OBJECTIVES: To explore the mechanism by which astragaloside IV (AS-IV) alleviates D-galactose (D-GAL)-induced senescence in human umbilical vein endothelial cells (HUVECs). METHODS: Cultured HUVECs were treated with D-GAL (40 g/L), AS-IV (200 μmol/L), D-GAL+AS-IV, or D-GAL+AS-IV+MTK458 (a mitochondrial autophagy agonist, 25 μmol/L) for 48 h, and the changes in cell proliferation, migration, and angiogenesis capacity were evaluated. Cell apoptosis, reactive oxygen species (ROS) levels, mitochondrial membrane potential, and expressions of autophagy-related proteins (LC3-II/LC3-I) and PINK1/Parkin pathway proteins in the treated cells were detected. RESULTS: AS-IV treatment significantly reduced the inhibitory effect of D-GAL on HUVEC viability, effectively alleviated D-GAL-induced impairment of tube-forming ability, and promoted angiogenesis and migration ability of the cells. AS-IV also significantly reduced the rate of D-GAL-induced HUVECs positive for senescence-associated β-galactosidase (SA-β-Gal) staining and inhibited the expression of senescence-related genes P21 and P53. AS-IV restored mitochondrial membrane potential and reduced intracellular ROS levels in D-GAL-induced HUVECs, and inhibited the fusion of autophagosomes and lysosomes to prevent the completion of autophagic flux. In HUVECs treated with both D-GAL and AS-IV, the application MTK458 significantly increased the number of yellow spots and enhanced the expressions of P21, P53, PINK1, Parkin, LC3, and Beclin proteins. CONCLUSIONS AS-IV alleviates D-GAL-induced endothelial cell senescence by inhibiting the PINK1/Parkin pathway to regulate mitochondrial autophagy.
Humans ; Human Umbilical Vein Endothelial Cells/drug effects* ; Cellular Senescence/drug effects* ; Autophagy/drug effects* ; Saponins/pharmacology* ; Ubiquitin-Protein Ligases/metabolism* ; Mitochondria/drug effects* ; Triterpenes/pharmacology* ; Protein Kinases/metabolism* ; Galactose/pharmacology* ; Reactive Oxygen Species/metabolism* ; Signal Transduction/drug effects* ; Cells, Cultured ; Apoptosis/drug effects* ; Membrane Potential, Mitochondrial ; Cell Proliferation/drug effects*

ObjectiveTo evaluate the effects of Tongnaoyin on the blood-brain barrier status and neurological impairment in acute ischemic stroke （AIS） patients with the syndrome of phlegm-stasis blocking collaterals by dynamic contrast-enhanced magnetic resonance imaging （DCE-MRI）. MethodsA total of 63 patients diagnosed with AIS in the Jiangsu Province Hospital of Chinese Medicine from October 2022 to December 2023 were enrolled in this study. According to random number table method，the patients were assigned into a control group （32 cases） and an observation group （31 cases）. The control group received conventional Western medical treatment，and the observation group took 200 mL Tongnaoyin after meals，twice a day from day 2 of admission on the basis of the treatment in the control group. After 7 days of treatment，the patients were examined by DCE-MRI. The baseline data for two groups of patients before treatment were compared. The National Institute of Health Stroke Scale （NIHSS） score and modified Rankin Scale （mRS） score were recorded before treatment and after 90 days of treatment for both groups. The rKtrans，rKep，and rVe values were obtained from the region of interest （ROI） of the infarct zone/mirror area and compared between the two groups. ResultsThere was no significant difference in the NIHSS or mRS score between the two groups before treatment. After 90 days of treatment，the NIHSS and mRS scores declined in both groups，and the observation group had lower scores than the control group （P<0.05）. After treatment，the rKtrans and rVe in the observation group were lower than those in the control group （P<0.01）. ConclusionCompared with conventional Western medical treatment alone，conventional Western medical treatment combined with Tongnaoyin accelerates the repair of the blood-brain barrier in AIS patients，thereby ameliorating neurological impairment after AIS to improve the prognosis.

Objective To use imaging features of pulmonary nodules to predict the risk of lymph node metastasis in patients with cT1-stage non-small cell lung cancer (NSCLC), providing a reference for clinical decision-making. Methods A retrospective analysis was conducted on the imaging features and postoperative pathological results of cT1 NSCLC patients who underwent surgical treatment at Linyi People’s Hospital from July 2019 to July 2022. Patients were grouped and analyzed according to lymph node metastasis status. Results A total of 1 123 patients were included, comprising 471 males and 652 females, with a median age of 59 (52, 66) years. Comparative analysis revealed that sex, age, nodule location, nodule size on imaging, solid component size, consolidation tumor ratio (CTR), average CT value, and tumor proximity to the pleura all influenced lymph node metastasis. A nomogram was constructed, indicating that the probability of lymph node metastasis in cT1 NSCLC was positively correlated with solid component size, CTR, and average CT value of the pulmonary nodule, and negatively correlated with patient age. The area under the receiver operating characteristic curve was 0.929. Conclusion For cT1 NSCLC patients, the probability of lymph node metastasis can be predicted by measuring the solid component size, CTR, and average CT value of the pulmonary nodule, in conjunction with patient age. However, relying solely on pulmonary nodule imaging characteristics is insufficient to determine a specific lymph node dissection strategy.

Objective To explore the impact of the TINCR-MAF:MAFB transcription factor network on the expression of proliferation and differentiation-related genes in keratinocytes,to verify the role of this network in the occurrence and development of psoriasis and its potential mechanisms.Methods Employed RNA interference technology to knock down TINCR gene expression,and the proliferation ability of keratinocytes was assessed using the CCK-8 method.Additionally,qRT-PCR and Western blot analyses were conducted to evaluate the RNA and protein expression levels of TINCR,MAFB,and KLF4 genes.Immunohistochemical methods were used to detect the expression of KLF4 protein in psoriasis tissues.Results After TINCR gene siRNA interference,the proliferation ability of keratinocytes significantly decreased at 24,48,and 72 hours(P<0.001),indicating that the TINCR gene plays a critical role in cell proliferation.The results of qRT-PCR and Western blot analyses showed that the RNA and protein expression levels of TINCR,MAFB,and KLF4 genes were significantly reduced(P<0.001),suggesting that TINCR may influence the differentiation of keratinocytes by regulating the expression of MAFB transcription factor and KLF4 differentiation-related genes.Furthermore,immunohistochemical results indicated that the expression of KLF4 protein was significantly elevated in psoriasis tissues compared to normal skin tissues,suggesting that KLF4 plays an important role in the pathogenesis of psoriasis.Conclusions The TINCR-MAF:MAFB transcription factor network may participate in the occurrence and development of psoriasis by affecting the proliferation and differentiation of keratinocytes.This finding provides a new perspective on the pathogenesis of psoriasis and potential targets for future therapeutic strategies.

Objective To explore the correlation between psoas major index(PMI)in patients with diffuse large B-cell lymphoma(DLBCL)and prognosis after rituximab,cyclophosphamide,doxorubicin,vincristine,prednisone(R-CHOP)chemotherapy.Methods Totally 148 DLBCL patients who received R-CHOP chemotherapy were retrospectively enrolled.Based on axis CT image at L3 vertebral transverse process,bilateral psoas muscles were delineated,and PMI was calculated.Then the patients were divided into muscle reduction group(≤lower quartile,n=37)and control group(＞lower quartile,n=111)according to the lower quartile of PMI.Clinical and imaging data were compared between groups,and the correlations of PMI with the above indexes were analyzed.The impact factors of prognosis of DLBCL were observed.Results Significant differences of body mass index(BMI),Eastern Cooperative Oncology Group(ECOG)score,international prognostic index(IPI),the proportion of B symptoms,mortality rate,3-year survival rate,β2-microglobulin,hemoglobin,albumin and PMI were found between groups(all P＜0.05).PMI of DLBCL patient was negatively correlated with age,ECOG score,IPI,β2-microglobulin and lactate dehydrogenase(LDH)(r=—0.126,-0.225,—0.262,—0.232,—0.229,all P＜0.05),but positively correlated with BMI and hemoglobin(r=0.282,0.376,both P＜0.05).The age,ECOG score,IPI and PMI were all independent impact factors of overall survival in DLBCL patients(all P＜0.05).Conclusion PMI was an independent impact factor of prognosis of DLBCL after R-CHOP chemotherapy.

Objective To explore the impact of the TINCR-MAF:MAFB transcription factor network on the expression of proliferation and differentiation-related genes in keratinocytes,to verify the role of this network in the occurrence and development of psoriasis and its potential mechanisms.Methods Employed RNA interference technology to knock down TINCR gene expression,and the proliferation ability of keratinocytes was assessed using the CCK-8 method.Additionally,qRT-PCR and Western blot analyses were conducted to evaluate the RNA and protein expression levels of TINCR,MAFB,and KLF4 genes.Immunohistochemical methods were used to detect the expression of KLF4 protein in psoriasis tissues.Results After TINCR gene siRNA interference,the proliferation ability of keratinocytes significantly decreased at 24,48,and 72 hours(P<0.001),indicating that the TINCR gene plays a critical role in cell proliferation.The results of qRT-PCR and Western blot analyses showed that the RNA and protein expression levels of TINCR,MAFB,and KLF4 genes were significantly reduced(P<0.001),suggesting that TINCR may influence the differentiation of keratinocytes by regulating the expression of MAFB transcription factor and KLF4 differentiation-related genes.Furthermore,immunohistochemical results indicated that the expression of KLF4 protein was significantly elevated in psoriasis tissues compared to normal skin tissues,suggesting that KLF4 plays an important role in the pathogenesis of psoriasis.Conclusions The TINCR-MAF:MAFB transcription factor network may participate in the occurrence and development of psoriasis by affecting the proliferation and differentiation of keratinocytes.This finding provides a new perspective on the pathogenesis of psoriasis and potential targets for future therapeutic strategies.

Objective To explore the correlation between psoas major index(PMI)in patients with diffuse large B-cell lymphoma(DLBCL)and prognosis after rituximab,cyclophosphamide,doxorubicin,vincristine,prednisone(R-CHOP)chemotherapy.Methods Totally 148 DLBCL patients who received R-CHOP chemotherapy were retrospectively enrolled.Based on axis CT image at L3 vertebral transverse process,bilateral psoas muscles were delineated,and PMI was calculated.Then the patients were divided into muscle reduction group(≤lower quartile,n=37)and control group(＞lower quartile,n=111)according to the lower quartile of PMI.Clinical and imaging data were compared between groups,and the correlations of PMI with the above indexes were analyzed.The impact factors of prognosis of DLBCL were observed.Results Significant differences of body mass index(BMI),Eastern Cooperative Oncology Group(ECOG)score,international prognostic index(IPI),the proportion of B symptoms,mortality rate,3-year survival rate,β2-microglobulin,hemoglobin,albumin and PMI were found between groups(all P＜0.05).PMI of DLBCL patient was negatively correlated with age,ECOG score,IPI,β2-microglobulin and lactate dehydrogenase(LDH)(r=—0.126,-0.225,—0.262,—0.232,—0.229,all P＜0.05),but positively correlated with BMI and hemoglobin(r=0.282,0.376,both P＜0.05).The age,ECOG score,IPI and PMI were all independent impact factors of overall survival in DLBCL patients(all P＜0.05).Conclusion PMI was an independent impact factor of prognosis of DLBCL after R-CHOP chemotherapy.

Aim To explore the mechanism of oxiracetam promoting neurogenesis and migration in rats with cer-ebral infarction through stromal cell-derived factor-1α(SDF-1α)/C-X-C chemokine receptor 4(CXCR4)pathway.Methods 100 SD rats were randomly divided into control group,cerebral ischemia(CI)group,oxiracetam(200 mg/kg)group,and oxiracetam(200 mg/kg)+AMD3100(5 mg/kg)group,with 25 rats in each group.Electrocoagulation was used to create rat model of local permanent cerebral infarction.After 1,7 and 14 days of modeling,neurological deficits were scored,TTC staining was used to detect the volume of cerebral infarction,Nissl staining was used to detect cell surviv-al in the infarcted area,Western blot was used to detect SDF-1α and CXCR4 protein levels in ischemic zone.After 1～7 days of modeling,BrdU(50 mg/kg)was continuously injected intraperitoneally.After 14 days,immunofluorescence double staining was used to detect the number of BrdU+Nestin+and BrdU+DCX+cells in the SVZ region.5 days before modeling,retroviruses carrying GFP were injected into the SVZ region.After 14 days,immunofluorescence double stai-ning was used to detect the number of GFP+DCX+,GFP+MAP-2+and GFP+GFAP+cells in infarction area.C17.2 cells were divided into control group,oxygen-glucose deprivation(OGD)group,oxiracetam(final concentration:200 mg/L)group,and oxiracetam(final concentration:200 mg/L)+AMD3100(final concentration:100 μmol/L)group.OGD was used to create cell CI model.After 12 hours,immunofluorescence double staining was used to detect the number of Br-dU+/Nestin+and BrdU+/MAP-2+cells,Transwell experiment was used to detect cell migration,Western blot was used to detect SDF-1α and CXCR4 protein levels in cell culture supernatant.Results Animal experiment results showed:compared with control group,mNSS score in CI group was increased,cerebral infarction volume was increased,the number of surviving cells in infarcted area was decreased,SDF-1α and CXCR4 protein levels were increased,the number of GFP+DCX+,GFP+MAP-2+and GFP+GFAP+cells in SVZ region were increased(P＜0.05);compared with CI group,mNSS score in oxiracetam group was decreased,cerebral infarction volume was decreased,the number of surviving cells in infarc-ted area was increased,SDF-1α and CXCR4 protein levels were increased,the number of GFP+DCX+,GFP+MAP-2+and GFP+GFAP+cells in SVZ region were increased,the number of GFP+DCX+,GFP+MAP-2+and GFP+GFAP+cells in in-farcted area were increased(P＜0.05);compared with oxiracetam group,mNSS score in oxiracetam+AMD3100 group was increased,cerebral infarction volume was increased,the number of surviving cells in infarcted area was decreased,CXCR4 protein level was decreased,the number of GFP+DCX+,GFP+MAP-2+and GFP+GFAP+cells in the SVZ region were de-creased(P＜0.05).Cell experiment results showed:compared with control group,the number of BrdU+/Nestin+and Br-dU+/MAP-2+cells in OGD group were increased,the number of cell migration,SDF-1α and CXCR4 protein levels in cell culture supernatant were increased(P＜0.05);compared with OGD group,the number of BrdU+/Nestin+and BrdU+/MAP-2+cells in oxiracetam group were increased,the number of cell migration,SDF-1α and CXCR4 protein levels in cell culture supernatant were increased(P＜0.05);compared with oxiracetam group,the number of BrdU+/Nestin+and BrdU+/MAP-2+cells in oxiracetam+AMD3100 group were decreased,the number of cell migration,CXCR4 protein level in cell culture supernatant were decreased(P＜0.05).Conclusion Oxiracetam may promote the migration of neural stem cells from the SVZ region to the ischemic zone,promoting neurogenesis and functional recovery in rats with cerebral infarction by activating SDF-1α/CXCR4 pathway.

ObjectiveTo investigate the changes in cerebral blood perfusion in patients with acute cerebral infarction after taking Tongnaoyin， a traditional Chinese medicine， based on head and neck computed tomography （CT） angiography （CTA） combined with brain CT perfusion imaging （CTP）. MethodA total of 240 patients with cerebral infarction of phlegm and blood stasis syndrome treated in Jiangsu Province Hospital of Traditional Chinese Medicine from March 2018 to September 2023 were randomly divided into a control group （99 cases） and a Tongnaoyin group （141 cases）. Based on the guidelines， the control group was treated with conventional treatment such as anti-aggregation， anticoagulation， lipid-lowering and plaque stabilization， brain protection， and supportive treatment. The Tongnaoyin group was treated with Tongnaoyin of 200 mL in warm conditions in the morning and evening on the basis of the control group. Both groups underwent CTA combined with CTP within 24 hours after admission， and they were reexamined by CTA and CTP in the sixth month after admission. The degree of intracranial artery stenosis was determined according to the North American Symptomatic Carotid Endarterectomy Trial （NASCET） method. The relative cerebral blood volume （rCBV）， relative cerebral blood flow （rCBF）， mean transit time （MTT）， and time to peak （TTP） of the lesion area before and after treatment were compared. The adverse outcomes of the two groups within six months after discharge were compared. ResultCompared with the group before treatment， the degree of vascular stenosis in the Tongnaoyin group was significantly reduced， and the difference was statistically significant （Z=105.369，P<0.05）. Compared with the control group after treatment， the improvement rate of vascular stenosis in the Tongnaoyin group was higher （χ²=84.179，P<0.01）， and the curative effect was better.After treatment， the rCBV and rCBF of patients in the Tongnaoyin group were significantly increased， and the difference was statistically significant （P<0.01）. MTT and TTP showed a trend of shortening， but the difference was not statistically significant. There was no statistically significant difference in rCBV， rCBF， MTT， and TTP in the control group. Compared with those in the control group after treatment， the rCBV and rCBF in the Tongnaoyin group were significantly increased， while MTT and TTP were significantly reduced （P<0.01）. After six months of discharge， the risk of poor prognosis in the Tongnaoyin group was significantly reduced compared with the control group （P<0.05）. ConclusionTongnaoyin has a good effect on improving cerebral blood perfusion in patients with acute cerebral infarction. It can be used as an effective supplement for the conventional treatment of ischemic stroke to improve clinical efficacy.

Objective To explore the optimal dose of twice daily premixed insulin regimen in Chinese patients with type 2 diabetes mellitus(T2DM). Methods A total of 361 T2DM patients admitted to the department of endocrinology in Huadong hospital affiliated to Fudan university from March 2017 to October 2021 were selected. All patients received stable doses of premixed insulin twice daily and had HbA1c<7％. Continuous blood glucose monitoring confirmed that TIR was>70％ and TBR was<1％. According to different insulin formulations,it was divided into Regular/NPH 30 group (n=161),Aspart 30 group (n=156),and Lispro 25 group (n=44). Clinical characteristics,total insulin dose,pre-breakfast insulin dose,pre-dinner insulin dose,and pre-breakfast/pre-dinner insulin ratio were compared. Pearson correlation analysis was used to analyze the correlation between total insulin dose and other indicators. Multiple linear regression analysis was used to analyze the influencing factors of the total insulin dose. Results The mean dose of total insulin,pre-breakfast insulin,and pre-dinner insulin was (30.6±8.6)U/d,(18.3±5.1)U/d,and (12.3±4.2)U/d,respectively. The median of insulin dose per kg was 0.4(0.4,0.5)U/d,and pre-breakfast/pre-dinner insulin ratio was 1.5(1.3,1.8). Pearson correlation analysis showed that total insulin dose was associated with BMI,HbA1c,FPG,and ALT. Multiple linear regression analysis showed that BMI,FPG,HbA1c,and ALT were independent factors of total insulin dose. Conclusions The initial dose of premixed insulin can be allocated at a ratio of 3∶2 before breakfast and before dinner in Chinese people with T2DM,not 1∶1.