Analysis of NK Cells and Receptors in Peripheral Blood of Patients with HBV Related Acute-on-chronic Liver Failure in Different Phases / 中山大学学报(医学科学版)

[Objective]To investigate the character of expression of NK cells and its receptors in peripheral blood of pa-tients with HBV-related acute-on-chronic liver failure(HBV-ACLF)in different phase.[Methods]There are thirteen pa-tients with HBV-ACLF in advanced phase group and thirty-three patients in plateau phase group,with thirteen healthy per-son in control group.The frequency of NK cells and expression of NK cell activating receptors(NKAR)including NKG2D, NKp30 and NKp46,NK cell inhibitory receptors(NKIR))cludingsd ate the character of including NKG2A,KIR2DL1, KIR2DL3 and KIR3DL1,NK cell killing function related factors including Perforin,GranzymeB and FasL in peripheral blood of all patients were detected by flow cytometry.[Results]The percentage of NK cells in advanced group was lower than plateau group and healthy control group(H=7.771,P=0.021).The expression of KIR3DL1 in healthy control group was high-er than advanced group and plateau group(Z=6.639,P=0.036)while the expression of FasL was significantly lower than ad-vanced group and plateau group(Z=22.5,P<0.001).[Conclusion]Patients in advanced group had lower frequency of NK cells,lower expression of inhibitory receptor KIR3DL1 and higher expression of FasL than patients in plateau group and healthy control group,which is associated with immune status of patients in different phase of liver failure.

To investigate the opportunity of entecavir treatment for HBeAg-negative acute-on-chronic liver / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore relations between the opportunities and effects of internal general treatment added Entecavir on acute-on-chronic liver failure (ACLF) of HBeAg-negative chronic hepatitis B in different score ranges of acute-on-chronic liver failure severity.</p><p><b>METHODS</b>A total of 108 ACLF of HBeAg-negative chronic hepatitis B patients with different ACLF severity score were treated with internal general treatment added Entecavir. The liver failure severity scores, HBV-DNA loads during the initiation of therapy, recovery phase and in deathbed phase, courses of Entecavir administration and mortalities were studied.</p><p><b>RESULTS</b>For 19 patients with high ACLF score (> or = 12), the difference in ACLF score between pre and post-treatment was not significant. The difference in HBV-DNA load between pre and post-treatment was not significant and the mortality was 18/19. For 30 patients with higher intermediate ACLF score (8-11), the difference in ACLF score between pre and post-treatment was not significant. The difference in HBV-DNA load between pre and post-treatment was significant, and the mortality was 66.67% (20/30). For 36 patients with lower intermediate ACLF score (5-7), the difference in ACLF score between pre and posttreatment was not significant. The difference in HBV-DNA load between pre and post-treatment was significant, and the mortality was 30.56% (11/36). For 23 patients with low ACLF score (< or = 4), the difference in ACLF score between pre and post-treatment was significant. The difference in HBV-DNA load between pre and post-treatment was significant, and the mortality was 8.70% (2/23).</p><p><b>CONCLUSIONS</b>A novel acute-on-chronic liver failure scoring system can syllabify differentiate the relations between the opportunities and efficacies on the Entecavir treatment for HBeAg-negative ACLF.</p>

Small interfering RNA targeting to hepatitis B virus X gene and 5-aza-2'-deoxycytidineon inhibited growth of the subcutaneous implanted tumor of hepatocellular carcinoma in nude mice / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the anti-tumor effect of small interfering RNA targeting to HBV X gene (X-siRNA) and 5-aza-2'-deoxycytidine (5-aza-dC) on HBV-related hepatocellular carcinoma.</p><p><b>METHODS</b>X-siRNA and control siRNA were synthesized. HepG2/GFP-HBx cells were treated with X-siRNA, and the levels of HBV X mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Nude mice were inoculated with HepG2/GFP and HepG2/GFP-HBx cells subcutaneous respectively to establish implant models of hepatocellular carcinoma, and were treated with X-siRNA, 5-aza-dC alone or in combination, and tumor growth was observed. The methylation of p16 gene promoter was detected by methylation specific polymerase chain reaction (MSP).</p><p><b>RESULTS</b>RT-PCR showed the expression of HBV X mRNA in HepG2/GFP-HBx cells was inhibited markedly by X-siRNA. The nude mice experiment showed that the gross tumor volume was much bigger in HepG2/GFP-HBx group than that in HepG2/GFP group (P < 0.05). The growth of palpable tumors in X-siRNA or 5-aza-dC treatment group notably decreased (P < 0.05). MSP analysis showed that p16 gene methylation was observed in HepG2/ GFP-HBx-caused palpable tumors, while no methylation was detected in HepG2/GFP group. However, after treatment with X-siRNA or 5-aza-dC, p16 gene methylation reduced.</p><p><b>CONCLUSIONS</b>HBV X-siRNA and methylation inhibitor can inhibit the growth of hepatoma cells via reversing p16 methylation.</p>

The relationship of HBeAg status with HBV DNA loads, MELD scores in patients with acute-on-chronic hepatitis B liver failure during terminal phases / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the relationship and clinical significances of HBeAg status with serum HBV DNA loads, model for end-stage liver disease (MELD) scores in patients with acute-on-chronic hepatitis B liver failure during terminal phase.</p><p><b>METHODS</b>120 fatal patients were enrolled. At three phases of 0 -14 d, 15-28 d and 29-90 d before death, they were detected serum HBeAg, HBV DNA loads order meanwhile MELD scores were calculated.</p><p><b>RESULTS</b>In 51 patients with HBeAg positive, HBV DNA levels were (5.25 +/- 1.99), (5.45 +/- 1.47) and (6.06 +/- 1.77) log10 copies/ml while MELD scores were (30.33 +/- 5.25), (26.36 +/- 6.43) and (20.13 +/- 6.47) respectively. In 69 patients with HBeAg negative,HBV DNA loads were (5.14 +/- 1.84), (5.49 +/- 1.75 ) and (4.62 +/- 1.65) log10 copies/ml while MELD scores were 32.38 +/- 9.95, 28.17 +/- 6.82 and 26.19 +/- 5.56 in sequence. Compared with the same phase between HBeAg-positive group and HBeAg-negative group, significant differences in both HBV DNA loads and MELD scores were found only at the phase of 29-90 d (P < 0.05). In multiple comparisons among three phases, regardless of the HBeAg status,there wasn't significant difference for HBV DNA loads (P > 0.05). But increasing MELD scores are associated with the disease exacerbation and significant differences were found (P < 0.05).</p><p><b>CONCLUSIONS</b>To initiate acute-on-chronic hepatitis B liver failure, serum HBV DNA loads of HBeAg-positive patients are higher than that of HBeAg-negative ones. Once ACLF has been initiated,sustained high HBV DNA loads may promote the disease worsened and be fatal regardless of the HBeAg status.</p>

Major constituent proteins in donkey hide and their interaction / 中国中药杂志

<p><b>OBJECTIVE</b>To analyze the constituent proteins in donkey hide, the key ingredient for Ejiao, an important traditional Chinese medicine for the blood-related conditions, in hope to eventually decipher the biochemical mechanism behind Ejiao's prominent medicinal efficacy.</p><p><b>METHOD</b>Two methods were employed to extract proteins in donkey skin. One used TriPure isolation reagent to extract the total proteins in donkey skin. Another used 1% sodium dodecyl sulfate (SDS) to heat the sample at 100 degrees C overnight. And then sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and capillary HPLC were used to analyze the component of proteins.</p><p><b>RESULT</b>There are not only collagen alpha1 (I) and collagen alpha2 (I), but also serum albumin in donkey skin. The content is over 25% in total proteins with the method of TriPure isolation reagent. The content of donkey serum albumin is up to 20% with the method of 1% SDS heating. And two bands, molecular weight are nearly 200 kDa,were found on 7.5% SDS-PAGE. Extracted these proteins to analyze with capillary HPLC, they were found to be the complex products of collagen and serum albumin of donkey.</p><p><b>CONCLUSION</b>Donkey serum albumin is a main protein component in the hide, which is a clue to expose is the effect of Ejiao on blood.</p>

Effects of glutamine granules on protein metabolism in trauma patients / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the effect of glutamine granules on protein metabolism in severe burns and trauma patients.</p><p><b>METHODS</b>120 patients with severe burns, multiple trauma and post operation who met the requirements of the protocol joined this double-blind randomized controlled, multi-center clinical trail. Patients were randomly divided into two groups: placebo control group (P group, 60 patients) and glutamine granules treatment group (GLN group, 60 patients). There was isonitrogenous and isocaloric intake in both groups, GLN and P group patents had been given glutamine granules or placebo (glycine) at 0.5 g.kg(-1).d(-1) for 7 days, respectively. The level of plasma glutamine, protein and urine nitrogen exclude were determined, wound healing rate of burn area and hospital stay were recorded, and then observed the complication and side effect.</p><p><b>RESULTS</b>After 7 days of taking glutamine granules orally, plasma GLN concentration was significant higher than that in P group (592.50 +/- 185.23 micro mol/L vs. 407.41 +/- 190.22 micro mol/L) (P < 0.01). Plasma prealbumin and transferrin in GLN group were significant higher than those in P group (P < 0.01), but the concentration of total protein and albumin were no marked changes compare with P group (P > 0.05). The capacity of urine nitrogen exclude in GLN group were significant lower than that in P group. Additional, the wound healing rate was faster and hospital stay days was shorter than P group (P < 0.05), and the occurrence of glutamine granules side effect was seldom.</p><p><b>CONCLUSION</b>Taking glutamine could promote protein synthesis, abate protein decompose, ameliorate wound healing rate and reduce hospital stay obviously.</p>