Value of Serum Procalcitonin and Percentage of Neutrophils in Early Diagnosis of Bacterial Infection in Decompensated Liver Cirrhosis / 中山大学学报(医学科学版)

[Objective] To explore the value of serum procalcitonin (PCT) and percentage of neutrophils (NE％) in early diagnosis of bacterial infection in decompensated liver cirrhosis.[Methods] The clinical data of 244 in the patients with decompensated liver cirrhosis in the Department of Infectious Diseases,The third Affiliated Hospital of Sun Yat-sen University from January 2016 to June 2017 were retrospectively analyzed.According to whether complicated with bacterial infection,patients were divided into infection group (n=120) and non-infection group (n=124).The infection group consisted of 60 cases with spontaneous bacterial peritonitis (SBP),25 cases with pulmonary infection,16 cases with bloodstream infection,19 cases with other kinds of infections.The levels of PCT,peripheral white blood cells (WBC),peripheral neutrophils (NEUT),percentage of neutrophils (NE％) and ratio of WBC to platelets (PLT)(WBC/PLT) in each group were compared.The receiver characteristic curve (ROC) was applied and area under curve (AUROC),sensitivity and specificity were computed.The parallel test was applied to explore the diagnostic value of PCT with NE％.[Results] The levels of PCT,WBC,NEUT,NE％ and WBC/PLT in infection group were higher than those in non-infection group (P＜0.05).The levels of PCT,NE％ and WBC/PLT in bloodstream infection group were higher than in non-blood-stream infection group (P＜0.05).The AUROC of the PCT was 0.947 (95％CI=0.922～0.971).When PCT was≥0.38 ng/mL,the sensitivity,specificity and Youden index(YDI) were 76.7％,97.6％,and 0.742.The AUROC of NE％ was 0.806 (95％CI=0.751～0.861).With a cutoff value of 75.0％ in NE％,the sensitivity and specificity were 83.3％ and 92.7％,respectively.[Conclusion] Serum PCT,with a suggesting cut-off value of 0.38 ng/mL,could be used as an marker for early diagnosis of bacterial infection in decompensated liver cirrhosis.PCT combined with NE％ can improve diagnostic accuracy.

Simultaneous separation of primary cardiomyocytes and cardiac fibroblasts from neonatal rats with density gradient centrifugation / 生理学报

To improve a fast and high-quality isolation method for culturing the primary cardiomyocyte and fibroblast in vitro, the neonatal Wistar rats were decapitated accordingly and left ventricles were isolated under the sterile condition. The ventricles were chopped and digested in the enzyme solution containing 0.5 mg/mL type II collagenase. During this process, the digesting time, frequency and stirring speed, centrifuging frequency and speed were strictly controlled. The cardiomyocytes were separated from the cardiac fibroblast by using the Percoll density gradient centrifugation. The cell viability was tested by staining with 0.2% trypan blue. The purity of cardiomyocytes and fibroblasts were determined by immunoflourescent staining with anti-cTnI, anti-Vimentin and anti-α-SMA antibodies. The results indicated that with this protocol, the viability and purity of cardiomyocytes were 92% and 95%. The automobile pulse of the adhered cardiomyocyte was visible. For fibroblasts, the cell viability and purity were 96% and 94%. Our results demonstrate that this advanced isolation method is reproducible, and can simultaneously produce high-quality primary cardiomyocytes and fibroblasts for the future study.

Evaluation of the efficacy and prognostic factors for colorectal liver metastases treated with transcatheter arterial chemoembolization / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to evaluate the therapeutic efficacy and to determine the prognostic factors of TACE in patients with colorectal liver metastases (CRLM).</p><p><b>METHODS</b>The clinical data of 183 patients with unresectable CRLM treated with TACE from Jan. 2002 to Dec. 2008 were retrospectively reviewed. Log-rank method was used for univariate analysis and Cox proportional hazard model was used for multivariate analysis of the prognostic factors.</p><p><b>RESULTS</b>The median survival time was 22 months, and the 0.5-, 1-, 2-, 3-, 5-year survival rates were 93.9%, 81.1%, 39.8%, 18.2%, and 3.9%, respectively. Multivariate analysis showed that tumor involved more than one lobe of the liver, and elevated CEA and CA19-9 levels were independent risk factors for the overall survival (P < 0.01). Females, more times of TACE, combination with regional therapy and received phase II resection were related with a good survival (P < 0.01) in CRLM patients after TACE treatment.</p><p><b>CONCLUSIONS</b>Transcatheter arterial chemoembolization is an effective therapy for unresectable colorectal liver metastases. Patients with tumor spread more than one lobe of the liver, high CEA and CA19-9 levels are independent poor prognostic factors. Females, patients received more times of TACE, combined with regional therapy and received phase II resection may have a good survival.</p>

TNFα induced IL-8 production through p38 MAPK- NF-kB pathway in human hepatocellular carcinoma cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To identify the role of p38 MAPK- NF-kB signaling pathway in TNF-α induced IL-8 production in human hepatocellular carcinoma cells.</p><p><b>METHODS</b>The concentrations of IL-8 from MHCC-97H cells were measured by an enzyme-linked immunosorbent assay (ELISA). The phosphorylation of p38 MAPK was analyzed by Western blot and immunofluorescence. NF-kB p65 protein nuclear translocation was determined by non-radioactive NF-kB p50 / p65 transcription factor activity kit and immunofluorescence.</p><p><b>RESULTS</b>The IL-8 production from MHCC-97H cells challenged with TNFa significantly increased in a time-dependent (F = 144.04, P < 0.01) and dose-dependent (F = 364.14, P < 0.01) manners, as compared with those without TNFa challenge. TNFa up-regulated the phosphorylation levels of p38 MAPK and increased the translocation of NF-kB p65 protein into the nucleus, also proved by immunofluorescence staining. p38 MAPK inhibitor (SB203580) could significantly inhibit IL-8 production in a dose-dependent manners (F = 65.47, P < 0.01), and partially inhibited NF-kB p65 nuclear translocation in a dose-dependent manner (F=141.20, P < 0.05).</p><p><b>CONCLUSION</b>TNF-α could increase the production of IL-8 in MHCC-97H cells and p38 MAPK- NF-kB pathways seem to play a central role in the regulation of IL-8 production.</p>

Involvement of ATP-sensitive potassium channels in proliferation and differentiation of rat preadipocytes / 生理学报

This paper was aimed to investigate the effects of ATP-sensitive potassium channels on the proliferation and differentiation of rat preadipocytes. We examined the expression of sulphonylurea receptor 2 (SUR2) mRNA in preadipocytes and adipocytes obtained by inducing for 5 d and the effects of the inhibitor (glibenclamide) and opener (diazoxide) of ATP-sensitive potassium channels on the expression of SUR2 mRNA in preadipocytes by real-time PCR. Preadipocyte proliferation and cell cycle were measured by MTT spectrophotometry and flow cytometer. The content of intracellular lipid was measured by oil red O staining, cell diameter was determined by Image-Pro Plus 5.0 software and the expression of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) mRNA was estimated by RT-PCR. SUR2 mRNA was expressed in both preadipocytes and adipocytes obtained by inducing for 5 d, and the expression in adipocytes was obviously higher than that in preadipocytes. Glibenclamide inhibited the expression of SUR2 mRNA in preadipocyte, promoted preadipocyte proliferation in a dose-dependent manner, increased the cell percentages in G(2)/M + S phase, increased lipid content, augmented adipocyte diameter, and promoted the expression of PPAR-gamma mRNA. But the actions of diazoxide were contrary to those of glibenclamide. These results suggest that ATP-sensitive potassium channels regulate the proliferation and differentiation of preadipocytes, and PPAR-gamma is probably involved in the effect of ATP-sensitive potassium channels.

Inhibition of vascular endothelial growth factor gene expression by T7-siRNAs in cultured human retinal pigment epithelial cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Retinal pigment epithelial (RPE) cells play an important role in the occurrence of choroidal neovascularization (CNV). Vascular endothelial growth factor (VEGF) as a positive regulatory growth factor is produced by the RPE in an autocrine or paracrine manner, promoting CNV development. Duplexes of 21 nt RNAs, known as short interfering RNAs (siRNAs), efficiently inhibit gene expression by RNA interference when introduced into mammalian cells. We searched for an efficient siRNA to interfere with VEGF expression in RPE cells and shed light on the treatment of CNV.</p><p><b>METHODS</b>Human primary RPE (hRPE) cells were cultured and identified. Three pairs of siRNAs were designed according to the sequence of VEGF 1-5 extrons and synthesized by T7 RNA polymerase transcription in vitro. To evaluate the inhibitory activity of T7-siRNAs, hRPE cells were transfected via siPORT Amine. The interfering effect of T7-siRNAs in hRPE cells was examined by semiquantitative reverse transcription-polymerase chain reaction and immunofluorescence.</p><p><b>RESULTS</b>Three pairs of T7-siRNAs synthesized by in vitro transcription with T7 RNA polymerase suppressed VEGF gene expression with efficiency from 65% to 90%. T7-siRNA (B), targeted region at 207 nt to 228 nt and double stranded for 21 nt with 2 nt UU 3' overhangs, was the most effective sequence tested for inhibition of VEGF expression in hRPE cells. Compared with nontransfected cells, the mean fluorescence in hRPE cells transfected with T7-sRNAs was significantly less (P < 0.01). siRNA with a single-base mismatch and ssRNA(+) did not show suppressing effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect (5 to 10 pmol).</p><p><b>CONCLUSION</b>T7-siRNA can effectively and specifically suppress VEGF expression in hRPE cells and may be a new way to treat CNV.</p>