Objective: To study the relation among inducible nitric oxide synthase(iNOS), endothelial nitric oxide synthase(eNOS) and vascular endothelial growth factor (VEGF)in oral squamous cell carcinoma(OSCC) and their relation to clinical pathology and angiogenesis.Methods:OSCC tissue specimens were surgically obtained from 64 patients,the expression of iNOS,eNOS and VEGF in the samples were studied by immunohistochemistry technique. Microvessel density(MVD) was measured by counting the endothelial cells stained with anti-FⅧRAg antibody. Results: ①Positive VEGF and iNOS and eNOS immunostaining was detected in 39(60.94%),42(65.63%)and 45( 70.31%)of the cases respectively.②Expression of VEGF was significantly related to that of iNOS(P05).③MVD in OSCC tissue was positively correlated with the expression of VEGF and iNOS(P05).④VEGF expression was positively correlated to the pathological grade of OSCC(P0.05).Conclusion:iNOS,rather than eNOS, plays a positive role in OSCC development.iNOS and VEGF have a positive correlation in angiogenesis of OSCC.

Objective To explore protective effects of panaxtrial saponins (PTS) on cognition and memory of diabetic rats and to reveal its mechanism by which might involve regulating activity of astrocytes. Methods SD rats (n=24) were ran?domly assigned into control, diabetic and PTS-treated groups (n=8 in each group). Rat diabetic model was induced through streptozotocin injection intraperitoneally. Rats in control group were native rats, and rats in PTS-treated group were diabetic rats that were administered with PTS. Body weight and blood glucose were monitored through the experiments. Three months later, state of cognition was examined by methods of water maze. Hippocampal astrocyte morphology were detected by immu?nohistochemistry, and the expression of glial cell line-derived neurotrophic factor (GDNF) and glial fibrillary acidic protein (GFAP) in hippocampus were revealed by Western blot. Results Compared with control group, diabetic group showed cog?nitive dysfunction, atrophic astrocyte soma, shrinked astrocyte processes, and down-regulation of hippocampal GFAP and GDNF (P<0.05). Compared with diabetic group, PTS-treated group exhibited improved cognition and morphology of hippo?campal astrocyte, and reversed expression of GFAP and GDNF in diabetic hippocampus (P<0.05). Conclusion PTS re?versed astrocytic reactivity as well as expression of GDNF and GFAP in diabetic hippocampus and ameliorated diabetic cog?nitive dysfunction.

This study examined the analgesic effect of diprospan in rats with trigeminal neuralgia. Rat model of trigeminal neuralgic pain was established by loosely ligating the left infraorbital branch of the trigeminal nerve. After allodynia developed, the rats were randomly divided into 2 groups (n=20 in each): diprospan group, in which the rats received diprospan (7 mg/mL, 0.1 mL) injected to the left infraorbital foramen area; control group, in which saline (0.1 mL) was administered as the same manner as the diprospan group. The pain threshold (PT) in the left infraorbital area was measured before and 2, 6, and 8 weeks after the administration. The expression of neuropeptides [substance P, preprotachykinin A (PPTA), calcitonin gene-related peptide (CGRP)] in the trigeminal nerve was detected at the same time points as the PT measurement by immunohistochemistry or in situ hybridization method. The results showed that in the diprospan group, the PT was 10.65±1.26, 10.77±1.19 and 14.13±1.34 g 2, 6, and 8 weeks after the administration respectively, significantly higher than that before the administration (PT value: 0.36±0.11) (P<0.05 for each). In the saline group, the PT was 0.37±0.13, 0.66±0.09, 4.45±1.29 and 13.72±1.72 g before and 2, 6, and 8 weeks after the administration respectively with differences being significant between before and 6, 8 weeks after the administration (P<0.01). No significant difference existed in the PT between the diprospan group and the saline group at pre-administration (P>0.05). The PT in the diprospan group was significantly greater than that in the saline group 2 and 6 weeks post-administration (P<0.05). In the diprospan group, the expression levels of neuropeptides were significantly reduced as compared with those in the saline group 2 and 6 weeks post-administration (P<0.05). It was concluded that diprospan has an obvious analgesic effect on the trigeminal neuropathic pain partly by reducing the expression of neuropeptides in the trigeminal ganglia.

Objective To investigate the effects of total flavonoids of litchi chinensis sonn (TFL) on cell proliferation and the molecular mechanism in rat hepatic stellate cells (HSC-T6) activated by growth factor-β1 (TGF-β1). Methods HSC-T6 cells were treated by 0.25%Trypsin-EDTA and then were digested into single cell suspension by DMEM (10%FBS included), which were mixed with TGF-β1 (5μg/L). (1) MTT method was used to detect the proliferation of HSC-T6 cells. Cells were cultured in 96-well plate and were treated by different concentrations of TFL including TGF-β1 group, the control group (5‰DMSO included), and different concentrations of TFL groups (80, 160, 320, 640 and 800 mg/L TFL). Each group has three wells. The absorbance (A) value was measured by enzyme standard meter at the 490 nm wavelength after 24 h, 48 h and 72 h treatment. The cell inhibitory rate was calculated. The subsequent experimental drug concentration and drug treatment time were determined according to half inhibitory concentration (IC50). (2) The expression levels of NF-κB andα-SMA mRNA were detected by PCR (for mRNA) and Western blot assay (for protein). Cells were cultured in the 10 cm culture dish and were divided into different TGF-β1 groups, including TGF-β1 group, the control group (5‰DMSO included), and different concentrations of TFL groups (125, 250 and 500 mg/L TFL). After 48 h, related indicators were measured. Results At the same treatment time point, with the increased concentrations of TFL, A values were gradually decreased, and the cell inhibitory rates were gradually increased. There were no significant differences in the expressions of NF-κB andα-SMA mRNA between TGF-β1 group and control group. And there were no significant differences in the expressions of NF-κB andα-SMA mRNA between TFL125 group, TGF-β1 group and control group. There was a gradually decrease in the expressions of NF-κB andα-SMA mRNA and protein with the increased concentrations of TFL. Conclusion TFL can inhibit TGF-β1-induced HSC-T6 cell proliferation, which is involved in the inhibited expressions of NF-κB andα-SMA to anti-fibrotic effects in liver fibrosis.

Objective To study the risk prediction for new intracerebral hemorrhage (ICH) with high sensitivity C-reactive protein ( hs-CRP) level. Methods In a retrospective, nested, case-controlled study, 323 cases of ICH were identified and matched with 646 controls. The hs-CRP levels at baseline were compared between the two groups. The relevance of different hs-CRP levels and the risk of ICH were analyzed. Results The ICH group had a higher median hs-CRP levels (1.10 mg/L) as compared with the control group (0. 66 mg/L) with significant difference ( P<0.01 ). In addition, the increase of risk associated with hs-CRP levels was primarily observed in the individuals with the highest quartile of hs-CRP levels(>2.12 mg/L). These patients had an increased risk of ICH (OR 2. 58, 95% CI 1. 77 to 3. 76) as compared with those in the lowest quartile(≤=0.30 mg/L). Individuals with basiline hs-CRP levels above the specified cut point of 3 mg/L ormore and those in the 80th percentile were at a markedly increased risk of ICH (for specified cut point of 3 mg/L,0R2.26, 95% CI 1.60-3.20, P<0.01; for 80th percentile, OR 2.24,95% CI 1.60-3.13, P <0.01, respectively). Conclusions Risk of ICH might be predicted with the level of hs-CRP. With the increase of hs-CRP level at baseline, the risk of ICH was increased.

Objective To observe the distribution and influence factors of serum high sensitivity C-reactive protein (hs-CRP) in general population. Methods In a cross-sectional population survey, a total of 101 510 subjects who were employed by Kailuan Group had been carried out a healthy examination in the period of 2006 to 2007. In the statistical analysis, we observed 91 123 subjects (males 72 805, females 18 318) who had full information and met the inclusion criteria of the study. Results ( 1 ) The geometric means of hs-CRP were 0. 70 mg/L, 0. 70 mg/L and 0. 73 mg/L in all subjects, males and females,respectively, the 95th percentiles were 6.28 mg/L, 6.20 mg/L and 6.49 mg/L, respectively. The concentrations of hs-CRP increased with age in both males and females (P trend = 0. 001 ). Serum hs-CRP geometric mean was 0. 54 mg/L and the 95th percentile was 5.40 mg/L in health group, while the geometric mean was 0. 80 mg/L and the 95th percentile was 6. 57 mg/L in non-health group. (2) Multiple linear regression analysis showed that concentrations of hs-CRP were positively associated with gender, age,systolic blood pressure, body mass index, total cholesterol, triglycerides, fasting blood glucose, smoking history, history of coronary heart disease and stroke history, but concentrations of hs-CRP were inversely related with diastolic blood pressure, high-density lipoprotein cholesterol and alcohol history. Conclusion Serum concentrations of hs-CRP level increased with age, concentrations of hs-CRP were higher in females than males; a variety of cardiovascular factors effected the concentrations of hs-CRP.

Objective: To investigate the impact of improved AHA cardiovascular health behavior score (CHS) on short-time systemic blood pressure variability (SBPV) in elder population.
Methods: A total of 2464 participants ≥ 60 years from 3 hospitals of Kailuan area were taken for cohort study. The participants had no cardiovascular disease, not taking anti-psychotic drug, Parkinson treatment drug, anti-depression drug and analgesic drug within 2 weeks. All participants received 24-hour ambulatory blood pressure monitoring (ABPM) and the 24-hour, day-time, night-time SBPV were deifned by the standard deviation of 24-hour, day-time, night-time systolic blood pressure. The influence of CHS on SBPV was studied by multi-linear regression analysis. Improved cardiovascular health behavior and factors implied as changing the vegetable intake amount to salt amount by American Humane Association, 2010; boundary of BMI based on《Guidelines for prevention of overweight and obesity in Chinese adults》; status of exercise was deifed as the ideal status: ≥80 min/week, general status: < 80 min/week and bad status: no exercise.
Results: Finally, 1812 participants were recruited for survey and they were divided into 3 groups according to improved CHS: Group①, CHS (0-4) points,n=56, Group②, CHS (5-9) pointsn=1600 and Group③, CHS (10-14) points,n=156. The 24-hour SBPV in Groups①,②and③were 16.02 mmHg, 14.91 mmHg and 13.18 mmHg; day-time SBPV were 15.42 mmHg, 14.50 mmHg and 13.22 mmHg; night-time SBPV were 12.68 mmHg, 11.44 mmHg and 10.16 mmHg, allP<0.05. Multi-linear regression analysis indicated that with adjusted confounding factors, with 1 point of CHS elevation, the 24 hour-, day-time, night-time SBPV would reduce for 0.20 mmHg, 0.19 mmHg and 0.37 mmHg respectively, allP<0.05.
Conclusion: CHS was negatively related to short-time SBPV in elder population.

Objective To investigate the effect of SP600125 on the proliferation, cell cycle, apoptosis and invasion of human cervical cancer HeLa cells. Methods CCK-8 method was used to detect the proliferation of HeLa cells treated with different concentrations of SP600125 at different time points. The 20 μmol/L of SP600125 was determined for subsequent experiments. Cell proliferation ability was detected using plate clone formation assay; nuclear morphology was observed by DAPI staining; cell cycle and apoptosis were measured by flow cytometry; cell migration and invasion were detected by cell scratch and Transwell methods; the mRNA and protein levels of p53, Mad2L1 and CDC20 were measured by qRT-PCR and Western blot after SP600125 treatment at different time points. Results Compared with control group (0.1%DMSO), cells proliferative activity were reduced by 10, 20, 30, 40 and 50 μmol/L SP600125 treatment for 24h. Compared with control group, the rate of apoptosis was significantly increased in SP600125 treatment groups, and the cell proportion in G₂/M phase increased (P < 0.001), while the cell proportion in G₀/G₁ phases cells was reduced after SP600125 treatment for 24h and 48h (P < 0.001), and the clonal number, migration and invasion ability of HeLa cells also decreased significantly (P < 0.001). qRT-PCR and Western blot results showed a significant decrease in Mad2L1 mRNA and protein expression (P < 0.05) and a significant increase in p53 and CDC20 mRNA and protein expression (P < 0.01). Conclusion SP600125 can induce cell cycle arrest of cervical cancer HeLa cells in G₂/M phase by upregulating p53 and CDC20 and downregulating Mad2L1 expression, and promote cell apoptosis to inhibit cell proliferation, migration and invasion.

This study was aimed to identify pET21b-HPV16E2/BL21(DE3) strain and to optimize the expression of human papillomavirus type 16 (HPV16) E2 protein by orthogonal analysis. Four influence factors on two levels were selected to increase the target protein quantity. The four factors were induction time, induction temperature, inductor concentration and cell density. The quantity of HPV16 E2 protein was used as the evaluation parameter. Induced by IPTG, HPV16 E2 protein was analyzed by SDS-PAGE and Western Blot. Target protein was analyzed by GIS imaging system to quantify the protein level. SPSS13. 0 software was applied to analyze the result. Data showed that the expression strain pET211rHPV16 E2/BL21(DE3) was identified correctly. HPV16 E2 protein expressed mainly at insoluble form. The 42KD protein band was identified by SDS-PAGE and Western blot. Orthogonal test was applied on influence factor analysis and expression optimization successfully. Main influence factors were inductor concentration and induction temperature. The optimimum condition of maximum expression quantity was 37 degrees C, 7h, 1.0 mmol/L IPTG and OD600 1.0. In this experiment, orthogonal test could not only be used to analyze the influential factors and promote the target protein expression, but also be used to provide a better experiment method for molecular biological study.
DNA-Binding Proteins ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Human papillomavirus 16 ; metabolism ; Humans ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Papillomavirus Infections ; virology ; Recombinant Proteins ; biosynthesis ; genetics

This study examined the analgesic effect of diprospan in rats with trigeminal neuralgia.Rat model of trigeminal neuralgic pain was established by loosely ligating the left infraorbital branch of the trigeminal nerve.After allodynia developed,the rats were randomly divided into 2 groups (n=20 in each):diprospan group,in which the rats received diprospan (7 mg/mL,0.1 mL) injected to the left infraorbital foramen area; control group,in which saline (0.1 mL) was administered as the same manner as the diprospan group.The pain threshold (PT) in the left infraorbital area was measured before and 2,6,and 8 weeks after the administration.The expression of neuropeptides [substance P,preprotachykinin A (PPTA),calcitonin gene-related peptide (CGRP)] in the trigeminal nerve was detected at the same time points as the PT measurement by immunohistochemistry or in siru hybridization method.The results showed that in the diprospan group,the PT was 10.65± 1.26,10.77± 1.19 and 14.13± 1.34 g 2,6,and 8 weeks after the administration respectively,significantly higher than that before the administration (PT value:0.36±0.11) (P＜0.05 for each).In the saline group,the PT was 0.37±0.13,0.66±0.09,4.45±1.29 and 13.72±1.72 g before and 2,6,and 8 weeks after the administration respectively with differences being significant between before and 6,8 weeks after the administration (P＜0.01).No significant difference existed in the PT between the diprospan group and the saline group at pre-administration (P＞0.05).The PT in the diprospan group was significantly greater than that in the saline group 2 and 6weeks post-administration (P＜0.05).In the diprospan group,the expression levels of neuropeptides were significantly reduced as compared with those in the saline group 2 and 6 weeks post-administration (P＜0.05).It was concluded that diprospan has an obvious analgesic effect on the trigeminal neuropathic pain partly by reducing the expression of neuropeptides in the trigeminal ganglia.