In order to investigate the effect of Polo-like kinase-1 (Plk1) depletion on cell cycle progression and cell growth in lung cancer cells, a recombinant plasmid containing antisense RNA targeting Plk1 (pcDNA3-Plk1) was transfected into A549 cells by lipofectine. RT-PCR and Western-blot were used to detect the Plk1 gene expression. Cell proliferation was evaluated by direct cell counting and bromodeoxyuridine (BrdU) labeling. Cell cycle distribution and apoptosis were examined by flow cytometry, and the inhibition rate (IR) by vinorebline (NVB) was determined by MTT assay. The results showed that after transfection of pcDNA3-Plk1 into A549 cells, the expression levels of Plk1 mRNA and protein were greatly decreased. In pcDNA3-Plk1 transfected groups, abnormal morphological changes of cells and growth inhibition were observed, and the BrdU labeling index was significantly lower than in the control groups (P<0.05). Cells in pcDNA3-Plk1 transfected groups were arresed in G₂/M phase and apoptosis was detectable 72 h post transfection. IR induced by vinorebline in pcDNA3-Plk1 transfected groups was significantly higher than in other groups. These data suggested that antisense RNA targeting Plk1 could suppress the Plk1 expression, and therefore, significantly inhibit cell proliferation and induce cell cycle arrest and apoptosis. Moreover, it sensitized lung cancer cells to chemotherapy.

In adipose tissue of rats on long-term high fat diet, the inflammatory changes the roles of angiotensin receptor blocker (ARB) in pimelitis and insulin resistance (IR) were observed. IR rat model was established by feeding high calorie and high fat diet. The change in insulin sensitivity was detected by euglycemic-hyperinsulinemic clamp technique 8 weeks after intervention by valsartan. The expression levels of CD68 and MCP-1 mRNA and proteins in adipose tissue were examined by RT-PCR and immunohistochemistry respectively. The parameters of blood glucose, insulin and blood lipid were analyzed. The results showed that in high fat diet group intra-abdominal obesity developed, the content of visceral fat and the number of inflammatory cells in local adipose tissue were significantly increased (P<0.01), the levels of serum triglyceride, free fatty acids and fasting serum insulin were markedly increased, the insulin sensitivity was significantly lowered (P<0.01), and the expression of CD68 and MCP-1 was significantly increased as compared with control group (P<0.01). In ARB interventional group, the content of visceral fat, the number of inflammatory cells and the expression of CD68 and MCP-1 in local adipose tissue were significantly reduced (all P<0.01), but the insulin sensitivity was significantly enhanced (P<0.01) as compared with high fat diet group. There were pimelitis and IR in rats with obesity induced by long-term high calorie and high fat diet. The ARB can significantly inhibit the infiltration of macrophages and the expression of MCP-1 in adipose tissue, thereby attenuating the inflammation and improving IR in rats.

The aim of this study was to determine if the potassium aspartate and magnesium (PAM) prevent reperfusion-induced ventricular arrhythmias (RIVA) in ischemia-reperfusion (IR) rabbit heart. Thirty rabbits were randomly divided into control, ischemia and PAM groups. Arterially-perfused rabbit left ventricular preparations were made, and transmural ECG as well as action potentials from both endocardium and epicardium were simultaneously recorded in the whole process of all experiments. In control group rabbit ventricular wedge preparations were continuously perfused with Tyrode’s solution, and in ischemia group and PAM groups the perfusion of Tyrode’s solution was stopped for 30 min. Then the ischemia group was reperfused with Tyrode’s solution and the PAM group with Tyrode’s solution containing 2.42 mg/L PAM, respectively. ECG, QT interval, transmural repolarization dispersion (TDR) and action potentials from epicardium and endocardium were simultaneously recorded, and the RIVA of the wedge preparation was observed. Compared with control group, TDR and incidence of RIVA were significantly increased in ischemia group (P<0.05). The incidence of RIVA in control, ischemia and PAM group was 0/10, 9/10 and 1/10, respectively. Compared with ischemia group, TDR and incidence of RIVA were significantly reduced in PAM group (P<0.05). Potassium aspartate and magnesium significantly reduce TDR and prevent ventricular arrhythmia in ischemic rabbit heart.

This study investigated the changes of CD4⁺ CD25⁺ regulatory T cells (Tregs) in peripheral blood of patients with hepatocellular carcinoma before and after transcatheter arterial chemoembolization (TACE). The proportion of CD4⁺ CD25⁺ Tregs among CD4⁺ T lymphocytes in peripheral blood of 33 patients with hepatocellular carcinoma was determined by flow cytometry before, 1 week and 1 month after TACE. And 25 healthy volunteers served as control. One month after TACE, the patients were divided into two groups: 22 in group A, who were in stable condition or getting better; and 10 in group B, who were deteriorating. One patient died and was excluded. The results showed that the percentage of CD4⁺CD25⁺ Tregs among CD4⁺ T lymphocytes did not significantly change in the 33 patients 1 week after TACE as compared with that before TACE, however, the difference was significant (P<0.01) between the patients with hepatocellular carcinoma and the healthy subjects. The percentage of CD4⁺ CD25⁺ Tregs among CD4⁺ T lymphocytes in group A 1 month after TACE was decreased significantly in comparison with that before and 1 week after TACE (P<0.01), whereas, that in group B was increased significantly 1 month after TACE (P<0.01). It was concluded that patients with hepatocellular carcinoma had a higher proportion of CD4⁺CD25⁺ Tregs in peripheral blood. TACE did not significantly affect the level of CD4⁺ CD25⁺ Tregs within short time (such as 1 week). The proportion of CD4⁺CD25⁺ Tregs in peripheral blood 1 month after TACE was related to the prognosis of hepatocellular carcinoma.

This study investigated the association of apolipoprotein A5 (apoA5) gene polymorphism at position −1131T>C with cerebral infarction in patients with type 2 diabetes. A total of 256 type 2 diabetic patients without cerebral infarction (T2DM), 220 type 2 diabetic patients with cerebral infarction (T2DMCI) and 340 healthy subjects were recruited from the same region (Hubei province, China). The genotype of apoA5 −1131T〉C was analyzed by polymerase chain reaction, followed by restriction fragment length polymorphism (PCR-RFLP). Total cholesterol, HDL cholesterol, LDL-cholesterol and triglycerides were quantitatively detected by using standard enzymatic techniques. The results showed that the prevalence of the apoA5 −1131C allele was significantly higher in T2DMCI group than that in control group (42.7% versus 31.2%, P<0.01). The carriers of rare C allele had higher TG levels as compared with carriers of common allele in the three groups (P<0.01). Logistic regression models, which were adjusted for age, gender, blood pressure, BMI, FBS, smoking, LDL-C and HDL-C, revealed that patients carrying the apoA5 −1131C allele and CC homozygotes were at high risk for T2DMCI. It was concluded that the apoA5 −1131C allele variant is an independent genetic risk factor for T2DMCI.

To examine the changes in number and function of endothelial progenitor cells (EPCs) from peripheral blood (PB) in hypertension disorder complicating pregnancy (HDCP), 20 women with HDCP and 20 normal pregnant women at the third trimester were studied. Mononuclear cells (MNCs) from PB were isolated by Ficoll density gradient centrifugation. EPCs were identified by positive expression of both CD34 and CD133 under fluorescence microscope and positive expression of factor VIII as shown by immunocytochemistry. The number of EPCs was flow-cytometrically determined. Proliferation and migration of EPCs were measured by MTT assay and modified Boyden chamber assay, respectively. The adhesion activity of EPCs was detected by counting the number of the adherent cells. The results showed that, compared with normal pregnant women, the number of EPCs was significantly reduced in HDCP (4.29%±1.21% vs 15.32%±2.00%, P<0.01), the functional activity of EPCs in HDCP, such as proliferation (13.45%±1.68% vs 18.45%±1.67%), migration (37.25±7.28 cells/field vs 67.10±9.55 cells/field) and adhesion activity (20.65±5.19 cells/field vs 34.40±6.72 cells/filed) was impaired (P<0.01). It is concluded that the number and function of EPCs are significantly decreased in HDCP.

The association between the single nucleotide polymorphisms (SNPs) in −174G/C and −634C/G of interleukin-6 (IL-6) promoter region and prostate cancer was examined in the population of Han people in Hubei region. TaqMan PCR was employed for the gene-typing of −174G/C and −634C/G in promoter region of IL-6 gene to compare the prostate cancer patients and normal controls in terms of genotype frequency, allele frequency and risk of prostate cancer. Enzyme-linked immunosorbent assay (ELISA) was used for the detection of IL-6 concentration in peripheral blood of the patients with prostate cancer and the relationship between the IL-6 level and the genotype was studied. Our results showed that in all the subjects, the genotype of genetic locus −174G/C was found to be GG and no CG and CC were observed. There was a significant difference in gene frequency of GG, CG and CC of −634C/G and allele frequency of G and C between prostate cancer patients and normal controls (P<0.05) and the gene frequency of GG+CG increased with the clinical stages and pathological grades of prostate cancer. The IL-6 level in GG+CG group was significantly higher than that in CC group. It was concluded that no SNP in −174G/C IL-6 promoter region was found in the population of Han people in Hubei region. The SNP in −634C/G was, to some extent, associated with the development and progression of prostate cancer. The population with GG+CG genetype has higher risk for prostate cancer.

This study investigated the effects of propofol on the mRNA expression of Toll-like receptor-4 (TLR4) in BV-2 cells during mimic ischemia-reperfusion (I/R) injury in vitro. BV-2 cells, a mouse microglia line, were cultured and divided into 4 groups at random: control group (group C), ischemia/reperfusion group (group I/R), low-dose propofol (25 μmol/L) intervention group (group PF₂₅) and high-dose propofol (100 μmol/L) intervention group (group PF₁₀₀). The mRNA expression of TLR4 and NF-κB was measured by means of RT-PCR. TNF-α levels in the supernatants of BV-2 cells were detected by ELISA. The results showed that the mRNA expression of TLR4 and NF-κB was significantly higher in groups I/R, PF₂₅ and PF₁₀₀ than in group C (P<0.01). And the TNF-α level in the supernatants was elevated in groups I/R, PF₂₅ and PF₁₀₀ as compared with that in group C (P<0.01). After pre-treatment with propofol, the mRNA expressions of TLR4 and NF-κ B and the TNF-α level were significantly decreased in groups PF₂₅ and PF₁₀₀ in comparison to those in group I/R (P<0.01). And the decrease in those indicators was more significant in group PF₁₀₀ than in group PF₂₅ (P<0.01). It was concluded that propofol exerted brain-protecting effects during I/R injury by suppressing the mRNA expressions of TLR4 and NF-κB and deceasing the TNF-α level.

This study examined the effect of different b values on diffusion-weighted MR imaging (DWI) of human prostate by using single-shot spin echo echo planar imaging (SE-EPI) sequences, observed the normal appearances and measured apparent diffusion coefficient (ADC) values in anatomical regions of normal prostate. Twenty-four healthy volunteers (mean age: 32 y) were studied by using a 1.5T system with a phased array surface multicoil. Two kinds of single-shot SE-EPI sequence were used to perform DWI in the prostate in volunteers, with five b values being 0, 30, 300, 500 to 1000 s/mm². The image quality with different imaging parameters was analyzed and the ADC values in anatomical regions of normal prostate were measured. DWI of prostate was successfully obtained in all volunteers. The images were of good quality, without artifacts containing pixels within the prostate. The contrast was good between the different anatomical regions of the prostatic gland, i.e., the peripheral zone (PZ), which exhibited higher signal intensity, and the central gland (CG). Signal intensity contrast was related to the magnitude of b values. The ADC values in PZ and CG were (1.27±0.22)×10⁻³ mm²/s and (1.01±0.17)×10⁻³ mm²/s, respectively. The ADC values were found to be significantly higher in PZ than in CG (P<0.05, paired t-test). Significant differences were found between the slice-selecting component and both the read-out and phase-encoding components of the ADC values. It is concluded that SE-EPI is a suitable DWI sequence for human prostate. The contrast between PZ and CG is good when b values are low, while the diffusion and ADC values are accurate when b values are high. ADC values are higher in PZ than in CG in normal prostate. Diffusional anisotropy is present in normal prostatic tissue.