LncRNA AFAP1-AS1 Promotes Oral Squamous Cell Carcinoma Development by Ubiquitin-Mediated Proteolysis.

BACKGROUND AND PURPOSE: Long noncoding RNA (lncRNA) dysregulation has been reported to play a pivotal role in the development of cancers. In this study, we aimed to screen the key lncRNA in oral squamous cell carcinoma (OSCC) via bioinformatics analysis and further validate the function of lncRNA in vitro and in vivo. METHODS: Bioinformatics analysis was conducted to identify differentially expressed lncRNAs between control and OSCC samples. Quantitative real-time-polymerase chain reaction was employed to detect the expression of differentially expressed lncRNAs in human tongue squamous cell carcinoma and human oral keratinocytes cell lines. The biological function of lncRNA and its mechanism were examined via the experimental assessment of the cell lines with the lncRNA overexpressed and silenced. Additionally, to further explore the function of lncRNA in the progression of OSCC, xenograft tumour mouse models were established using 25 mice (5 groups, each with 5 mice). Tumour formation was observed at 2 weeks after the cell injection, and the tumours were resected at 5 weeks post-implantation. RESULTS: Two lncRNAs, LINC00958 and AFAP1-AS1, were found to be correlated with the prognosis of OSCC. The results of the quantitative real-time-polymerase chain reaction indicated that the 2 lncRNAs were highly expressed in OSCC. In combination with the previous literature, we found AFAP1-AS1 to be a potentially important biomarker for OSCC. Thus, we further investigated its biological function and found that AFAP1-AS1 silencing inhibited cell proliferation, migration, and invasion whereas AFAP1-AS1 overexpression reversed the effect of AFAP1-AS1 silencing (P < .05). Mechanism analysis revealed that AFAP1-AS1 regulated the development of OSCC through the ubiquitin-mediated proteolysis pathway. CONCLUSIONS: AFAP1-AS1 is an oncogene that aggravates the development of OSCC via the ubiquitin-mediated proteolysis pathway. It also provides a novel potential therapy for OSCC.

Transanal total mesorectal excision combined with intersphincteric resection has similar long-term oncological outcomes to laparoscopic abdominoperineal resection in low rectal cancer: a propensity score-matched cohort study.

Background: Transanal total mesorectal excision (taTME) or intersphincteric resection (ISR) has recently proven to be a valid and safe surgical procedure for low rectal cancer. However, studies focusing on the combination of these two technologies are limited. This study aimed to evaluate perioperative results, long-term oncologic outcomes, and anorectal functions of patients with low rectal cancer undergoing taTME combined with ISR, by comparing with those of patients undergoing laparoscopic abdominoperineal resection (laAPR). Methods: After 1:1 propensity score matching, 200 patients with low rectal cancer who underwent laAPR (n = 100) or taTME combined with ISR (n = 100) between September 2013 and November 2019 were included. Patient demographics, clinicopathological characteristics, oncological outcomes, and anal functional results were analysed. Results: Patients in the taTME-combined-with-ISR group had less intraoperative blood loss (79.6 ± 72.6 vs 107.3 ± 65.1 mL, P = 0.005) and a lower rate of post-operative complications (22.0% vs 44.0%, P < 0.001) than those in the laAPR group. The overall local recurrence rates were 7.0% in both groups within 3 years after surgery. The 3-year disease-free survival rates were 86.3% in the taTME-combined-with-ISR group and 75.1% in the laAPR group (P = 0.056), while the 3-year overall survival rates were 96.7% and 94.2%, respectively (P = 0.319). There were 39 patients (45.3%) in the taTME-combined-with-ISR group who developed major low anterior resection syndrome, whereas 61 patients (70.9%) had good post-operative anal function (Wexner incontinence score ≤ 10). Conclusion: We found similar long-term oncological outcomes for patients with low rectal cancer undergoing laAPR and those undergoing taTME combined with ISR. Patients receiving taTME combined with ISR had acceptable post-operative anorectal function.

Effect of COVID-19 outbreak on the treatment time of patients with acute ST-segment elevation myocardial infarction.

OBJECTIVE: To explore the effect of COVID-19 outbreak on the treatment time of patients with ST-segment elevation myocardial infarction (STEMI) in Hangzhou, China. METHODS: We retrospectively reviewed the data of STEMI patients admitted to the Hangzhou Chest Pain Center (CPC) during a COVID-19 epidemic period in 2020 (24 cases) and the same period in 2019 (29 cases). General characteristics of the patients were recorded, analyzed, and compared. Moreover, we compared the groups for the time from symptom onset to the first medical contact (SO-to-FMC), time from first medical contact to balloon expansion (FMC-to-B), time from hospital door entry to first balloon expansion (D-to-B), and catheter room activation time. The groups were also compared for postoperative cardiac color Doppler ultrasonographic left ventricular ejection fraction (LVEF),the incidence of major adverse cardiovascular and cerebrovascular events (MACCE),Kaplan-Meier survival curves during the 28 days after the operation. RESULTS: The times of SO-to-FMC, D-to-B, and catheter room activation in the 2020 group were significantly longer than those in the 2019 group (P < 0.05). The cumulative mortality after the surgery in the 2020 group was significantly higher than the 2019 group (P < 0.05). CONCLUSION: The pre-hospital and in-hospital treatment times of STEMI patients during the COVID-19 epidemic were longer than those before the epidemic. Cumulative mortality was showed in Kaplan-Meier survival curves after the surgery in the 2020 group was significantly different higher than the 2019 group during the 28 days.The diagnosis and treatment process of STEMI patients during an epidemic should be optimized to improve their prognosis.

Overview on new progress of hereditary diffuse gastric cancer with CDH1 variants.

Hereditary diffuse gastric cancer (HDGC), comprising 1%-3% of gastric malignances, has been associated with CDH1 variants. Accumulating evidence has demonstrated more than 100 germline CDH1 variant types. E-cadherin encoded by the CDH1 gene serves as a tumor suppressor protein. CDH1 promoter hypermethylation and other molecular mechanisms resulting in E-cadherin dysfunction are involved in the tumorigenesis of HDGC. Histopathology exhibits characteristic signet ring cells, and immunohistochemical staining may show negativity for E-cadherin and other signaling proteins. Early HDGC is difficult to detect by endoscopy due to the development of lesions beneath the mucosa. Prophylactic gastrectomy is the most recommended treatment for pathogenic CDH1 variant carriers. Recent studies have promoted the progression of promising molecular-targeted therapies and management strategies. This review summarizes recent advances in CDH1 variant types, tumorigenesis mechanisms, diagnosis, and therapy, as well as clinical implications for future gene therapies.

Extraovarian Brenner tumor in the uterus: a case report and review of literature.

BACKGROUND: Extraovarian Brenner tumors (EOBTs) are extremely rare and can be observed incidentally in both female and male patients, raising concerns regarding the origin of Brenner tumors. CASE PRESENTATION: A 53-year-old postmenopausal woman presented with a nodular lesion in the left side of the corpus uteri, which was found at a routine health check. Macroscopically, the lesion appeared as a solid nodule with a yellowish-gray cut surface, approximately 6 cm in greatest diameter. Microscopically, the lesion consisted of well-defined epithelial nests and spindled stromal cells. Parenchymal cells expressed CK7, GATA3, CK5/6, 34ßE12, and p63. A single layer of cavity-lined cells with umbrella-like shape showed apical Uroplakin III positivity. Stromal cells were positive for SMA, ER, and PR. The final diagnosis was EOBT and the patient was followed for 2 months with no recurrence. CONCLUSIONS: We report here the third case of EOBTs in the uterus. The combination of morphologic and immunohistochemical results supported the involvement of urothelial metaplasia in the development of EOBTs. The similarities between EOBTs and Walthard nests made Müllerian epithelium an attractive candidate as the cellular origin. Changes of tissue structure or sex hormones imbalance may lead to the translocation of Müllerian remnants to distant organs, explaining the pathogenesis of EOBTs.

Epithelioid sarcoma of the scalp: a case report and literature review.

Epithelioid sarcoma (ES) is an aggressive mesenchymal tumor exhibiting bi-directional phenotypes. According to its proclivity for different sites, ES can be subdivided into two categories: distal and proximal variants. Proximal ES often affects the truncal tissue, thighs, head, and neck. Accumulating evidence indicates that several locations in the head, including the orbital, gingival, and nasal cavities, are involved in ES. However, the underlying mechanisms of ES carcinogenesis and progression are largely unknown, including and especially the reason why the tumor cells are positive for both epithelial and mesenchymal classical markers. Thus, we wish to share a rare case of ES in the scalp and its clinical and molecular features. Only 9 cases to date have been reported. An 80-years-old man had sustained a painful swollen mass in his scalp for three months. A diagnosis of epithelioid sarcoma was established based on the combination of the histopathological and immunohistochemical findings. The tumor cells were positive for both mesenchymal (vimentin and S100) and epithelial markers (pan-cytokeratin). This case suggests that ES can be derived from the soft tissue of the scalp. The tumor cells co-expressed biomarkers of epithelial and mesenchymal cells, suggesting the mesenchymal-epithelial transition (MET) may be involved.

Comprehensive assessment of the association between genes on JAK-STAT pathway (IFIH1, TYK2, IL-10) and systemic lupus erythematosus: a meta-analysis.

Previous studies have reported that genes relating to JAK-STAT pathway (IFIH1, TYK2 and IL-10) conferred the susceptibility to SLE. In this study, we performed a meta-analysis (including 43 studies) to evaluate the association between IFIH1 (9288 patients and 24,040 controls), TYK2 (4928 patients and 11,536 controls), IL-10 (3623 patients and 4907 controls) polymorphisms and systemic lupus erythematosus (SLE) in a comprehensive way. We found that IFIH1 rs1990760_T allele was associated with risk of SLE in overall population under three models (allelic: P = 2.56 × 10-11, OR 1.135, 95% CI 1.094-1.179, dominant: P = 1.8 × 10-8, OR 1.203, 95% CI 1.128-1.284, recessive: P = 2.6 × 10-7, OR 1.163, 95% CI 1.098-1.231). A strong association had been observed between TYK2 polymorphism rs2304256_C allele and SLE in Europeans (P = 5.82 × 10-5, OR 1.434, 95% CI 1.203-1.710). When coming to overall population, TYK2 rs2304256_C showed a significant association with SLE under recessive model (P = 8.05 × 10-3, OR 1.314, 95% CI 1.074-1.608). However, the other two SNPs (rs12720270, rs280519) of TYK2 were not significant. The results also indicated an association between IL-10 rs1800896_G allele and SLE in Asians under recessive model (P = 4.65 × 10-3, OR 2.623, 95% CI 1.346-5.115), while, IL-10 rs1800896_G had a trend of association with SLE in European population in dominant model (P = 1.21 × 10-2, OR 1.375, 95% CI 1.072-1.764). In addition, we found IL-10 rs1800896 GG homozygote might be associated with increased susceptibility to SLE (GG vs AA, P = 4.65 × 10-3, OR 1.539, 95% CI 1.142-2.072). We concluded that IFIH1 rs1990760_T and TYK2 rs2304256_C alleles were significantly associated with SLE, and IL-10 rs1800896 GG homozygote might have an enhancement effect on SLE risk.

Transanal total mesorectal excision as a surgical procedure for diffuse cavernous hemangioma of the rectum: A case report.

BACKGROUND: The safety and feasibility of transanal total mesorectal excision (TaTME) were demonstrated in the management of rectal cancer. However, its role in the management of patients with diffuse cavernous hemangioma of the rectum (DCHR) has not been evaluated. METHODS: A female patient with DCHR was admitted to our hospital. Colonoscopy, magnetic resonance imaging (MRI), abdominal computed tomography (CT) and arteriography were performed. Lesions were detected in mesorectum and rectal wall extending from the dentate line to 5cm proximally. TaTME with a protecting loop ileostomy were performed. The research work has been reported in line with the SCARE criteria Agha et al., 2016 [1]. RESULTS: TaTME and a protecting loop ileostomy were safely performed, with an intact mesorectal specimen being harvested. The entire procedure took 348min. The estimated blood loss was 100ml. The patient recovered uneventfully. Her symptom of painless rectal bleeding was resolved satisfactorily following the surgery. The histopathological evaluation confirmed the diagnosis of DCHR. CONCLUSIONS: TaTME appears to be a safe and feasible procedure for patients with DCHR in experienced hands.

Pneumomediastinum and pneumoperitoneum on computed tomography after peroral endoscopic myotomy (POEM): postoperative changes or complications?

BACKGROUND: Many of the acute alterations after peroral endoscopic myotomy (POEM) may be of little clinical significance, while others may herald major clinical problems. The question whether pneumomediastinum/pneumoperitoneum is a normal postoperative finding after POEM, or should be regarded as a sign of a complication needs to be evaluated. Familiarity with these findings in computed tomography (CT) is essential for radiologists. PURPOSE: To evaluate whether or not pneumomediastinum/pneumoperitoneum detected by chest CT is a sign of a complication after POEM using CO2 insufflation for esophageal achalasia. MATERIAL AND METHODS: One hundred and eight patients with esophageal achalasia who underwent chest CT within 30 hours after POEM were included. CT findings were retrospectively reviewed by two radiologists in consensus. The correlation between pneumomediastinum and/or pneumoperitoneum shown on CT and the development of complications was analyzed. RESULTS: Abnormal findings were identified on post-treatment CT, including pneumomediastinum and/or pneumoperitoneum (53.7%, 58/108), pneumothorax (0.9%, 1/108), subcutaneous emphysema (29.6%, 32/108), pleural effusion (69.4%, 75/108), segmental atelectasis of lung tissue (29.6%, 32/108), minor inflammation of lungs (69.4%, 75/108), and ascites (0.9%, 1/108). Pneumomediastinum and pneumoperitoneum were observed simultaneously in 29 cases. The incidence rate of mild complications was high (79.6%, 86/108), while the rate of severe complications was low (2.8%, 3/108). There was no significant correlation between the occurrence of pneumomediastinum and/or pneumoperitoneum on CT and the development of complications (P = 0.542), or the development of severe complications including delayed hemorrhage, esophageal perforation, and retroperitoneal abscess. CONCLUSION: Pneumomediastinum and pneumoperitoneum detected by CT occur frequently after POEM and may be regarded as normal postoperative changes.

Meta-analysis of the therapeutic effects of antibiotic versus appendicectomy for the treatment of acute appendicitis.

Appendicectomy has been the gold standard treatment of acute appendicitis for more than a century, while nonoperative therapies, including antibiotics, have acquired increased interest in recent years. The present meta-analysis aimed to compare the therapeutic effects of antibiotics versus appendicectomy for the treatment of acute appendicitis. Medline, Embase and The Cochrane Library databases were searched. Prospective randomized controlled trials that compared antibiotic treatment with surgery were included. The outcomes evaluated included the time of hospital stay, complications and time to work. There were no statistically significant differences between the antibiotic and appendicectomy groups with regard to the time of hospital stay and complications. However, the time to work was significantly longer in the appendicectomy group when compared with the antibiotic group. In addition, the therapeutic effects of antibiotics and appendicectomy were comparable for the treatment of acute appendicitis.

[Expression and clinicopathological significance of serine-257/threonine-261 phosphorylated MKK4 in colorectal carcinoma].

OBJECTIVE: As an activator of JNK and p38, phosphorylated MKK4 is considered to be associated with tumor progression and prognosis. This study was to examine the expression of pMKK4 and evaluate its prognostic significance in colorectal carcinoma. METHODS: A total of 343 cases of colorectal cancer were followed up to analyze the associations between the expression of pMKK4 and various clinicopathological factors. The expression of Serine-257/Threonine-261 pMKK4 was detected immunohistochemically by tissue microarray. RESULTS: The staining of pMKK4 was present in cytoplasm of colorectal carcinoma. And the expression of pMKK4 was correlated with invasion depth (P = 0.003), differentiation (P = 0.018), lymph node metastasis (P < 0.001), metastasis (P < 0.001), hepatic metastasis (P = 0.039) and TNM stage (P < 0.001). The patients with strong pMKK4 staining had a better overall survival than those with lowered levels (Log rank = 4.531, P = 0.033). Univariate analysis indicated that the expression of pMKK4 was correlated with either overall survival (HR = 0.785, P = 0.035) or relapse-free survival (HR = 0.788, P = 0.044). In multivariate analysis, there was no prognostic significance of pMKK4 after adjusting for invasion depth, differentiation, lymph node metastasis, metastasis, liver metastasis and TNM stage. CONCLUSIONS: The down-regulation of S257/T261 pMKK4 is associated with more advanced stages and it plays an important role in tumor progression. A high-level expression of pMKK4 indicates favorable clinical outcomes, but it is not an independent predictor.

The effects of perioperative probiotic treatment on serum zonulin concentration and subsequent postoperative infectious complications after colorectal cancer surgery: a double-center and double-blind randomized clinical trial.

BACKGROUND: Zonulin is a newly discovered protein that has an important role in the regulation of intestinal permeability. Our previous study showed that probiotics can decrease the rate of infectious complications in patients undergoing colectomy for colorectal cancer. OBJECTIVE: The objective was to determine the effects of the perioperative administration of probiotics on serum zonulin concentrations and the subsequent effect on postoperative infectious complications in patients undergoing colorectal surgery. DESIGN: A total of 150 patients with colorectal carcinoma were randomly assigned to the control group (n = 75), which received placebo, or the probiotics group (n = 75). Both the probiotics and placebo were given orally for 6 d preoperatively and 10 d postoperatively. Outcomes were measured by assessing bacterial translocation, postoperative intestinal permeability, serum zonulin concentrations, duration of postoperative pyrexia, and cumulative duration of antibiotic therapy. The postoperative infection rate, the positive rate of blood microbial DNA, and the incidence of postoperative infectious complications-including septicemia, central line infection, pneumonia, urinary tract infection, and diarrhea-were also assessed. RESULTS: The infection rate was lower in the probiotics group than in the control group (P < 0.05). Probiotics decreased the serum zonulin concentration (P < 0.001), duration of postoperative pyrexia, duration of antibiotic therapy, and rate of postoperative infectious complications (all P < 0.05). The p38 mitogen-activated protein kinase signaling pathway was inhibited by probiotics. CONCLUSIONS: Perioperative probiotic treatment can reduce the rate of postoperative septicemia and is associated with reduced serum zonulin concentrations in patients undergoing colectomy. We propose a clinical regulatory model that might explain this association. This trial was registered at http://www.chictr.org/en/ as ChiCTR-TRC-00000423.

Impact of image integration on catheter ablation for atrial fibrillation using three-dimensional electroanatomic mapping: a meta-analysis.

BACKGROUND: Studies comparing the procedural and clinical outcomes of catheter ablation for atrial fibrillation (AF) guided by CartoMerge and that by Carto have achieved mixed results (Carto, Biosense Webster, Diamond Bar, CA, USA). We collected these studies and conducted a meta-analysis to determine whether CartoMerge results in better procedural and clinical outcomes. METHODS AND RESULTS: Three randomized controlled trials and two controlled observational studies were collected for analysis. The clinical and procedural outcomes of interest were AF recurrence after catheter ablation, major complications, procedure durations, and fluoroscopy time. Meta-analysis was performed using RevMan 5.0.18 software (The Cochrane Collaboration, Copenhagen, Denmark) and pooled estimates of effect were reported as risk ratios with 95% confidence intervals (CI). The overall results of this meta-analysis indicate that catheter ablation for AF guided by CartoMerge is insignificantly associated with a decreased risk of recurrences (RR = 0.76; 95% CI: 0.55-1.04; P = 0.09) and major complications (RR = 0.73; 95% CI: 0.37-1.45; P = 0.37) compared with that by Carto. CONCLUSION: The image integration using CartoMerge guiding catheter ablation for AF does not improve the main clinical outcomes significantly compared with that by Carto in centers with experienced operators.

[Meta-analysis of randomized controlled trials comparing laparoscopic with open surgery for rectal cancer on oncologic clearance and long-term oncologic outcomes].

OBJECTIVE: To compare oncologic adequacy of resection and long-term oncologic outcomes between laparoscopic-assisted surgery (LS) and open surgery (OS) in the treatment of rectal cancer. METHODS: Literature searches of electronic databases (PubMed, Embase, Web of Science,and Cochrane Library) and manual searches up to June 30, 2010 were performed to identify RCTs comparing values of oncologic adequacy of resection, recurrence and survival following LS and OS. Fixed and random effects models were used. RESULTS: Six RCTs enrolling 1033 participants (LS group:577 cases, OS group:456 cases)were included in the meta-analysis. Number of lymph node harvested was similar(WMD=-0.38, 95%CI:-1.35-0.58, P=0.43). LS had a slightly higher circumference resection margin(CRM) positive rate with no statistical significance[7.94% vs. 5.37%; risk ratio(RR)=1.13; 95%CI:0.69-1.85, P=0.63]. There was no significant difference between the two groups in local recurrence (RR=0.55; 95%CI:0.22-1.40, P=0.21). The 3-year overall survival [Hazard ratio(HR)=0.76; 95%CI:0.54-1.07, P=0.11] and 3-year disease-free survival(HR=1.16; 95%CI:0.61-2.20, P=0.64) were not significantly different between the two groups. CONCLUSION: Compared with open surgery, laparoscopic surgery of rectal carcinoma offers similar oncological clearance and long-term oncological outcomes.

[Laparoscopic versus open appendectomy in patients with chronic appendicitis].

OBJECTIVE: To compare the advantages and disadvantages of laparoscopic versus open appendectomy in patients with chronic appendicitis. METHODS: Two hundred twenty- four patients were divided into laparoscopic group (n=98) and open appendectomy group (n=126) according to individual willing. Prospective non- randomized study was performed to compare the operative time, operative bleeding, hospitalization time, the discovery and management concerned in operation. Abdominal pain in these chronic appendicitis cases was followed up. RESULTS: The operative time was (54.8+/-21.8) min in open group and (51.8+/-18.0) min in laparoscopic group (t=0.80,P > 0.05). The operative bleeding was (18.6+/-23.3) ml in open group and (9.8+/-4.7) ml in laparoscopic group (t=3.13, P < 0.05). The hospitalization time was (8.9+/-5.3) d in open group and (6.8+/-3.0) d in laparoscopic group (t=2.66,P < 0.05). Twenty- five cases had abdominal adhesion in laparoscopic group, including 9 cases of adhesion around appendix, 6 cases of adhesion between ileocecum and anterior or lateral abdominal wall, 4 cases of adhesion between epiploon and abdominal wall or intestines, 6 cases of adhesion around colon and others. All adhesion had been dissected. Fourteen cases adhesion around appendix had been discovered in 126 cases of open group and dissected (chi(2) =7.95,P < 0.05). In follow- up research, 24 cases still had chronic abdominal pain in 98 case of open group, and 9 cases had chronic abdominal pain in 87 of laparoscopic group, the difference was significant (chi(2)=6.29,P < 0.05). CONCLUSION: The laparoscopic appendectomy possesses more advantages in treating chronic appendicitis and can decrease the incidence of chronic abdominal pain after operation.

[C-reactive protein induced human endothelium cells apoptosis is associated with Bcl-2/Bax gene expression changes].

OBJECTIVE: In the present study, we examined the expression changes of Bcl-2/Bax in C-reactive protein (CRP) treated human endothelium cells in vitro. METHODS: The human umbilical vein endothelial cells (HUVEC) were cultured by digest method for 2 - 3 posterities and incubated with human CRP (0, 1, 5, 25 mg/L for 24 hours) and analyzed by flow cytometer for apoptosis ratio. The effects of CRP in various concentrations on Bcl-2/Bax mRNA and protein expression were examined by RT-PCR and Western Blotting. RESULTS: Apoptosis ratio increased, downregulated Bcl-2 (gene promoting cell survival) and upregulated Bax (gene promoting apoptosis) at mRNA and protein levels in proportion to increased CRP concentrations. CONCLUSION: These results demonstrate that Bcl-2/Bax could be regulated by CRP in human HUVECs and might play a causal role in CRP-induced apoptosis.

[Effects of puerarin on proliferation of vascular smooth muscle cells induced by thrombin].

OBJECTIVE: To investigate the effects of puerarin on the proliferation of vascular smooth muscle cells (VSMC) induced by thrombin and the mechanism thereof. METHODS: VSMCs were isolated from the thoracic aorta of a SD rat and cultured, then co-cultured with thrombin of the concentration 0.1, 0.3, 1.0, 3.0, and 10 U/L for 24 h, thrombin of the concentration of 1 U/L for 0, 6, 12, 24, 36, and 48 h respectively, or thrombin of the concentration of 1 U/L combined with puerarin of the concentrations of 1.5 x 10(-5), 1.5 x 10(-4), or 1.5 x 10(-3) mol/L for 24 h. Flow cytometry was used to detect the cell number and cell cycle. Western blotting was used to indicate the protein expression of the oncogenes c-fos and bcl-2 RT-PCR was used to evaluate the thrombin receptor (TR) mRNA expression. RESULTS: he numbers of the groups of VSMCs stimulated by 0.1, 0.3, 1.0, 3.0, and 10 U/L thrombin for 24 hours were 4.82 x 10(4)/ml +/- 0.11 x 10(4)/ml, 6.37 x 10(4)/ml +/- 0.09 x 10(4)/ml, 8.78 x 10(4)/ml +/- 0.08 x 10(4)/ml, 7.37 x 10(4)/ml +/- 0.07 x 10(4)/ml, and 5.28 x 10(4)/ml +/- 0.12 x 10(4)/ml respectively, all significantly higher than that of the control group (4.08 +/- 0.054 x 10(4)/ml, all P < 0.05). The effect of thrombin was in a dose-dependent manner within a concentration range of 0.1 - 1.0 U/L. The suppression rates of VSMC proliferation in the combination groups with puerarin of the concentrations of 1.5 x 10(-5), 1.5 x 10(-4), and 1.5 x 10(-3) mol/L were 10.9% +/- 1.6%, 32.1% +/- 3.3%, and 42.6% +/- 5.2% respectively in comparison with the thrombin group (all P < 0.05). The c-fos protein expression of the VSMCs after thrombin stimulation for 24 h increased by 156.0% +/- 11.3% (P < 0.05), and the bcl-2 protein expression of the VSMCs pretreated with puerarin of the concentrations of 1.5 x 10(-5), 1.5 x 10(-4), and 1.5 x 10(-3) mol/L, and then stimulated by thrombin was significantly lower than that of the VSMCs only stimulated by thrombin with the suppression rates of 20.7% +/- 2.1%, 31.6% +/- 5.2%, and 44.5% +/- 7.5% respectively (all P < 0.05). The bcl-2 protein expression of the VSMCs after thrombin stimulation for 24 h increased by 96.7% +/- 8.3% (P < 0.05), and the bcl-2 protein expression of the VSMCs pretreated with puerarin of the concentrations of 1.5 x 10(-5), 1.5 x 10(-4), and 1.5 x 10(-3) mol/L, and then stimulated by thrombin was significantly lower than that of the VSMCs only stimulated by thrombin with the suppression rates of 7.1% +/- 0.8%, 18.8% +/- 1.2%, and 39.6% +/- 6.4% respectively (all P < 0.05). The stimulation of thrombin increased the TR mRNA expression by 183.9% +/- 9.4%. The puerarin of the concentrations of 1.5 x 10(-5) mol/L and 1.5 x 10(-4) mol/L decreased the increase of TR mRNA expression induced by thrombin, however, without significant differences (both P > 0.05), and puerarin of the concentration of 1.5 x 10(-3) mol/L significantly suppressed the increase of TR mRNA expression induced by thrombin by 17.6% +/- 1.7% (P < 0.05). CONCLUSION: Puerarin suppresses the proliferation and DNA synthesis of VSMC induced by thrombin. The inhibitory effect of puerarin is closely related with the suppression of the protein expression of c-fos and bcl-2n, and partly related with the suppression of the TR mRNA expression.