Objective To explore the proliferation property of vascular smooth muscle cells (VSMCs) in the stable CMKLR1 gene knock-down mouse VSMCs line and explore related mechanism.Methods The short hairpin RNA sequence targeting to knockdown the coding regions of mouse CMKLR1 mRNA was synthesized and subsequently employed to construct recombinant lentivirus vector.Mouse VSMCs were cultured and infected with the recombinant lentivirus (knockdown VSMCs).mRNA and protein CMKLR1 expression in Knockdown VSMCs was measured by real-time PCR and Western blot and compared with those in normal VSMCs (vehicle VSMCs) and lentivirus control VSMCs (control VSMCs).The proliferation of normal,knockdown and control VSMCs was induced by platelet-derived growth factor-BB (PDGF VSMCs) and measured by cell number counting and BrdU.The phosphorylated c-Jun N-terminal kinase (p-JNK) protein was investigated by Western blot.Results The relative level of CMKLR1 mRNA in knockdown VSMCs (0.23 ± 0.04) was significantly downregulated compared with which in vehicle VSMCs (1.05 ± 0.05) as well as control VSMCs (0.99 ± 0.04) (P ＜ 0.01).The relative level of CMKLR1 protein in knockdown VSMCs (0.29 ± 0.04) was also significantly decreased,compared with which in vehicle VSMCs (1.06 ±0.04) as well as control VSMCs (0.95 ±0.02) (P ＜0.01).The VSMCs number ((50.33 ± 1.20) × 103/cm2) and BrdU A450 value (1.80 ±0.05) in PDGF VSMCs were significantly increased in vehicle VSMCs ((42.02 ± 1.53) × 103/cm2,1.55 ± 0.04) (both P ＜ 0.05).Compared with those in vehicle VSMCs,the VSMCs number ((23.33 ± 2.03) × 103/cm2) and BrdU A450 value (1.32 ± 0.02) in knockdown VSMCs were significantly decreased.The proliferation property between PDGF VSMCs and control VSMCs was similar(P ＞0.05).Compared with the relative level of p-JNK protein (1.03 ±0.03) in vehicle VSMCs,the p-JNK protein level was significantly increased in PDGF VSMCs (1.36 ± 0.02,P ＜ 0.05) and significantly downregulated in knockdown VSMCs (0.79 ± 0.05,P ＜ 0.05).Conclusion Knockdowning CMKLR1 gene can reduce the proliferation of mouse vascular smooth muscle cells,which was related with the down-regulation of p-JNK expression.

OBJECTIVES: To explore the relevant protective and risk factors that affect spontaneous supratentorial large volume intracerebral hemorrhage. METHODS: Clinical data of hospitalized patients with spontaneous supratentorial intracerebral hemorrhage in Xiangya Hospital of Central South University from January 2014 to December 2018 were retrospectively analyzed. According to the amount of intracerebral hemorrhage (≥30 mL), the patients were divided into a large volume intracerebral hemorrhage group (255 cases) and a non-large volume intracerebral hemorrhage group (397 cases). Univariate and multivariate logistic regression analysis for the clinical data from the two groups of patients were performed. RESULTS: Systolic blood pressure, diastolic blood pressure, white blood cell count, neutrophil count, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein cholesterol, calcium ion concentration, and international standard ratio between the large volume intracerebral hemorrhage group and the non-large volume intracerebral hemorrhage group were significantly different (all CONCLUSIONS The occurrence of spontaneous supratentorial large volume intracerebral hemorrhage is the result of combination of multiple factors. The increased white blood cell count, the increased systolic blood pressure, the increased high-density lipoprotein cholesterol, and the increased international standard ratio may increase its risk, while the increased LDL cholesterol and calcium concentration may reduce its risk.
Blood Pressure ; Cerebral Hemorrhage ; Cholesterol, HDL ; Humans ; Retrospective Studies ; Risk Factors