Objective To evaluate the effect of arsenic trioxide(As_(2)O_(3)) on the transgenic TNF-? promoter activity in cultured vascular smooth muscle cells(VSMCs) and macrophages(M?s).Methods Human TNF-? promoter was constructed by reporter gene system and was transiently transfected into VSMCs and M?s.Promoter activity was tested by luciferase activity with or without LPS and Ang Ⅱ stimulation,before or after different dosage of As_(2)O_(3) treatment.Results The results showed: TNF-? promoter stably expressed in VSMCs and M?s compared with CMV promoter(58.3% and 55.6%,respectively).Both LPS and Ang Ⅱ up-regulated TNF-? promoter activity (P

Objective To study the anti-proliferation,pro-apoptosis and cell cycle blocking effects of arsenic trioxide(As_(2)O_(3)) on rat vascular smooth muscle cells(VSMCs).Methods The effects of As_(2)O_(3) on VSMCs viability,growth and proliferation were assayed by MTT,trypan blue exclusion and()~3H-thymidine incorporation methods.Change of cell cycle and apoptosis of VSMCs induced by As_(2)O_(3) were observed by flow cytometry and DNA laddering methods.Western blot was applied to detect the expression changes of P53 and P21()~(waf1/cip1) proteins.As_(2)O_(3) inhibited the growth and DNA synthesis of VSMCs both time-and dose-dependently,while had no obvious cytotoxic effects on cell viability at the concentrations from 1 to(16 ?mol/L).(8 ?mol/L) of As_(2)O_(3) significantly blocked the cell cycle progression,decreased the S phase and increased G_(0)G_(1) phase partition with sub-G_(1) apoptotic distribution. Results Typical DNA fragmentation of VSMCs induced by(16 ?mol/L) of As_(2)O_(3) was observed at different time points.(8 ?mol/L) of As_(2)O_(3) significantly up-regulated P53and P21()~(wif1/cip1) expression in a time-dependent manner.Conclusion These results suggest that As_(2)O_(3) inhibited the proliferation,accelerated the apoptosis and blocked the cell cycle progression in VSMCs which may relate to P53 and P21~(waf1/cip1) pathway.

Objective To investigate the expression of vascular endothelial growth factor C(VEGF-C ) in tissue, serum and lymph fluid in esophageal cancer patients, and the relationship between its expression and tumor lymphatic metastasis. Methods Expressions of VEGF-C in 76 cases of ESC was detected by immunohisto-chemical SP method and VEGF-C levels in lymph fluid and serum were assessed by enzyme-linked immunosorbent assay. Results Positive immunostaining of VEGF-C in esophageal cancer tissue was 63.1%,The expression of VEGF-C in tissue was significantly associated with lymph node metastasis and TNM stage ( P<0.01, P<0.05 respectively), but not to patient age, tumor length or differentiation; VEGF-C in lymph fluid is significantly higher than in serum( P<0.05) ; The expression of VEGF-C in lymph fluid end serum were significantly associated with lymph node metastasis( P<0.05). Conclusion There are cliuical significance between VEGF-C expression and lymph node metastasis、TNM stage in tissue of esophageal cancer. The level of VFGF-C in lymph fluid is higher than that in serum, both associate with lymph-node metastasis closely.

Objective To investigate the expression of vascular endothelial growth factorC(VEGF-C),its receptor3(VEGFR-3) and podoplanin in tissue of esophageal cancer,and the relationship between their expression and tumor lymphatic metastasis.Methods Expression of VEGF-C,VEGFR-3 and podoplanin in 76 cases of ESC was detected by immunohistochemical SP method and lymphatic vessel density(LVD) was calculated.Results Positive immunostaining of VEGF-C in esophageal cancer tissue was 63.1%;the expression of VEGF-C was significantly associated with the depth of lymph node metastasis and tumor invasion(P

Objective: To explore the effect of resveratrol (Res) on angiogenesis of human umbilical vein endothelial cells (HUVEC) with the possible mechanisms in vitro.
Methods: The HUVECs were cultured in 6 groups.①Control group, HUVEC were cultured with high glucose in DMEM,②Res group, the cell were cultured with Res at different concentrations,③Res with PI3K blocker LY294002 (Res+Blocker 1) group, ④Blocker 1 group, HUVEC were cultured with LY294002 alone, ⑤Res with eNOS blocker L-NAME (Res +Blocker 2) group and ⑥Blocker 2 group. The effect of Res on HUVEC proliferation was detected by CCK-8 kit, the protein expressions of p-Akt, p-eNOS were examined by Westin blot analysis, nitric oxide (NO) level was measured by nitrate reduction method and the endothelial cell migration was assayed by transwell chamber method.
Results: ① Compared with Control group, HUVEC proliferation increased in Res (1, 5μmol/L ) group, P<0.01, the proliferation in Res (5μmol/L) group was higher than those in Res (0.2, 10, 20μmol/L) group, P<0.01, while Res (20 μmol/L) group could inhibit the proliferation P<0.01. ②Compared with Control group, Res (5μmol/L) group had the higher protein expressions of p-Akt, p-eNOS, P<0.05-0.01, higher NO level, P<0.05.③Compared with Res group, Res+Blocker 1 group had lower expressions of p-Akt, p-eNOS, P<0.01, lower NO, P<0.05; the expressions of p-Akt, p-eNOS and NO level were similar between Res+Blocker 1 group and Blocker 1 group, all P>0.05.④Compared with Control group, the cell migration and tubing formation were higher in Res (5μmol/L) group, P<0.01;compared with Res group, the cell migration and tubing formation were lower in Res+Block2 group, P<0.01.
Conclusion: Res could up-regulate NO level via activating PI3-K/Akt/eNOS signaling and therefore, improving the proliferation, migration and tubing formation of HUVEC in vitro.

Objective: To explore the effect of atorvastatin (Atv) on high glucose-induced oxidative stress injury in human umbilical vein endothelial cells (HUVECs) by SIRT1/NADPH oxidase pathway with the possible mechanisms.
Methods: HUVECs were cultured in low glucose medium and then divided into 6 experimental groups:①Normal group,②Osmotic pressure control group,③High glucose (HG) group,④HG+Atv (0.1, 1.0, 10.0)μmol/L group,⑤HG+sirtinol (SIRT1 inhibitor) group,⑥HG+apocynin (NOX4 inhibitor) group, and HUVECs were further cultured for 24 hours. The cell proliferation was examined by CCK-8 kit, ROS level was detected by lfow cytometry method, protein expressions of SIRT1 and NOX4 were measured by Western blot analysis.
Results: ① Compared with Normal group, HG group had decreased HUVECs proliferation, Atv improved the HG inhibited proliferation in a does dependent manner. ② HG group had the higher level of ROS, increased NOX4 protein expression and decreased SIRT1 protein expression. ③ In HG condition, Atv up-regulated SIRT1 expression and down-regulated ROS and NOX4 expressions in a does dependent manner.④In HG condition, sirtinol decreased SIRT1 expression, increased NOX4 expression, and apocynin decreased NOX4 expression, while it had no inlfuence on SIRT1 expression.
Conclusion: Atorvastatin could resist HG-induced oxidative stress injury in HUVECs, which might be related to up-regulated SIRT1 expression, and SIRTI plays the role in NADPH oxidase at upstream.

BACKGROUND:Currently, antisense oligonucleotides (AS-ODN) have a good prospect in gene therapy, but AS-ODN with smal molecular weight cannot easily enter into the cels, which is susceptible to nuclease degradation. Therefore, there is stil a lack of fundamental understanding about how to improve their transfection efficiency, and target-based transferring. OBJECTIVE:To investigate whether a weak cationic and phosphorylcholine-containing diblock copolymer (MPC30-DEA70) can act as a carrier system to deliver a chemicaly synthesized transforming growth factor-β1 (TGF-β1) AS-ODN into myocardial cels. METHODS: MPC30-DEA70 was compounded with TGF-β1 AS-ODN at various N/P ratios and the MPC30-DEA70/TGF-β1 AS-ODN complexes were characterized by DNA electrophoresis. MTT assay was used to observe the biocompatibility. Confocal laser scanning microscope was used to observe the distribution and location of MPC30- DEA70/TGF-β1 AS-ODN in cells. Flow cytometry was used to detect the transfection efficiency and fluorescence intensity of MPC30-DEA70/TGF-β1 AS-ODN in cells. Western blot and RT-PCR methods were employed to measure the expression of TGF-β1 in cells. RESULTS AND CONCLUSION: Cell growth inhibition showed that the MPC30-DEA70 had low cytotoxicity to myocardial cells within the effective transfection dosage range (< 20 mg/L). Data from the flow cytometry test indicated a clear trend of increasing transfection efficiency with the increasing of N/P ratios. At high N/P ratios, the expression levels of TGF-β1 mRNA and protein in myocardial cells were significantly lower. This study shows that MPC30-DEA70 can work as an effective transgenic vector in myocardial cells. TGF-β1 AS-ODN can silence the expression of TGF-β1 gene efficiently and specially, and may antagonize TGF-β1-mediated biological function.

AIM: To detect the effects of resveratrol (RSV) on the expression of microRNA-21 (miR-21) in primarily cultured neonatal rat atrial myocytes with electric remodeling induced by rapid electrical stimulation (RES).Furthermore, to find out the possible mechanism of miR-21 regulating electrical remodeling.METHODS: The neonatal rat atrial myocytes were isolated by double-enzyme (trypsin and collagenase I) digestion and differential adhesion method.The atrial fibrillation (AF) model was induced by RES.Atrial myocytes were randomly divided into 4 groups: control group, RSV group, RES group, and RSV+RES group.To further detect whether RSV regulated electric remodeling by miR-21, except the 4 groups, we add miR-21 over-expression group and miR-21 inhibitor group: RES+negative control (NC) group, RES+miR-21 mimics group, RES+miR-21 mimics+RSV group, RES+miR-21 inhibitor group, and RES+miR-21 inhibitor+RSV group.The optimal concentration and pretreatment time of resveratrol were determined by CCK-8 assay.The expression of miR-21 and the mRNA expression of L-type calcium channels CACNA1C and CACNB2 in atrial myocytes were detected by qPCR.The protein expression of L-type calcium channels Cav1.2 and Cavβ2 in the atrial myocytes was analyzed by Western blot.RESULTS: The expression of miR-21 in RES group was significantly increased compared with control group, while preconditioning with RSV decreased the expression of miR-21.Compared with RES+miR-21 mimics group, the expression of miR-21 in RES+miR-21 mimics+RSV group was significantly decreased.Meanwhile, the mRNA expression of CACNA1C and CACNB2, and the protein levels of Cav1.2 and Cavβ2 were increased (P<0.05).Compared with RES group, the expression of miR-21 in RES+miR-21 inhibitor group and RES+miR-21 inhibitor+RSV group was decreased, while the mRNA expression of CACNA1C and CACNB2, and the protein levels of Cav1.2 and Cavβ2 were increased.However, no difference of the expression of miR-21, the mRNA expression of CACNA1C and CACNB2, and the protein levels of Cav1.2 and Cavβ2 among RSV+RES, RES+miR-21 inhibitor and RES+miR-21 inhibitor+RSV groups was observed (P<0.05).CONCLUSION: In AF model induced by RES, RSV may reduce electric remodeling by inhibiting the expression of miR-21 and regulating the downstream target genes.

AIM:To investigate the effects of Akt1 gene transfection into myocardium after ischemia-reperfusion (I/R) on mitochondrial permeability transition. METHODS:Forty adult male SD rats were divided randomly into five groups with 8 rats each:control group,I/R group,Ad-gene group,Ad-blank group and Ad-inhibitor group. The rats in Ad-gene group were injected with 30 μL Lipofectamine 2000 solution including Akt1 gene to the myocardium 48 h before ischemia while those in control group and I/R group were injected with PBS of the same volume. Rats in Ad-blank group were injected with Lipofectamine 2000 of the same volume into myocardium. In Ad-inhibitor group 30 μL Lipofectamine 2000 and gene complexes with LY294002 were injected. Hemodynamics,apoptotic index,the concentrations of lactate dehydrogenase,creatine kinase,the expression of Akt1,cytosolic,mitochondrial cytochrome C and MPT were also measured. RESULTS:The lowest level of Akt1 protein expression was observed in control group. The protein expression of Akt1 in Ad-gene group was higher than that in I/R group,Ad-blank group and Ad-inhibitor group. The AI,LDH and CK in Ad-gene group were significantly lower than those in other groups except control group. Transfection of Akt1 markedly reduced the loss of mitochondrial cytochome C after I/R injury. Ad-gene transfection led to a significant increase in absorbance at 540 nm compared to I/R group,Ad - black group and Ad-inhibitor group (P<0.05). CONCLUSION:Akt1 gene prevents myocardial apoptosis after I/R injury. Akt1 gene also inhibits the opening of mitochondria permeability transition and protects mitochondrial functions of myocardium in I/R injury.

Objective:To analyze the hospitalization costs of delivery for postpartum women with different delivery methods, ages and comorbidities or complications, for references for medical institutions and medical insurance management departments to develop payment and compensation standards for inpatient delivery.Methods:The first page of medical records of hospitalized delivery women admitted to 8 large tertiary hospitals in Beijing from January 2018 to December 2021 were selected. Descriptive analysis was made on the mode of delivery, age, hospitalization expenses and cost structure of puerpera, as well as the hospitalization expenses of puerpera with different complications or complications. Wilcoxon rank sum test, Kruskal-Wallis H test, and Chi-squared test were used to statistically compare the level and structure differences between groups. Results:A total of 23 320 pregnant women were included, with an average age of 32.3 years. There were 13 605 cases of natural delivery and 9 715 cases of caesarean section. The median cesarean section rate in the right age group (<35 years old) and the elderly group (35-50 years old) were 36.73% and 56.58%, respectively, and the median hospital expenses were 5 865 yuan and 7 042 yuan, respectively. The median hospital expenses for natural delivery and caesarean section were 4 452 yuan and 10 033 yuan, respectively. The highest proportion of hospitalization expenses for natural delivery and cesarean section were treatment expenses (23.45%) and medicine expenses (29.19%), respectively. The median of hospitalization cost for women with≥2 comorbidities or complications (6 736 yuan) was higher than that for women with 1 comorbidities or complications (5 794 yuan).Conclusions:The hospitalization cost of cesarean section was significantly higher than that of natural delivery and the rate of cesarean section and the cost of delivery in women aged 35 and above were higher than those under 35 years old. The structure of hospitalization cost was different in different delivery modes, and the complications or complications had a greater impact on the average hospitalization expenses.