Gingipain and oncostatin M synergistically disrupt kidney tight junctions in periodontitis-associated acute kidney injury.

BACKGROUND: Acute kidney injury (AKI) is characterized by rapid renal decline. Periodontitis, a chronic oral inflammatory disease, is increasingly associated with renal dysfunction. Although periodontitis is recognized as a contributor to kidney damage, the mechanisms linking it to AKI remain unclear. METHODS: This study explored the effects of Porphyromonas gingivalis (P. gingivalis) W83-infected periodontitis on AKI in C57BL/6J mice, using ischemia-reperfusion injury 55 days post-infection. Gingipain inhibitors, KYT-1 and KYT-36, were applied. Detection of P. gingivalis was performed using quantitative real-time polymerase chain reaction (qRT-PCR) and PCR, while transcriptome sequencing, qRT-PCR, immunohistochemistry, and immunofluorescence staining assessed renal damage. In vitro, HK-2 cells were exposed to P. gingivalis at a multiplicity of infection of 10 for 48 h, with inhibition by gingipain or oncostatin M (OSM). Disruption of tight junctions (TJs) was quantified using qRT-PCR, transepithelial electrical resistance, and cell counting kit-8 assays. RESULTS: Periodontitis worsened AKI, linked to P. gingivalis infection and renal TJ disruption in the kidney. P. gingivalis infection activated OSM expression, which correlated positively with gingipain. Significantly, OSM and gingipain might collaboratively contribute to the damage of renal TJs, with the reduced expression of TJ proteins. Suppressing gingipain activity presented itself as a protective strategy against the destruction of TJs and the attendant worsening of AKI due to periodontitis. CONCLUSIONS: Our study enhances the understanding of the interplay between periodontitis and AKI, highlighting the harmful impact of P. gingivalis in AKI.

Bromelain supplementation and inflammatory markers: A systematic review of clinical trials.

BACKGROUND AND AIMS: The inflammatory process is a response mechanism to any stressor agent. Emerging novel therapeutic options derived mainly from natural products such as bromelain have been used to reduce the significant side effects of available anti-inflammatory drugs. Bromelain is an enzyme complex derived from Ananas comosus, known for its anti-inflammatory potential and good tolerance. Therefore, the aim was to assess whether bromelain supplementation exerts anti-inflammatory effects in adults. METHODS: The systematic review was registered in PROSPERO (n° CRD42020221395), and the search was performed in MEDLINE, Scopus, Web of Science, and Cochrane Library. The terms used in the search were: "bromelains", "bromelain", "randomized clinical trial", and "clinical trial". Eligibility criteria were: randomized clinical trials with participants aged 18 years or over, of both sexes, who received supplementation with bromelain alone or in combination with other oral compounds, with an evaluation of inflammatory parameters as primary and secondary outcomes, published in English, Portuguese or Spanish. RESULTS: 1375 studies were retrieved, of which 269 were duplicates. Seven (7) randomized controlled trials were eligible for the systematic review. In most studies, supplementation with bromelain, isolated or in combined therapy, reduced inflammatory parameters. Regarding the reduction of inflammatory parameters among studies with associated bromelain, two presented reduction of inflammatory parameters, while in the evaluation of bromelain treated alone, two studies also showed reduction. In relation to doses supplemented, the studies with associated bromelain ranged from 99.9 to 1200 mg/day and the supplementation time ranged from 3 to 16 weeks. Moreover, the inflammatory parameters evaluated were: IL-12, PGE-2, COX-2, IL-6, IL-8, TNF-α, IL-1ß, IL-10, CRP, NFγ B1, PPAR-α, TNF, TRAF, MCP-1 and adiponectin. In studies with isolated bromelain supplementation, it ranged from 200 to 1050 mg/day for 1 week to 16 weeks. Markers associated with inflammation varied between studies, including IL-2, IL-5, IL-6, IL-8, IL-10, IL-13, IFNγ and MCP-1, PGE-2, CRP and fibrinogen. Eleven (11) participants experienced side effects, and two discontinued treatment in the studies. The reported adverse effects were mainly gastrointestinal but well tolerated. CONCLUSION: The general effect of bromelain supplementation on inflammation is inconsistent because of population heterogeneity, doses used, treatment duration, and parameters evaluated. The observed effects are punctual and isolated, and further standardization is needed to establish doses, supplementation time, and which type of inflammatory condition is indicated.

Raw walnut kernel: A natural source for dietary proteases and bioactive proteins.

Walnut kernels are health-promoting nuts, which are mainly attributed to polyunsaturated fatty acids, phenolics, and phytosterols. However, the information concerning benefits of walnut proteins are limited. In this study, endopeptidases, aminopeptidases, carboxypeptidases, superoxide dismutases, catalases, and phospholipases with respective relative abundance of 2.730, 1.728, 0.477, 3.148, 0.743, and 0.173‰ were identified by liquid chromatography tandem mass spectrometry. These endogenous proteases exhibited activity in a broad pH range of 2-6.5, and optimal at pH 4.5 and 50 °C. Aspartic endopeptidases were predominant endopeptidases, followed by cysteine ones. There were two types of aspartic endopeptidases, one (not inhibited by pepstatin A) exerted activity at pH 2-3 and the other (inhibited by pepstatin A) optimal at pH 4.5. Carboxypeptidases were optimal at pH 4.5, and aminopeptidases exerted activity at pH near 6.5. These endogenous proteases assisted the digestion of walnut proteins, and soaking, especially peeling, greatly improved the in vitro digestibility.

Localized probiotic-guided pocket recolonization in the treatment of chronic periodontitis: a randomized controlled clinical trial.

PURPOSE: This randomized clinical placebo-controlled trial was conducted to evaluate the effectiveness of Lactobacillus reuteri as a probiotic in guided pocket recolonization (GPR) for the treatment of chronic periodontitis (CP) adjunctive to scaling and root planing (SRP). METHODS: Forty-eight CP patients were randomly assigned to 3 treatment groups: group 1 (SRP+placebo), group 2 (SRP+single application of probiotic), and group 3 (SRP+incremental application of probiotic). Clinical parameters were evaluated at baseline and at 8, 12, and 24 weeks, whereas biochemical parameters were measured at baseline and 12 weeks. RESULTS: At 24 weeks, the probing pocket depth and clinical attachment level improved in all 3 groups from baseline with no significant intergroup differences; however, a statistically significant difference was observed in localized plaque and gingival scores between groups 1 and 3 (P<0.05). At 12 weeks, matrix metalloproteinase-8 (MMP-8), nitric oxide (NO), and gingipains-R (Rgps) levels improved in all 3 groups, with statistically significant differences between groups 1 and 3 for MMP-8 and NO (P<0.05), but no difference for Rgps levels. CONCLUSIONS: Within its limitations, the results of this study show that incremental 3-time application of L. reuteri as a probiotic led to improvements in clinical and biochemical parameters. This protocol can be a useful adjunct to SRP in the non-surgical management of CP. TRIAL REGISTRATION: Clinical Trials Registry - India Identifier: CTRI/2017/03/008231.

Role of Mechanotransduction in Periodontal Homeostasis and Disease.

Novel findings broaden the concept of mechanotransduction (MT) in biophysically stimulated tissues such as the periodontium by considering nuclear MT, convergence of intracellular MT pathways, and mechanoresponsive cotranscription factors such as Yes-associated protein 1 (YAP1). Regarding periodontal disease, recent studies have elucidated the role of bacterial gingipain proteases in disturbing the barrier function of cadherins, thereby promoting periodontal inflammation. This leads to dysregulation of extracellular matrix homeostasis via proteases and changes the cell's biophysical environment, which leads to alterations in MT-induced cell behavior and loss of periodontal integrity. Newest experimental evidence from periodontal ligament cells suggests that the Hippo signaling protein YAP1, in addition to integrin-FAK (focal adhesion kinase) mechanosignaling, also regulates cell stemness. By addressing mechanosignaling-dependent transcription factors, YAP1 is involved in osteogenic and myofibroblast differentiation and influences core steps of autophagy. Recent in vivo evidence elucidates the decisive role of YAP1 in epithelial homeostasis and underlines its impact on oral pathologies, such as periodontitis-linked oral squamous cell carcinogenesis. Here, new insights reveal that YAP1 contributes to carcinogenesis via overexpression rather than mutation; promotes processes such as apoptosis resistance, epithelial-mesenchymal transition, or metastasis; and correlates with poor prognosis in oral squamous cell carcinoma. Furthermore, YAP1 has been shown to contribute to periodontitis-induced bone loss. Mechanistically, molecules identified to regulate YAP1-related periodontal homeostasis and disease include cellular key players such as MAPK (mitogen-activated protein kinase), JNK (c-Jun N-terminal kinase), Rho (Ras homologue) and ROCK (Rho kinase), Bcl-2 (B-cell lymphoma 2), AP-1 (activator protein 1), and c-myc (cellular myelocytomatosis). These findings qualify YAP1 as a master regulator of mechanobiology and cell behavior in human periodontal tissues. This review summarizes the most recent developments in MT-related periodontal research, thereby offering insights into outstanding research questions and potential applications of molecular or biophysical strategies aiming at periodontal disease mitigation or prevention.

Alzheimer's Disease-Like Neurodegeneration in Porphyromonas gingivalis Infected Neurons with Persistent Expression of Active Gingipains.

BACKGROUND: Porphyromonas gingivalis (P. gingivalis) and its gingipain virulence factors have been identified as pathogenic effectors in Alzheimer's disease (AD). In a recent study we demonstrated the presence of gingipains in over 90% of postmortem AD brains, with gingipains localizing to the cytoplasm of neurons. However, infection of neurons by P. gingivalis has not been previously reported. OBJECTIVE: To demonstrate intraneuronal P. gingivalis and gingipain expression in vitro after infecting neurons derived from human inducible pluripotent stem cells (iPSC) with P. gingivalis for 24, 48, and 72 h. METHODS: Infection was characterized by transmission electron microscopy, confocal microscopy, and bacterial colony forming unit assays. Gingipain expression was monitored by immunofluorescence and RT-qPCR, and protease activity monitored with activity-based probes. Neurodegenerative endpoints were assessed by immunofluorescence, western blot, and ELISA. RESULTS: Neurons survived the initial infection and showed time dependent, infection induced cell death. P. gingivalis was found free in the cytoplasm or in lysosomes. Infected neurons displayed an accumulation of autophagic vacuoles and multivesicular bodies. Tau protein was strongly degraded, and phosphorylation increased at T231. Over time, the density of presynaptic boutons was decreased. CONCLUSION: P. gingivalis can invade and persist in mature neurons. Infected neurons display signs of AD-like neuropathology including the accumulation of autophagic vacuoles and multivesicular bodies, cytoskeleton disruption, an increase in phospho-tau/tau ratio, and synapse loss. Infection of iPSC-derived mature neurons by P. gingivalis provides a novel model system to study the cellular mechanisms leading to AD and to investigate the potential of new therapeutic approaches.

Intracellular Porphyromonas gingivalis Promotes the Proliferation of Colorectal Cancer Cells via the MAPK/ERK Signaling Pathway.

Porphyromonas gingivalis (P. gingivalis) is a keystone pathogen in periodontitis. However, several clinical studies have revealed an enrichment of P. gingivalis in the stool samples and colorectal mucosa of colorectal cancer patients. Thus, the goal of this study was to determine whether P. gingivalis can promote colorectal cancer progression in vitro. We established an acute infection model (24 h, multiplicity of infection =100) of P. gingivalis invasion of colorectal cancer cells to study the alterations induced by P. gingivalis in the proliferation and cell cycle of colorectal cancer cells. We observed that P. gingivalis can adhere and invade host cells a few hours after infection. Once invaded, P. gingivalis significantly promoted colorectal cancer cell proliferation, and the percentage of S phase cells was increased in the cell cycle assay. However, KDP136, a gingipain-deficient mutant of P. gingivalis 33277, showed a decreased ability to promote colorectal cancer cell proliferation, indicating that gingipain is associated with colorectal cancer cell proliferation. Furthermore, we extracted RNA from colorectal cancer cells for high-throughput sequencing analysis and reconfirmed the results by quantitative polymerase chain reaction and western blot analyses. The results suggested that the MAPK/ERK signaling pathway is significantly activated by P. gingivalis, while these changes were not observed for KDP136. In conclusion, P. gingivalis can invade cells and promote the proliferation of colorectal cancer cells by activating the MAPK/ERK signaling pathway. Gingipain is an essential virulence factor in this interaction.

Filling the Gaps to Solve the Extensin Puzzle.

Extensins (EXTs) are highly repetitive plant O-glycoproteins that require several post-translational modifications (PTMs) to become functional in plant cell walls. First, they are hydroxylated on contiguous proline residues; then they are O-glycosylated on hydroxyproline and serine. After secretion into the apoplast, O-glycosylated EXTs form a tridimensional network organized by inter- and intra-Tyr linkages. Recent studies have made significant progress in the identification of the enzymatic machinery required to process EXTs, which includes prolyl 4-hydroxylases, glycosyltransferases, papain-type cysteine endopeptidases, and peroxidases. EXTs are abundant in plant tissues and are particularly important in rapidly expanding root hairs and pollen tubes, which grow in a polar manner. Small changes in EXT PTMs affect fast-growing cells, although the molecular mechanisms underlying this regulation are unknown. In this review, we highlight recent advances in our understanding of EXT modifications throughout the secretory pathway, EXT assembly in cell walls, and possible sensing mechanisms involving the Catharanthus roseus cell surface sensor receptor-like kinases located at the interface between the apoplast and the cytoplasmic side of the plasma membrane.

The role of UCH-L3 and LC3Ⅱin the diagnosis of acute myeloid leukemia / 中华检验医学杂志

Objective Todetect the mechanism of acute myeloid leukemia ( AML ) and the relationship between autophagy andubiquitin c-terminal hydrolases L3( UCH-L3) for providing new targets and guiding clinical management in AML .Methods 84 cases of AML patients and 25 controls were chosen from the First Affiliated Hospital of Dalian Medical University from 2014 to 2015,including 47 males and 37 females.The AML patients were divided into 3 groups :initialdiagnosis group [40 cases including 24 males and 16 females,with an median age of 54 (23 -85)];complete remission group [30 cases including 14 males and 16 females,with an median age of 45 (22-74)]and refractory group[14 cases including 9 males and 5 females,with an median age of 50 (18-80)].Among those patients, there were 3 cases of M1, 42 cases of M2, 18 cases of M3, 3 cases of M4 and 18 cases of M5 subtypes.The expression of UCH-L3 and LC3II were detected by Real Time PCR and Western blot after the HL-60 cells were treated by TCID.The expression of UCH-L3 and LC3IIin the PBMC of AML patients and controls were detected with Real Time PCR.The expression levels of UCH-L3 and LC3II in different types of AML were analyzed .The relationship between the expression of LC 3Ⅱand clinical features , clinical stages , laboratory results and therapeutic effects of patients were investigated .It further detected the expression of UCH-L3 and LC3IImRNA in post-induction status.t test were used for measurement data , χ2 test were performed for rate comparison; rank correlation test were performed for correlation analysis .Non-parametric test were performed for non-normal distribution data.Results Both the UCH-L3 and LC3 II were down-regulated after TCID treatment in HL-60 cellsat gene and protein levels (t=-29.435, t=-8.105,P<0.05).The expression of UCH-L3 in initial diagnosis group was lower than control group ( Z=-3.87,P<0.05),butit was higher in remission group than initial diagnosis group and refractory group ( Z=-6.70 , Z=-4.09, P<0.05 ) .The expression level of LC3Ⅱin diagnosis group was significantly higher than control group , remission group and refractory group(Z=-6.96,Z=-5.32, Z=-3.52,P<0.05).Cytogenetic abnormalities in patients with high expression of LC3II were more common(χ2 =6.510,P<0.05).The relative expression of UCH-L3 and LC3ⅡmRNA in the same patient before and after treatment were significantly different ( P<0.05 ) . During the remission, the expression of UCH-L3 was up-regulated compared with that before primary treatment, while the expression of LC3II was down-regulated.There was no statistically significant in the relative expression of UCH-L3 and LC3Ⅱamong the FAB type.Conclusion The UCH-L3 and autophagy are associated with pathogenesis of AML .

Study on expression of ASC and Caspase-1 in peripheral blood of patients with primary biliary cirrhosis / 重庆医学

Objective To explore the influence of apoptosis-associated speck-like protein containing a caspase recruitment do-main(ASC)and Caspase-1 on the pathogenesis of primary biliary cirrhosis(PBC).Methods The real-time PCR,Western blot,py-rophosphate sequencing and ELISA were adopted to respectively detect the relative expressions of mRNA and protein of peripheral blood mononuclear cells(PBMS)Caspase-1 and ASC as well as the methylation status of ASC promoter region and plasma Caspase-1 and IL-18 expression levels in 30 cases of PBC and healthy controls.Results The mRNA and protein expressions of PBMC Caspase-1 and ASC in the PBC group were significantly higher than those in the control group(P<0.05).The methylation rate of ASC promoter Island1(ISI)was significantly lower than that of the control group(P<0.05),which of Island 2(IS2)was smaller than the background value and had no methylation occurrence.The levels of plasma Caspase-1 and IL-18 in the PBC group were sig-nificantly higher than those in the control group(P<0.05).The ASC mRNA in the PBC group was significantly correlated with the Caspase-1 mRNA expression(P<0.05);the methylation rates at loci 1,2,4,5 of ASC promoter region CpG island were nega-tively correlated with ASC mRNA expression(P< 0.05),and which at loci 3,6 had no correlation with their expressions(P>0.05);plasma Caspase-1 and IL-18 levels showed the obviously positive correlation.Conclusion ASC and Caspase-1 are involved in the immune inflammatory response in PBC.

Expression analysis of KDEL-CysEPs programmed cell death markers during reproduction in Arabidopsis.

KEY MESSAGE: CEP cell death markers. Programmed cell death (PCD) is essential for proper plant growth and development. Plant-specific papain-type KDEL-tailed cysteine endopeptidases (KDEL-CysEPs or CEPs) have been shown to be involved in PCD during vegetative development as executors for the last step in the process. The Arabidopsis genome encodes three KDEL-CysEPs: AtCEP1, AtCEP2 and AtCEP3. With the help of fluorescent fusion reporter lines, we report here a detailed expression analysis of KDEL-CysEP (pro)proteins during reproductive processes, including flower organ and germline development, fertilization and seed development. AtCEP1 is highly expressed in different reproductive tissues including nucellus cells of mature ovule and the connecting edge of anther and filament. After fertilization, AtCEP1 marks integument cell layers of the seeds coat as well as suspensor and columella cells of the developing embryo. Promoter activity of AtCEP2 is detected in the style of immature and mature pistils, in other floral organs including anther, sepal and petal. AtCEP2 mainly localizes to parenchyma cells next to xylem vessels. Although there is no experimental evidence to demonstrate that KDEL-CysEPs are involved in PCD during fertilization, the expression pattern of AtCEPs, which were previously shown to represent cell death markers during vegetative development, opens up new avenues to investigate PCD in plant reproduction.

Effect of mild hypothermia on expression of small ubiquitin-like modifier-specific proteases 3 in brain tissues during cerebral ischemia-reperfusion in mice / 中华麻醉学杂志

Objective To investigate the effect of mild hypothermia on the expression of small ubiquitin-like modifier-specific proteases 3 (SENP3) in the brain tissues during cerebral ischemia-reperfusion (I/R) in mice.Methods Ninety-six male C57/BL6 mice,aged 10-12 weeks,weighing 22-30 g,were randomly divided into 3 groups (n =32 each) using a random number table:sham operation group (group S),group I/R,and mild hypothermia group (group H).Cerebral I/R was produced by occlusion of bilateral common carotid arteries for 15 min followed by reperfusion.The surface cooling was started immediately after reperfusion,and the rectal temperature was maintained at 32-34 ℃ for 3 h.At 6,12,24 and 72 h of reperfusion,8 mice were selected from each group and sacrificed.The hippocampi were removed for examination of the pathological changes (with light microscope) and for determination of cell apoptosis (using TUNEL) and expression of SENP3 (by Western blot).The apoptosis rate was calculated.Results Compared with group S,the apoptosis rate in hippocampal CA1 region was significantly increased,and the expression of SENP3 was significantly up-regulated at each time point of reperfusion in group I/R (P＜0.05).Compared with group I/R,the apoptosis rate in hippocampal CA1 region was significantly decreased,and the expression of SENP3 was significantly down-regulated at each time point of reperfusion (P＜0.05),and the pathological changes were significantly reduced in group H.Conclusion The mechanism by which mild hypothermia reduces cerebral I/R injury is associated with inhibition of SENP3 expression in the brain tissues of mice.

Liensinine promotes apoptosis of bladder cancer 5637 cells and enhances Caspase-7 expression and activation / 天津医药

Objective To study the effects of Liensinine on apoptosis of 5637 cells, and its mechanism thereof. Meth?ods CCK-8 method and the colony formation test were used to detect cell viabilities, and then inhibition rates were calcu?lated. Flow cytometry was used to detect the effects of Liensinine on apoptosis of 5637 cells. Western blot assay was used to detect Caspase-7 protein expression. Results CCK-8 assay and colony formation test indicated that Liensinine inhibited the cell proliferation significantly. Results of flow cytometry indicated that Liensinine induced early apoptosis of 5637 cells. Western blot assay showed that Liensinine improved the expression of Caspase-7 and enhanced the activation of Caspase-7 in 5637 cells. Conclusion Liensinine could inhibit the proliferation of 5637 cells, induce early apoptosis, which may be re?lated with the enhanced expression of Caspase-7 and its activation.

Effects and mechanisms of trophoblast cells autophagy in preeclampsia / 中华妇产科杂志

Objective To investigate the occurrence law of autophagy in trophoblast cells from preeclampsia and its underlying mechanisms.Methods Twenty cases of placenta tissues were collected from women suffered from preeclampsia and normal pregnant women respectively.Autophagosome of trophoblast cells were observed by transmission electron microscope.The expressions of LC3-Ⅱ/Ⅰ and Atg4B in placenta tissues were detected by western blot and real-time PCR.Results Compared with the control group,typical autophagosomes of trophoblast cells were observed by transmission electron microscope.The ratio of LC3-Ⅱ / Ⅰ in placenta of PE patients was increased (1.43 ± 0.23) compared with control group (0.59 ±0.12),and the expression of Atg4B was up-regulated in both mRNA [(1.73 ±0.16) folds] and protein levels (0.71 ± 0.13) compared with control group (P ＜ 0.05).Conclusions Autophagy was significantly up-regulated in trophoblast cells from patients suffered from preeclampsia.Thus,all the data suggest that autophagy might be involved in the generation of preeclampsia.

Expression and significance of LMP2 and PPM1A in gestational trophoblastic disease / 中华妇产科杂志

Objective To investigate the expression of low molecular mass polypeptide-2 (LMP2)and protein phosphatase 1A (PPM1A) in gestational trophoblastic disease and elucidate their predictive value in malignant transformation of hydatidiform mole. Methods The expressions of LMP2 and PPM1A protein in 196 complete hydatidiform moles (in which 28 cases with malignant transformation) , 7 invasive moles, 5 choriocarcinomas and 20 normal chorionic villus were detected with the method of En Vision immunohistochemistry. Their clinicopathologic data were retrospectively analyzed. Results LMP2 and PPM1A protein expressed in cytotrophocytes, syncytiotrophoblast and extravillous trophoblast. The level of LMP2 expression in deteriorative hydatidiform mole was significantly higher than that in non-deteriorative hydatidiform mole or normal chorionic villus (6. 79 ±2. 38, 5.26 ±2.63 and 3. 10 ±1.65, all P 0. 05). Conclusions High expression of LMP2 and low expression of PPM1A might play an important role in the motility and invasiveness of trophohlast cells and malignant transformation of hydatidiform mole. Testing the expression of LMP2 and PPM1A in hydatidiform mole tissues of initial uterine evacuation might be have some reference significance in judging outcomes of hydatidiform mole.

The effect of pingyangmycin on vascular endothelial cells cultured in vitro / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the inhibiting effect of Pingyangmycin on cultured vein endothelial cells in vitro and illustrate the mechanism of Pingyangmycin on treatment cavernous hemangioma of head and neck. METHODS By MTT assay,the inhibiting rates of cultured Ecv-304 cells which were managed with Pingyangmycin for 24h and 48h were compared respectively. The changes in the cell cycle,apoptosis,and Caspase 3 protein expression of the cells managed with Pingyangcin for 24h were examined by flow cytometry(FCM). RESULTS The results showed that the inhibiting rate of Ecv-304 was dependent on the concentration of Pingyangmycin. However,in the same concentration,the inhibiting effect for 24h was stronger than that for 48h. The percentage of G1 phase cells increased while the S phase decreased,but the percentage of G2 phase cells remained unchanged after managed with Pingyangcin for 24h. An apoptosis peak appeared before G1 phase. The apoptosis rate was also concentration-dependant. The expression of caspase 3 increased in every group of different concentration. CONCLUSION Pingyangmycin can possibly arrest the growth of Ecv-304 in the G1 phase through the mechanism of inducing apoptosis. Caspase 3 may play an important role in the induced apoptosis.