BACKGROUND:Forskolin,a diterpenoid natural compound extracted from Coleus forskohlii,has a crucial regulatory role in skeletal muscle repair.However,the regulatory role of forskolin on myogenic differentiation of C2C12 skeletal muscle cells has not been fully explored.OBJECTIVE:To explore the effects of forskolin on the differentiation of C2C12 myoblast cell line and probe into the underlying molecular mechanisms.METHODS:C2C12 cells were treated with 0,0.1,0.25,0.5,1,5,10 and 20 μmol/L forskolin during growth,and cell proliferation was detected by cell counting kit-8 and qRT-PCR.C2C12 cells were treated with 0,0.25,0.5 and 1 μmol/L forskolin during the induction of myogenic differentiation.Immunofluorescence staining and qRT-PCR were used to detect C2C12 cells differentiation.Western blot was used to detect the expression level of myogenic differentiation-related signaling pathway proteins.RESULTS AND CONCLUSION:(1)The viability of C2C12 cells was decreased and cell proliferation was inhibited after treatment with high concentrations(＞1 μmol/L)of forskolin.(2)The qRT-PCR results showed that forskolin up-regulated the expression of Myh2,Myh4,Myomaker,but down-regulated the expression of Myh7 compared with the 0 μmol/L group,when C2C12 cells were differentiated for 4 days.Immunofluorescence staining results showed that the fusion index and myotube diameter of C2C12 cells were increased after forskolin treatment,and the number of myotubes was also increased.(3)Western blot results showed that the phosphorylated extracellular signal-regulated kinase 1/2 expression was inhibited;however,the phosphorylated protein kinase B was promoted after treatment with forskolin.The protein expression level of the myogenic differentiation transcription factor Myogenin was significantly up-regulated after treatment with forskolin.The above results demonstrate that forskolin may promote myogenic differentiation of C2C12 skeletal muscle cells through the extracellular signal-regulated kinase 1/2 and protein kinase B signaling pathway.

BACKGROUND:Forskolin,a diterpenoid natural compound extracted from Coleus forskohlii,has a crucial regulatory role in skeletal muscle repair.However,the regulatory role of forskolin on myogenic differentiation of C2C12 skeletal muscle cells has not been fully explored.OBJECTIVE:To explore the effects of forskolin on the differentiation of C2C12 myoblast cell line and probe into the underlying molecular mechanisms.METHODS:C2C12 cells were treated with 0,0.1,0.25,0.5,1,5,10 and 20 μmol/L forskolin during growth,and cell proliferation was detected by cell counting kit-8 and qRT-PCR.C2C12 cells were treated with 0,0.25,0.5 and 1 μmol/L forskolin during the induction of myogenic differentiation.Immunofluorescence staining and qRT-PCR were used to detect C2C12 cells differentiation.Western blot was used to detect the expression level of myogenic differentiation-related signaling pathway proteins.RESULTS AND CONCLUSION:(1)The viability of C2C12 cells was decreased and cell proliferation was inhibited after treatment with high concentrations(＞1 μmol/L)of forskolin.(2)The qRT-PCR results showed that forskolin up-regulated the expression of Myh2,Myh4,Myomaker,but down-regulated the expression of Myh7 compared with the 0 μmol/L group,when C2C12 cells were differentiated for 4 days.Immunofluorescence staining results showed that the fusion index and myotube diameter of C2C12 cells were increased after forskolin treatment,and the number of myotubes was also increased.(3)Western blot results showed that the phosphorylated extracellular signal-regulated kinase 1/2 expression was inhibited;however,the phosphorylated protein kinase B was promoted after treatment with forskolin.The protein expression level of the myogenic differentiation transcription factor Myogenin was significantly up-regulated after treatment with forskolin.The above results demonstrate that forskolin may promote myogenic differentiation of C2C12 skeletal muscle cells through the extracellular signal-regulated kinase 1/2 and protein kinase B signaling pathway.

With the continuous deepening of the concept of precision diagnosis and treatment for lung cancer, how to achieve higher efficiency and accuracy in the screening, diagnosis, and treatment pathways in clinical practice has become an important issue that urgently needs to be overcome. The current clinical difficulty lies in the fact that despite continuous advancements in imaging and molecular diagnostic technologies, there are still limitations in manual efficiency and subjective experience when it comes to massive data analysis and multi-scale feature extraction. Artificial intelligence (AI), especially algorithm systems based on deep learning, is an innovative technology capable of deeply empowering medical big data. This method utilizes algorithms such as convolutional neural networks, combined with radiomics, pathomics, and multi-modal data fusion analysis, demonstrating immense potential in early precise detection and benign-malignant differentiation of pulmonary nodules, digital pathological subtype recognition and non-invasive prediction of driver genes, precise 3D surgical planning and automatic delineation of radiotherapy target volumes, as well as dynamic risk warning during follow-up. This innovative technology provides a brand-new solution for realizing intelligent and individualized lung cancer diagnosis and treatment models. This consensus, based on the latest evidence from evidence-based medicine and combined with the development trends in the AI field and real-world clinical needs, was ultimately formed by gathering the consensus opinions of multidisciplinary experts in radiology, pathology, thoracic surgery, and other fields. The main content covers the application specifications of AI in the three core scenarios of lung cancer screening, diagnosis, and treatment, the technical standards for data collection and algorithm validation, as well as the ethical and regulatory challenges faced at the current stage. It aims to clarify the applicable boundaries of AI as a clinical auxiliary decision support tool, providing scientific guidance and standardized exploration directions for peers currently engaged in or planning to carry out AI-assisted clinical diagnosis, treatment, and translation of lung cancer.

Severe intra-abdominal infections are life-threatening conditions and a significant challenge for surgeons. This article presents a case of an elderly patient with a severe intra-abdominal infection complicated by an anastomotic leak. This patient had experienced prolonged sepsis and multiple surgical traumas. Upon admission to our department, exploratory surgery revealed extensive bowel edema and adhesions, an anastomotic leak, and abdominal contamination with infection. In accordance with the principles of damage control surgery, the anastomotic leak was exteriorized, the abdomen was left open, and continuous intra-abdominal lavage with dual-lumen catheters was implemented to effectively control the infection. Negative pressure wound therapy was used to manage the open abdomen, and a negative pressure-assisted drainage device was used to manage the enteroatmospheric fistula. After granulation of the abdominal wound, split-thickness skin grafting was performed. The enteroatmospheric fistula was converted into an enterocutaneous fistula. A 3D-printed stoma baseplate was used to manage the digestive fistula. Concurrently, enhanced parenteral and enteral nutritional support was provided. Six months later, the patient successfully underwent definitive fistula resection and abdominal wall defect repair.

Objective:To evaluate the efficacy of vacuum sealing drainage (VSD) in the comprehensive management of full-thickness perineal wound dehiscence following pelvic exenteration (PE).Methods:This study employed a descriptive case series design. We retrospectively analyzed the clinical data of 29 patients who developed postoperative perineal wound infections with full-thickness dehiscence after PE. These cases included 16 patients from the Department of General Surgery at Jiangyin People's Hospital (Jiangsu Province) and 13 patients from the Department of Colorectal Surgery at the Second Affiliated Hospital of Naval Medical University (Shanghai Changzheng Hospital). VSD was applied to manage the dehisced wounds, with outcomes assessed based on wound healing time, complications, and follow-up data.Results:A total of 29 patients were included in the study. The operative time for PE was (498 ± 83) minutes. Among them, 23 patients underwent combined sacrococcygeal resection. The median number of VSD devices used was 28 (22, 39). The postoperative perineal wound healing time was 95 (82, 110) days in patients who underwent combined sacrococcygeal resection, 74 (63, 89) days in those without sacrococcygeal resection, 93 (79, 102) days in those treated with simple pelvic-abdominal isolation using a biological basement membrane mesh and 76 (60, 91) days in those who received combined pelvic packing with a pedicled omental flap. All patients uniformly developed Clavien-Dindo grade III complications at 2 weeks postoperatively, manifesting as perineal wound infection and dehiscence, which were successfully managed with VSD therapy. Subsequent evaluation identified delayed (>30 days) grade III complications, including enterocutaneous (3 cases) and urinary (2 cases) fistulae, all requiring surgical revision. All patients completed the follow-up at 6 months postoperatively. Three patients still presented with minimal exudate from the perineal wound, which resolved after standardized wound care and packing with alginate silver ion dressings. Four cases (13.8%) developed stoma high-output syndrome, which improved after oral medication. Eight patients (27.6%) developed adhesive intestinal obstruction, which improved with conservative treatment.Conclusions:VSD demonstrates unique advantages in managing complex wounds. For full-thickness perineal wound dehiscence after PE, VSD is a safe and effective therapeutic strategy.

Objective:To explore the value of contrast enhanced ultrasonography(CEUS) combined with shear wave elastography (SWE) in the diagnosis of urethral stricture.Methods:One hundred and twenty patients with urethral stricture were recruited, and preoperative retrograde urography, common urethral ultrasound, and CEUS were used to measure the length of the stricture site, and SWE was used to detect the elasticity value of the stricture site and its nearby spongiosum. Intraoperative urethral stricture lesion site measurements were performed to confirm the actual lesion location and length during surgery and to compare them with the lengths measured in the preoperative examination;4 months after surgery, SWE was used to detect the stiffness of the surgical site and to assess the surgical efficacy and prognosis.Results:The stenosis length measured by CEUS and intraoperative measurement had no statistical difference: (4.41 ± 1.47) cm vs. (4.37 ± 1.36) cm, t = 0.22, P>0.05. The elastic value in urethra stenosis in SWE was higher than that in normal urethra: (33.1 ± 6.7) kPa vs. (23.3 ± 4.1) kPa, there was statistical difference ( t = 13.67, P<0.05). The elastic value of unobstructed urethra and that of normal urethra after reconstruction had no statistical difference ( t = 0.95, P>0.05). Conclusions:CEUS combined with SWE is a safe, accurate and effective method to evaluate urethral stricture. SWE can be used to measure postoperative outcomes and recurrence.

Objective To explore the clinical characteristics and treatment methods of female primary enteric adenocarcinoma of the urethra.Methods The diagnosis and treatment of one case were reported.The origin,etiology,imaging findings,diagnosis,treatment,and prognosis were discussed in combination with relevant literature.Results The patient is a 61-year-old female,visited our hospital due to vaginal odor complicated with urethral bleeding for 2 months,and was diagnosed with urethral adenocarcinoma after examination.She underwent partial urethral resection,urethral meatus shaping and temporary bladder fistula surgery,with negative resection margins.However,local recurrence occurred 7 months after surgery.The patient was readmitted for a second time,undergoing laparoscopic total urethral resection,anterior vaginal wall resection,bladder neck resection,and permanent bladder fistulation,followed by chemotherapy.At the 19th month after the first surgery,the patient was admitted due to tumor recurrence,and the third surgery consisted of total bladder and uterus resection,ileal neobladder,and perineal closure.Postoperative pathology indicated urethral adenocarcinoma originating from the intestines.During the 39-month follow-up,the tumor recurred twice.No recurrence or metastasis was observed after the final total pelvic exenteration.Conclusion Enteric adenocarcinoma of the urethra is extremely rare.Comprehensive treatment including surgery,radiotherapy,and chemotherapy may prolong survival,but the overall prognosis is poor,requiring long-term follow-up.

Objective To construct mice hepatocellular carcinoma models with different tumor immune microenvironments(TIME)and explore the differences.Methods H22 and hepa1-6 were used to construct subcutaneous transplantation tumor model of C57 mice as homologous hepatocellular carcinoma cell lines(denoted as H22 group and hepal-6 group,each n=8),and the differences of TIME were evaluated.Immunohistochemistry was used to detect and quantify the infiltration of T cells,CD4+T cells,CD8+T cells,regulatory T cells and B cells in TIME.Flow cytometry was performed to detect the differences of composition of immune cell subpopulations in peripheral blood and tumor parenchyma.Gene expression profile characteristics of tumor tissue were analyzed based on high-throughput transcriptome sequencing technology,and enrichment analyses of immune-related signaling pathways were evaluated combined with gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG).Results H22 group showed cold and hepa1-6 group showed hot TIME characteristics.The number of T cells,CD4+T cells and CD8+T cells in tumor tissue of H22 group were all lower,while the proportion of T cells,CD4+T cells and CD8+T cells in peripheral blood were all higher than those of hepa1-6 group(all P＜0.05).Compared with H22 group,up-regulated genes of tumor tissue in hepa1-6 group expressed significantly enriched in tumor immune activation-related signaling pathways.Conclusion H22 and hepa1-6 hepatocellular carcinoma models showed distinct TIME characteristics of cold and hot tumors,respectively,and the amount of immune cells in tumor tissue of the former were significantly lower than those in the latter.

Objective:To investigate the correlation between postoperative spinal sagittal parameters and reinforced vertebral recompression fractures in patients with osteoporotic vertebral compression fractures (OVCFs) who have undergone percutaneous kyphoplasty (PKP).Methods:Data on patients with OVCFs treated with PKP at the Department of Orthopaedics, Second Affiliated Hospital of Soochow University, from August 2020 to August 2024, were collected. Among these, 31 patients who underwent single-segment PKP experienced postoperative reinforced vertebral recompression fractures (recompression fracture group), comprising 8 males and 23 females, with a mean age of 73.74±8.76 years, a body mass index (BMI) of 23.83±1.87 kg/m 2, and a bone mineral density T-value of -2.29±0.55. The remission rate of the visual analogue scale (VAS) after surgery was 80.14%±4.86%, with a mean volume of bone cement used at 5.37±0.69 ml. The surgical segments involved included T 5 (1 case), T 8 (1 case), T 10 (1 case), T 11 (4 cases), T 12 (9 cases), L 1 (7 cases), L 2 (4 cases), L 3 (2 cases), and L 4 (2 cases). Following a 1∶1 matching principle, 31 patients whose vertebrae did not experience reinforced recompression fractures during the same period (non-recompression fracture group) were included. This group also comprised 8 males and 23 females, with a mean age of 74.88±8.31 years, a BMI of 23.15±2.04 kg/m 2, a bone mineral density T-value of -2.76±0.64, and a VAS remission rate of 79.75%±5.01%. The mean volume of bone cement used in this group was 5.41±0.72 ml. The surgical segments involved included T 8 (1 case), T 10 (1 case), T 11 (4 cases), T 12 (8 cases), L 1 (7 cases), L 2 (5 cases), L 3 (2 cases), L 4 (2 cases), and L 5 (1 case). There were no statistically significant differences in the aforementioned indicators between the two patient groups ( P>0.05). A comparison of the postoperative spinal sagittal parameters between the two groups was conducted, focusing on the local kyphosis angle (LKA), lumbar lordosis (LL), thoracic kyphosis (TK), pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS), and the lumbar-pelvic matching value (PI-LL). Indicators that exhibited statistically significant differences were included in the binary logistic regression analysis to evaluate the impact of spinal sagittal parameters following PKP on the recompression of the reinforced vertebral. Results:The time to reinforced vertebral recompression fractures after PKP ranged from 35 to 184 d, with a median of 69 d. The TK in the recompression fracture group (46.56°±7.02°) was significantly greater than that in the non-recompression fracture group (41.95°±5.76°). Additionally, the LKA, PI and SS were all smaller in the recompression fracture group (9.84°±2.13°, 41.36°±4.27°, 22.69°±5.53°, respectively) compared to the non-recompression fracture group (12.37°±2.64°, 48.09°±6.33°, 28.41°±7.64°), with all differences being statistically significant ( P<0.05). However, no significant differences were observed between the LL, PT, and PI-LL values ( P>0.05). TK, LKA, PI, and SS were included in the binary logistic regression analysis, which indicated that TK [ OR=1.533, 95% CI(1.47, 1.59)] after PKP was positively correlated with the occurrence of reinforced vertebral recompression fractures. Conversely, LKA [ OR=0.882, 95% CI(0.80, 0.96)], PI [ OR=0.815, 95% CI(0.71, 0.91)], and SS [ OR=0.833, 95% CI(0.73, 0.93)] were negatively correlated. Conclusions:The incidence of reinforced vertebral recompression fractures following PKP is associated with spinal sagittal parameters, including TK, LKA, PI, and SS. Specifically, a larger TK and smaller values of LKA, PI, and SS are correlated with an elevated risk of reinforced vertebral recompression fractures.

Objective:To investigate the clinical effect of the transfer of lateral femoral fascia or combined with anteriolateral thigh flap (ALTF) on treatment of defects of Achilles tendon and surrounding soft tissue.Methods:From May 2004 to December 2021, 15 patients with Achilles tendons defects were treated with the transfer of either lateral femoral fascia or combined with ALTF in the Department of Hand and Foot Microsurgery, Xi'an Fengcheng Hospital. Two of the patients had postoperative infection after the primary treatment for Achilles tendon rupture and 13 patients had Achilles tendon defect together with various grade of defects of surrounding soft tissues. According to the size of wound, ALTFs and lateral superior knee flaps were used for the reconstructions. Various complex tissue flaps were used in reconstructive surgery: ALTF in 10 patients, the lateral thigh muscular flap in 3 patients and the lateral knee artery flap in 5 patients. The length of Achilles tendon defect was 3.0-8.0 cm and the length of fascia was 4.0-9.0 cm. The sizes of the flaps were 4.0 cm×3.0 cm-30.0 cm×18.0 cm. After the fascia and the flaps were harvested, the fascia was rolled up or wrapped up around the muscle with the knee at 30° flexion and the ankle at 30° flexion. Then end-to-end or end-to-side blood vessel anastomoses were carried out. The ALTFs were used to cover the wounds.Results:All the 15 flaps survived. After 2 to 5 years of follow-up, the heel lift test was found negative, without re-broken of Achilles tendon. All patients were in normal walking gait, except 2 patients who had mild lameness. According to the Amer-Lindholm scoring criteria, the results were excellent in 13 patients and good in 2 patients.Conclusion:Lateral thigh composite tissue transfer is an effective method to reconstruct Achilles tendon and the defects of its surrounding tissues.