Prevention and treatment strategy of hypertension based on etiology and pathogenesis of "lower deficiency and upper excess" / 国际中医中药杂志

The etiology and pathogenesis of patients with hypertension are lower deficiency and upper excess. The deficiency below the etiology comes first, and then leads to upper excess. Lower deficiency is manifested by the yin deficiency of liver and kidney. The clinical symptoms are lumbar and knee weakness, or low back pain, or lower limb weakness, or systemic fatigue, or dry throat. In fact, it is mostly manifested as excess of liver yang, manifesting Qi inversion symptoms of high blood pressure, dizziness, dry throat, or bitter mouth, or headache, or tinnitus and deafness, or palpitations, or chest pain, or gastroesophageal reflux, or heartburn acid reflux, which are also named like hyperactivity of liver yang, liver yang heat, liver defencing spleen, or spleen deficiency leading to phlegm. Deficiency of liver and kidney leads to yin failing to restrain yang, resulting in hyperactivity of liver yang, spleen deficiency generating phlegm, and phlegm blocking orifices. Hypertension is due to deficiency of liver and kidney, and its sign is hyperactivity of liver yang. Therefore, lower deficiency and upper excess process and guides the treatment and prevention of hypertension. In the treatment, with the principle of lower deficiency and upper excess, we can nourish liver and kidney Yin to defence liver Yang for hypertension.

PRKCDBP Methylation is a Potential and Promising Candidate Biomarker for Non-small Cell Lung Cancer / 中国肺癌杂志

BACKGROUND@#The occurrence and development of lung cancer are closely linked to epigenetic modification. Abnormal DNA methylation in the CpG island region of genes has been found in many cancers. Protein kinase C delta binding protein (PRKCDBP) is a potential tumor suppressor and its epigenetic changes are found in many human malignancies. This study investigated the possibility of PRKCDBP methylation as a potential biomarker for non-small cell lung cancer (NSCLC).@*METHODS@#We measured the methylation levels of PRKCDBP in the three groups of NSCLC tissues. Promoter activity was measured by the dual luciferase assay, with 5'-aza-deoxycytidine to examine the effect of demethylation on the expression level of PRKCDBP.@*RESULTS@#The methylation levels of PRKCDBP in tumor tissues and 3 cm para-tumor were higher than those of distant (>10 cm) non-tumor tissues. Receiver operating characteristic (ROC) curve analysis between tumor tissues and distant non-tumor tissues showed that the area under the line (AUC) was 0.717. Dual luciferase experiment confirmed that the promoter region was able to promote gene expression. Meanwhile, in vitro methylation of the fragment (PRKCDBP_Me) could significantly reduce the promoter activity of the fragment. Demethylation of 5'-aza-deoxycytidine in lung cancer cell lines A549 and H1299 showed a significant up-regulation of PRKCDBP mRNA levels.@*CONCLUSIONS@#PRKCDBP methylation is a potential and promising candidate biomarker for non-small cell lung cancer.

Regulation of autophagy by store-operated calcium channel in arterial smooth muscle cells / 中国药理学通报

Aim To investigate the effect of store-oper-ated calcium channel( SOCC) on autophagy in rat arte-rial smooth muscle cells A7 R5 . Methods Lentiviruses containing STIM1 or Orai1 gene were packaged in 293 T cells and then were used to infect rat arterial smooth muscle cells A7 R5 . The expression levels of STIM1 , Orai1 and Beclin 1 , a critical autophagy-regu-lating protein, of lentivirus-infected A7R5 cells, were detected by Western-blot. Autophagy in lentivirus-in-fected A7 R5 cells was induced by starvation or rapamy-cin, an inhibitor of mammalian target of rapamycin ( mTOR ) . Autophagy marker LC3 of these cells was detected by Western-blot. Results The constructions of vector pLV-STIM1 and pLV-Orai1 were confirmed by restriction enzymes digestion analysis. Compared with the control group, expressions of STIM1 or Orai1 protein was significantly increased after lentivirusLV-STIM1 and LV-Orai1infection, whereas the expressions of autophagy related protein Beclin-1 were down-regu-lated. Starvation or rapamycin stimulated A7R5 auto-phagy but overexpression of STIM1 or Orai1 significant-ly inhibited starvation or rapamycin induced autoph-agy. Conclusion Overexpression of store-operated calcium channel components STIM1 and/or Orai1 in rat arterial smooth muscle cells A7 R5 inhibit autoph-agy. This mechanism might contribute to the develop-ment of pulmonary arterial hypertension.

STIM1 promotes arterial smooth muscle cells proliferation by regulating Akt/mTOR pathway / 中国药理学通报

Aim To investigate the expression of stro-mal interaction molecule 1 (STIM1) in rat pulmonary arterial hypertension ( PAH ) tissues and effects of STIM1 on arterial muscle cells proliferation. Methods PAH was induced by a single intraperitoneal injec-tion of MCT at a dose of 60 mg·kg - 1 . The mRNA or protein expressions of STIM1 in monocrotaline-induced pulmonary hypertensive rats were measured by real-time PCR or Western blot, respectively. The arterial smooth muscle cells A7R5 were transiently transfected with STIM1 plasmids to prepare STIM1 overexpressed cells. Cell proliferations were detected by using CCK-8 kits. The expressions of Akt/ mTOR pathway molecules of A7R5 were measured by Western blot. Results The right ventricular systolic blood pressure ( RVSP) and right ventricular mass index ( RVMI ) were markedly elevated in MCT-treated rats (P < 0. 01) in comparison to control rats. The mRNA and protein ex-pression levels of STIM1 in monocrotaline-induced pul-monary hypertensive rats were 2. 19 and 1. 66 folds of control rats, respectively. STIM1 were transiently over-expressed in cultured A7R5. Cells transfected with STIM1 grew more quickly than non-transfected control. Overexpression of STIM1 significantly increased the phosphorylation of Akt, mTOR, p70-S6K, and 4E-BP1, but did not change their protein expression lev-els. Conclusion STIM1 are over-expressed in rat PAH tissues. Overexpression of STIM1 can promote ar-terial smooth muscle cells proliferation by regulating Akt/ mTOR pathway.

Clinical significance of thrombospondin 4 expression in gastric carcinoma / 解剖学报

Objective The aim of this study was to measure quantitatively the TSP-4 mRNA expression and its significance in gastric carcinoma was evaluated by correlating its expression with clinicopathological features, microvessel density(MVD) and MMP-9.Methods Eighty-two patients with gastric carcinoma were recruited in this study. TSP-4 mRNA expression in gastric carcinoma and adjacent tissue was detected by real-time PCR. CD34 and MMP-9 protein expression were examined by immunohistochemistry. MVD was determined according to CD34-positive tubular structures. Results The expression level of TSP-4mRNA in gastric carcinoma was obviously higher than those of adjacent tissue(P=0.03) and it was associated with tumor size, histological type, lymph node metastasis, MVD and MMP-9(P=0.002, P=0.031, P=0.014,r＝0.67 P<0.01, P=0.008),but not sex, age and depth of invasion. (P=0.53,P=0.57,P=0.15).Conclusion TSP-4 may play an important role in occurrence and progression of gastric carcinoma and that the effect of TSP-4 on tumor growth and metastasis may be exerted through regulation of angiogenesis and MMP-9 expression in gastric carcinoma.

Inhibition Effect of a Snake Venom Metalloproteinase Inhibitor on Experimental Pulmonary Metastasis in Mice Inoculated with B16 Melanoma Cells / 中国生物工程杂志

Although there is emerging evidence that BJ46a can function as potent inhibitor of the SVMPs proteolytic activities,its anticancer effect on invasion and metastasis has not yet been evaluated.Inhibition effect of BJ46a on experimental pulmonary metastasis in mice inoculated with B16 melanoma cells at the protein level was investigated. First,BJ46a was produced in baculovirus expression system. SDS-PAGE and Western blot analysis confirmed that BJ46a recombinant protein was produced by Sf9 cells infected with high-titer viral stock. Then,recombinant fusion protein was purified by ProBondTM at the point of maximal expression. B16 cells pre-treated with recombinant BJ46a injected into C57BL/6 mice via the tail lateral vein to form experimental pulmonary metastasis model. The numbers of metastatic lesions in C57BL/6 mice changed dramatically:BJ46a different concentrations of recombinant protein group were 1.1 ? 0.83,0.9 ? 0.7,significantly lower than the control group (6.3?3.00,P