[Multi-omics research contributes to early screening, diagnosis and treatment of liver cancer].

In 2016, the World Health Organization set an ambitious goal of reducing viral hepatitis-related deaths by 65% by 2030. The key to this goal is to reduce viral hepatitis-related HCC deaths. Liver cancer is the fourth most common malignant tumor and the second leading cause of cancer death in China. The onset of HCC is insidious, and most patients are already in the middle and late stage when diagnosed. Despite the great progress on management of HCC, the therapeutic effect and prognosis of HCC are still unsatisfactory. Therefore, multi-dimensional and comprehensive analysis of the mechanism of liver cancer, improving the early screening, diagnosis and treatment rate of liver cancer are the key points of reducing the harm of liver cancer in China. In recent years, multi-omics studies have been widely applied in the field of liver cancer, providing a basis for the pathogenesis of liver cancer, early detection and diagnosis, development of individual treatment strategies and prognosis assessment. This issue will focus on the application of genomics, proteomics, metabolomics and imaging omics in early screening, diagnosis and treatment of liver cancer.

[Proteomic-driven precision medicine research in hepatocellular carcinoma].

Proteomics is the study of proteins-the direct executor of life activities. Protein plays a vital role in the development and growth of human body and the genesis and development of diseases. It is the most widely used clinical marker type and the direct drug target. In recent years, the advance of proteomic core technology with chromatography and mass spectrometry has promoted the rapid development in the depth and breadth of proteomic research, and its application in the large cohort of hepatocellular carcinoma opens a new era of proteomics-driven precision medicine (PDPM). This review highlights the proteomic research in new techniques, directions and discoveries of hepatocellular carcinoma research in recent years, providing new ideas and references for clinicians to understand proteomics, and to use proteomics to assist in the diagnosis of diseases and the development of personalized therapies.

[Combination of PP242 and dasatinib suppresses the progression of acute myeloid leukemia in a mouse model].

Objective: To establish the acute myeloid leukemia (AML) mouse model, and to preliminarily investigate the efficiency of dasatinib, a tryosine kinase inhibitor, and PP242, an inhibitor of PI3K/Akt/mTOR signaling pathway in the development of AML. Methods: The lineage(-) (Lin(-)) cells of C57BL/6J were transduced with retrovirus carrying MSCV-MLL-AF9-IRES-GFP fusion gene. The transduced cells were transplanted into lethally irradiated recipient mice to induce AML, and then the AML mouse model were established. The leukemia mice were treated with vehicle, dasatinib, PP242, dasatinib+ PP242, separately. The survival of the recipient mice was observed and the percentage of leukemia cells in peripheral blood (PB) and bone marrow (BM) was examined every four days. The apoptosis rates and cell cycle status of leukemia cells were also examined by FLOW. The leukemia cells in different group were sorted, the mRNA of these leukemia were extracted and reverse transcripted for related gene expression by qRT-PCR. The molecular mechanism of supression of leukemia cells growth was studied via RNA-Seq experiments. Results: Compared with control group, either PP242 or dasatinib could prolong the survival rate of recipient mice, however, the combination treatment AML mice with PP242 and dasatinib prolonged the life span of AML mice more significantly. The combination of PP242 and dasatinib could decrease the percentage of leukemia cells in PB and BM more significantly, arresting more leukemia cells in G0 phase, inducing more apoptosis of leukemia cells. Conclusion: Combination of tryosine kinase inhibitor-dasatinib and PI3K/Akt/mTOR signaling pathway inhibitor- PP242 could delay the progression of AML by inducing more leukemia to apoptosis and arrest more leukemia cells in the cell cycle G0 phase, it may be provied a new method for clinical therapy.

Exploring rat plasmatic proteomes: what triggered the liver regeneration?

To further clarify the priming mechanism of liver regeneration, proteins and protein complexes from rat plasma (normal group, partial hepatectomy (PHx) group and sham-operation group) were comparatively studied by two-dimensional gel electrophoresis and two-dimensional blue native gel electrophoresis. Our results suggested that Kupffer cell--NF-kappaB/ROS might trigger the liver regeneration.

[Coding region cDNA sequence cloning of rat neuroglobin gene, its polymorphism feature and tissue expression profile analysis].

The coding region cDNA sequence of rat neuroglobin (NGB) was obtained by RT-PCR technique using a degeneracy PCR primer pair based on previously reported cDNA sequence of human and mouse NGB gene. Result demonstrated that the coding region cDNA sequence of rat NGB gene is 456 bp in length, which could encode a protein of 151 amino acids. The rat NGB gene is highly homology with mouse (96%) and human (88%) NGB gene. However, several polymorphism sites were also detected in the rat NGB coding region: 113 t/c [L38P], 133 a/g [N45D], 388 a/g[R130G], 417 t/c. The cDNA sequence of rat NGB gene has been registered in GenBank under the accession number AF333245. Moreover, highly expression level of rat NGB in brain, liver, kidney, heart and skeletal muscle was detected by using multiple tissue RT-PCR technique, indicating the functional importance of this novel gene.

[A new melanoma antigen-encoding gene subfamily in human chromosome X].

A lot of studies have been focused on the tumor-related genes. We have cloned a new melanoma antigen-encoding gene (MAGE) from human fetal liver of third trimester and subsequently found that it represented a new MAGE gene subfamily, named MAGE-D. The MAGE-D subfamily contained three orthologs including human MAGE-D1, rat SNERG-1 and mouse DLXIN-1, and two paralogs including human MAGE-D and human KIAA1114. All human members of MAGE-D subfamily are located in human chromosome Xp11.21-p11.23. Moreover, MAGE-D subfamily stands out from other known subfamilies MAGE-A, -B and -C of MAGE family in view of typical features such as exon/intron organization, absence of tumorspecific antigens, evolutionarily divergent in sequences. The identification of MAGE-D subfamily will be helpful in understanding the genesis of tumor.

[cDNA cloning and sequencing of human urokinase receptor].

Human urokinase receptor (uPAR), a 55 kD glycoprotein linked to the cell membrane by a glycosylphosphatidylinositol anchor, plays a central role in cell migration and tissue remodeling. The human uPAR cDNA was cloned from a highly metastatic human lung giant cell line PG by RT-PCR and then subcloned into pGEM-T vector and sequenced. The data indicate that there are three bases substitution (705, 746, 755) which subsequently leads to two amino acid mutation (249, 252) compared to that of previously reported. The cDNA sequence of uPAR was registered in GenBank with accession number AF257789.

[Cloning of human TRAIL cDNA and its expression in COS-7 cells].

The human TRAIL cDNA was amplified with the total RNA from the human acute promyelocytic leukemia cell line HL-60 by means of RT-PCR, and was cloned into the pGEM-T vector. The DNA sequence analysis showed that it was consistent with the published sequence. Then, the insert of human TRAIL cDNA was subcloned into the mammalian expression vector pcDNA3. The hybrid plasmid pcDNA3-hTRAIL was transformed into COS-7 cells, and transiently expressed in the COS-7 cells. The activity of the expressed product could induce apoptosis in U937 cell line.

[Construction of fusion recombinant plasmid coding for RGD peptide and urokinase B chain, its expression in Escherichia coli and preliminary characterization of its biological activity].

A synthetic RGD (Arg-Gly-Asp) peptide-coding sequence was fused with urokinase B-chain cDNA and then cloned into the prokaryotic expression vector pBV220. The fused gene was expressed in E. coli DH5 alpha under the control of PRPL promoter by 42 degrees C induction. The expression level of the fusion protein was over 9.2% of the total bacterial proteins as a or of inactive inclusion body. The purified fusion protein was obtained with similar antigenicity as urokinase shown by Western blotting. Its in vitro fibrinolysis and anti-platelet aggregation activity was also evaluated by bioassay.

Assessment of tissue blood flow following small artery welding with an intraluminal dissolvable stent.

Using the technique of radioactive 51Cr-labeled biological microspheres, this study evaluated arterial blood flow following small vessel anastomosis by CO2 laser welding and a dissolvable stent in the lumen. A total of 30 Sprague-Dawley rats were divided into two groups. Group A: 11 rats had their femoral arteries ligated on one side. The contralateral side served as a control, with the artery transected and repaired using conventional microsuturing. Group B: 19 rats had their femoral arteries transected and repaired using CO2 laser welding and an intraluminal dissolvable stent technique. The contralateral side was again used as a control using conventional microsuturing. At 1 hr postoperatively, 51Cr-labeled biological microspheres were injected centripetally into the left common carotid artery and the legs and thighs immediately harvested for measurement of radioactivity. All repaired arteries were patent (30/30 in the microsuturing group and 19/19 in the stented welding group), with no detectable stenosis or dilation at the repaired site. Statistical analysis showed that tissue radioactivity (cpm/g) in the ligated group (3,972 +/- 384 in thighs and 3,142 +/- 742 in legs) was significantly lower than in the microsuturing group (7,132 +/- 1,723 in thighs and 6,557 +/- 1,469 in legs) (P < 0.01). In the ligated group, a significant reduction of blood flow was seen in the legs when compared with the thighs (P < 0.05). There was no significant difference in radioactivity when comparing the microsuturing control with the stented welding group, in both thighs (7,064 +/- 2,599 and 7,006 +/- 2,406, respectively; P > 0.05) and legs (6,386 +/- 1,703 and 6,288 +/- 1,757, respectively; P > 0.05). This study provided evidence that the dissolvable stent placed intraluminally does not impair blood circulation and that when coupled with CO2 laser welding offers a high-quality alternative to conventional small vessel anastomosis.

[Biological activity of recombinant human hepatopoietin].

We examined the effects of rhHPO on the cell growth and DNA synthesis of both rat primarily cultured hepatocytes and hepatic carcinoma cell line in vitro by MTS and 3H-TdR in corporation methods. It was indicated that rhHPO is an important stimulating factor of regeneration, which may be developed as a potential drug for the treatment of severe hepatic diseases. We also found an inhibitory effect of rhHPO on the DNA synthesis of lung cancer cell lines GLC-82 in vitro, which might provide a valuable indicator for the study of its specificity and mechanisms.

[Studies on characteristics of proliferation and differentiation of hematopoietic stem cells in mice].

By using Y chromosome specific sex-determining region (Sry) as a new cytogenetic marker and PCR technique, the characteristics of proliferation and differentiation of hematopoietic stem cells in mice were studied. Bone marrow cells from male mice were injected into lethally-irradiated female mice, PCR results indicated that all of the CFU-S were originated from donor. It was found that CFU-S was able to proliferate and differentiate into various hematopoietic cells in vivo during its transplantation. Whereas the fibroblasts within donor CFU-S and the fibroblasts from bone marrow in recipient mouse reestablished by donor CFU-S were shown to be originated from the recipient. The above data demonstrated that CFC-S from bone marrow in mice possessed the characteristics of hematopoietic stem cells but was not prone to differentiate into fibroblasts.

[Studies of a 35 KDa substance from human fetal liver on the regulation of hematopoiesis].

About 30%-40% of hematopoietic stem cells in human fetal liver of 3-5 months are in S phase of cell cycle, much higher than the ratio of 10% of that in adult bone marrow. The existance of highly active hematopoietic stem cell proliferation stimulators is probably its molecular basis. CFU-S "suicide rate" in rats was adopted to detect the effective substance. Through several steps of separation, we obtained a relatively purified substance of 35 kDa, termed it as FLS-4. CD 34 positive cord blood cells were sorted and assayed for their response to FLS-4 in 3H-TdR incorporation assay. The response to FLS-4 alone was approximately 1 times the background response seen with no factor added. In combination with IL-6 and IL-3 produced response that was 2.9 and 6.5 fold respectively greater than that observed with no factor added, but was weakly in comparison with the effects of SCF. In combination with GM-CSF or IL-3, FLS-4 can stimulate the formation of blast-colonies. The results indicate that the FLS-4 is very likely to be a novel hematopoietic stem cell proliferation stimulator. In physical or biological characteristics, it exhibited a unique character different from IL-3, IL-6, GM-CSF, SCF or FLT3 ligand those are known to have hematopoietic stem cell proliferation stimulating activity. During the period of active hematopoiesis in fetal liver, FLS-4 might be the candidate in triggering hematopoietic stem cells from resting G0 to S phase.

[In vitro stimulation of HTC hepatoma cell growth by recombinant human augmenter of liver regeneration (ALR)].

It was demonstrated that biologically active ALR could be expressed from its cDNA in transient expression experiments in cos-7 cells. The results showed that the cytosolic fraction from the transfected cells with constructed plasmids DNA stimulated of DNA synthesis of in vitro HTC cells in a dose dependent manner. This finding suggests that the HTC hepatoma cell line may be used as a target for bioassay of human ALR in vitro.

[Increase in the level of augmenter of liver regeneration mRNA in the rat regenerating liver after partial hepatectomy].

Augmenter of liver regeneration (ALR) is a novel hepatic stimulator. After 70% of the rat liver was removed, ALR-like activity in the remnant liver began to increase within 24 h. In parallel with the activity, the ALR mRNA level in the remnant liver increased 12 h after the operation and reached a maximum in 24 h. These findings indicate that the liver itself produces ALR, which may be a hepatotropic factor acting as a trigger for liver regeneration.