[Associations between gene polymorphisms of signal transducer and activator of transcription 3 and the susceptibility to hepatitis B virus related liver cirrhosis].

To investigate the associations between gene polymorphisms of signal transducer and activator of transcription 3 (STAT3) and liver cirrhosis (LC) after hepatitis B virus (HBV) infection. A case-control study was conducted in 243 patients with hepatitis B cirrhosis (HBV-LC, case group) and 486 HBV-infected subjects without LC (non-LC, control group) collected from January 2018 to September 2020 at the Changsha Central Hospital Affiliated to Nanhua University. Three single nucleotide polymorphisms (SNPs) of STAT3 gene, including rs4796793C>G, rs2293152C>G, and rs1053004T>C were selected through literature and biological information database, and the genotypes were detected by real-time fluorescent quantitative PCR (RFQ-PCR). The distribution differences of STAT3 SNPs genotypes between the two groups were compared using Chi-square test and haplotype analysis was conducted by Shesis online. The proportion of HBV C genotype in HBV-LC patients was significantly higher than that in the control group (80.91% vs. 70.79%, χ2=7.109, P=0.008), while the logarithm of ALT was significantly lower than that of the control group (1.78±0.43 vs. 1.95±0.54, t=3.801, P=0.000). The genotypes distributions of rs4796793, rs2293152, and rs1053004 were not significantly different between HBV-LC and non-LC in overall analysis and stratified analysis by gender (χ²=2.610, 1.505, 0.586, 2.653, 2.685, 1.583, 0.351, 5.388, 0.339, respectively, P>0.05 for each). Among the subjects infected with HBV genotype C, rs1053004 CC (vs. TT) significantly increased the risk of HBV-LC [odds ratio (OR) = 1.40, 95% confidence interval (CI): 1.03-1.91]. Among the HBV-infected subjects with HBeAg negative, rs4796793 GG genotype (vs. CC) and G allele (vs. C) significantly increased the risks of HBV-LC (OR = 2.17, 95%CI: 1.11-4.23; OR = 1.45, 95%CI: 1.06-1.97, respectively). Haplotypes analysis showed that the frequency of haplotype C-G-T composed of rs4796793, rs2293152, and rs1053004 was significantly lower in HBV-LC than that in the control group (non-LC) (27.3% vs. 35.6%, χ²=9.949, P = 0.001). The correlation between STAT3 and HBV-LC is different in HBV-infected subjects with different infection status. The HBV-infected subjects carrying haplotype rs4796793C-rs2293152G-rs1053004T of STAT3 gene have significantly decreased risk of LC.

The effect of platelet-derived microparticles in stored apheresis platelet concentrates on polymorphonuclear leucocyte respiratory burst.

BACKGROUND AND OBJECTIVES: Platelet-derived microparticles (PMPs) and other proinflammatory mediators, which are accumulated during the storage process, might induce transfusion adverse events. We hypothesized that the PMP primed polymorphonuclear neutrophil (PMN) respiratory burst after the transfusion, which could be linked to the transfusion-related acute lung injury (TRALI). MATERIALS AND METHODS: The PMPs were isolated by centrifugation of the platelet-free plasma from 10 apheresis platelet concentrates (A-PLTs) at 20,000×g for 1 h. The PMPs were counted by flow cytometric analysis, followed by Western blotting, that were performed on isolated PMPs. The soluble CD40 ligand (sCD40L, sCD154) was assayed with ELISA. The priming of the formyl-Met-Leu-Phe (fMLP)-activated PMN respiratory burst was measured with the hydrogen peroxide production. RESULTS: The PMP counts increased by 1·7-folds after 3 days of storage. Meanwhile, sCD40L also significantly increased in PMP fraction isolated from the 3-day stored A-PLTs. Furthermore, Western blotting indicated that sCD40L was involved and concentrated in PMP. The PMPs were able to effectively prime the fMLP-activated PMN respiratory burst, which was partly inhibited by CD154 monoclonal antibody or by filtration with 0·1 µM membrane. Significant relativity was existed between the PMP counts, sCD40L and priming activity during the A-PLT storage. CONCLUSION: The platelet-derived microparticles, which carried the sCD40L, accumulated in the apheresis platelet concentrates during the 5 days of storage. They primed the fMLP-activated PMN respiration burst, which might be relative to TRALI.

Introducing a dexamethasone mouse model for monitoring in vivo viability of stored human platelets.

OBJECTIVES: The aim of this study is to establish an available animal model which can evaluate in vivo viability of stored human platelets (HuPLTs). BACKGROUND: The viability in vivo of HuPLTs was usually evaluated by transfusing HuPLTs into animals before clinical trials. It is necessary to develop a method which may slow down rapid clearance of HuPLTs from circulation of the animal. METHODS: Carbon clearance tests were performed by treating mice with dexamethasone (DEX) to determine the phagocytic ability of the mice macrophages. HuPLTs in mice whole blood were detected by flow cytometric analysis with mouse anti-human CD41-fluorescein isothiocyanate monoclonal antibody. Recovery and survival of the HuPLTs stored at 22 °C for 1 day were evaluated after transfusing these HuPLTs into DEX-treated mice, and compared with those either stored at 22 °C for 5 days or at 4 °C for 1 day. RESULTS: Corrected phagocytic indexes of DEX-treated mice decreased significantly compared with those of control mice (P < 0.05). The recovery after 24 h and survival time of fresh HuPLTs in DEX-treated mice were much higher than those in control mice (P < 0.01). After transfused into the DEX-treated mice, HuPLTs stored either at 22 °C for 5 days or at 4 °C for 1 day showed decrease in recovery and survival compared with those stored at 22 °C for 1 day (P < 0.05). CONCLUSION: Dexamethasone slows down the rate of HuPLTs clearance efficiently in mouse circulation. And the DEX-treated mouse model was able to evaluate the in vivo viability of stored HuPLTs.

Active oxygen radicals produced by leukocytes of malignant lymphoma.

The electron spin resonance (ESR) spectra of active oxygen radicals produced during the respiratory burst of PMA-stimulated leukocytes and oxygen consumption are studied by ESR spin trapping and spin probe oxymetry for 31 times in 17 cases of malignant lymphoma. The results showed that the spectra produced from PMA-stimulated patients' leukocytes were predominantly spin adducts of DMPO-hydroxyl radicals (DMPO-OH). The decrease in oxygen consumption during respiratory burst suggested that respiratory burst function was inhibited. Kinetic observations of patient condition showed that respiratory burst and oxygen consumption were improved and even approached normal, when the patient was in remission or the condition was ameliorated. This paper is the first of its type and helps clarify the mechanism of malignant proliferation and metastasis.

[Effect of cyclophosphamide on the bone marrow hematopoiesis in the mouse].

This paper reports the effect of cyclophosphamide on the bone marrow hematopoiesis in the mouse. Cyclophosphamide 0.12 mg/g body weight was injected into the mice once and the observation lasted for 2 weeks. After the injection, peripheral leukocytes were reduced to the lowest level on day 4 and then increased higher than the control on day 7 to 14. The number of nucleated cell in the bone marrow was the lowest at the 48th hour and gradually became normal within two weeks. The pluripotent hemopoietic stem cells--CFU-S (colony forming unit-spleen) were depleted abruptly in 24 hours, then reproliferated exponentially to a peak on day 3, followed by a second decrease and came back to normal level on day 11 to 14. The changes of granulocytic progenitor cell CFU-D (colony forming unit-diffusion chamber) and CFU-C (colony forming unit-culture) were quite similar to that of CFU-S but their proliferation peak was on day 4. The peripheral leukocyte drop was slower and the return to normal was earlier than the hemopoietic cells. So the recovery of leukocyte count does not mean a real reconstruction of hematopoiesis. The bone marrow stroma observed by CFU-F (colony forming unit-fibroblastoid) assay and marrow microcirculation were also damaged and did not recover to normal during the observation. The bone marrow stroma and microcirculation showed a more serious damage.

[Role of prostaglandin E in modulation of granulopoiesis by marrow stroma].

The stroma layer (SL) cultured from mouse bone marrow cells for one week, had an inhibitory effect on the granulocyte-macrophage progenitor cells (CFU-GM). The inhibitory effect decreased in the 2nd week SL, while the 3rd week SL promoted CFU-GM growth greatly. When indomethacin (1 x 10(-7) mol/L) was added into the CFU-GM culture system, the score of CFU-GM on the 1st week SL was raised, and on the 2nd week SL was further increased significantly, but it leveled off on the 3rd week SL. The addition of exogenous 10(-8) mol/L PGF1 suppressed the CFU-GM growth on all the 1st to 3rd week SLs. When 1 x 10(-8) mol/L PGE1 was added with 2 x 10(-7) mol/L indomethacin, 1st week SL-CFU-GM increased to 42.61%, 2nd week SL-CFU-GM nearly to 100% of the control. For the 3rd week SL-CFG-GM, 1 x 10(-7) mol/L indomethacin was enough to reverse the inhibition induced by exogenous 1 x 10(-8) mol/L PGE1. It is suggested that definite amount of PGE was produced from cells in the 1st week SL. The secretion of PGE from SL was reduced during 2nd week, and almost stopped in the 3rd week SL. The results indicate that the modulation of granulopoiesis by marrow stroma is, at least partly, mediated by PGE.