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.

Medicinal plants， with diverse species， high heterozygosity， and special breeding objectives， can be hardly bred with conventional hybridization techniques. Plant genetic transformation is highly selective and can specifically change the traits of plants， serving as an important technical means for the breeding of medicinal plants. The commonly used plant genetic transformation technologies include Agrobacterium-mediated transformation and particle bombardment. Agrobacterium-mediated transformation is the most widely used method， while it is not applicable to all medicinal plants due to the high specificity. Although not specific， particle bombardment is limited in application due to the low conversion efficiency and external force damage to cells and tissue. With the rise and development of nanotechnology， the emerging nanomaterial-mediated transformation has solved the problems of the above two technologies. However， limited by its late development， the mechanism of nanomaterial-mediated introduction of genetic materials into plant cells remains unclear， and thus this technology is rarely used in medicinal plants. This article summarizes the development status of several commonly used or emerging plant genetic transformation technologies such as Agrobacterium-mediated transformation， particle bombardment， and nanomaterial-mediated transformation， as well as their application in different medicinal plants. Furthermore， this article looks forward to the development trend of genetic transformation technologies for plants and their application prospects in medicinal plants and Chinese materia medica resources， aiming to provide new technical ideas for the genetic improvement and germplasm innovation of medicinal plants and inject new impetus into the sustainable development of Chinese materia medica resources.

Aim To investigate the high expression of LINC00626 in colorectal cancer,and explore the effects of LINC00626 on the proliferation,apoptosis,and drug sensitivity of colorectal cancer SW480 cells,as well as its underlying mechanisms.Methods Flu-orescence in situ hybridization(FISH)was used to de-tect the expression levels of LINC00626 in 38 colorec-tal cancer tissues and their corresponding adjacent nor-mal tissues.The JASPAR database was utilized to pre-dict co-expressed genes and their possible binding sites.Cell transfection technology was employed to knockdown LINC00626.Western blot and qRT-PCR techniques were used to verify the transfection efficien-cy.CCK-8 assay,cell apoptosis and necrosis staining,and Western blot were used to detect the changes in the proliferation,apoptosis,drug sensitivity,and ap-optotic proteins of SW480 cells,respectively.Results The FISH results indicated that LINC00626 was highly expressed in colorectal cancer tissues(P＜0.05).The expression of LINC00626 was not associat-ed with the age or gender of patients,but was related to the TNM stage and the presence of lymph node me-tastasis($ P＜0.05 $).The results of CCK-8 assay and cell apoptosis and necrosis staining showed that af-ter knockdown of LINC00626,the proliferation ability of SW480 cells decreased,the apoptosis level in-creased,and the drug resistance decreased(P＜0.05).Western blot results showed that with the de-crease in the expression level of LINC00626,the ex-pression of caspase-3 protein decreased,the expression of cleaved caspase-3 protein increased,and the expres-sion of Bcl-2 protein decreased(P＜0.05).Conclu-sions LINC00626 is highly expressed in colorectal cancer and is associated with the TNM stage and the presence of lymph node metastasis.LINC00626 can af-fect the proliferation,apoptosis,and drug sensitivity of SW480 cells and alter the expression of apoptotic pro-teins.

Aim To investigate the expression levels of cytoplasmic FMR1-interacting protein-1(CYFIP1)in colorectal cancer and assess the impact of CYFIP1 interaction on the proliferation and apoptosis of colorec-tal cancer cell HT29,along with its potential mecha-nisms.Methods Immunohistochemistry was em-ployed to assess CYFIP1 expression in 32 colorectal cancer tissues and adjacent tissues.Coexpressed genes were identified using the GEPIA2 website to predict potential correlations and binding sites.Following the construction of a siRNA-CYFIP1,alterations in cell proliferation,apoptosis,and levels of apoptosis-related proteins were evaluated through CCK-8 assay,Hoechst 33342/PI double staining assay,and Western blot a-nalysis,respectively.Results The immunohisto-chemical findings revealed a significantly elevated level of CYFIP1 expression in colorectal cancer tissues com-pared to paracancer tissues(P＜0.05).The expres-sion of CYFIP1 did not show any correlation with age and gender,but exhibited associations with TNM stage and lymph node metastasis(P＜0.05).A conserved TP53 binding site was predicted in the 3kbps DNA re-gion upstream of the CYFIP1 gene using GEPIA2,JASPAR databases,and rVista 2.0 promoter prediction software.Following transfection of HT29 cells with siRNA-CYFIP1,the clonogenesis and proliferation of cells significantly decreased(P＜0.05).Additional-ly,the levels of cleaved caspase-3 were elevated,while the expression levels of caspase-3 and Bcl-2 were reduced after transfection with siRNA-CYFIP1(P＜0.05),which might be related to the interaction be-tween CYFIP1 and TP53.Conclusions The upregu-lation of CYFIP1 in colorectal cancer is associated with TNM stage and lymph node metastasis.Upon silen-cing,CYFIP1 demonstrates the ability to suppress pro-liferation in HT29 cells and modulate the expression of apoptotic proteins.

Objective:To investigate the role of mRNA expression in the pathogenesis of gout by analyzing the difference of gene expression in peripheral blood mononuclear cells(PBMCs)among acute gouty(AG),intermittent gouty(IG)and health con-trols(HC).Methods:AG patients,IG patients and HC were enrolled in this study.RNA-seq and bioinformatics techniques were used to observe the differences of mRNA expression in PBMCs of different groups,and to explore the genes and signaling pathways as-sociated with gout attack.GO and KEGG databases were used to investigate the biological functions of differentially expressed genes and the relationship between genes and signaling pathways.Genes involved in the KEGG enriched PPAR signaling pathway(CYP27A1,ACSL1,CD36,PPARG,ANGPTL4,PLIN2)were validated in PBMCs from 35 patients with AG,35 patients with IG,and 35 normal healthy subjects using real-time PCR.Results:Compared with HC group,there were 222 significant differential genes in AG group,including 193 up-regulated genes and 29 down-regulated genes.GO analysis showed that the genes differentially ex-pressed in AG group were mainly enriched in the regulation of multicellular biological processes such as inflammation,trauma,and stress response and immune response regulation compared with HC group.However,KEGG analysis showed that the up-regulated genes in AG group were enriched in"Toll-like receptor signaling pathway""complement and coagulation cascades""PPAR signaling pathway""lipid and atherosclerosis""osteoclast differentiation""cytokine-cytokine receptor interaction""chemokine signaling path-way""IL-17 signaling pathway",and"cholesterol metabolism"compared with HC group.The results of PCR validation of related genes in the PPAR signaling pathway showed that the expression levels of ACSL1,CD36,PPARG,and PLIN2 in the AG group were higher than those in the control group(P<0.05),and the expression levels of CYP27A1 and ANGPTL4 in the AG group were not dif-ferent from those in the HC group(P>0.05).Conclusion:PPAR signaling pathway may be involved in the pathogenesis of AG,of which ACSL1,CD36,PPARG,and PLIN2 may be used as potential therapeutic targets for acute gouty arthritis;CYP27A1 and ANG-PTL4 may not be associated with the pathogenesis of AG.

Objective:To evaluate the treatment outcome of liver transplantation for patients with polycystic liver disease (PLD).Methods:Clinical data of 28 PLD patients admitted to the Affiliated Hospital of Qingdao University from May 2014 to November 2023 were retrospectively analyzed, including 10 males and 18 females, aged (50.4±6.6) years. Patients were divided into liver transplantation group ( n=15) and non-liver transplantation group ( n=13). In the liver transplantation group, we analyzed seve-ral critical parameters including methods of liver transplantation, intra-abdominal fluid volume, intraoperative blood loss, intraoperative red blood cell transfusion requirements, and postoperative complications. The prognosis of the two groups were also compared. Results:Among the 28 patients with PLD, 15 underwent liver transplantation, including 11 classic in situ liver transplantations, one modified back-to-back liver transplantation, and three liver-kidney combined transplantations. The 15 patients had 2 000 (300, 4 000) ml of abdominal fluid, 1 000 (600, 2 000) ml of intraoperative blood loss, and 8.0 (6.0, 17.0) U of red blood cells transfused during the operation. Postoperative complications occurred in eight cases, with four of which were managemed successfully, and the other four died. The 1-, 5-, and 10-year survival rates of after liver transplantation were 80.0%, 80.0%, and 73.3%, respectively. The 1-, 5-, and 10-year survival rates of patients with PLD without liver transplantation were 69.2%, 46.2%, and 38.5%, respectively. The difference between the two groups was statistically significant ( χ2=3.91, P=0.048). Conclusion:Liver transplantation is a treatment option for patients with PLD, with a better long-term survival compared to patients without liver transplantation.

Objective:To analyze the prognostic factors of patients with liver cirrhosis and portal vein thrombosis (PVT), and to construct a prognostic prediction model based on machine learning methods.Methods:The clinical data of 388 patients with liver cirrhosis and PVT admitted to the Fifth Medical Center of PLA General Hospital from January 2022 to April 2024 were retrospectively collected and analyzed, including 243 males and 145 females, aged (56.9±10.9) years. A total of 388 patients were randomly divided into the training set ( n=310) and the testing set ( n=78) in a 4∶1 ratio. The Boruta algorithm was used to screen the key features in the training set, and then four machine learning algorithms, including random forest, support vector machine, generalized linear model and Bayesian, were used to establish a survival prediction model. Model performance was evaluated by the receiver operating characteristic (ROC) curves of the test set and the training set. The patients were followed up for 1 year for survival. Sort the importance of features based on the SHAP value. Results:There were 250 patients (80.6%) who survived and 60 (19.4%) who died. The model for end-stage liver disease score, total bilirubin, serum creatinine, prothrombin time, international normalized ratio, D-dimer, white blood cell count, severe ascites ratio, and Child-Pugh grade C ratio of liver function in the death group were higher than those in the survival group, and the red blood cell count and hematocrit were lower than those in the survival group, and the differences were statistically significant (all P<0.05). The areas under the ROC curve for predicting survival by random forest, support vector machine, generalized linear model and Bayesian model were 0.92, 0.78, 0.81 and 0.71 in the training set, and the area under the ROC curve in the testing set were 0.81, 0.72, 0.67 and 0.68, respectively. Random forest had the best prediction performance, with an accuracy of 81.7%, a sensitivity of 84.6%, and a specificity of 76.9% in the testing set. In the analysis of the importance of characteristic parameters of the random forest model, total bilirubin, red blood cells, hematocrit, serum creatinine, ascites classification, etc. had a relatively high contribution to the model. Conclusion:In the survival prediction model of patients with liver cirrhosis and PVT based on machine learning algorithm, the random forest model had high prediction performance, and total bilirubin may be the most important factor affecting the survival prognosis of patients.

Objective:To evaluate the treatment outcome of liver transplantation for patients with polycystic liver disease (PLD).Methods:Clinical data of 28 PLD patients admitted to the Affiliated Hospital of Qingdao University from May 2014 to November 2023 were retrospectively analyzed, including 10 males and 18 females, aged (50.4±6.6) years. Patients were divided into liver transplantation group ( n=15) and non-liver transplantation group ( n=13). In the liver transplantation group, we analyzed seve-ral critical parameters including methods of liver transplantation, intra-abdominal fluid volume, intraoperative blood loss, intraoperative red blood cell transfusion requirements, and postoperative complications. The prognosis of the two groups were also compared. Results:Among the 28 patients with PLD, 15 underwent liver transplantation, including 11 classic in situ liver transplantations, one modified back-to-back liver transplantation, and three liver-kidney combined transplantations. The 15 patients had 2 000 (300, 4 000) ml of abdominal fluid, 1 000 (600, 2 000) ml of intraoperative blood loss, and 8.0 (6.0, 17.0) U of red blood cells transfused during the operation. Postoperative complications occurred in eight cases, with four of which were managemed successfully, and the other four died. The 1-, 5-, and 10-year survival rates of after liver transplantation were 80.0%, 80.0%, and 73.3%, respectively. The 1-, 5-, and 10-year survival rates of patients with PLD without liver transplantation were 69.2%, 46.2%, and 38.5%, respectively. The difference between the two groups was statistically significant ( χ2=3.91, P=0.048). Conclusion:Liver transplantation is a treatment option for patients with PLD, with a better long-term survival compared to patients without liver transplantation.

Objective:To analyze the prognostic factors of patients with liver cirrhosis and portal vein thrombosis (PVT), and to construct a prognostic prediction model based on machine learning methods.Methods:The clinical data of 388 patients with liver cirrhosis and PVT admitted to the Fifth Medical Center of PLA General Hospital from January 2022 to April 2024 were retrospectively collected and analyzed, including 243 males and 145 females, aged (56.9±10.9) years. A total of 388 patients were randomly divided into the training set ( n=310) and the testing set ( n=78) in a 4∶1 ratio. The Boruta algorithm was used to screen the key features in the training set, and then four machine learning algorithms, including random forest, support vector machine, generalized linear model and Bayesian, were used to establish a survival prediction model. Model performance was evaluated by the receiver operating characteristic (ROC) curves of the test set and the training set. The patients were followed up for 1 year for survival. Sort the importance of features based on the SHAP value. Results:There were 250 patients (80.6%) who survived and 60 (19.4%) who died. The model for end-stage liver disease score, total bilirubin, serum creatinine, prothrombin time, international normalized ratio, D-dimer, white blood cell count, severe ascites ratio, and Child-Pugh grade C ratio of liver function in the death group were higher than those in the survival group, and the red blood cell count and hematocrit were lower than those in the survival group, and the differences were statistically significant (all P<0.05). The areas under the ROC curve for predicting survival by random forest, support vector machine, generalized linear model and Bayesian model were 0.92, 0.78, 0.81 and 0.71 in the training set, and the area under the ROC curve in the testing set were 0.81, 0.72, 0.67 and 0.68, respectively. Random forest had the best prediction performance, with an accuracy of 81.7%, a sensitivity of 84.6%, and a specificity of 76.9% in the testing set. In the analysis of the importance of characteristic parameters of the random forest model, total bilirubin, red blood cells, hematocrit, serum creatinine, ascites classification, etc. had a relatively high contribution to the model. Conclusion:In the survival prediction model of patients with liver cirrhosis and PVT based on machine learning algorithm, the random forest model had high prediction performance, and total bilirubin may be the most important factor affecting the survival prognosis of patients.

AIM To study the chemical constituents from the sticks and leaves of Croton cascarilloides Raeusch.and their biological activities.METHODS The 95％ethanol extract from the sticks and leaves of C.cascarilloides was isolated and purified by MCI,silica gel,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.LPS-induced NO RAW264.7 cell model induced by LPS was used to evaluate its anti-inflammatory activity in vitro.GES-1 injury model induced by taurocholic acid was used to screen the gastric mucosal protection activity.RESULTS Fourteen compounds were isolated and identified as bullatantriol(1),(-)-boscialin(2),(+)-dehydrovomifoliol(3),3-(hydroxylacetyl)-indole(4),pinoresinol(5),3,7-dimethyl-octa-1,7-diene-3,6-ol(6),(+)-syringaresinol(7),curcasinlignan B(8),cleomiscosin C(9),cleomiscosinD(10),2,6-dimethyl-octa-1,7-dien-3,6-diol(11),vanillin(12),vanillic acid(13),methyl vanillate(14).Compound 4 had certain anti-inflammatory activity,with IC50 values of 73.62 μmol/L.The protective rates of 25 μmol/L compounds 1-4,6,9-12 and 14 on gastric mucosal epithelial cells were 30.07％,34.18％,23.91％,30.92％,17.51％,19.69％,31.76％,22.46％,30.56％and 14.49％,respectively.CONCLUSION Compounds 1-14 are isolated from this plant for the first time.Compound 4 shows anti-inflammatory activity,1-4,6,9-12 and 14 show different degrees of gastric mucosal epithelial cell protective activity.