Association of mRNA expression level of IP-10 in peripheral blood mononuclear cells with HBV-associated acute-on-chronic liver failure and its prognosis.

HBV-associated acute-on-chronic liver failure is prevalent in mainland China. The prognosis of HBV-ACLF is poor. The mortality of HBV-ACLF is approximately 80%. Therefore, a prognostic indicator was needed in order to allow us to intervene as soon as possible. The model for end-stage liver disease (MELD) scoring system is widely used to predict the prognosis of liver failure. However, the assessment is too complex to restrict its application. This study aimed to investigate the expression of IP-10 in peripheral blood mononuclear cells (PBMC), in order to explore the relationship between the expression and prognosis of patients with HBV-ACLF. The mRNA level of IP-10 in PBMCs were analyzed in 80 patients with HBV-ACLF, 40 patients with chronic hepatitis B (CHB) and 40 healthy people by fluorescent quantitative PCR. IP-10 mRNA level was significantly higher in the HBV-ACLF group than in the other two groups (P<0.01). Group with MELD score below 30 had lower IP-10 mRNA level than group with MELD score over 30 (P<0.05). The IP-10 mRNA level in PBMCs in positive group was higher than that in negative group (P<0.01). With a threshold of 0.925, the area under the receiver operating characteristic (ROC) curves was 0.815. These findings suggest that assessment of IP-10 mRNA level in the PBMCs would be helpful for evaluating the disease severity and prognosis in patients with HBV-ACLF.

Association of mRNA Expression Level of IP-10 in Peripheral Blood Mononuclear Cells with HBV-associated Acute-on-chronic Liver Failure and Its Prognosis / 华中科技大学学报（医学）（英德文版）

HBV-associated acute-on-chronic liver failure is prevalent in mainland China.The prognosis of HBV-ACLF is poor.The mortality of HBV-ACLF is approximately 80％.Therefore,a prognostic indicator was needed in order to allow us to intervene as soon as possible.The model for end-stage liver disease (MELD) scoring system is widely used to predict the prognosis of liver failure.However,the assessment is too complex to restrict its application.This study aimed to investigate the expression ofIP-10 in peripheral blood mononuclear cells (PBMC),in order to explore the relationship between the expression and prognosis of patients with HBV-ACLF.The mRNA level of IP-10 in PBMCs were analyzed in 80 patients with HBV-ACLF,40 patients with chronic hepatitis B (CHB)and 40 healthy people by fluorescent quantitative PCR.IP-10 mRNA level was significantly higher in the HBV-ACLF group than in the other two groups (P＜0.01).Group with MELD score below 30 had lower IP-10 mRNA level than group with MELD score over 30 (P＜0.05).The IP-10 mRNA level in PBMCs in positive group was higher than that in negative group (P＜0.01).With a threshold of 0.925,the area under the receiver operating characteristic (ROC) curves was 0.815.These findings suggest that assessment of IP-10 mRNA level in the PBMCs would be helpful for evaluating the disease severity and prognosis in patients with HBV-ACLF.

[Correlation of -31G/C polymorphisms of survivin promoter to tumorigenesis of gastric carcinoma].

BACKGROUND & OBJECTIVE: Survivin, a bifunctional protein that regulates cell division and suppresses apoptosis, may play an important role in tumorigenesis. This study was to determine the correlations of survivin gene 31-GC polymorphisms to the occurrence of gastric carcinoma and survivin mRNA expression. METHODS: The -31G/C single nucleotide polymorphism (SNP) of survivin promoter in peripheral blood samples from 96 gastric carcinoma patients and 67 healthy subjects was analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and gene sequencing. Survivin mRNA expression in gastric carcinoma tissues was detected by real-time quantitative reverse transcription-polymerase chain reaction (RQ-RT-PCR). RESULTS: The genotype frequencies for -31G/G, -31G/C and -31C/C were 20.84%, 39.58% and 39.58% in gastric carcinoma patients, and 46.26%, 41.80% and 11.94% in healthy subjects, respectively. The frequencies of survivin-31C allele and C/C genotype were significantly higher in gastric carcinoma patients than in healthy subjects [59.37% vs. 32.84%, Chi2 = 22.26, P<0.005, odds ratio (OR)=2.98, 95% confidence interval (CI)=1.96-4.51; 39.58% vs. 11.94%, Chi2 =14.88, P<0.005, OR=4.83, 95% CI=2.91-8.03], but survivin mRNA was overexpressed with no significant difference in gastric cancer tissues subgrouped by genotypes. CONCLUSION: The -31C genotype of survivin promoter is associated with gastric cancer, and may be a risk factor of gastric carcinoma.

Rapid quantification of hepatitis B virus DNA by direct real-time PCR from serum without DNA extraction.

The purpose of this study was to quantify hepatitis B virus DNA by direct real-time PCR from serum without the need for DNA extraction. Crossing point (Cp) values were determined automatically using the second derivative maximum mode. Since serum samples from patients are inevitably haemolysed, lipaemic or icteric, the interference of endogenous substances from the serum in real-time PCR was evaluated. The result showed that, although serum protein quenched the intensity of fluorescence, the Cp value adopted to calculate the quantity of DNA copies remained unchanged. Importantly, real-time PCR from serum with or without DNA extraction reached a high level of concordance. This direct serum PCR method without the DNA extraction and gel electrophoresis allows for substantial labour and cost savings. In addition, it is also suitable for rapid DNA quantification during clinical diagnosis.