BACKGROUND:Mitochondrial reactive oxygen bursts have been shown to play a key role in skeletal muscle ischemia-reperfusion injury.3-Nitro-N-methylsalicylamide(3-NNMS)can effectively reduce the electron transport rate and has a potential protective effect on limb ischemia-reperfusion injury,but there is no clear research and clinical application. OBJECTIVE:To investigate the protective effect of 3-NNMS on the skeletal muscle after limb ischemia-reperfusion injury in rats and its mechanism. METHODS:Forty healthy 8-week-old Sprague-Dawley rats were randomly divided into control group,0,25 and 125 μg/mL 3-NNMS groups,with 10 rats in each group.Animal models of limb ischemia-reperfusion injury were prepared in the latter three groups.3-NNMS was injected into the injury site 30 minutes before reperfusion.The animals were sacrificed 2 hours after reperfusion.Blood from the apical part of the heart,and the tissue of the rectus femoris muscle of the right lower limb were taken for testing.The pathological morphology of the rectus femoris muscle was detected by hematoxylin-eosin staining.Serum levels of creatine kinase found in the skeletal muscle(CK-MM),lactate dehydrogenase,and myeloperoxidase were detected using ELISA;the levels of nuclear factor κB,tumor necrosis factor α,interleukin 1β,cyclooxygenase 2,malondialdehyde,reactive oxygen species,superoxide dismutase,catalase and glutathione peroxidase in the rectus femoris muscle were measured;and adenosine triphosphate(ATP)level,ATPase activity,and mitochondrial respiratory control rate were tested. RESULTS AND CONCLUSION:Compared with the control group,the model rats with ischemia-reperfusion injury had increased serum levels of CK-MM,lactate dehydrogenase,and myeloperoxidase,increased levels of nuclear factor κB,tumor necrosis factor α,interleukin 1β,cyclooxygenase 2,malondialdehyde and reactive oxygen species in the rectus femoris muscle,decreased levels of catalase and glutathione peroxidase in the rectus femoris muscle,and reduced ATPase activity and mitochondrial respiratory control rate.Moreover,cell morphology was irregular,inflammatory cell infiltration was obvious,and the cells were swollen in rats after ischemia-reperfusion injury.Compared with the 0 μg/mL group,the serum CK-MM and lactate dehydrogenase levels decreased,the levels of nuclear factor κB and cyclooxygenase 2 in the rectus femoris muscle decreased,reactive oxygen species level decreased,and superoxide dismutase activity increased in the 25 μg/mL group;cell morphology was more regular,inflammatory cell infiltration was lighter,and cell swelling was alleviated.Compared with the 0 μg/mL group,the 125 μg/mL group had a reduction in the serum levels of CK-MM,lactate dehydrogenase,and myeloperoxidase and the levels of nuclear factor κB,tumor necrosis factor α,cyclooxygenase 2,malondialdehyde and reactive oxygen species in the rectus femoris muscle,as well as an increase in the levels of superoxide dismutase and glutathione peroxidase in the rectus femoris muscle,and mitochondrial respiratory control rate.Moreover,the cells were arranged neatly,the outline was clear and complete,and the inflammatory cell infiltration was light.To conclude,3-NNMS can alleviate the functional impairment of the skeletal muscle caused by limb ischemia-reperfusion,and its mechanism of action may be through improving mitochondrial function,reducing reactive oxygen species production,decreasing oxidative stress and inflammatory response,and thus reducing tissue damage and repairing skeletal muscle function.

Objective To explore the feasibility and accuracy of using indocyanine green fluorescence (ICGF) to identify the intersegmental plane after ligation of the target pulmonary vein during thoracoscopic segmentectomy. Methods From December 2022 to June 2023, the patients with pulmonary nodules undergoing video-assisted thoracoscopic anatomical segmentectomy with intersegmental plane displayed using ICGF after ligation of the target pulmonary vein by the same medical team in our hospital were collected. Preoperative three-dimensional reconstruction was used to identify the target segment where the pulmonary nodule was located and the anatomical structure of the arteries, veins, and bronchi in the target segment. The intersegmental plane was first determined by the inflation-deflation method after the target pulmonary vein was ligated during the operation. During the waiting period, the target artery and bronchus could be separated but not cut off. The inflation-deflation boundary was marked by electrocoagulation, and then ICGF was injected via peripheral vein to identify the intersegmental plane again, and the consistency of the two intersegmental planes was finally evaluated. Results Finally 32 patients were collected, including 14 males and 18 females, with an average age of 58.69±11.84 years, ranging from 25 to 76 years. The intersegmental plane determined by inflation-deflation method was basically consistent with ICGF method in all patients. All the 32 patients successfully completed uniportal thoracoscopic segmentectomy without ICGF-related complications or perioperative death. The average operation time was 98.59±20.72 min, the average intraoperative blood loss was 45.31±35.65 mL, and the average postoperative chest tube duration was 3.50±1.16 days. The average postoperative hospital stay was 4.66±1.29 days, and the average tumor margin width was 26.96±5.86 mm. Conclusion The ICGF can safely and accurately identify the intersegmental plane by target pulmonary venous preferential ligation in thoracoscopic segmentectomy, which is a useful exploration and important supplement to the simplified thoracoscopic anatomical segmentectomy.

Objective:To discuss the effect of temperature-controlled shrinkage polytrimethylene carbonate(PTMC)/polyvinylpyrrolidone(PVP)nanofiber membrane on the biological behavior of mouse fibroblasts and the repairment effect on full-thickness skin defects in the rats,and to clarify the potential mechanism.Methods:The murine L929 fibroblast cells were used in the in vitro experiments and were divided into control group and experimental group(treated with PTMC/PVP nanofiber membranes).The proliferation activities of the cells in two groups were detected by CCK-8 assay;the numbers of live/dead cells in two groups were observed by live/dead cell staining;the morphology of the cells was observed by cytoskeletal staining.A total of 12 six-week-old male SD rats were selected in the in vivo experiment,and were randomly divided into control group and experimental group,and there were six rats in each group.The full-thickness skin defect model was established,and the rats in experimental group were treated with PTMC/PVP nanofiber membranes.The photographs were taken after operation,and the wound healing rates of the cells in two groups were calculated on the 0,3rd,6th,and 12th days.On the 6th and 12th days after operation,the skin samples around the wound of the rats in two groups were taken,and the histopathology of the would skin and adjacent tissue was detected by HE staining;the collagen deposition in wound skin tissue of the rats in two groups was observed by Masson trichrome staining;the numbers of angiogenesis in wound skin tissue of the rats were detected by CD31 immunohistochemical staining.Results:The CCK-8 assay results showed that the proliferation activity of the cells in experimental group showed an increasing trend on the 1st,3st,and 5st days,and there was no significant difference in the proliferation activities of the cells bewteen experimental group and control group(P＞0.05).The live/dead cell staining experiment results showed that compared with control group,the cell density and number of the cells in experimental group had no significant changes,and were predominantly live cells.The cytoskeletal staining results showed that the cells in experimental and control groups appeared spindle-shaped and well-spread.In the in vivo experiments,on the 3rd,6th,and 12th days,compared with control group,the wound healing rates of the cells in experimental group were increased(P＜0.01),and the wound healing rate of the cells was 95.45％on the 12th day,indicating nearly complete healing of the wound.The HE staining showed that on the 12th day,the wound skin structure of the cells in experimental group was more similar to the normal skin,and there was abundant granulation tissue,regular epidermal structure,and new blood vessel formation.The Masson trichrome staining results showed that compared with control group,the collagen deposition in wound tissue of the rats in experimental group was increased.The immunohistochemical staining results showed that the expression of CD31 in wound tissue of the rats in experimental group was increased,indicating the increasing of the number of angiogenesis.Conclusion:The PTMC/PVP thermoresponsive nanofiber membranes exhibit good biocompatibility and can promote the repairment of full-thickness skin defects in the rats;its mechanism may be related to the enhancement of proliferation activity of the basal cells.

Chitooligosaccharides(COS),as the degradation products of the natural polysaccharide chitosan(CS),retain the good biocompatibility,non-toxicity,and biodegradability of CS.Additionally,due to the shortened sugar chains and reduced molecular weight,their water solubility and bioactivity are improved,making them more easily absorbed and utilized by organisms.In recent years,COS has received increasing attention.While there are numerous studies on the biological functions of CS and its applications in the biomedical field,the research on the functions and applications of COS is relatively limited.This study summarizes and analyzes the main biological functions of COS(anti-inflammatory,anti-tumor,antibacterial,and tissue regeneration promotion)and their possible mechanisms.It also discusses the research progress in the applications of COS in the biomedical field,in order to provide the theoretical basis and reference for the further research and broader application of COS in biomedicine.

Objective    To analyze the effect of indocyanine green (ICG) fluorescence dual-visualization technique on evaluating tumor margins during the thoracoscopic segmentectomy. Methods    A total of 36 patients who underwent thoracoscopic anatomical segmentectomy using ICG fluorescence dual-visualization technique in our hospital from December 2020 to June 2021 were retrospectively included. There were 15 males and 21 females aged from 20 to 69 years. The clinical data of the patients were retrospectively analyzed. Results    The ICG fluorescence dual-visualization technique clearly showed the position of lung nodules and the plane boundary line between segments during the operation. There was no ICG-related complication. The average operation time was 98.6±21.3 min, and the average intraoperative bleeding amount was 47.1±35.3 mL, the average postoperative drainage tube placement time was 3.3±2.8 d, the average postoperative hospital stay was 5.4±1.8 d, and the average tumor resection distance was 2.6±0.7 cm. There was no perioperative period death, and one patient suffered a persistent postoperative air leak. Conclusion    The ICG fluorescence dual-visualization technique is safe and feasible for evaluating the tumor margins during thoracoscopic segmentectomy. It simplifies the surgical procedure, shortens the operation time, ensures sufficient tumor margins, and reserves healthy pulmonary parenchyma to the utmost extent, providing reliable technical support for thoracoscopic anatomical segmentectomy.

MicroRNAs(miRNAs)are important regulators in gene expression.The dysregulation of miRNA expression is widely reported in the transformation from physiological to pathological states of cells.A large number of differentially expressed miRNAs(DEMs)have been identified in various human cancers by using high-throughput technologies,such as microarray and miRNA-seq.Through mining of published studies with high-throughput experiment information,the data-base of DEMs in human cancers(dbDEMC)was constructed with the aim of providing a systematic resource for the storage and query of the DEMs.Here we report an update of the dbDEMC to version 3.0,which contains two-fold more data entries than the second version and now includes also data from mice and rats.The dbDEMC 3.0 contains 3268 unique DEMs in 40 different cancer types.The current datasets for differential expression analysis have expanded to 9 generalized cat-egories.Moreover,the current release integrates functional annotations of DEMs obtained by using experimentally validated targets.The annotations can be of great benefit to the intensive analysis of the roles of DEMs in cancer.In summary,dbDEMC 3.0 provides a valuable resource for charac-terizing molecular functions and regulatory mechanisms of DEMs in human cancers.

BACKGROUND: Segmentectomy has gradually become one of the standard surgical methods for small pulmonary nodules with early lung cancer on imaging. This study aimed to investigate the perioperative outcomes of patients who underwent uniport video-assisted thoracoscopic surgery (VATS) segmentectomy for identifying the intersegmental boundary line (IBL) by the near-infrared fluorescence imaging with intravenous indocyanine green (ICG) method or the modified inflation-deflation (MID) method and assess the feasibility and effectiveness of the ICG fluorescence (ICGF)-based method. METHODS: We retrospectively analyzed the perioperative data in total 198 consecutive patients who underwent uniport VATS segmentectomy between February 2018 and August 2020. With the guidance of preoperative intelligent/interactive qualitative and quantitative analysis-three dimensional (IQQA-3D), the targeted segment structures could be precisely identified and dissected, and then the IBL was confirmed by ICGF-based method or MID method. Clinical effectiveness and postoperative complications of the two methods were evaluated. RESULTS: An IBL was visible in 98% of patients by the ICGF-based group, even with the low-doses of ICG. The ICGF-based group was significantly associated with the shorter IBL clear presentation time [(23.59±4.47) s vs (1,026.80±318.34) s] (P<0.01) and operative time [(89.3±31.6) min vs (112.9±33.3) min] (P<0.01), compared to the MID group. The incidence of postoperative prolonged air leaks was higher in the MID group than in the ICGF-based group (8.0% vs 26.5%, P=0.025). There were no significant differences in bleeding volume, chest tube duration, postoperative hospital stays, surgical margin width and other postoperative complications (P>0.05). CONCLUSIONS The ICGF-based method could highly accurately identify the IBL and make anatomical segmentectomy easier and faster, and therefore has the potential to be a feasible and effective technique to facilitate the quality of uniport VATS segmentectomy.
Feasibility Studies ; Humans ; Indocyanine Green ; Ion Transport ; Lung Neoplasms/surgery* ; Pneumonectomy ; Postoperative Complications ; Retrospective Studies ; Thoracic Surgery, Video-Assisted

Objective:To investigate the efficacy and feasibility of radiofrequency resection of supraglottic laryngeal carcinoma under a multifunctional opener.Methods:The clinical data of four cases of supraglottic laryngeal carcinoma (type T1N1M0 in two cases, T2N1M0 in one case, and T1N0M0 in one case) who received treatment in the Affiliated Hospital of Jining Medical University during January-June 2019 were retrospectively analyzed. Radiofrequency resection of supraglottic laryngeal carcinoma under a multi-functional opener combined with bilateral neck II-IV region lymph node dissection was performed. Swallowing, breathing and phonation were observed and analyzed based on references.Results:Among the four cases, two cases had a normal diet at 3 days after surgery, one case had a normal diet at 7 days after surgery, and one case had a normal diet at 16 days after surgery. Tracheotomy was not performed in any case. After surgery, breathing and speech communication were not affected.Conclusion:Radiofrequency surgery under a multifunctional opener can be used for treatment of early supraglottic laryngeal cancer. It is an effective treatment with minimal trauma, mild postoperative pain and promotes the early recovery of normal swallowing function.

Objective:To explore the antagonistic effect of Fengzhecao extract against human red blood cell (RBC) hemolysis induced by wasp venom.Methods:Water extract method was used to extract dried Fengzhecao and vacuum-dried to obtain Fengzhecao extract. It was diluted into 1 g/L for next use. Wasp venom was collected from the wasp workers. A, B, O, AB type healthy blood donors' suspended RBC solution was obtain to make washed RBC solutions and adjust the RBCs count (4.0-80.0)×10 9/L (the number of RBC counted on the hemocytometer is 1-20 cells/small checker). According to treatment factors, they were divided into the normal saline controlled group (NS group; 200 μL RBC solution+20 μL normal saline), Fengzhecao extract group (FZC group; 200 μL RBC solution+10 μL Fengzhecao extract+10 μL normal saline), wasp venom group (FD group; 200 μL RBC solution+10 μL wasp venom+10 μL normal saline), and Fengzhecao extract+wasp venom group (FCD group; 200 μL RBC solution+10 μL Fengzhecao extract+10 μL wasp venom), with 10 blood samples per group of every blood type. The solutions were put into the glass test tube respectively, and then into 37 ℃ water bath thermostat. After 10 minutes, the blood cell counting plate was directly observed under the microscope and the RBCs was counted. Differences in RBC count was compared between the same treatment factors of different blood types and between different treatment factor groups of the same blood type. Results:There was no statistically significant difference in RBC count between blood types under the same treatment factors. The RBC count (×10 9/L) of the type A, B, O, AB in the NS group were 5.567±1.368, 5.146±1.690, 4.577±0.774, 5.197±1.587 ( F = 0.852, P = 0.475), the FZC group were 5.751±1.489, 5.268±1.418, 4.727±1.174, 5.298±1.229 ( F = 0.987, P = 0.410), the FD group were 0.546±0.450, 0.804±0.428, 0.679±0.283, 0.846±0.453 ( F = 1.089, P = 0.366), and the FCD group were 5.532±1.330, 5.051±1.596, 4.589±0.879, 5.140±1.492 ( F = 0.820, P = 0.492), respectively. Comparison of RBC count between groups with different treatment factors of the same blood type was done. There was no significant difference between the FZC group and the NS group, indicating that the extract of Fengzhecao extract had no effect on hemolysis of RBC; in the FD group, it was significantly lower than the NS group (all P < 0.05), indicating that wasp venom had a significant hemolytic effect on RBC; but there was no statistically significant difference in RBC count between the FCD group and the NS group, indicating that the Fengzhecao extract antagonizes the hemolytic effect of wasp venom without affecting the RBC count; however, the RBC count in the FCD group was significantly higher than that in the FD group (all P < 0.05), further indicating that the Fengzhecao extract antagonizes the hemolytic effect of wasp venom. Conclusion:Wasp venom has a significant hemolytic effect which can be effectively antagonized by Fengzhecao extract and has nothing to do with the human ABO blood type.

Objective    To analyze the feasibility, advantages and disadvantages of the fluorescence method and the inflation-deflation method in defining the intersegmental plane during thoracoscopic lung segmental resection. Methods    From February to October 2018, 60 patients underwent thoracoscopic anatomical segmentectomy in Thoracic Surgery Department of Nanjing Chest Hospital, with 28 males and 32 females, aged from 25 to 82 years. Three-dimension computed tomography bronchography and angiography was used to reconstruct pulmonary vessels, bronchus and virtual intersegmental plane. Among them, 20 patients used the fluorescence method to define the intersegmental plane, and the other 40 patients used the traditional inflation-deflation method to define the intersegmental plane. Results    Fluorescent injection of indocyanine green (ICG) showed a clear intersegmental line with a duration sufficient to complete the label. With the fluorescence method, the intersegmental plane occurrence time was significantly shortened (10.75±3.78 s vs. 988.00±314.24 s, P<0.001) and had satisfactory repeatability. The lungs did not need to be inflated, which was convenient for the operation. And the operation time was shortened (108.75±31.28 min vs 138.00±32.47 min, P=0.002). No obvious ICG injection-related concurrency symptoms was found. Conclusion    Compared with the traditional inflation-deflation method, the fluorescence method can display the intersegmental line quickly, accurately and clearly, reduce the difficulty of surgery, shorten the operation time, and provide reliable technical support for thoracoscopic anatomical segmentectomy. The fluorescence is a safe and effective method that is worthy of clinical application.