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.

BACKGROUND: Meningeal metastasis (MM) is a form of malignant metastasis where tumor cells spread from the primary site to the pia mater, dura mater, arachnoid, subarachnoid space, and other cerebrospinal fluid compartments. Lung cancer is one of the most common malignant tumor types with MM. MM not only signifies that the lung cancer has progressed to an advanced stage but also leads to a range of severe clinical symptoms due to meningeal involvement. Currently, the risk factors associated with the development of MM are not fully elucidated. The aim of this study was to investigate the risk factors for MM in patients with lung adenocarcinoma (LUAD) who underwent non-surgical interventions, in order to identify LUAD patients at high risk for MM. METHODS: This retrospective study analyzed the clinical data of patients diagnosed with LUAD at the First Affiliated Hospital of Zhengzhou University from January 2020 to July 2024. Missing data were imputed using multiple imputation methods, and risk factors were identified through LASSO, univariate, and multivariate Logistic regression analyses. RESULTS: A total of 170 patients with LUAD were included in this study and divided into two groups: 87 patients with MM and 83 patients without MM. Univariate and multivariate Logistic regression analyses revealed that younger age at diagnosis (P=0.004), presence of the epidermal growth factor receptor (EGFR) L858R gene mutation (P=0.008), and concurrent liver metastasis at baseline (P=0.004) were independent risk factors for developing MM in LUAD patients who did not undergo surgical intervention. Conversely, higher baseline globulin levels (P=0.039) and the presence of the anaplastic lymphoma kinase (ALK) gene mutation (P=0.040) were associated with a reduced risk of MM development. CONCLUSIONS Age at diagnosis, EGFR L858R mutation status, ALK gene mutation status, concurrent liver metastasis, globulin levels at baseline were significantly associated with the risk of developing MM in patients with LUAD patients who did not undergo surgical intervention. For patients diagnosed at a younger age, carrying the EGFR L858R mutation, or presenting with baseline liver metastasis, early implementation of tertiary prevention strategies for MM is crucial. Regular monitoring of MM status should be conducted in these high-risk groups.
Humans ; Male ; Adenocarcinoma of Lung/therapy* ; Female ; Middle Aged ; Risk Factors ; Lung Neoplasms/therapy* ; Retrospective Studies ; Aged ; Meningeal Neoplasms/genetics* ; Adult

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 investigate the clinical effect of simultaneous use of three endoscopes(laparoscope,choledochoscope,and duodenoscope)combined with holmium laser in the treatment of gallstones complicated with common bile duct stones.Methods A prospective randomized controlled study was carried out.A total of 80 patients with cholecystolithiasis combined with choledocholithiasis in our hospital from August 2022 to January 2024 were selected and divided into three-endoscope group(n=40)and two-endoscope group(n=40)according to random number table method.Patients in the three-endoscope group underwent laparoscopic cholecystectomy(LC)and laparoscopic common bile duct exploration(LCBDE)combined with holmium laser lithotripsy,primary closure of the common bile duct,endoscopic retrograde cholangiopancreatography(ERCP),and endoscopic nasobiliary drainage(ENBD),while the two-endoscope group underwent LC+LCBDE+T-tube drainage.The intraoperative bleeding volume,operation time,hospitalization time,hospitalization costs,and postoperative complications between the two groups were observed and analyzed.Results There was no significant difference in intraoperative bleeding volume between the two groups[(55.6±14.3)ml vs.(53.2±16.3)ml,t=0.703,P=0.484].The operation time of the three-endoscope group was significantly longer than that in the two-endoscope group[214.5(171.5,246.8)min vs.178.0(151.0,227.8)min,Z=-2.069,P=0.039].The length of hospital stay in the three-endoscope group was significantly shorter than that in the two-endoscope group[(13.2±3.3)d vs.(16.2±3.7)d,t=3.864,P=0.001].The hospitalization costs were significantly higher in the three-endoscope group than those in the two-endoscope group[39 316.0(32 338.5,43 421.0)yuan vs.33 717.0(30 873.3,37 813.3)yuan,Z=-3.272,P=0.001].There were no significant differences in the incidence of postoperative pancreatitis[2.5%(1/40)vs.2.5%(1/40),χ2=0.000,P=1.000],the incidence of bile leakage[7.5%(3/40)vs.2.5%(1/40),χ2=0.263,P=0.615],and residual stone rate[7.5%(3/40)vs.0.0%(0/40),P=0.241]between the two groups.Forty cases in the three-endoscope group were followed up for 2-20 months,with a median of 13 months.One case of stone recurrence occurred at 8 months after surgery.Forty patients in the two-endoscope group were followed up for 1-21 months,with a median of 15 months.There was 1 case of stone recurrence at 6 and 10 months postoperatively,respectively.There was no statistically significant difference in stone recurrence rate between the two groups[2.5%(1/40)vs.5.0%(2/40),χ2=0.000,P=1.000].Conclusions LC+LCBDE combined with holmium laser lithotripsy,primary closure of the common bile duct,ERCP,and ENBD in the treatment of cholecystolithiasis and choledocholithiasis can shorten the hospitalization time.The replacement of T-tube with nasobiliary duct protects the function of the Oddi sphincter,which is more in line with the concept of minimally invasive surgery and worthy of clinical application.

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 investigate the safety and efficacy of trans-breast endoscopic approach for lateral neck dissection (LND) in the treatment of thyroid cancer.Methods:A retrospective analysis was conducted on 80 patients who underwent trans-breast endoscopic LND at the Second Affiliated Hospital of Zhejiang University School of Medicine from Dec. 2023 to Aug. 2024. Demographic characteristics, operative duration, postoperative complications, and pathological results were analyzed, with data presented using descriptive statistics.Results:The cohort included 7 males and 73 females, with a mean age of (30.0 ± 6.7) years. The mean diameter of thyroid cancer lesion was (1.3 ± 0.7) cm. The average operative duration was (4.6 ± 1.5) hours, with a 0.0% (0/80) rate of conversion to open surgery. Intraoperative recurrent laryngeal nerve (RLN) "loss of signal" occurred in 6.3% (5/80) of cases. The mean hospital stay was (2.3 ± 1.0) days. Postoperative complications included transient hoarseness in 3.8% (3/80) and transient hypoparathyroidism in 10.0% (8/80), with no cases of permanent hoarseness or hypoparathyroidism. The mean total number of dissected lymph nodes was 47.8 ± 18.6, with 8.3 ± 6.2 metastatic lymph nodes. The incidence of postoperative lymphatic leakage was 3.8% (3/80), and no cases of incision infection were observed. The mean visual analog scale (VAS) score for pain was 4.2 ± 0.4. The mean follow-up duration was 10.3 months, at 3-month follow-up, the cosmetic satisfaction score was 4.5 ± 0.4.Conclusions:The trans-breast endoscopic approach for LND is safe and effective, with high patient cosmetic satisfaction. With appropriate patient selection, it can be considered a viable surgical option for lateral neck dissection in the treatment of thyroid cancer.

Objective To investigate the effects of imatinib mesylate and sunitinib malate on the migration and invasion of gastroin-testinal stromal tumor(GIST)cells.Methods After identifying primary-cultured GIST cells,their morphology was characterized using atomic force microscopy(AFM).Changes in the expression of genes related to the PI3K/AKT signaling pathway were analyzed using quan-titative real-time PCR following drug treatments.Changes in the binding of related molecules were detected using AFM,and alterations in cell migration,invasive ability,and apoptosis were determined using scratch assay,Transwell assay,and flow cytometry,respectively.Results AFM imaging showed that pseudopods were flatly spread around the GIST cells,indicating characteristics consistent with easy metastasis.Administration of either imatinib or sunitinib significantly reduced the expression of genes related to the PI3K/AKT signaling pathway,the density of epidermal growth factor receptor(EGFR)on the surface of GIST cells,and molecular binding force with EGF.These changes were more pronounced with the combination treatment.Correspondingly,the invasive and migratory abilities of GIST cells were significantly reduced when either drug was administered alone and the inhibitory effect was more significant when the drugs were combined.Conclusion Both imatinib and sunitinib can significantly inhibit the expression of genes related to the PI3K/AKT signaling pathway,reduce the density of EGFR on the surface of GIST cells,and attenuate their molecular binding to EGF,thereby reducing the migration and invasion of GIST cells.However,the combination of these two drugs has a more significant effect.

Objective To compare the effect of different intensity of neuromuscular training(NMT)on muscle strength and knee joint function of patients after anterior cruciate ligament reconstruction(ACLR).Methods From January,2023 to January,2024,60 ACLR patients in Changhai Hospital were selected,and they received the same intensity of NMT from one to eight weeks after surgery.Eight weeks after surgery,they were randomly divided into low intensity group(n=30)and high intensity group(n=30),and then they received different inten-sities of NMT from nine to 16 weeks after surgery,each training session lasted one hour,with three sessions per week,totaly 48 sessions.The Lysholm score,knee flexor and extensor muscle strength and muscle endurance-were compared at eight weeks and 16 weeks after surgery.Results After group training,the Lysholm score significantly increased in both groups(|t|>13.739,P<0.001),and was higher in the high intensity group than in the low intensity group(t=-2.574,P<0.05);in the high intensity group,the relative peak torque and endurance of the extensor and flexor muscles improved at angular velocities of 60°/s,120°/s and 180 °/s(|t|>2.320,P<0.05);in the low intensity group,the flexor peak torque improved at all the three angular velocities(t>2.177,P<0.05),the extensor peak torque improved at angular velocities of 60°/s and 180°/s(|t|>1.715,P<0.05),and the extensor endurance improved at angular velocity of 60°/s(t=-2.293,P<0.05).However,there was no significant difference in the relative peak torque and endurance of the extensor and flexor muscles at all the three angular velocities(P>0.05).Conclusion Both high and low intensity NMT could improve the muscle strength,muscle endurance and knee joint func-tion.Maybe,high intensity is superior to low intensity.Further verification is still needed.

Objective:To investigate the safety and efficacy of trans-breast endoscopic approach for lateral neck dissection (LND) in the treatment of thyroid cancer.Methods:A retrospective analysis was conducted on 80 patients who underwent trans-breast endoscopic LND at the Second Affiliated Hospital of Zhejiang University School of Medicine from Dec. 2023 to Aug. 2024. Demographic characteristics, operative duration, postoperative complications, and pathological results were analyzed, with data presented using descriptive statistics.Results:The cohort included 7 males and 73 females, with a mean age of (30.0 ± 6.7) years. The mean diameter of thyroid cancer lesion was (1.3 ± 0.7) cm. The average operative duration was (4.6 ± 1.5) hours, with a 0.0% (0/80) rate of conversion to open surgery. Intraoperative recurrent laryngeal nerve (RLN) "loss of signal" occurred in 6.3% (5/80) of cases. The mean hospital stay was (2.3 ± 1.0) days. Postoperative complications included transient hoarseness in 3.8% (3/80) and transient hypoparathyroidism in 10.0% (8/80), with no cases of permanent hoarseness or hypoparathyroidism. The mean total number of dissected lymph nodes was 47.8 ± 18.6, with 8.3 ± 6.2 metastatic lymph nodes. The incidence of postoperative lymphatic leakage was 3.8% (3/80), and no cases of incision infection were observed. The mean visual analog scale (VAS) score for pain was 4.2 ± 0.4. The mean follow-up duration was 10.3 months, at 3-month follow-up, the cosmetic satisfaction score was 4.5 ± 0.4.Conclusions:The trans-breast endoscopic approach for LND is safe and effective, with high patient cosmetic satisfaction. With appropriate patient selection, it can be considered a viable surgical option for lateral neck dissection in the treatment of thyroid cancer.

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.