BACKGROUND:Spleen has the functions of blood storage,hematopoiesis,and immunity.With the increase of age,the structural degeneration and functional decline of spleen lead to the impairment of immune system function,thus accelerating the aging process of the body.The treatment of spleen aging in tree shrews with highly active umbilical cord mesenchymal stem cells has not been reported. OBJECTIVE:To explore the intervention effect and mechanism of highly active umbilical cord mesenchymal stem cells on spleen aging in tree shrews. METHODS:Highly active umbilical cord mesenchymal stem cells were isolated,cultured,and obtained from the umbilical cord tissue of newborn tree shrews by caesarean section.The differentiation abilities of adipogenesis,osteogenesis,and chondrogenesis were detected by three-line differentiation kit.Cell cycle and surface markers were detected by flow cytometry.The second generation of highly active umbilical cord mesenchymal stem cells were transfected with Genechem Green Fluorescent Protein with infection complex values of 100,120,140,160,180,and 200,respectively,to screen the best transfection conditions.After transfection,the fourth generation of highly active umbilical cord mesenchymal stem cells was injected into the tail vein of tree shrews in the elderly treatment group.The young control group and the aged model group were not given special treatment.After 4 months of treatment,the spleen tissue was taken and the structure of the spleen was observed by hematoxylin-eosin staining.β-Galactosidase staining was used to detect the activity of aging-related galactosidase.Immunohistochemical staining was used to detect the expression levels of p21 and p53 proteins.Ki67 and PCNA immunofluorescence staining was used to detect cell proliferation activity.Immunofluorescence staining was used to detect the expression levels of spleen autophagy protein molecules Beclin 1 and APG5L/ATG5.Reactive oxygen species fluorescence staining was used to detect the content of reactive oxygen species in spleen tissue.CD3 immunofluorescence staining was used to detect the change of the proportion of total T lymphocytes.The secretion levels of interleukin 1β and transforming growth factor β1 in spleen were detected by enzyme linked immunosorbent assay.The distribution of highly active umbilical cord mesenchymal stem cells labeled with green fluorescent protein in spleen tissue was observed by DAPI double staining of nucleus. RESULTS AND CONCLUSION:(1)Highly active umbilical cord mesenchymal stem cells grew in a short spindle shape with fish-like growth,with a large proportion of G0/G1 phase,and had the potential to differentiate into adipogenesis,osteogenesis,and chondrogenesis.(2)Multiplicity of infection=140 and transfection for 72 hours were the best conditions for labeling tree shrews highly active umbilical cord mesenchymal stem cells with Genechem Green Fluorescent Protein.(3)Compared with the aged model group,in the aged treatment group,the spleen tissue cells of tree shrews were arranged closely,and the area of white pulp was increased(P<0.01);the boundary between red pulp and white pulp was clear;the proportion of germinal centers did not show statistically significant difference(P>0.05).The activity level of galactosidase related to spleen tissue aging was decreased(P<0.001),and the expression levels of aging protein molecules p21 and p53 were down-regulated(P<0.001).The expression levels of proliferation-related molecules Ki67 and PCNA were up-regulated(P<0.001,P<0.05);expression levels of autophagy-related molecules Beclin 1 and APG5L/ATG5 were up-regulated(P<0.001),and the content of reactive oxygen species decreased(P<0.001),and the proportion of CD3+T cells increased(P<0.05).The secretion level of interleukin 1β in the aging-related secretion phenotype decreased(P<0.001);no significant difference was found in transforming growth factor β1 level(P>0.05).Compared with the young control group,the above indexes were significantly different in the elderly treatment group(P<0.05).(4)Green fluorescent cells labeled with green fluorescent protein were observed in spleen tissue of tree shrews the elderly treatment group by frozen tissue section observation.The results show that intravenous infusion of highly active umbilical cord mesenchymal stem cells can migrate to spleen tissue,inhibit the production of reactive oxygen species,down-regulate the expression of aging-related proteins,induce autophagy,promote cell proliferation,reduce chronic inflammation,and then improve the structure and function of spleen tissue.

Objective:To evaluate the application of the reaming-irrigation-aspirator (RIA) technique in autologous bone grafting for non-union of lower limb fracture.Methods:A retrospective analysis was conducted of the data of 16 patients with non-union of lower limb fracture who had undergone autologous bone grafting using the RIA technique at Department of Lower Limb Orthopaedics Ⅰ, Zhengzhou Orthopaedic Hospital from July 2023 to March 2024. There were 12 males and 4 females with an age of (47.2±14.3) years. Sites of non-union: the femur in 14 cases and the tibia in 2 cases. Initial surgical fixation: intramedullary nail fixation in 5 cases, plate fixation in 10 cases, and intramedullary nail plus plate fixation in 1 case. The time for fracture non-union was (21.0±12.0) months. Intraoperatively, autologous bone reaming was conducted using a drill bit of RIA system, and simultaneous flushing using irrigation fluid. The bone debris carried out by the irrigation fluid was collected into a filtration device through negative pressure aspiration. After the filter was opened to squeeze out congestion, bone paste was taken out for standby. Internal fixation methods were adjusted according to the non-union condition of the fractures. Finally, the bone paste was implanted into bone defects. The time for autologous bone harvest, amount of bone harvested, intraoperative blood loss, postoperative pain at a donor site, fracture healing, functional recovery of a lower limb and occurrence of complications were recorded.Results:In 2 of the 16 patients, fracture of the RIA drill bit occurred when bone marrow was harvested from the tibia, and their operation was completed only after the drill bits were replaced. In the 16 patients, time for bone harvest was (31.4±6.1) min, bone harvest volume (42.6±9.5) mL, and intraoperative blood loss (815.6±238.6) mL. The incisions in this cohort healed primarily after operation, and 4-week follow-up revealed that no patients had pain symptoms at a donor site. The 16 patients were followed up for (8.0±2.0) months after operation. All patients achieved bone union after (4.4±0.5) months. Follow-ups showed that no patients had complications like delayed infection, donor site fracture, or loosening or fracture of an internal fixator. According to the rating scale for lower limb function at the last follow-up, the function of affected limb was evaluated as excellent in 7 cases, as good in 8 cases, and as fair in 1 case.Conclusion:Application of RIA technique for autologous bone grafting to treat non-union of lower limb fracture can lead to fine fracture healing and good lower limb function, but no long-term pain symptoms in the donor area for the patients.

Objective:To compare the clinical efficacy of the reamer-irrigator-aspirator (RIA) technique and iliac crest bone grafting (ICBG) in the treatment of adult noninfectious long bone nonunion of the lower extremity, and to evaluate the safety and effectiveness of the RIA technique.Methods:A retrospective review was conducted of 42 adult patients with noninfectious long bone nonunion of the lower extremity treated surgically at Zhengzhou Orthopaedic Hospital from January 2021 to June 2024. The cohort included 33 men and 9 women with a mean age of 47.0±14.4 years (range, 18-62 years). Twenty-three patients had nonunion on the left side and 19 on the right. Initial surgical fixation methods included plate fixation (19 cases), intramedullary nail (16 cases), external fixator (5 cases), and combined plate-and-nail fixation (2 cases). Revision procedures included debridement and freshening of the nonunion ends, revision or adjustment of internal fixation, and autologous bone grafting. Nineteen patients underwent graft harvesting with the RIA technique (RIA group), and 23 received autograft harvested from the iliac crest (iliac group). Baseline and perioperative variables compared between the two groups included sex, age, body-mass index, preoperative comorbidities, anatomical site of the nonunion (tibial or femoral), duration from the index fracture operation to the current revision, operative time, intraoperative blood loss, volume of blood transfusion and rate of perioperative complications, nonunion fracture sites, visual analogue scale (VAS) pain score at the fracture site, donor-site morphology and functional appearance scores, and radiographic union score for tibial fractures (RUST).Results:There was no significant difference in age between the RIA and ICBG groups (45.1±14.1 vs. 48.5±14.7 years, t=0.746, P=0.449). However, intraoperative blood loss was higher in the RIA group [300 (150, 720) ml, Z=-3.072, P=0.002] and the transfusion rate was higher (14/19 vs. 8/23, χ 2=6.313, P=0.012). The harvested bone volume was very similar [40(35, 60) cm 2vs. 40(35, 40) cm 2, Z=-1.077, P=0.281]. All patients were followed for a mean of 16.5±2.8 months (range, 7-40 months). VAS scores at the surgical site showed no significant difference between groups at 3 months (3.2±1.0 vs. 3.3±1.4, t= 4.754, P=0.720) and 6 months (1.6±0.8 vs. 1.8±1.2, t=6.841, P=0.542). Donor-site morphology and functional scores were significantly better in the RIA group at 3 months [14.5(13.0, 15.5) vs. 9.0(7.0, 10.0), Z=-3.748, P<0.001] and 6 months [15.5(15.0, 16.0) vs. 11.0(10.0, 13.0), Z=-3.782, P<0.001]. At 12 months postoperatively, RUST scores did not differ significantly between groups (10.6±1.2 vs. 10.4±1.5, t=0.483, P=0.632). The incidence of perioperative complications was comparable [42%(8/19) vs. 30%(7/23), χ 2=0.617, P=0.432]. Conclusions:Both RIA and ICBG achieved favorable clinical outcomes in the treatment of adult noninfectious lower-extremity long bone nonunion. Although RIA was associated with greater intraoperative blood loss, it resulted in less donor-site morbidity and better cosmetic and functional outcomes. Bone healing capacity and overall complication rates did not differ significantly between the two groups. These findings suggest that the RIA technique is safe and effective for use in the Chinese patient population.

Objective To explore the application value of three dimensions(3D)visualization combined with ultrasound(US)in percutaneous transhepatic cholangial drainage(PTCD).Methods A retrospective analysis was conducted on clinical data from 44 PTCD procedures performed by our team between January 2020 and March 2025.Twenty-one cases before January 2023 underwent conventional US-guided PTCD(US group),while 23 subsequent cases received 3D visualization-assisted US-guided PTCD(3D+US group).Both groups had comparable baseline characteristics.For the3D+US group,preoperative upper abdominal CT(non-contrast+contrast-enhanced)data were imported into the open-source 3D software 3D Slicer 5.6.2 to reconstruct individualized 3D models for preoperative puncture planning.Outcomes including puncture success rate,procedure time,times of puncture,therapeutic efficacy,and complications were compared between the two groups.Results Compared with the US group,the 3D+US group required fewer puncture times(1/2/3 times:19/4/0 cases vs.10/9/2 cases,Z=-2.602,P=0.009)and shorter procedure time[(38.8±12.2)min vs.(56.6±14.8)min,t=-4.372,P=0.000].No significant differences were observed in puncture success rate[100.0%(23/23)vs.90.5%(19/21),P=0.222],therapeutic efficacy(markedly effective/effective/ineffective:13/9/1 cases vs.10/9/2 cases,Z=-0.688,P=0.492),or complication rate[8.7%(2/23)vs.19.0%(4/21),χ2=0.313,P=0.576].Conclusion 3D visualization combined with US guidance for PTCD is safe and effective,facilitating reduced procedure time and fewer puncture times.

Objective To detect the expression of cytochrome c oxidase assembly factor 6(COA6)in breast cancer,study its clinical significance,and analyze the effect of COA6 on immune infiltration in breast cancer.Methods Differential genes were screened by the whole transcriptome sequencing and the expression of COA6 was explored with TCGA(The Cancer Genome Atlas Program)database.The tissues of 125 breast cancer and adjacent tissues were stained by immunohistochemistry to detect COA6 protein expression and analyze the correlation with clinical features.The COA6 mRNA and protein expression in breast cancer cells and tissues were determined by qRT-PCR and Western blot,respectively.The TIMER(Tumor Immune Estimation Resource)database was used to analyze the correlation between high COA6 gene expression and immune cell infiltration.Results The positive expression rate of COA6 in breast cancer tissues was 88％(110/125),which was higher than that of paracarinoma tissues(7.2％,9/125)(P＜0.05)and was positively correlated with tumor size and histological grade.In fresh breast cancer tissues,COA6 protein expression was significantly higher than that in adjacent tissues.Both COA6 mRNA and protein expression were significantly increased in the breast cancer cell lines.COA6 was positively cor-related with the infiltration of helper T cells,NK cells,CD8+T cells,M1 type macrophages,regulatory T cells,dendritic cells,and memory CD4+T cells in the tumor microenvironment.Conclusions The expression level of COA6 is increased in breast cancer and is positively correlated with tumor immune infiltration,which provides a po-tential therapeutic target for breast cancer treatment.

Objective:To compare the clinical efficacy of the reamer-irrigator-aspirator (RIA) technique and iliac crest bone grafting (ICBG) in the treatment of adult noninfectious long bone nonunion of the lower extremity, and to evaluate the safety and effectiveness of the RIA technique.Methods:A retrospective review was conducted of 42 adult patients with noninfectious long bone nonunion of the lower extremity treated surgically at Zhengzhou Orthopaedic Hospital from January 2021 to June 2024. The cohort included 33 men and 9 women with a mean age of 47.0±14.4 years (range, 18-62 years). Twenty-three patients had nonunion on the left side and 19 on the right. Initial surgical fixation methods included plate fixation (19 cases), intramedullary nail (16 cases), external fixator (5 cases), and combined plate-and-nail fixation (2 cases). Revision procedures included debridement and freshening of the nonunion ends, revision or adjustment of internal fixation, and autologous bone grafting. Nineteen patients underwent graft harvesting with the RIA technique (RIA group), and 23 received autograft harvested from the iliac crest (iliac group). Baseline and perioperative variables compared between the two groups included sex, age, body-mass index, preoperative comorbidities, anatomical site of the nonunion (tibial or femoral), duration from the index fracture operation to the current revision, operative time, intraoperative blood loss, volume of blood transfusion and rate of perioperative complications, nonunion fracture sites, visual analogue scale (VAS) pain score at the fracture site, donor-site morphology and functional appearance scores, and radiographic union score for tibial fractures (RUST).Results:There was no significant difference in age between the RIA and ICBG groups (45.1±14.1 vs. 48.5±14.7 years, t=0.746, P=0.449). However, intraoperative blood loss was higher in the RIA group [300 (150, 720) ml, Z=-3.072, P=0.002] and the transfusion rate was higher (14/19 vs. 8/23, χ 2=6.313, P=0.012). The harvested bone volume was very similar [40(35, 60) cm 2vs. 40(35, 40) cm 2, Z=-1.077, P=0.281]. All patients were followed for a mean of 16.5±2.8 months (range, 7-40 months). VAS scores at the surgical site showed no significant difference between groups at 3 months (3.2±1.0 vs. 3.3±1.4, t= 4.754, P=0.720) and 6 months (1.6±0.8 vs. 1.8±1.2, t=6.841, P=0.542). Donor-site morphology and functional scores were significantly better in the RIA group at 3 months [14.5(13.0, 15.5) vs. 9.0(7.0, 10.0), Z=-3.748, P<0.001] and 6 months [15.5(15.0, 16.0) vs. 11.0(10.0, 13.0), Z=-3.782, P<0.001]. At 12 months postoperatively, RUST scores did not differ significantly between groups (10.6±1.2 vs. 10.4±1.5, t=0.483, P=0.632). The incidence of perioperative complications was comparable [42%(8/19) vs. 30%(7/23), χ 2=0.617, P=0.432]. Conclusions:Both RIA and ICBG achieved favorable clinical outcomes in the treatment of adult noninfectious lower-extremity long bone nonunion. Although RIA was associated with greater intraoperative blood loss, it resulted in less donor-site morbidity and better cosmetic and functional outcomes. Bone healing capacity and overall complication rates did not differ significantly between the two groups. These findings suggest that the RIA technique is safe and effective for use in the Chinese patient population.

Objective To explore the application value of three dimensions(3D)visualization combined with ultrasound(US)in percutaneous transhepatic cholangial drainage(PTCD).Methods A retrospective analysis was conducted on clinical data from 44 PTCD procedures performed by our team between January 2020 and March 2025.Twenty-one cases before January 2023 underwent conventional US-guided PTCD(US group),while 23 subsequent cases received 3D visualization-assisted US-guided PTCD(3D+US group).Both groups had comparable baseline characteristics.For the3D+US group,preoperative upper abdominal CT(non-contrast+contrast-enhanced)data were imported into the open-source 3D software 3D Slicer 5.6.2 to reconstruct individualized 3D models for preoperative puncture planning.Outcomes including puncture success rate,procedure time,times of puncture,therapeutic efficacy,and complications were compared between the two groups.Results Compared with the US group,the 3D+US group required fewer puncture times(1/2/3 times:19/4/0 cases vs.10/9/2 cases,Z=-2.602,P=0.009)and shorter procedure time[(38.8±12.2)min vs.(56.6±14.8)min,t=-4.372,P=0.000].No significant differences were observed in puncture success rate[100.0%(23/23)vs.90.5%(19/21),P=0.222],therapeutic efficacy(markedly effective/effective/ineffective:13/9/1 cases vs.10/9/2 cases,Z=-0.688,P=0.492),or complication rate[8.7%(2/23)vs.19.0%(4/21),χ2=0.313,P=0.576].Conclusion 3D visualization combined with US guidance for PTCD is safe and effective,facilitating reduced procedure time and fewer puncture times.

Objective:To evaluate the application of the reaming-irrigation-aspirator (RIA) technique in autologous bone grafting for non-union of lower limb fracture.Methods:A retrospective analysis was conducted of the data of 16 patients with non-union of lower limb fracture who had undergone autologous bone grafting using the RIA technique at Department of Lower Limb Orthopaedics Ⅰ, Zhengzhou Orthopaedic Hospital from July 2023 to March 2024. There were 12 males and 4 females with an age of (47.2±14.3) years. Sites of non-union: the femur in 14 cases and the tibia in 2 cases. Initial surgical fixation: intramedullary nail fixation in 5 cases, plate fixation in 10 cases, and intramedullary nail plus plate fixation in 1 case. The time for fracture non-union was (21.0±12.0) months. Intraoperatively, autologous bone reaming was conducted using a drill bit of RIA system, and simultaneous flushing using irrigation fluid. The bone debris carried out by the irrigation fluid was collected into a filtration device through negative pressure aspiration. After the filter was opened to squeeze out congestion, bone paste was taken out for standby. Internal fixation methods were adjusted according to the non-union condition of the fractures. Finally, the bone paste was implanted into bone defects. The time for autologous bone harvest, amount of bone harvested, intraoperative blood loss, postoperative pain at a donor site, fracture healing, functional recovery of a lower limb and occurrence of complications were recorded.Results:In 2 of the 16 patients, fracture of the RIA drill bit occurred when bone marrow was harvested from the tibia, and their operation was completed only after the drill bits were replaced. In the 16 patients, time for bone harvest was (31.4±6.1) min, bone harvest volume (42.6±9.5) mL, and intraoperative blood loss (815.6±238.6) mL. The incisions in this cohort healed primarily after operation, and 4-week follow-up revealed that no patients had pain symptoms at a donor site. The 16 patients were followed up for (8.0±2.0) months after operation. All patients achieved bone union after (4.4±0.5) months. Follow-ups showed that no patients had complications like delayed infection, donor site fracture, or loosening or fracture of an internal fixator. According to the rating scale for lower limb function at the last follow-up, the function of affected limb was evaluated as excellent in 7 cases, as good in 8 cases, and as fair in 1 case.Conclusion:Application of RIA technique for autologous bone grafting to treat non-union of lower limb fracture can lead to fine fracture healing and good lower limb function, but no long-term pain symptoms in the donor area for the patients.

Tubular aggregates(TA)are ultrastructural abnormalities in muscle biopsies,which can be detected in muscle biopsy specimens from patients with a variety of hereditary and acquired disorders.A 34-year-old male patient diagnosed with systemic lupus erythematosus(SLE)presented with intermittent muscle weakness localized to the proximal extremities of both lower limbs during prolonged oral administration of methylprednisolone,hydroxychloroquine and tacrolimus.Laboratory findings indicated normal creatine kinase levels,and anti-U1-RNP/Sm antibodies were elevated up to 49.45 RU/mL.Electromyography revealed myogenic lesions in the left iliopsoas muscle and muscle pathology demonstrated TA within the muscle fibers.Genetic testing excluded the possibility of hereditary disorders with tubular aggregas.Combined with literature review,the etiology and clinical characteristics of TA were discussed to increase the understanding of the diagnosis of diseases with TA.This case report demonstrates that SLE patients can have fluctuating muscle weakness and TA in muscle pathology.The symptoms of SLE can be partially relieved by adjusting SLE medications.