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Objective: To explore the role of calpain in pulmonary vascular remodeling in hypoxia-induced pulmonary hypertension and the underlying mechanisms.
, Methods: Sprague-Dawley rats were randomly divided into the hypoxia group and the normoxia control group. Right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP) were monitored by a method with right external jugular vein cannula. Right ventricular hypertrophy index was presented as the ratio of right ventricular weight to left ventricular weight (left ventricle plus septum weight). Levels of calpain-1, -2 and -4 mRNA in pulmonary artery were determined by real-time PCR. Levels of calpain-1, -2 and -4 protein were determined by Western blot. Primary rat pulmonary arterial smooth muscle cells (PASMCs) were divided into 4 groups: a normoxia control group, a normoxia+MDL28170 group, a hypoxia group and a hypoxia+MDL28170 group. Cell proliferation was detected by MTS and flow cytometry. Levels of Ki-67 and proliferating cell nuclear antigen (PCNA) mRNA were determined by real-time PCR.
, Results: RVSP, mPAP and right ventricular remodeling index were significantly elevated in the hypoxia group compared to those in the normoxia group. In the hypoxia group, pulmonary vascular remodeling was significantly developed, accompanied by up-regulation of calpain-1, -2 and -4. MDL28170 significantly inhibited hypoxia-induced proliferation of PASMCs concomitant with the suppression of Ki-67 and PCNA mRNA expression.
, Conclusion: Calpain mediates vascular remodeling via promoting proliferation of PASMCs in hypoxia-induced pulmonary hypertension.

Objective: To investigate the effects and the underlying molecular mechanisms of curcumin on pulmonary artery smooth muscle cells in rat model with chronic obstructive pulmonary disease (COPD). Methods: A total of 75 male Wistar rats were randomly divided into control group (group CN), model group (group M), low-dose curcumin group (group CL), medium-dose curcumin group (group CM) and high-dose curcumin group (group CH). HE staining was used to observe the morphology of pulmonary artery. Proliferating cell nuclear antigen (PCNA), apoptosis-related protein Bcl-2 and Bax were detected by immunohistochemical staining. TUNEL kit was used to analyze the effects of curcumin on apoptosis of smooth muscle cells, and the protein expressions of SOCS-3/JAK2/STAT pathway in lung tissues were determined by western blot. Results: Right ventricular systolic pressure (RVSP) and right ventricular hypertrophy index (RVMI) in group M were significantly higher than those in group CN, group CH and group CM (all P <0.05). HE staining and TUNEL kit test showed that the number of pulmonary artery smooth muscle cells had a significant increase in group M, while the pulmonary artery tube became thin, and the smooth muscle cells shrinked in group CM and group CH. Immunohistochemistry showed that PCNA and Bcl-2 in group M were significantly higher than those in group CN (all P <0.05), while Bax expression was significantly lower than that in group CN ( P <0.05). PCNA in group CM and group CH were significantly lower than that in group M (all P <0.05), while Bax expression was significantly higher than that in group M ( P <0.05). Western blot showed that SOCS-3 protein was significantly decreased in group M, while the p-JAK2, p-STAT1, p-STAT3 were significantly increased (all P <0.05). Compared with group M, SOCS-3 protein in group CM and group CH were significantly increased (all P <0.05), while the p-JAK2, p-STAT3 were significantly reduced (all P <0.05). Conclusion: Curcumin could promote the apoptosis of smooth muscle cells in rats with COPD, and improve the mean pulmonary artery pressure and RVMI through stimulating SOCS-3/JAK2/STAT signaling pathway.

Aim: To explore the effects of different doses of nano silver of proliferation of cultured vascular endothelial cells in vitro., Methods: The hearts of three newborn SD rats 5 day old were mechanically minced the enzymatically digested with collagenase and trypsin, then vascular endothelial cell were counted, washed and resuspended in Dulbecco's minimumes sential medium (DMEM) added with 20% heat in activated fetal calf serum, then inoculated d in 2% gelatin coated tissue culture flasks. Vascular endothelial cells at passage 3 were used in the experiment. Except for the normal control group, the vascular endothelial cells were cultured with nano silver in various concentrations (0.5, 0.25, 0.125, 0.0625, 0.03125 g/l) for 24 hours, and the morphology and the number of the cultured endothelial cells were observed. Methly thiazolyl tetrazolium (MTT) colorimetry was used to determine the proliferation of the cultured vascular endothelial cells. Flow cytometry (FCM) were used to detect the proliferation index (PI) of the vascular endothelial cells and the expressions of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemical method., Results: The cell morphology was normal under the inverted microscoped in each group. The number and proliferation activities of vascular endothelial cells were significantly decreased by 0.5, 0.25, 0.125, 0.0625, 0.03125 g/l nano silver compared with those of the blank control group, especially the 0.25 g/l nao silver group, and there were no remarkable changes in with 0.5 g/L, 0.125 g/L, 0.0625 g/L and 0.03125 g/L nano silver groups compared to each other. The same results were seen in the positive rate of PCNA expression and PI., Conclusion: Nano silver has dose-dependent effects on the proliferation activity of vascular endothelial cells. It inhibited the proliferation of vascular endothelial cells.

Objectives: This study is to investigate the effects of tea polyphenols and tea pigments on cell cycle regulators in rat liver precancerous lesions., Methods: The modified Solt-Farber precancerous liver rat model was used. Rats were given water, tea polypheol solution (0.1%) or tea pigment solution (0.1%) throughout the whole experiment (56 days). Cyclin D1, P21(WAF1/CIP1), GADD45 and PCNA protein expression were detected by Western blotting and the RT-PCR method was applied to study the expression of Cdk4., Results: Cyclin D1, Cdk4 and PCNA expressions were significantly inhibited, and the expression of P21(WAF1/CIP1) and GADD45 were significantly induced by tea polyphenols and tea pigments treatments., Conclusion: Tea polyphenols and tea pigments induced cell cycle arrest and inhibited cell proliferation by regulating cell cycle regulators.

Background & Objectives: The high expression of proliferation cell nuclear antigen were found in the patients with esophageal carcinoma in poor prognosis. The aim of this study was to investigate the effect in vitro of antisense oligodeoxynucleotide (ASODN) of PCNA on its gene expression and cell proliferation in human esophageal squamous cell cancer T. Tn cell line., Methods: Phosphorothioate PCNA ASODN and sense oligodeoxynucleotide (SODN) were synthesized and transfected to T. Tn cells mediated by dosper liposomal. Their effects on cell proliferation were examined by MTT method, 3H-TdR method, immunohistochemical study, and flow cytometer (FCM)., Results: PCNA ASODN could significantly inhibit the growth of T. Tn cell lines. The SODN and control lipsome showed no such effect. The inhibition peaked at 24 hour after transfection, the inhibition rate reached 63.3% at 0.2 mumol/L by MTT method, the inhibition effect was dose-dependent. ASODN could downregulate the expression levels of PCNA protein. And PCNA ASODN transfection could inhibit the transition period of G0/G1 phase to S phase., Conclusions: The data suggested that ASODN could specific inhibit the expression of PCNA protein and regulate cell cycle and cell proliferation in T. Tn cells and antisense therapy could be an useful method to improve prognosis in ESCC.

Objective: To study the inhibitory effect of PCNA antisense oligonucleotides or/and VEGF antisense oligonucleotides on growth of human hepatocellular carcinoma in nude mice., Methods: Human hepatocellular carcinoma transplanted subcutaneously in nude mice model was established, and subsequently was divided into four groups: PCNA-ASODN group, VEGF-ASODN group, combined group and control group. Different treatments were given respectively. The weigh and volume of subcutaneous tumors was measured, and the morphological changes of tumor cells was observed under microscope., Results: The growth inhibitory rate in the PCNA-ASODN group was 72.8%, 44.9% in the VEGF-ASODN group, and 87.2% in the combined group. The combined group which was treated with PCNA-ASODN and VEGF-ASODN showed more effective inhibitory than the PCNA-ASODN group or the VEGF-ASODN group respectively., Conclusions: The growth of human hepatocellular carcinoma in nude mice can be inhibited by PCNA antisense oligonucleotides and VEGF antisense oligonucleotides respectively. Better inhibitory effect can be obtained if PCNA antisense oligonucleotides and VEGF antisense oligonucleotides are combined in use.