ObjectiveTo explore effect of Xianlian Jiedu prescription on the recurrence of colorectal cancer （CRC） and investigate the related mechanisms. MethodsA postoperative recurrence model was established in 25 Balb/c mice by injecting CT26 cells subcutaneously into the armpit， followed by surgical removal of 99% of the subcutaneous tumor. The mice were randomly divided into model group， low-dose Xianlian Jiedu prescription （XLJDP-L） group （6.45 g·kg^-1·d^-1）， medium-dose Xianlian Jiedu prescription （XLJDP-M） group （12.9 g·kg^-1·d^-1）， high-dose Xianlian Jiedu prescription （XLJDP-H） group （25.8 g·kg^-1·d^-1）， and 5-fluorouracil （5-FU） group （1×10^-3 g·kg^-1·d^-1）. The mice were euthanized after 14 days of continuous intervention， and recurrent tumor tissue was harvested. Hematoxylin and eosin （HE） staining was used to observe pathological and morphological changes in the recurrent tumor tissue. Immunohistochemistry （IHC） was employed to assess the expression of proliferating cell nuclear antigen （Ki67）， vascular endothelial growth factor （VEGF）， and platelet-endothelial cell adhesion molecule （CD31） in recurrent tumor tissue. The Western blot was used to detect the protein expression levels of angiopoietin-2 （ANG-2）， VEGF， phosphorylated-protein kinase B （p-Akt）， protein kinase B （Akt）， phosphorylated-phosphatidylinositol 3-kinase （p-PI3K）， and phosphatidylinositol 3-kinase （PI3K） in recurrent tumor tissue. ResultsBefore treatment， there were no statistical differences in tumor volume， tumor weight， and body mass among the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group compared to the model group， indicating model stability. After treatment， compared with those in the model group， the tumor volume and tumor weight in the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly reduced （P<0.01）， showing dose dependency. Meanwhile， there were no significant differences in body weight among the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group compared to the model group. HE staining showed that compared with that in the model group， tumor tissue in the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group had loosely arranged cells， increased intercellular spaces， small and shriveled nuclei， light staining， fewer mitotic figures and atypical nuclei， and increased necrotic areas. IHC showed that compared with those of the model group， the positive rates of Ki67， VEGF， and CD31 in the recurrent tumor tissue of the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly reduced （P<0.01） in a dose-dependent manner. Western blot results showed that compared with those of the model group， the protein expression levels of ANG-2 and VEGF in the recurrent tumor tissue of the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly downregulated （P<0.05， P<0.01）， and the p-Akt/Akt and p-PI3K/PI3K ratios were significantly decreased in a dose-dependent manner （P<0.05， P<0.01）. ConclusionXianlian Jiedu prescription significantly inhibits the recurrence of CRC in mice after subcutaneous tumor surgery. The mechanism may involve regulating the PI3K/Akt pathway and downregulating key angiogenic proteins such as ANG-2， VEGF， and CD31.

Objective To compare the differences in alkaloids between Nelumbinis Semen and Nelumbinis Plumula and their inhibitory effects on the proliferation of HepG2 cells, and investigate the material basis for their anti-cancer activity differences. Methods Simultaneous Thermal Analysis was used to preliminarily compare the component differences between Nelumbinis Semen and Nelumbinis Plumula. Alkaloids were extracted from them by both using reflux extraction, and their contents were measured by UV and HPLC methods. The CCK-8 method was used to assess the in vitro inhibitory effects of the alkaloids on the HepG2 cells, and to verify pharmacological differences. Results Simultaneous thermal analysis revealed distinct peak shapes, positions, and sizes in the thermal analysis curves of Nelumbinis Semen and Nelumbinis Plumula at respective temperature stages. The contents of total alkaloids showed as follows: the total alkaloids in Nelumbinis Plumula > total extract of Nelumbinis Plumula > the total alkaloids in Nelumbinis Semen. The total alkaloids in Nelumbinis Plumula effectively inhibited HepG2 cell proliferation, while the total alkaloids in Nelumbinis Semen showed no impact. Conclusion Differences in the composition and content of alkaloids may be key factors underlying the biological activities differences between Nelumbinis Semen and Nelumbinis Plumula. This study provided a basis for exploring the material foundation of the differential efficacy and properties of Nelumbinis Semen and Nelumbinis Plumula, which could support their rational clinical application.

Premature ovarian insufficiency(POI)is a complex endocrine disorder that significantly affects the physiological and repro-ductive functions of women,and it has become one of the main causes of infertility in women of childbearing age.The clinical features of POI include amenorrhea or oligomenorrhea,low estrogen or estrogen deficiency,and an increase in the level of gonadotropin.The in-cidence rate of POI tends to increase,yet its etiology and pathogenesis remain unclear.At present,the conventional treatment methods for POI have limited efficacy in fundamentally improving ovarian function or addressing the fertility issue.With the development of re-generative medicine,mesenchymal stem cells(MSCs)have become one of the research hotspots in POI therapy,and their exosomes have attracted wide attention as a promising"cell-free therapy".A number of animal experimental studies have shown that MSCs and their exosomes can exert a therapeutic effect on POI by affecting granulosa cell proliferation and apoptosis,promoting ovarian angiogen-esis,reducing oxidative stress and fibrosis,enhancing follicular development,and regulating immunity.In addition,related clinical studies have also made some progress.This article reviews the mechanisms and clinical effect of MSCs and their exosomes in the treatment of POI,in order to provide a reference for further research and bench-to-bedside translation of POI therapies.

Glycosylation modification is an important post-translational modification of proteins, which participates in regulating protein half-life, biological activity, and immunogenicity, thereby affecting their functions. Glycoproteins expressed in Pichia pastoris predominantly carry high-mannose type glycans, primarily composed of mannose residues, which starkly contrasts with the complex-type glycans synthesized by mammalian cells. This study aims to transform the high mannose glycosylation modification of P. pastoris into a hybrid glycosylation modification similar to that of mammalian cells through genetic engineering technology. We introduced the mannosidase Ⅰ gene (MDSⅠ) from Trichoderma viride and the human β-1,2-N-acetylglucosaminyltransferase I gene (GnTⅠ) into a previously constructed P. pastoris strain (∆och1) capable of producing Man₈GlcNAc₂ glycans. To precisely regulate the expression of MDSⅠ and GnTⅠ, we designed various promoter combinations, including the strong inducible AOX promoter and the constitutive GAP promoter. The receptor-binding domain (RBD, residues 377-588) of the spike protein from the Middle East respiratory syndrome coronavirus (MERS-CoV) was selected as the reporter protein for this investigation (MERS-RBD). The N-glycosylation profile of MERS-RBD was systematically analyzed using PNGase F digestion coupled with mass spectrometry. The results showed that after the knockout of och1 and the introduction of MDSⅠ and GnTⅠ genes with different promoter combinations, P. pastoris strains capable of producing GlcNAcMan₅GlcNAc₂ glycans were successfully generated. When the AOX promoter was used to control the MDSⅠ gene and the GAP promoter was used to control the GnTⅠ gene, the engineered strain exhibited the highest proportion of hybrid-type GlcNAcMan₅GlcNAc₂ glycans, which accounted for 68.38% of the total N-glycosylation. In conclusion, we successfully engineered a P. pastoris strain capable of synthesizing hybrid-type GlcNAcMan₅GlcNAc₂ glycans, establishing a foundation for subsequent research on the biosynthesis of complex-type N-glycans in P. pastoris.
Glycosylation ; Glycoproteins/genetics* ; Polysaccharides/metabolism* ; N-Acetylglucosaminyltransferases/metabolism* ; Pichia/metabolism* ; Humans ; Mannosidases/metabolism* ; Genetic Engineering ; Trichoderma/genetics* ; Recombinant Proteins/genetics* ; Saccharomycetales

Objective:To evaluate the predictive value of whole blood cell derived inflammatory marker (including systemic immunoinflammatory index (SII), systemic inflammatory response index (SIRI), neutrophil count/lymphocyte count (NLR), platelet count/lymphocyte count (PLR), and monocyte count/lymphocyte count (MLR)) and in combination with N-terminal pro-B-type natriuretic peptide (NT-proBNP) on the prognosis of patients with chronic heart failure.Methods:This study was a retrospective cohort study. Patients with chronic heart failure hospitalized in the Department of Cardiovascular Medicine, Nanfang Hospital, Southern Medical University from January 2019 to August 2022 were enrolled. Patients were followed up and were divided into survival group and death group according to the follow-up results. Clinical characteristics of the two groups were compared. Receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value of each whole blood cell derived inflammatory marker for predicting all-cause death in patients with chronic heart failure. Kaplan-Meier survival curve was drawn, and log-rank test was used to compare the difference in survival of chronic heart failure patients with different levels of whole blood cell derived inflammatory markers. Univariate and multivariate Cox proportional hazards models were used to analyze the effects of whole blood cell derived inflammatory markers and NT-proBNP on the all-cause death of patients with chronic heart failure. ROC curve was used to analyze the predictive value of whole blood cell derived inflammatory markers combined with NT-proBNP on the prognosis of patients with chronic heart failure.Results:A total of 324 patients with heart failure aged (64.76±13.78) years were enrolled, with 212 males (65.43%). 297 patients (91.67%) completed follow-up, 27 patients (8.33%) were lost to follow-up. The follow-up time was 24.0 (18.0, 41.8) months. There were 258 patients in the survival group and 66 patients in the death group. The optimal cut-off values of SII, SIRI, NLR, PLR and MLR determined by ROC curve were 739.83, 1.65, 3.14, 151.95 and 0.37, respectively. Kaplan-Meier survival curve analysis showed that patients with chronic heart failure with high levels of SII (≥739.83), SIRI (≥1.65), NLR (≥3.14), PLR (≥151.95) and MLR (≥0.37) had higher incidence of all-cause death than patients with low levels of inflammatory markers (all P<0.001). Multivariate Cox proportional hazard regression analysis showed that age ( HR=1.04, 95% CI 1.01-1.06, P=0.002), NT-proBNP ( HR=2.93, 95% CI 1.64-5.23, P<0.001), SII≥739.83 ( HR=3.27, 95% CI 1.18-9.02, P=0.022) and PLR≥151.95 ( HR=2.67, 95% CI 1.02-6.96, P=0.045) were independent predictors of all-cause death in patients with chronic heart failure. ROC curve analysis showed that the predictive value of SII and PLR combined with NT-proBNP ( AUC=0.850) for the prognosis of patients with chronic heart failure was better than that of SII ( AUC=0.779)、PLR ( AUC=0.782)、NT-proBNP ( AUC=0.727) and CRP ( AUC=0.668) alone (all P<0.001). Conclusions:Whole blood cell derived inflammatory markers——SII, PLR, and NT-pro BNP were independently associated with all-cause death in patients with chronic heart failure. SII and PLR can independently predict the prognosis of patients with chronic heart failure, combination of SII and PLR with NT-pro BNP has better predictive value for the prognosis of patients with chronic heart failure.

Purpose To evaluate the diagnostic value of an abbreviated protocol MRI(AP-MRI)based on first post-contrast subtracted(FAST)images for breast cancer detection.Materials and Methods This study included imaging data from 160 female patients with solid breast lesions who underwent breast MRI in the First Affiliated Hospital of Henan University of Chinese Medicine from April 2021 to January 2024.Two AP-MRI protocols were extracted from the full diagnostic protocol(FDP),including:dynamic contrast-enhanced MRI(DCE-MRI)A protocol:FAST and maximum-intensity projection(MIP)images,and DCE-MRI B protocol:FAST+MIP+diffusion-weighted imaging(DWI).Lesions categorized as breast imaging reporting and data system(BI-RADS)1-3 were classified as negative,and those categorized as BI-RADS 4-5 were classified as positive.Pathological findings served as the diagnostic gold standard.Two radiologists independently evaluated the lesions as negative/positive and compared with the gold standard.The sensitivity,specificity and accuracy of the three protocols were compared.Receiver operating characteristic curves were generated for each protocol,and the area under the curve(AUC)was compared.Results The accuracy of the three protocols showed statistically significant differences(Cochran's Qreader1=6.000,P=0.050;Cochran's Qreader2=10.909,P=0.012).The accuracy of the DCE-MRI A protocol was significantly lower than that of the FDP protocol(Z=2.449,Preader1=0.043;Z=2.858,Preader2=0.013).There were no statistically significant differences in sensitivity(Cochran's Qreader1=3.000,P=0.223;Cochran's Qreader2=2.667,P=0.264)or specificity(Cochran's Qreader1=3.000,P=0.223;Cochran's Qreader2=2.800,P=0.247)between the two AP-MRI protocols and the FDP protocol.There were no statistically significant differences in AUC between the DCE-MRI B protocol and the FDP protocol(Z=1.390-1.719,all P>0.05),while the AUC of the DCE-MRI A protocol had lower AUCs than the FDP protocol(Z=1.980,2.441;both P<0.05).Conclusion The AP-MRI protocol combining FAST,MIP and DWI shows diagnostic accuracy comparable to that of the FDP and greatly saves time and cost,suggesting its potential as an alternative imaging strategy for women with dense breasts and as a new diagnostic approach for high-risk populations.

Objective:To investigate the relationship between nerve growth factor (NGF) concentration in follicular fluid and abnormal menstrual cycle in infertile patients with polycystic ovary syndrome (PCOS).Methods:A retrospective cohort study was conducted on 100 infertile patients with PCOS who underwent in vitro fertilization and embryo transfer (IVF-ET) at Department of Obstetrics and Gynecology, Peking University Third Hospital from March 2017 to June 2019. For comparison, the 100 patients with PCOS were divided into low NGF group ( n=50) and high NGF group ( n=50) based on the median NGF concentration (1 644.03 ng/L) in follicular fluid. Baseline characteristics, menstrual status and clinical outcomes of assisted reproductive technology were compared. We performed multiple linear regression analysis to examine the effect of NGF in follicular fluid on menstrual cycle length for multivariate analysis. Results:1) PCOS patients in the low NGF group had significantly higher body mass index [(27.24±5.17) kg/m 2] and white blood cell count [7.31(5.99, 8.43)×10 9/L ] than those in the high NGF group [(25.03±4.46) kg/m 2, P=0.024; 5.95(5.08,7.01)×10 9/L, P=0.001], while high-density lipoprotein cholesterol [1.15 (0.98, 1.36) mmol/L] and basic follicle-stimulating hormone level [6.51 (5.10,7.95) U/L] in the low NGF group were significantly lower than those in the high NGF group [1.36 (1.09,1.52) mmol/L, P=0.039;6.51 (5.10,7.95)U/L, P=0.040]. 2) PCOS patients in the low NGF group had significantly higher menstrual cycle length [60.00 (35.00, 180.00) d] than the high NGF group [32.50 (27.00,67.50) d, P=0.001]. 3) Multiple linear regression analysis revealed that after adjustment for body mass index, age, infertility duration, infertility type, and glucose and lipid metabolic parameters, the NGF concentration in the follicular fluid independently and negatively correlated with menstrual cycle length ( P<0.05). 4) The NGF concentration in follicular fluid was not correlated with assisted reproductive outcomes. Conclusion:NGF concentration in follicular fluid is closely related to the degree of menstrual cycle abnormalities in patients with PCOS.

Objective To observe the effects of Yuebi Decoction on adriamycin-induced nephrotic syndrome(NS)and adhesion molecule levels in rats;To explore the possible mechanism of its inhibition of NS edema.Methods A NS rat model was established by a one-time tail vein injection of 6.5 mg/kg adriamycin.The rats were randomly divided into normal group,model group,prednisone group and Yuebi Decoction group.The prednisone group and Yuebi Decoction group were given 5 mg/kg prednisone and 22.2 g/kg Yuebi Decoction by gavage,respectively.The normal group and the model group were given an equal amount of normal saline by gavage,once a day for 2 consecutive weeks.The wet and dry weight ratios of skin and kidney were detected,24 hour urine total protein quantification(24 hUTP)and the contents of blood urea nitrogen(BUN),serum creatinine(SCr),total cholesterol(TC),albumin(ALB),hemoglobin(HB)were detected,HE and PAS staining was used to observe the morphology of renal tissue,ELISA was used to detect vascular cell adhesion molecule-1(VCAM-1)and intercellular adhesion molecule-1(ICAM-1)contents in renal tissue,the positive expressions of VCAM-1 and ICAM-1 were detected by immunohistochemistry,the expressions of VCAM-1,Rac1,signal transducer and activator of transcription 3(STAT3)and p-STAT3 proteins were detected by Western blot.Results Compared with the normal group,the wet dry weight ratio of the skin and kidney of the model group rats significantly increased(P<0.01),and the 24 hUTP,SCr,BUN and TC contents significantly increased(P<0.01),while the ALB and HB contents significantly decreased(P<0.01);with glomerular balloon atrophy,the renal tubules exhibit protein casts,inflammatory cell infiltration in renal interstitium,and significant increase in the contents of VCAM-1,ICAM-1 and protein expressions of VCAM-1,Rac1,p-STAT3 in renal tissue(P<0.01).Compared with the model group,the wet dry weight ratio of the skin and kidneys of rats in the prednisone group and Yuebi Decoction group was significantly reduced(P<0.05,P<0.01),and the 24 hUTP,SCr,BUN and TC were significantly reduced(P<0.01),while the contents of ALB and HB significantly increased(P<0.01);the infiltration of inflammatory cells in renal tissue was reduced,the protein casts decreased,and the contents of VCAM-1 and ICAM-1 in renal tissue,as well as the expressions of VCAM-1,Rac1 and p-STAT3 proteins significantly decreased(P<0.05,P<0.01).Conclusion Yuebi Decoction can alleviate edema,reduce urine protein,and improve capillary permeability in NS rats.Its mechanism is related to inhibiting the Rac1/STAT3 signaling pathway and reducing the expression of renal tissue adhesion molecules.

Objective:To investigate the relationship between nerve growth factor (NGF) concentration in follicular fluid and abnormal menstrual cycle in infertile patients with polycystic ovary syndrome (PCOS).Methods:A retrospective cohort study was conducted on 100 infertile patients with PCOS who underwent in vitro fertilization and embryo transfer (IVF-ET) at Department of Obstetrics and Gynecology, Peking University Third Hospital from March 2017 to June 2019. For comparison, the 100 patients with PCOS were divided into low NGF group ( n=50) and high NGF group ( n=50) based on the median NGF concentration (1 644.03 ng/L) in follicular fluid. Baseline characteristics, menstrual status and clinical outcomes of assisted reproductive technology were compared. We performed multiple linear regression analysis to examine the effect of NGF in follicular fluid on menstrual cycle length for multivariate analysis. Results:1) PCOS patients in the low NGF group had significantly higher body mass index [(27.24±5.17) kg/m 2] and white blood cell count [7.31(5.99, 8.43)×10 9/L ] than those in the high NGF group [(25.03±4.46) kg/m 2, P=0.024; 5.95(5.08,7.01)×10 9/L, P=0.001], while high-density lipoprotein cholesterol [1.15 (0.98, 1.36) mmol/L] and basic follicle-stimulating hormone level [6.51 (5.10,7.95) U/L] in the low NGF group were significantly lower than those in the high NGF group [1.36 (1.09,1.52) mmol/L, P=0.039;6.51 (5.10,7.95)U/L, P=0.040]. 2) PCOS patients in the low NGF group had significantly higher menstrual cycle length [60.00 (35.00, 180.00) d] than the high NGF group [32.50 (27.00,67.50) d, P=0.001]. 3) Multiple linear regression analysis revealed that after adjustment for body mass index, age, infertility duration, infertility type, and glucose and lipid metabolic parameters, the NGF concentration in the follicular fluid independently and negatively correlated with menstrual cycle length ( P<0.05). 4) The NGF concentration in follicular fluid was not correlated with assisted reproductive outcomes. Conclusion:NGF concentration in follicular fluid is closely related to the degree of menstrual cycle abnormalities in patients with PCOS.

Objective To observe the protective effect of Yuebi Decoction on adriamycin(ADR)-induced mouse glomerular endothelial cells(MGEC),and to explore the possible mechanism of action.Methods MGEC were stimulated with ADR at a concentration of 1.25 μmol·L-1 for 12 h.MGEC were divided into five groups:normal group,model group,and Yuebi Decoction low,medium,and high dosage groups(5%,10%,and 20%of drug-containing serum);the viability of cells was detected by CCK-8 assay,the cell mobility was detected by Transwell assay.The relative expression levels of MGEC VCAM-1,Rac1,STAT3,and p-STAT3 were detected by Western blot.The expression of endothelial marker CD31,VCAM-1 and ICAM-1 was detected by immunofluorescence.Results Compared with the normal group,the endothelial cell migration rate was decreased in the model group(P<0.01),the protein expression levels of VCAM-1,ICAM-1,Rac1 and p-STAT 3 were significantly increased(P<0.05,P<0.01).The endothelial cell migration rate of the high-dose group of Yuebi Decoction was significantly increased compared with that of the model group(P<0.05).VCAM-1,ICAM-1,Rac1 and p-STAT3 protein expression levels of the drug-containing serum of Yuebi Decoction each group were significantly reduced(P<0.05,P<0.01).Conclusion Yuebi Decoction inhibited ADR-induced adhesion molecule expression in MGEC,alleviated endothelial injury,and improved endothelial cell migration rate.Its mechanism may be related to the inhibition of the Rac1/STAT3 signaling pathway.