ObjectiveTo study the mechanism by which Shexiang Tongxin dropping pills （STDP） ameliorate the dysfunction of coronary microvascular endothelial cells in the rat model of heart failure. MethodsThe heart failure model was established by ligation of the left anterior descending coronary artery in rats， which were then allocated into sham， model， STDP， and telmisartan （TLM） groups and treated for 21 days. The heart function was detected by echocardiography， and the levels of myocardial injury markers， nitric oxide （NO）， endothelin-1 （ET1）， and angiotensinⅡ （AngⅡ） were determined by enzyme-linked immunosorbent assay （ELISA）. The protein levels of endothelial nitric oxide synthase （eNOS） and inducible nitric oxide synthase （iNOS） were determined by Western blot. The model of cardiac microvascular endothelial cell injury was established by AngⅡ induction and then treated with the STDP-containing serum （5%， 10%， and 20%） for 24 h. The levels of NO and ET1 were measured by ELISA. Western blot was employed to determine the protein levels of eNOS， iNOS， angiotensin-converting enzyme 2 （ACE2）， and angiotensinⅡ receptor 2 （AT2）. MLN-4760， an ACE2 inhibitor， was used to explore the mechanism underpinning the regulatory effect of STDP on the ACE2-AT2/MAS pathway. ResultsCompared with the sham group， the model group showed decreases in left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （LVFS） （P<0.05）， a decline in serum NO level， elevations in serum AngⅡ and ET1 levels， a reduction in p-eNOS/eNOS ratio， and up-regulation in iNOS expression （P<0.05）. Compared with the model group， STDP increased LVEF， LVFS， and cardiac output （P<0.05）， raised the level of NO and lowered the levels of AngⅡ and ET1 in the serum （P<0.05）， increased the p-eNOS/eNOS value， and inhibited iNOS expression （P<0.05）. Compared with the AngⅡ group， STDP increased the NO content and decreased the ET1 content in endothelial cells （P<0.05）， increased the p-eNOS/eNOS ratio， and inhibited the iNOS expression （P<0.05）. The ACE2 inhibitor MLN-4760 reversed the regulatory effects of STDP on p-eNOS， eNOS， and iNOS. ConclusionSTDP improves the cardiac function in the rat model of heart failure， enhances the synthesis and release of NO in cardiac microvascular endothelial cells， reduces AngⅡ and ET1 levels， and regulates the expression of p-eNOS and eNOS， thereby ameliorating the dysfunction of microvascular endothelial cells in heart failure. This mechanism is related to the upregulation of the expression of proteins in the ACE2-AT2/MAS pathway.

ObjectiveTo study the mechanism of Shexiang Tongxin Dripping pills-containing serum （STDP） in ameliorating angiotensinⅡ （AngⅡ）-induced cell phenotype transformation， proliferation， migration， and dysfunction of vascular smooth muscle cells. MethodsAn AngⅡ-induced proliferation and migration model of vascular smooth muscle cells was established. The cells were treated with STDP at 5%， 10%， and 20% for 24 h. The immunofluorescence assay was employed to detect the expression of α smooth muscle actin （α-SMA） and matrix metalloproteinase-2 （MMP-2）. The cell-counting kit-8 （CCK-8） assay and 5-ethynyl-2'-deoxyuridine （EdU） staining were employed to detect the proliferation of vascular smooth muscle cells， and the scratch assay was employed to detect the migration of the cells. Western blot was employed to determine the expression levels of pathway proteins such as angiotensin-converting enzyme 2 （ACE2）， angiotensin Ⅱ type 2 （AT2）， angiotensin Ⅱ type 1 （AT1）， as well as proliferation and migration proteins such as typeⅠ collagen （ColⅠ） and osteopontin （OPN）. ResultsCompared with the model group， STDP increased the expression of α-SMA， reduced the expression of MMP-2， and inhibited the proliferation and migration of vascular smooth muscle cells （P<0.05）. Furthermore， STDP up-regulated the expression levels of ACE2， AT2， and MAS1， while down-regulating the expression level of AT1， PCNA， ColⅠ， MMP-9， Rock1， Rock2， and SRF （P<0.05）. Compared with the STDP group， the ACE2 inhibitor reversed the regulatory effects of STDP. ConclusionSTDP inhibits the phenotype transformation， proliferation， and migration of vascular smooth muscle cells and regulates the expression of cell proliferation and migration-related proteins to ameliorate the dysfunction of vascular smooth muscle cells. It exerts the effects by up-regulating the expression of proteins in the ACE2-AT2/MAS pathway and down-regulating the expression of proteins in the AT1-Rock signaling pathway.

Objective:To explore the efficacy and safety of the ballistic-ultrasound-negative pressure three-in-one energy platform (Trilogy) for micro-channel percutaneous nephrolithotomy (mini-PCNL).Methods:A retrospective analysis was conducted on the clinical data of 140 patients with upper urinary tract stones treated at Tianjin Medical University Second Hospital from February to October 2024. All patients underwent mini-PCNL and were divided into the holmium laser group and the Trilogy group based on the stone fragmentation equipment used during the procedure. There were 69 patients in the holmium laser group and 71 in the Trilogy group. The two groups had similar mean ages (55.1±10.2 years vs. 53.4±10.8 years), male patient proportions (50 cases, 72.5% vs. 50 cases, 70.4%), body mass indices (25.2±3.6 kg/m 2 vs. 25.3±4.0 kg/m 2), incidence rates of hypertension (29 cases, 42.0% vs. 31 cases, 43.7%), diabetes (15 cases, 21.7% vs. 12 cases, 16.9%), mean cumulative stone lengths (39.2±12.6 mm vs. 35.9±14.8 mm), total stone volumes preoperatively (6 184.3±3 653.5 mm 3 vs. 5 644.9±4 173.8 mm 3), mean CT values for stones (1 138.2±264.3 HU vs. 1 151.3±208.0 HU), stone locations (ureter 14 cases, 20.3% vs. 22 cases, 31.0%; kidney 48 cases, 69.6% vs. 39 cases, 54.9%; both ureter and kidney 7 cases, 10.1% vs. 10 cases, 14.1%), preoperative mean urinary white blood cell counts [9.6(3.6, 31.2) cells/HPF vs. 11.9(3.8, 34.5) cells/HPF], proportions of patients with preoperative urinary white blood cells (+ + + ; 23 cases, 33.3% vs. 25 cases, 35.2%), nitrite positivity rates (4 cases, 5.8% vs. 3 cases, 4.2%), and urine culture positivity rates (12 cases, 17.4% vs. 18 cases, 25.4%) showed no statistically significant differences. The proportion of patients with moderate or higher hydronephrosis in the holmium laser group was lower than that in the Trilogy group (32 cases, 46.4% vs. 47 cases, 66.2%, P=0.018). The holmium laser group utilized holmium laser lithotripsy, where stone fragments were either flushed out with a vortex or retrieved with a stone basket. The Trilogy group employed a three-in-one energy platform to break the stones. This device incorporated pneumatic ballistic, ultrasound, and negative pressure suction capabilities within the same metallic probe, allowing the stone to be fragmented into small pieces while simultaneously performing ultrasonic negative pressure stone clearance. The parameters for the three-in-one energy platform were adjusted based on intraoperative conditions, typically setting negative pressure at 30%-50%, ultrasound power at 80%-100%, ballistic power at 80%, and frequency at 8 Hz. During the stone fragmentation process, the ballistic device fragmented the stones while ultrasound further reduced larger fragments and removed them. Some fragments that were difficult to break could also be flushed out or retrieved with a stone basket. The efficiency of stone clearance (volume of stones cleared per unit time) was compared between the two groups, as well as the stone-free rates on postoperative day 1 and day 30. Stone clearance time was defined as the duration from the start of fragmentation to the placement of the nephrostomy tube. Changes in postoperative white blood cells, hemoglobin, and albumin levels compared to preoperative levels, as well as the incidence of Clavien-Dindo complications, were compared between the two groups. Equipment failure incidents were recorded (fiber fracture in the holmium laser group indicating it could not be used; probe fracture in the Trilogy group). Patients were sub-grouped based on stone CT values into CT ≥ 1 000 HU and CT < 1 000 HU categories to compare stone clearance efficiency between the two devices within each sub-group. In the CT≥1 000 HU sub-group, there were 51 cases in the holmium laser group and 54 in the Trilogy group, there were no significant differences in preoperative total stone volume (6 785.0±3 902.3 mm 3 vs. 5 678.1±4 297.7 mm 3). In the CT < 1 000 HU sub-group, there were 18 cases in the holmium laser group and 17 in the Trilogy group. There were no significant differences between the groups in preoperative total stone volume (4 482.2±2 110.6 mm 3 vs. 5 530.9±3 845.3 mm 3). Results:The overall stone clearance efficiency in the Trilogy group was higher than that in the holmium laser group (87.9±35.7 mm 3/min vs. 77.1±24.3 mm 3/min, P=0.038). There were no significant differences in residual stone volume before discharge [5.5(0, 84.0) mm 3 vs. 5.3(0, 175.0) mm 3], stone clearance time (79.4±43.2 min vs. 66.6±49.7 min), or the proportion of patients using stone baskets during the procedure (33 cases, 47.8% vs. 36 cases, 50.7%). Postoperative changes in white blood cells, hemoglobin, and albumin compared to preoperative levels were not significantly different [(4.1±2.9)×10 9/L vs. (3.3±2.2)×10 9/L; (-2.9±10.5) g/L vs. (-1.6±9.3) g/L; (-2.5±3.6) g/L vs. (-1.8±5.0) g/L] Furthermore, there were no statistically significant differences in equipment failure rates (1 case, 1.4% vs. 4 cases, 5.6%), stone-free rates (postoperative day 1: 43 cases, 62.3% vs. 47 cases, 66.2%; postoperative day 30: 50 cases, 72.5% vs. 53 cases, 74.6%), or Clavien-Dindo complication rates (grade Ⅰ: 11 cases, 15.9% vs. 8 cases, 11.3%; grade Ⅱ: 2 cases, 2.8% vs. 0 cases; grade Ⅲ: 1 case, 1.4% vs. 0 cases). In the CT ≥ 1 000 HU sub-group, the clearance time for the holmium laser was longer than that for Trilogy (93.3±41.0 min vs. 74.6±51.9 min, P=0.044), there were no significant differences in residual stone volume before discharge [6.3(1.6, 173.8) mm 3 vs. 4.5(0, 69.0) mm 3] between the two groups. In the CT < 1 000 HU sub-group, the overall stone clearance efficiency of the Trilogy group exceeded that of the holmium laser group (134.2±38.0 mm 3/min vs. 105.5 ± 7.1 mm 3/min, P=0.004), there were no significant differences between the groups in residual stone volume before discharge [0(0, 51.1) mm 3 vs. 16.3(0, 957.2) mm 3], or stone clearance time (40.2±18.1 min vs. 39.1±27.5 min). Conclusions:In mini-PCNL surgery, the stone fragmentation efficiency of the three-in-one lithotripsy energy platform is superior to that of the holmium laser, while the overall complication rate is comparable to that of the holmium laser.

BACKGROUND:Due to the aging population and the increase in elderly diseases,research and utilization of epidermal stem cells in the elderly have become increasingly important.However,due to the weak proliferation and drying ability of epidermal stem cells in the elderly,their application in biomedical and clinical research is limited.OBJECTIVE:To investigate the ability of rotating cell culture system to promote the proliferation and maintenance of epidermal stem cells in elderly individuals.METHODS:Epidermal stem cells were obtained from children (7-12 years old) and the elderly (over 60 years old) using traditional planar culture methods.After identification by cell morphology and cell immunofluorescence,the differences in proliferation ability of epidermal stem cells between the children and the elderly were analyzed by flow cytometry,CCK-8 assay,and cell clone formation assay.Elderly epidermal stem cells were cultured using a rotating cell culture system and 3D TableTrix? microcarriers.The cell proliferation and expression of stemness markers were detected by Live/Dead staining,cell doubling efficiency,cell doubling time,and real-time quantitative polymerase chain reaction.RESULTS AND CONCLUSION:(1) Under planar culture conditions,the children group had stronger proliferative ability than the elderly group.(2) Compared with planar culture,the dynamic cultivation of elderly epidermal stem cells using a rotating cell culture system had a higher population doubling (P<0.01) and a shorter population doubling time (P<0.01).(3) The dynamic cultivation of rotating cell culture system promoted the self-assembly of elderly epidermal stem cells to form 3D human epidermal stem cell spheres based on microcarrier scaffolds,which had similar epidermal stem cell proliferation as the children group.(4) After dynamic culture,the expression of stemness marker genes (ITGA6 and ITGB1),basal cell marker genes (K5) and differentiation marker genes (K10 and K1) of elderly epidermal stem cells reached the same level as that of children.The results showed that the dynamic cultivation of rotating cell culture system can not only improve the efficiency of elderly epidermal stem cell culture,promote cell proliferation and self-assembly into 3D cell spheres,but also maintain a certain stemness of elderly epidermal stem cells without complete terminal differentiation.

BACKGROUND:Due to the aging population and the increase in elderly diseases,research and utilization of epidermal stem cells in the elderly have become increasingly important.However,due to the weak proliferation and drying ability of epidermal stem cells in the elderly,their application in biomedical and clinical research is limited.OBJECTIVE:To investigate the ability of rotating cell culture system to promote the proliferation and maintenance of epidermal stem cells in elderly individuals.METHODS:Epidermal stem cells were obtained from children (7-12 years old) and the elderly (over 60 years old) using traditional planar culture methods.After identification by cell morphology and cell immunofluorescence,the differences in proliferation ability of epidermal stem cells between the children and the elderly were analyzed by flow cytometry,CCK-8 assay,and cell clone formation assay.Elderly epidermal stem cells were cultured using a rotating cell culture system and 3D TableTrix? microcarriers.The cell proliferation and expression of stemness markers were detected by Live/Dead staining,cell doubling efficiency,cell doubling time,and real-time quantitative polymerase chain reaction.RESULTS AND CONCLUSION:(1) Under planar culture conditions,the children group had stronger proliferative ability than the elderly group.(2) Compared with planar culture,the dynamic cultivation of elderly epidermal stem cells using a rotating cell culture system had a higher population doubling (P<0.01) and a shorter population doubling time (P<0.01).(3) The dynamic cultivation of rotating cell culture system promoted the self-assembly of elderly epidermal stem cells to form 3D human epidermal stem cell spheres based on microcarrier scaffolds,which had similar epidermal stem cell proliferation as the children group.(4) After dynamic culture,the expression of stemness marker genes (ITGA6 and ITGB1),basal cell marker genes (K5) and differentiation marker genes (K10 and K1) of elderly epidermal stem cells reached the same level as that of children.The results showed that the dynamic cultivation of rotating cell culture system can not only improve the efficiency of elderly epidermal stem cell culture,promote cell proliferation and self-assembly into 3D cell spheres,but also maintain a certain stemness of elderly epidermal stem cells without complete terminal differentiation.

Objective:To explore the efficacy and safety of the ballistic-ultrasound-negative pressure three-in-one energy platform (Trilogy) for micro-channel percutaneous nephrolithotomy (mini-PCNL).Methods:A retrospective analysis was conducted on the clinical data of 140 patients with upper urinary tract stones treated at Tianjin Medical University Second Hospital from February to October 2024. All patients underwent mini-PCNL and were divided into the holmium laser group and the Trilogy group based on the stone fragmentation equipment used during the procedure. There were 69 patients in the holmium laser group and 71 in the Trilogy group. The two groups had similar mean ages (55.1±10.2 years vs. 53.4±10.8 years), male patient proportions (50 cases, 72.5% vs. 50 cases, 70.4%), body mass indices (25.2±3.6 kg/m 2 vs. 25.3±4.0 kg/m 2), incidence rates of hypertension (29 cases, 42.0% vs. 31 cases, 43.7%), diabetes (15 cases, 21.7% vs. 12 cases, 16.9%), mean cumulative stone lengths (39.2±12.6 mm vs. 35.9±14.8 mm), total stone volumes preoperatively (6 184.3±3 653.5 mm 3 vs. 5 644.9±4 173.8 mm 3), mean CT values for stones (1 138.2±264.3 HU vs. 1 151.3±208.0 HU), stone locations (ureter 14 cases, 20.3% vs. 22 cases, 31.0%; kidney 48 cases, 69.6% vs. 39 cases, 54.9%; both ureter and kidney 7 cases, 10.1% vs. 10 cases, 14.1%), preoperative mean urinary white blood cell counts [9.6(3.6, 31.2) cells/HPF vs. 11.9(3.8, 34.5) cells/HPF], proportions of patients with preoperative urinary white blood cells (+ + + ; 23 cases, 33.3% vs. 25 cases, 35.2%), nitrite positivity rates (4 cases, 5.8% vs. 3 cases, 4.2%), and urine culture positivity rates (12 cases, 17.4% vs. 18 cases, 25.4%) showed no statistically significant differences. The proportion of patients with moderate or higher hydronephrosis in the holmium laser group was lower than that in the Trilogy group (32 cases, 46.4% vs. 47 cases, 66.2%, P=0.018). The holmium laser group utilized holmium laser lithotripsy, where stone fragments were either flushed out with a vortex or retrieved with a stone basket. The Trilogy group employed a three-in-one energy platform to break the stones. This device incorporated pneumatic ballistic, ultrasound, and negative pressure suction capabilities within the same metallic probe, allowing the stone to be fragmented into small pieces while simultaneously performing ultrasonic negative pressure stone clearance. The parameters for the three-in-one energy platform were adjusted based on intraoperative conditions, typically setting negative pressure at 30%-50%, ultrasound power at 80%-100%, ballistic power at 80%, and frequency at 8 Hz. During the stone fragmentation process, the ballistic device fragmented the stones while ultrasound further reduced larger fragments and removed them. Some fragments that were difficult to break could also be flushed out or retrieved with a stone basket. The efficiency of stone clearance (volume of stones cleared per unit time) was compared between the two groups, as well as the stone-free rates on postoperative day 1 and day 30. Stone clearance time was defined as the duration from the start of fragmentation to the placement of the nephrostomy tube. Changes in postoperative white blood cells, hemoglobin, and albumin levels compared to preoperative levels, as well as the incidence of Clavien-Dindo complications, were compared between the two groups. Equipment failure incidents were recorded (fiber fracture in the holmium laser group indicating it could not be used; probe fracture in the Trilogy group). Patients were sub-grouped based on stone CT values into CT ≥ 1 000 HU and CT < 1 000 HU categories to compare stone clearance efficiency between the two devices within each sub-group. In the CT≥1 000 HU sub-group, there were 51 cases in the holmium laser group and 54 in the Trilogy group, there were no significant differences in preoperative total stone volume (6 785.0±3 902.3 mm 3 vs. 5 678.1±4 297.7 mm 3). In the CT < 1 000 HU sub-group, there were 18 cases in the holmium laser group and 17 in the Trilogy group. There were no significant differences between the groups in preoperative total stone volume (4 482.2±2 110.6 mm 3 vs. 5 530.9±3 845.3 mm 3). Results:The overall stone clearance efficiency in the Trilogy group was higher than that in the holmium laser group (87.9±35.7 mm 3/min vs. 77.1±24.3 mm 3/min, P=0.038). There were no significant differences in residual stone volume before discharge [5.5(0, 84.0) mm 3 vs. 5.3(0, 175.0) mm 3], stone clearance time (79.4±43.2 min vs. 66.6±49.7 min), or the proportion of patients using stone baskets during the procedure (33 cases, 47.8% vs. 36 cases, 50.7%). Postoperative changes in white blood cells, hemoglobin, and albumin compared to preoperative levels were not significantly different [(4.1±2.9)×10 9/L vs. (3.3±2.2)×10 9/L; (-2.9±10.5) g/L vs. (-1.6±9.3) g/L; (-2.5±3.6) g/L vs. (-1.8±5.0) g/L] Furthermore, there were no statistically significant differences in equipment failure rates (1 case, 1.4% vs. 4 cases, 5.6%), stone-free rates (postoperative day 1: 43 cases, 62.3% vs. 47 cases, 66.2%; postoperative day 30: 50 cases, 72.5% vs. 53 cases, 74.6%), or Clavien-Dindo complication rates (grade Ⅰ: 11 cases, 15.9% vs. 8 cases, 11.3%; grade Ⅱ: 2 cases, 2.8% vs. 0 cases; grade Ⅲ: 1 case, 1.4% vs. 0 cases). In the CT ≥ 1 000 HU sub-group, the clearance time for the holmium laser was longer than that for Trilogy (93.3±41.0 min vs. 74.6±51.9 min, P=0.044), there were no significant differences in residual stone volume before discharge [6.3(1.6, 173.8) mm 3 vs. 4.5(0, 69.0) mm 3] between the two groups. In the CT < 1 000 HU sub-group, the overall stone clearance efficiency of the Trilogy group exceeded that of the holmium laser group (134.2±38.0 mm 3/min vs. 105.5 ± 7.1 mm 3/min, P=0.004), there were no significant differences between the groups in residual stone volume before discharge [0(0, 51.1) mm 3 vs. 16.3(0, 957.2) mm 3], or stone clearance time (40.2±18.1 min vs. 39.1±27.5 min). Conclusions:In mini-PCNL surgery, the stone fragmentation efficiency of the three-in-one lithotripsy energy platform is superior to that of the holmium laser, while the overall complication rate is comparable to that of the holmium laser.

Objective:To investigate the feasibility and safety of a treatment system for stentless discharge after flexible ureteroscopic lithotripsy assisted by flexible negative pressure sheath.Methods:The clinical data of 72 patients with upper urinary calculi admitted to the Second Hospital of Tianjin Medical University from November 2022 to February 2023 were retrospectively analyzed. All patients achieved stentless discharge after flexible ureteroscopic lithotripsy assisted by flexible negative pressure sheath. There were 50 males and 22 females. The average age was (54.7±12.1) years. Preoperative urine culture was positive in 14 cases, negative in 3 cases (4.2%)with nitrite positive, and 11 cases were negative for urine culture and nitrites but positive of white blood cells (+ + + ). There were 29 cases of renal calculi, 33 cases of upper ureteral calculi, and 10 cases of upper ureteral calculi combined with renal calculi.The mean stone diameter was 17.0(14.0, 24.0)mm. CT value was (1 049.3±258.6)HU. Twenty-four patients carried ureteral stents before operation. A total of 42 cases used ureteral sheaths with diameters of F11/13, and 30 cases used sheaths with diameters of F12/14.During the operation, an infusion pump was used to provide sufficient irrigation pressure. The negative pressure suction was attached to the distal end of the sheath. The flexible head of the sheath was guided to the target renal calyx, to completely aspirate stone fragments. Stone baskets was used in 11 cases during the procedure. The level of ureteral injury was assessed according to the Traxer grading system at the end of the operation. A ureteral stent with extraction string was retained.On the first day postoperatively, CT scanning was performed to evaluate the residual stone fragments. Patients were discharged on postoperative day 2-3 after the removal of the ureteral stent and catheter. Follow-up was conducted for 30 days postoperatively, during which the Ureteral Stent Symptom Questionnaire (USSQ) was used to assess voiding symptoms and pain. Painkiller usage and emergency revisit situations were recorded. CT scans were performed to evaluate the stone-free rate on postoperative day 30.Results:The average operation time was 30.0 (20.0, 44.5) minutes. A total of 70 cases had no ureteral injuries, and 2 cases had Grade 1 ureteral injuries (minor mucosal damage). Three cases developed fever within 72 hours postoperatively, with no cases of septic shock or fever after stent removal. Eight patients reported waist and abdominal discomfort after discharge and took oral pain medication. Among them, one patient returned to the emergency department for pain treatment. Five patients reported moderate or severe genitourinary symptoms (including voiding frequency, nocturia, urgency/incontinence, dysuria, hematuria, and incomplete emptying) based on subjective evaluation. All patients could work and recovered a normal daily life after discharge and there was no readmission or additional surgical procedures. There were 61 patients achieved immediate stone-free status on the first day after surgery, and 66 patients achieved stone-free status during follow-up at 30 days postoperatively.Conclusions:Stentless discharge after flexible ureteroscopic lithotripsy assisted by flexible negative pressure sheath is safe and feasible.

Objective:This study aimed to explore the ″special″ review points and ethical strategies in the ethical review of medical research involving children, providing references for the medical research on this group.Methods:Combining the physical and mental characteristics of children, and comparing clinical research ethics review points, this study discussed the key aspects to be concerned about in the ethical review of medical research involving children, and proposed " special protection" strategies to be implemented.Results:Ethics committees should fully recognize the uniqueness of child development. The definition of minimal risk in the child population needs to be fully understood at the risk-benefit assessment stage, to develop ethical responses to the risk-benefit ratios in different situations, and to avoid special risks. Informed consent procedures should adhere to basic principles while respecting dissenting voices by designing multiple versions of informed consent for research, and developing informed consent countermeasures for vulnerable children groups.Conclusions:Medical research involving children should follow more rigorous and scientific procedures to protect rights, standardize clinical research in pediatric healthcare, and promote the healthy development of pediatric health endeavors.

BACKGROUND:Intervertebral disk degeneration is a pathological change caused by a series of complex molecular mechanisms that result in the aging and damage of intervertebral discs,ultimately leading to severe clinical symptoms.Traditional Chinese medicine has unique advantages in the treatment of intervertebral disk degeneration due to its low cost,non-addictive nature,multi-target effects,minimally toxic and side effects,and high patient acceptance. OBJECTIVE:To review the latest research results of traditional Chinese medicine monomer intervention-related signaling pathways in the treatment of intervertebral disk degeneration,describe and analyze the action mechanism of traditional Chinese medicine monomer on intervertebral disk degeneration,and provide a new approach and theoretical basis for future basic research and clinical treatment. METHODS:The first author searched for relevant literature from January 2018 to February 2023 in CNKI,PubMed,VIP,and WanFang using the search terms"intervertebral disc,signal pathway".The articles that did not meet the criteria were excluded after preliminary screening of the titles and abstracts.Finally,72 articles were selected for review and analysis. RESULTS AND CONCLUSION:Traditional Chinese medicine monomers can regulate multiple classical signaling pathways such as Wnt/β-catenin,PI3K/Akt,mTOR,NF-κB,and MAPK.They achieve this by regulating oxidative stress,adjusting the expression of pro/anti-apoptotic proteins in cells,stimulating cellular autophagy function,reducing stimulation of cell inflammatory factors,increasing the expression of extracellular matrix markers,reducing the production of matrix-degrading enzymes,maintaining the synthesis and stability of extracellular matrix,inducing differentiation of mesenchymal stem cells in the nucleus pulposus into nucleus pulposus cells,promoting endogenous repair and reconstruction,controlling apoptosis and aging of nucleus pulposus cells,and increasing the activity of nucleus pulposus cells.These actions improve the microenvironment within the intervertebral disc,maintain the normal physiological function of the intervertebral disc,and delay intervertebral disc degeneration.

BACKGROUND:At present,bone marrow mesenchymal stem cells are the main seed cells used in cell therapy for the treatment of intervertebral disc degeneration.However,the use of bone marrow mesenchymal stem cells as seed cells for the regeneration of fibrous rings is at risk of heterotopic ossification and teratoma at the repair site.Therefore,it is of great economic and social significance to find a new kind of seed cells for tissue engineering of annulus fibrosus for the treatment of intervertebral disc degeneration. OBJECTIVE:To isolate and purify rat annulus fibrosus-derived stem cells by adherent method combined with fibronectin differential adhesion screening method,and to observe its purification effect and biological characteristics. METHODS:Annulus fibrosus tissue was obtained from a SD rat intervertebral disc.Primary annulus fibrosus-derived stem cells were obtained by the mechanical-enzymatic digestion method.Annulus fibronectin differential adhesion method was used to purify annulus fibrosus-derived stem cells.Morphological changes and proliferation of cells were observed through a microscope.The expression levels of stem cell markers were detected by immunofluorescence technique and qPCR.The screened cells were subjected to multi-lineage cell differentiation and characteristic gene detection. RESULTS AND CONCLUSION:(1)The purified cells grew well,and most of them were angular and star-shaped multi-process cells,which had good proliferation ability.(2)Cells were positive for cell membrane surface antigens CD73,CD90 and CD105,while negative for CD45 and CD34.(3)After specific induction,cells could successfully differentiate into osteoblasts,chondroblasts and lipoblasts.(4)Collagen-I,Runx-2 after osteogenic induction,Collagen Ⅱ,Sox-9 after chondrogenic induction,and PPAR-γ and LPL after lipogenic induction were highly expressed in cells,and the difference was significant compared with that before induction(P<0.05).(5)These findings confirm that the adherent method combined with fibronectin differential adhesion method is effective enough to screen,isolate and purify rat annulus fibrosus-derived stem cells,and has good cell biological properties,good proliferation ability and multiple differentiation potential.