment, the fasting blood glucose levels in patients in the 2 groups were lower than those before treatment, with statistically significant differences [10.2 (8.8, 12.5) mmol/L vs. 7.5 (6.6, 8.7) mmol/L, Z=-19.443, P<0.001; 10.0 (8.6, 11.7) mmol/L vs. 7.8 (6.6, 9.0) mmol/L, Z=-15.449, P<0.001], but the difference in fasting blood glucose levels between the 2 groups after treatment was not statistically significant ( Z=-1.427, P>0.05). The incidence of hypoglycemia in the insulin degludec group was lower than that in the insulin glargine U100 group [1.09% (6/552) vs. 2.83% (17/600), Z=4.481, P=0.032]. The intraday blood glucose standard deviation, maximum blood glucose fluctuation range, postprandial blood glucose fluctuation range, and average blood glucose fluctuation range in patients with complete blood glucose monitoring data in the insulin degludec group were significantly lower than those in the insulin glargine U100 group [(1.7±0.6) mmol/L vs. (2.4±1.0) mmol/L, (4.5±1.6) mmol/L vs. (6.7±2.9) mmol/L, (1.8±1.0) mmol/L vs. (3.3±1.2) mmol/L, (2.9±1.3) mmol/L vs. (4.6±2.1) mmol/L; all P<0.001]. Conclusion:The efficacy of insulin degludec in the treatment of type 2 diabetes mellitus is equivalent to that of insulin glargine U100, but the risk of hypoglycemia and blood glucose fluctuation is lower.