Construction and validation of a machine learning model for the diagnosis of juvenile idiopathic arthritis based on fecal microbiota.

Purpose: Human gut microbiota has been shown to be significantly associated with various inflammatory diseases. Therefore, this study aimed to develop an excellent auxiliary tool for the diagnosis of juvenile idiopathic arthritis (JIA) based on fecal microbial biomarkers. Method: The fecal metagenomic sequencing data associated with JIA were extracted from NCBI, and the sequencing data were transformed into the relative abundance of microorganisms by professional data cleaning (KneadData, Trimmomatic and Bowtie2) and comparison software (Kraken2 and Bracken). After that, the fecal microbes with high abundance were extracted for subsequent analysis. The extracted fecal microbes were further screened by least absolute shrinkage and selection operator (LASSO) regression, and the selected fecal microbe biomarkers were used for model training. In this study, we constructed six different machine learning (ML) models, and then selected the best model for constructing a JIA diagnostic tool by comparing the performance of the models based on a combined consideration of area under receiver operating characteristic curve (AUC), accuracy, specificity, F1 score, calibration curves and clinical decision curves. In addition, to further explain the model, Permutation Importance analysis and Shapley Additive Explanations (SHAP) were performed to understand the contribution of each biomarker in the prediction process. Result: A total of 231 individuals were included in this study, including 203 JIA patients and Non-JIA individuals. In the analysis of diversity at the genus level, the alpha diversity represented by Shannon value was not significantly different between the two groups, while the belt diversity was slightly different. After selection by LASSO regression, 10 fecal microbe biomarkers were selected for model training. By comparing six different models, the XGB model showed the best performance, which average AUC, accuracy and F1 score were 0.976, 0.914 and 0.952, respectively, thus being used to construct the final JIA diagnosis model. Conclusion: A JIA diagnosis model based on XGB algorithm was constructed with excellent performance, which may assist physicians in early detection of JIA patients and improve the prognosis of JIA patients.

Using machine learning techniques to predict the risk of osteoporosis based on nationwide chronic disease data.

Osteoporosis is a major public health concern that significantly increases the risk of fractures. The aim of this study was to develop a Machine Learning based predictive model to screen individuals at high risk of osteoporosis based on chronic disease data, thus facilitating early detection and personalized management. A total of 10,000 complete patient records of primary healthcare data in the German Disease Analyzer database (IMS HEALTH) were included, of which 1293 diagnosed with osteoporosis and 8707 without the condition. The demographic characteristics and chronic disease data, including age, gender, lipid disorder, cancer, COPD, hypertension, heart failure, CHD, diabetes, chronic kidney disease, and stroke were collected from electronic health records. Ten different machine learning algorithms were employed to construct the predictive mode. The performance of the model was further validated and the relative importance of features in the model was analyzed. Out of the ten machine learning algorithms, the Stacker model based on Logistic Regression, AdaBoost Classifier, and Gradient Boosting Classifier demonstrated superior performance. The Stacker model demonstrated excellent performance through ten-fold cross-validation on the training set and ROC curve analysis on the test set. The confusion matrix, lift curve and calibration curves indicated that the Stacker model had optimal clinical utility. Further analysis on feature importance highlighted age, gender, lipid metabolism disorders, cancer, and COPD as the top five influential variables. In this study, a predictive model for osteoporosis based on chronic disease data was developed using machine learning. The model shows great potential in early detection and risk stratification of osteoporosis, ultimately facilitating personalized prevention and management strategies.

Porphyromonas gingivalis promotes the progression of oral squamous cell carcinoma by stimulating the release of neutrophil extracellular traps in the tumor immune microenvironment.

BACKGROUND: The aim of this study was to investigate the impact of Porphyromonas gingivalis (P. gingivalis) on the progression of oral squamous cell carcinoma (OSCC) through neutrophil extracellular traps (NETs) in the tumor immune microenvironment. METHODS: The expression of NETs-related markers was identified through immunohistochemistry, immunofluorescence, and Western blotting in different clinical stages of OSCC samples. The relationship between NETs-related markers and clinicopathological characteristics in 180 samples was analyzed using immunohistochemistry data. Furthermore, the ability to predict the prognosis of OSCC patients was determined by ROC curve analysis and survival analysis. The effect of P. gingivalis on the release of NETs was identified through immunofluorescence and immunohistochemistry, both in vitro and in vivo. CAL27 and SCC25 cell lines were subjected to NETs stimulation to elucidate the influence of NETs on various cellular processes, including cell proliferation, migration, invasion, and metastasis in vitro. Furthermore, the impact of NETs on the growth and metastatic potential of OSCC was assessed using in vivo models involving tumor-bearing mice and tumor metastasis mouse models. RESULTS: Immunochemistry analysis revealed a significant correlation between the NETs-related markers and clinical stage, living status as well as TN stage. P. gingivalis has demonstrated its ability to effectively induce the release of NETs both in vivo and in vitro. NETs have the potential to facilitate cell migration, invasion, and colony formation. Moreover, in vivo experiments have demonstrated that NETs play a pivotal role in promoting tumor metastasis. CONCLUSION: High expression of NETs-related markers demonstrates a strong correlation with the progression of OSCC. Inhibition of the NETs release process stimulated by P. gingivalis and targeted NETs could potentially open up a novel avenue in the field of immunotherapy for patients afflicted with OSCC.

PLAU and LAMC2 can predict a poor prognosis in patients with HNSCC.

Objectives: Head and neck squamous cell carcinoma (HNSCC) is the most common malignancy of the head and neck. However, the molecular mechanisms governing the development of HNSCC have not been fully elucidated. Materials and Methods: Differentially expressed genes (DEGs) were screened out from The Cancer Genome Atlas (TCGA) and GSE23036 datasets. Weighted gene coexpression network analysis (WGCNA) was used to reveal the correlations among genes and to search for significantly correlated gene modules. The expression levels of genes in HNSCC and normal samples according to antibody-based detected methods was assessed by utilizing the Human Protein Atlas (HPA). The impact of the selected hub genes on the prognosis of HNSCC patients was assessed by analysing immunohistochemistry (IHC) and immunofluorescence (IF) expression levels and clinical data. Results: Twenty-four genes positively correlated with tumour status and 15 genes negatively correlated with tumour status were screened out by WGCNA. PLAU and LAMC2 were associated with a poor prognosis in patients with HNSCC and were finally screened out and verified by GEPIA and HPA database analysis. Immunohistochemistry of samples collected from 175 patients with HNSCC and subsequent statistical analysis also showed that PLAU and LAMC2 were associated with a poor prognosis in patients with HNSCC, and the levels of these two factors were positively correlated. The expression and co-localization of PLAU and LAMC2 in HNSCC tissues were confirmed by double immunofluorescence labeling. Conclusions: There was a positive correlation between PLAU and LAMC2 expression in HNSCC samples, and PLAU and LAMC2 might be independent prognostic biomarkers for HNSCC.

[Application of 3D printing in the treatment of unilateral comminuted zygomatic bone fracture].

To explore the validity of 3D printing technique in the treatment of unilateral comminuted zygomatic bone fracture. METHODS: Twenty-one patients with unilateral comminuted zygomatic bone fracture were included in the present study, which were treated from hospital January 2014 to April 2017. All patients underwent CT scan and the data were imported in Mimics 10.01 software. The zygomatic bone of healthy side was mirrored to the fracture side to rebuild a "perfect" reduction model. Bone fixation plates were pre-modeled on the model printed by a 3D printing machine and used for bone reduction and fixation during operation. Three dimensional measurements were performed to evaluate the validity of 3D printing based on pre- and post-operative three dimensional CT model. SPSS25.0 software package was used to perform paired t test on the measured data. RESULTS: No significant difference were observed between postoperative CT model and preoperative "perfect" reduction model. All patients were satisfied with their facial appearance. CONCLUSIONS: 3D printing technique is helpful to improve the accuracy of reduction of unilateral comminuted zygomatic bone fracture via preoperative pre-modeling.

Propranolol induces hemangioma endothelial cell apoptosis via a p53­BAX mediated pathway.

The use of propranolol for the treatment of infantile hemangioma (IH) has been widely investigated in recent years. However, the underlying therapeutic mechanism of propranolol for the treatment of IH remains poorly understood. The aim of the present study was to investigate the expression of proteins regulated by cellular tumor antigen p53 (p53) in associated apoptosis pathways in IH endothelial cells (HemECs) treated with propranolol. Furthermore, the present study aimed to investigate the exact apoptotic pathway underlying the therapeutic effect of propranolol against IH. In the present study, HemECs were subcultured and investigated using an inverted phase contrast microscope, immunocytochemical staining and a scanning electron microscope (SEM). Experimental groups and blank control groups were prepared. All groups were subjected to drug treatment. A high p53 expression model of HemECs was successfully established via transfection, and a low p53 expression model of HemECs was established using pifithrin­α. The apoptosis rate of each group was determined using Annexin V­fluorescein isothiocyanate/propidium iodide double staining and flow cytometry. The expression levels of downstream proteins regulated by p53 [tumour necrosis factor receptor superfamily member 6 (FAS), p53­induced death domain­containing protein (PIDD), death receptor 5 (DR5), BH3­interacting domain death agonist (BID), apoptosis regulator BAX (BAX), p53 unregulated modulator of apoptosis (PUMA), phosphatidylinositol­glycan biosynthesis class S protein (PIGS), and insulin­like growth factor­binding protein 3 (IGF­BP3)] were revealed in the experimental and control groups via western blotting. Microscopic observation revealed the growth of an adherent monolayer of cells, which were closely packed and exhibited contact inhibition. Immunocytochemical staining demonstrated increased expression of clotting factor VIII. SEM analysis revealed presence of Weibel­Palade bodies. The results of the analyses verified that the cultured cells were HemECs. The staining of the samples resulted in a significantly increased rate of apoptosis in experimental groups compared with the blank control group. This result suggested that there is an association between p53 expression and the rate of apoptosis of propranolol­treated HemECs. The results of the western blot analysis demonstrated an upregulation of BAX expression and a downregulation of IGF­BP3 expression in the HemECs treated with propranolol. There were no significant differences in the expression levels of FAS, DR5, PIDD, BID, PUMA and PIGS between experimental and control groups. This result suggests that p53 has an important role in HemEC apoptosis. The results of the present study additionally suggest that the propranolol­induced HemEC apoptosis pathway is a mitochondrial apoptosis pathway and is regulated by p53­BAX signaling.

Efficacy of modified endoscopic release in treatment of carpal tunnel syndrome / 中国内镜杂志

Objective To explore a new technology system with single-portal and without special equipment to complete the arthroscopic treatment of carpal tunnel syndrome. Methods All the patients with CTS were randomly divided into two groups, modified endoscopic carpal tunnel release (MECTR) group (22 cases) and open carpal tunnel release (OCTR) group (24 cases). Nine parameters were evaluated, which including operation time, intraoperative complications, two-point discrimination 3 months after the operation, hospitalization time, scar pain score, incision infection rate, each patient's symptom amelioration (Kelly grading), the time needed to resume normal lifestyle and activity, symptoms of sympathetic dystrophy. Results No significant difference was observed between the MECTR group and OCTR group in regard to incision infection rate, intraoperative complications, two-point discrimination,symptoms of sympathetic dystrophy and clinical symptoms amelioration. In comparison to OCTS, MECTR significantly decreased operation time, hospitalization time, scar pain score and the time needed to resume normal lifestyle and activity. Conclusion MECTR for treatment of carpal tunnel syndrome has higher patient satisfaction, shorter operation time and hospitalization time, earlier return to work or normal lifestyle, less postoperative scar pain, so it is an effective method for the treatment of idiopathic carpal tunnel syndrome.

Pingyangmycin stimulates apoptosis in human hemangioma-derived endothelial cells through activation of the p53 pathway.

Pingyangmycin (also known as Bleomycin A5) is produced by Streptomyces verticillus var. pingyangensis n.sp., and has anti­tumor activities against a variety of tumor cells. The aim of the present study was to determine the molecular mechanism(s) underlying the therapeutic effects of pingyangmycin against infantile hemangiomas. Human hemangioma­derived endothelial cells (HemECs) were treated with pingyangmycin at varying concentrations (100, 200 or 300 µg/ml), and the morphological changes and apoptosis levels were assessed. The gene expression changes were determined by cDNA microarray technology. Transmission electron microscopy examination revealed that the pingyangmycin­treated HemECs exhibited typical apoptotic characteristics, including chromatin condensation and the formation of apoptotic bodies. Annexin­V staining demonstrated that pingyangmycin caused a significant and dose­dependent induction of apoptosis in the HemECs. In the pingyangmycin­treated HemECs, 4,752 genes demonstrated at least 2­fold expression changes at the mRNA level. Quantitative polymerase chain reaction confirmed that pingyangmycin significantly upregulated the expression of p53, p53­induced protein with death domain, Bax, p53 upregulated modulator of apoptosis and p53 inducible gene 3, and downregulated the expression of murine double minute 2. The data demonstrated that the pro­apoptotic activity of pingyangmycin against infantile hemangiomas involves p53 pathway activation.

Induction of apoptosis in infantile hemangioma endothelial cells by propranolol.

Propranolol, a non-selective ß-blocker, is emerging as an effective treatment for complicated hemangiomas. The aim of this study was to investigate the molecular mechanism(s) underlying the therapeutic effects of propranolol against hemangiomas, using primary infantile hemangioma endothelial cells (IHECs). IHECs were treated with various concentrations of propranolol and morphological changes and apoptosis were assessed. Changes in the expression levels of apoptosis-related genes were examined. Annexin-V staining revealed that propranolol at 40, 50 and 60 µg/ml caused a concentration-dependent increase in the apoptosis of IHECs. Morphological analyses revealed that exposure to 50 µg/ml propranolol resulted in typical apoptotic changes, including shrinkage, the formation of apoptotic bodies and retention of plasma membrane integrity. Gene expression analyses revealed that propranolol treatment led to a marked increase in the expression of caspase-8, cytochrome c, apoptosis-inducing factor, caspase-3 and poly (ADP-ribose) polymerase 1, as well as a concomitant reduction in lamin B1 expression. Our data collectively demonstrate that propranolol induces apoptosis of IHECs through activation of the intrinsic and extrinsic apoptotic pathways, which represents an important mechanism for its therapeutic effects against infantile hemangiomas.

Proteomic analysis of mitochondria from infantile hemangioma endothelial cells treated with sodium morrhuate and its liposomal formulation.

Hemangioma is the most common benign tumor of infancy. The aim of this study is to evaluate the biological effects of sodium morrhuate (SM) and its liposomal formulation on infantile hemangioma endothelial cells (IHECs). Morphological analysis revealed that exposure to liposomal sodium morrhuate (LSM) preferentially caused apoptotic death in IHECs, manifested as shrunken configuration and formation of apoptotic bodies. In contrast, necrotic death was prominent in IHECs treated with an equal concentration of SM. By means of proteomic analysis and confirmation experiments, we revealed that the apoptosis-inducing effects of LSM were associated with an upregulation of a set of genes involved in mitochondrial death pathway, including apoptosis-inducing factor, cytochrome c1, caspase-8, and lamin B1. In conclusion, our data highlight the proapoptotic activity of LSM in IHECs through the mitochondrial apoptotic pathway and may provide a promising avenue to treat hemangiomas of infancy.

Cartilage tissue engineering with demineralized bone matrix gelatin and fibrin glue hybrid scaffold: an in vitro study.

To develop a cartilage-like tissue with hybrid scaffolds of demineralized bone matrix gelatin (BMG) and fibrin, rabbit chondrocytes were cultured on hybrid fibrin/BMG scaffolds in vitro. BMG scaffolds were carefully soaked in a chondrocyte-fibrin suspension, which was polymerized by submerging the constructs into thrombin-calcium chloride solution. Engineered cartilage-like tissue grown on the scaffolds was characterized by histology, immunolocalization, scanning electron microscopy, biochemical assays, and analysis of gene expression at different time points of the in vitro culture. The presence of proteoglycan in the fibrin/BMG hybrid constructs was confirmed by positive toluidine blue and alcian blue staining. Collagen type II exhibited intense immunopositivity at the pericellular matrices. Chondrogenic properties were further demonstrated by the expression of gene-encoded cartilage-specific markers, collagen type II, and aggrecan core protein. The glycosaminoglycan production and hydroxyproline content of tissue grown on the fibrin/BMG hybrid scaffolds were higher than that of the BMG group. In conclusion, the fibrin/BMG hybrid scaffolds may serve as a potential cell delivery vehicle and a structural basis for cartilage tissue engineering.

[Effects of RNA interference against aggrecanase 1 gene on extracellular matrix metabolism of cultured chondrocytes in vitro].

OBJECTIVE: To study the effect of RNA interference (RNAi)-mediated aggrecanase-1 gene silencing on extracellular matrix metabolism of cultured rat costochondral chondrocytes. METHODS: Rat costochondral chondrocyte monolayers were obtained by microdissection and digestion. The growth and morphological changes of the chondrocytes were observed after RNAi of aggrecanase-1 gene. The mRNA expression of aggrecanase-1 was detected by RT-PCR method, and aggrecan content was determined by Western blotting. RESULTS: The specific inhibition of aggrecanase-1 by RNAi produced no adverse effect on the morphology and growth of the chondrocytes. The mRNA of aggrecanase-1 decreased and aggrecan content increased significantly after transfection of the chondrocytes. CONCLUSION: Inhibition of aggrecanase-1 decreases aggrecan degradation in cultured rat chondrocytes. RNAi technique can be a useful means for studying extracellular matrix metabolism in the cartilage.

Effects of RNAi-mediated inhibition of aggrecanase-1 and aggrecanase-2 on rat costochondral chondrocytes in vitro.

AIM: Failure of transplanted cartilage or allogenic chondrocytes is attributed mainly to immunological rejection and cartilage degradation. A major feature is the loss of aggrecan from the cartilage matrix, primarily due to the action of the specific proteinases aggrecanase-1 and aggrecanase-2. The aim of this in vitro study was to determine whether the specific inhibition of aggrecanase-1 and aggrecanase-2 by RNAi would mitigate aggrecan loss from cultured chondrocytes. METHODS: Expression plasmid vectors of shRNA targeting aggrecanase-1 and aggrecanase-2 were constructed and transfected into cultured rattus costochondral chondrocytes. The transfected cells were induced with interleukin-1beta (IL-1beta). Gene mRNA levels were analyzed by RT-PCR. Aggrecan and collagen II content were measured by immunohistochemistry and Western blotting. RESULTS: As the chondrocytes underwent dedifferentiation, aggrecanase-1 increased significantly. The specific inhibition of aggrecanase-1 and aggrecanase-2 by RNAi had no negative effect on the morphology and growth velocity of the chondrocytes. The mRNA of aggrecanase-1 and aggrecanase-2 decreased significantly. The alpha-2-macroglobulin expression level was increased by the shRNA specific for aggrecanase-1. Other genes of the chondrocytic extracellular matrix were not affected. RNAi significantly increased the aggrecan and collagen II content of chondrocytes treated with IL-1beta. CONCLUSION: The results suggest that inhibition of aggrecanase-1 and aggrecanase-2 by RNAi can mitigate aggrecan degradation, without interfering with chondrocytic gene phenotype recovery. RNAi technology can be a useful tool for studying degenerative processes in cartilage.

[Study of immunolipo-sodium morrhuate applied on human hemangioma endothelial cells in vitro].

PURPOSE: To explore the method of preparing immunolipo-sodium morrhuate and evaluate its effect on human hemangioma endothelial cells in vitro. METHODS: Using SPDP((N-Succinimidyl-3-(2-pyridyldithio)) propionate) as cross-linker, anti-VEGFR2/KDR monoclone antibody was combined to the liposome surface to prepare immunolipo-sodium morrhuate by extruding method, and then its effect on human hemangioma endothelial cells in vitro was observed by laser scanning confocal microscope, inverted microscope, Gimsa staining, transmission electron microscope, MTT and flow cytometry. RESULTS: The average diameter of the immunoliposome was 122.9 nm, which had a very good stability when compared with normal liposome, it had stronger and faster combining ability, its potential to induce apoptosis was much more prominent, and its toxic effect on human hemangioma endothelial cells was gentle, which was similar to normal liposome. CONCLUSION: We have prepared immunolipo-sodium morrhuate successfully, which has very good specific initiative targetting ability in vitro and can induce pervasive apoptosis of human hemangioma endothelial cells.

[The effect of aloesin on melanocytes in the pigmented skin equivalent model].

OBJECTIVE: To observe the effect of aloesin, tea polyphenols, arbutin on melanocytes in the pigmented skin equivalent model. METHODS: First, we constructed the pigmented skin equivalent model in vitro. And then we detected the effect of aloesin, tea polyphenols and arbutin on the cells' shape, tyrosinase activity and formation of melanin in the constructed pigmented skin equivalent. RESULTS: Three depigmenting agents showed an inhibition effect on the tyrosinase activity of melanocytes and reduced significantly melanin content in the pigmented skin equivalent model, in which the tea polyphenols had the strongest effect, and then was the aloesin. But the tea polyphenols showed the strongest toxicity, while the aloesin and arbutin had a much lower toxicity. CONCLUSIONS: All the three depigmenting agents showed a concentration dependent suppression effect on the tyrosinase activity and formation of melanin, in which the tea polyphenols was the strongest effect( P <0.05). Aoesin has a good suppression effect on the tyrosinase activity and formation of melanin, but has a much lower toxicity, which could be used as a safe depigmenting agent.

[Construction of an in vitro equivalent of the pigmented skin].

OBJECTIVE: To construct an in vitro equivalent of the pigmented skin using tissue engineering methods. METHODS: Surgically removed foreskins was used as the source of keratinocytes and melanocytes harvested by routine tissue digestion. The fibroblasts were enriched by tissue block culture and seeded into the scaffold constructed using mouse tail collagens to construct the pigmented skin equivalent model. The general structure and the melanocyte distribution and growth status in this model were observed with HE staining and Fontana Masson staining. The ultrastructure of the constructed pigmented skin equivalent was observed by transmission electron microscope. RESULTS AND CONCLUSION: The pigmented skin equivalent model was structurally intact, and allowed optimal cell growth. Fontana Masson staining identified in the basal layer numerous melanocytes in normal growth, and the constructed model was structurally similar to normal skin tissue, suggesting successful construction of the pigmented skin equivalent model.

[The experimental study of lipo-sodium morrhuate promoting apoptosis of ECV-304 cells].

OBJECTIVE: To evaluate the effect of lipo-sodium morrhuate on ECV-304 cell line. METHODS: The effect lipo-sodium morrhuate was evaluated by toxicology trial (MTT), electron microscope, DNA electrophoresis and flow cytometer. RESULTS: The toxicology results showed, that the number of vital cells in lipo-sodium morrhuate group decreased slowly. The electron microscope exhibited apoptosis in the lipo-sodium morrhuate group. And there were typical DNA ladder in DNA electrophoresis and typical apoptosis peak in flow cytometer. The apoptosis rate was 22.23%. CONCLUSIONS: Unlike the normal preparation of sodium morrhuate, lipo-sodium morrhuate could induce apoptosis of ECV-304 cell line.

[Apoptotic study on the effect of fluorine and selenium on the human hair follicle in vitro].

OBJECTIVE: The aim of this study was to observe the human hair follicle apoptosis status affected by fluorine and the antagonism effect by selenium in vitro. METHODS: The single hair follicles were separated and cultured, then they were added in different concentrations of sodium fluoride and sodium selenite. Chosen the appropriate concentrations, they were divided into 7 groups. The TUNEL was used to investigate the apoptotic cells of different parts. The morphous of hair follicles was observed consecutively and electron microscope was used. RESULTS: We found that in 1 mmol/L and 10 mmol/L sodium fluoride groups, when the human hair follicles in vitro were cultured on the 5th day, the apoptotic cells of outer root sheath (ORS), dermal sheath and hair papilla, hair bulb were obviously increased. But 0.01 mmol/L sodium selenite weakened the toxicity of 1 mmol/L sodium fluoride at the outer root sheath and hair bulb (P < 0.05). CONCLUSIONS: Different concentrations of sodium fluoride had different effect on the growth of human hair follicle in vitro which were cultured on 5th day. Sodium fluoride of certain concentration could accelerate the apoptosis of human hair follicle in vitro. Sodium selenite of certain concentration could act antagonism to the toxicity of sodium fluoride.

[Compare the effect of lipo- and normal prepared Sodium Morrhuate on ECV-304 cell line].

OBJECTIVE: To compare the effect of Sodium Morrhuate on ECV-304 between its lipo- and normal preparation. METHODS: The ECV-304 cell line was supplemented with Sodium Morrhuate and lipo-Sodium Morrhuate in order, and the result on morphology (microscope, Giemsa Staining and electron microscope), cell activity (MTT), and flow cytometer between the two preparation were compared. RESULTS: In normal preparation group, cell's edema occurred. Chromatin was like catkins. Tumefaction and degeneration of mitochondrion and endoplasmic reticulum appeared. In lipo-Sodium Morrhuate group, the membrane was creased and processus appeared. Chromatin aggregates to the membrane of nucleus was like crescent, and then broken. The apoptotic body was formed. MTT changes showed that the curve of the normal preparation group was steep and the change time was short relatively, which cues the vital cells decreased sharply. The curve of lipo-Sodium Morrhuate group was gentle and the change time was long relatively, which cues the vital cells decreased slowly. The flow cytometer showed that typical apoptosis peak appeared. CONCLUSION: The normal preparation group shows an acute toxic effect on ECV-304 cell line, which result in a necrosis course, while lipo-Sodium Morrhuate shows a gradual releasing process, which may indicate a apoptosis course.