Abstract:The endless emergence of new theories and scientific instruments aiming to genome sequencing boosts the large ship of history advance of biology in the post-genomic era．With the genome sequences of major infectious disease-causing microorganisms being determined one by one, the subsequent annotation of gene function and the reconstruction work about protein functions are being carried out．All the efforts are to get a breakthrough understanding of biological characteristics, diagnostic strategies and treatment methods of pathogenic microorganisms．Mycobacterium tuberculosis has been a serious threat to human health globally．All genetic events occurring in the genome evolution play an important role in all aspects such as M． tuberculosis's biological properties, pathogenicity and drug resistance．This review will summarize the origins and genomic features of M．tuberculosis, and discuss the current progress about its genome evolution．

Abstract:Influenza virus is one of global constant research highlights, because it can cause the most severe disease in humans and animals as well as the most likely to trigger a pandemic. The surface glycoprotein hemagglutinin (HA) is critical determinants of the host specificity, virulence and infectivity of the influenza virus. The genetic mutations and glycosylation of HA can affect the biological properties of HA. The binding of HA to sialylated glycan receptors on host epithelial cells is the critical initial step in the infection and transmission of the virus. Understanding these components is important in comprehending the infection and the transmission of both existing human influenza viruses and newly emerging avian influenza viruses. This review summarizes studies how influenza virus and receptor components might act as determinants for successful viral replication and transmission and new progress for understanding the role of the structure of sialylated glycan receptors in influenza virus pathogenesis.

Abstract:Ubiquitin is best known for targeting protein degradation by the 26 S proteasome. In recent years, however, roles of ubiquitin are found far more than these. Ubiquitin not only has "traditional function" participating proteins degradation, but also plays a more varied and decisive role in cellular regulation than previously imagined. It is a multilayer regulator of important cellular processes and has a great many nonproteolytic functions including DNA damage repair, DNA replication, signal transduction, transcriptional regulation, membrane trafficking, endocytosis, protein kinase activation, chromatin remodeling and virus budding. These functions involve polyubiquitylation, monoubiquitylation and multiubiquitylation. Therefore, abnormities in ubiquitylation involves occurrence and development of diseases. Understanding these functions will provide further insights into the repertoire of ubiquitin, help to understand diverse cellular processes and facilitate our development of related new drugs.

Abstract:Starving tumor therapy is a new approach to treat cancers by blocking the function of angiogenic factors and inhibiting angiogenesis of tumor tissues. Endocrine-gland-derived vascular endothelial growth factor (EG-VEGF) was firstly identified as a novel tissue-selective angiogenic factor in 2001. During the past ten years, it has been demonstrated that EG-VEGF could play a wide range of other biological functions, including inducing differentiation of haematopoiteic stem cells, stimulating contraction of gastrointestinal smooth muscle and regulating development of enteric nervous system (ENS). In addition, the abnormal expression of EG-VEGF is involved in the occurrence and development of several kinds of angiogenesis-dependent diseases, such as tumors and polycystic ovary syndrome. EG-VEGF has been considered as a potential target for the development of diagnostic and therapeutic agents. Herein, the recent progress on biological function, related diseases and potential application of EG-VEGF is reviewed.

Abstract:Biotechnology is a rapidly expanding field of science and technology to change our life. It has been realized that strategic alliance is an important mode for technology transfer from biomedical research to the industry. Appropriate mechanisms of Industry-Academe-Research Institute strategic alliance have been built in major developed countries, significantly propelling the growth of biotechnology industry. Such advanced cooperation mode has been recently gained increasing concerns by China government. However, strategic alliance in China is in its preliminary stage with very small scale and inadequate functionality, its expected benefits are still far from significant. In this paper, we first review the development and management of effective strategic alliances in some leading countries; we then examine the successful mode of strategic alliances that is performing in USA, and identify the difficulties of China’s strategic alliance development based on its current progress. The future development of biomedical strategic alliances in China is discussed.

Abstract:Utilizing the typeⅥ secretion system (T6SS), Pseudomonas aeruginosa secretes the effector protein Tse2, a toxin to other competitive bacteria. It is a newly identified molecular mechanism for P. aeruginosa to win a survival advantage. To avoid being poisoned itself, P. aeruginosa synthesizes the specific immunity protein, Tsi2 to inhibit the toxin. Sequence analysis shows that Tsi2 is a novel antitoxin-like protein which is specific for P. aeruginosa. Using the SAD method, we have successfully resolved the crystal structure of Tsi2 at 1.8Å resolution. Our crystallographic studies reveal that Tsi2 adopts a novel coiled coil conformation which is not found in the antitoxins family previously. Meanwhile, Tsi2 is a well-assembled protein instead of an unfolded protein as an antitoxin in the toxin-free state. Tsi2 functions as a stable dimer and assembles as a unique "clamp" structure through extensive hydrophobic interactions. Two grooves on the dimerization interface combining with one helix of two symmetric molecules respectively imply the potential region interacting with Tse2. This research not only offers comprehensive insights into the molecular essence of Tsi2 as an antitoxin, but also reveals its structural basis of antitoxin activity. Furthermore, the apo Tsi2 structure provides a good framework for further researches on the structure and function of the complex Tse2-Tsi2.

Abstract:Dvl (Dishevelled) is a key effector molecule of the Wnt signaling pathway. The DIX domain of Dvl (Dvl-DIX) can homo-oligomerize into cytoplasmic puncta via the intra-filament SiteⅠ, SiteⅡ and the inter- filament SiteⅢ, and form hetero-complex with the DIX domain of Ccd1 (Ccd1-DIX) through SiteⅠ and SiteⅡ. Since all efforts to solve the wildtype Dvl2-DIX structure were unsuccessful, we carried out crystallization trials with the SiteⅢ mutants of Dvl2-DIX that display impaired homo-oligomerization and Wnt activity. Crystals of Dvl2-DIX(G65A) protein were obtained, but the diffraction data encountered an unexpected lattice-translocation defect. These SiteⅢ mutations of Dvl2-DIX retained the ability to hetero-interact with Ccd1-DIX, and we thus successfully generated various complexes between Dvl2-DIX mutants and wildtype Ccd1-DIX. After extensive trials, crystals of the Dvl2-DIX(G65A)-Ccd1-DIX complex were produced, but of poor quality and unsuitable for diffraction studies. Optimization is under way.

Abstract:In this study, we investigate the efffects of curcumin trathecal injection on pain threshold and the expressions of TLR4, TNF-α, IL-1β and IL-10 in the spinal cord of rats with chronic constrictive injury (CCI). 120 males Sprague-Dawley (SD) rats fitted with intrathecal catheters were randomly divided into four groups: sham group (Sham), CCI group (CCI), solvent control group (SC), and curcumin treated group (Cur, 100 μg/d). Neuropathic pain was produced by CCI of right sciatic nerve as described previously. On the 1st, 3rd, 7th, 10th，and 14th day after surgery, 100 μg curcumin dissolved in DMSO 10 μl was administered intrathecally once daily, and then pain threshold was measured. The expressions of TLR4, HMGB1 mRNA, and protein in lumbar spinal cord 4～6(L4～L6) were assessed by RT-PCR and Western blotting, respectively. The levels of TNF-α, IL-1β and IL-10 in the spinal cord were detected by ELISA. We found that the paw withdrawl threshold (PWT) and paw withdrawl latency (PWL) were significantly decreased after CCI (P<0.05), and the expressions of TLR4, HMGB1 mRNA and protein were significantly increased (P<0.05); the levels of TNF-α and IL-1β in the spinal cord were also significantly increased after CCI compared with those in the sham goup (P<0.05). Curcumin markedly attenuated CCI- induced mechanical allodynia and thermal hyperalgesia through inhibiting the activation of TLR4 pathway and production of inflammatory cytokines. These results demonstrated that curcumin alleviated neuropathic pain may be by decreasing the expression inflammatory cytokines produced by TLR4 pathway.

Abstract:Effect of oxymatrine on the H₂O₂ of L₆ rat myoblast cells was investigated in this study. The hydrogen peroxide(H₂O₂) was used to establish the H₂O₂-induced apoptosis model by L₆ rat myoblast cells that were treated with 0.3, 0.15, 0.75 g/L oxymatrine. The survival rate was measured by MTT method，cell cycles and apoptosis rate of L₆ rat myoblast cells were detected by flow cytometry, the injury degree was determined by apoptosis cytochemistry fluorescent antibody of Bcl-2 and Bax, DAPI staining and HE staining. The protein differences were examined by Western blot. Results showed that survival rate of L₆ myoblast cells was degraded and the apoptosis rate was increased by damaging H₂O₂. The survival rate of L₆ myoblast cells was heightened by oxymatrine of each dose. They could induce the increase in expression of Bcl-2 and the decrease in expression of Bax. Its degree of protection was increased with raising the dosage of oxymatrine and the protective effect of 0.3 g/L dosage is obvious. The protective effect of 0.15, 0.75 g/L oxymatrine were secondary. The main physiological and biochemical effects of oxymatrine was related to protect rat myoblast apoptosis model through NFκB signal transduction. These results implied that oxymatrine may have the potential as a new antioxidant treatment medicine.

Abstract:To investigate the expression and the relationship of P16^INK4a and sonic hedgehog signal pathway in cervical squamous cell carcinoma and its precursor lesions. The expression of P16^INK4a, Smo, Ptch and Gli in different HPV types positive cell lines were detected by Western-blot. A tissue microarray constructed with 20 normal cervical tissues and 100 uterine cervical cancers and related lesions (28 squamous cell carcinomas, 26 cervical intraepithelial neoplasia (CIN)Ⅲ, 16 CINⅡ, 12 CINⅠ, 18 tumor-adjacent tissue specimens) was immunohistochemically analyzed with anti- P16^INK4a, Shh, Patched (Ptch), Smoothened (Smo), Gli antibodies. The correlation between their expressions was analyzed. There was no significant difference among different HPV type cell lines regarding the expression of P16^INK4a and Shh, Ptch and Gli proteins(P>0.05). The expression of P16^INK4a and the Hh-signaling molecules was greatly enhanced in cervical carcinoma tissues, compared with that in normal epithelium and tumor-adjacent tissues (P<0.05). There was no significant difference between CINⅠ and normal epithelium(P>0.05), whereas, in case of P16^INK4a, Shh, Smo, and Gli, the differences among CINⅠ, CINⅡ and CINⅢ were significant (P<0.05). The expression of P16^INK4a protein was significantly correlated with that of Shh, Smo and Gli protein in CINⅡ-CINⅢ and cervical carcinoma and was correlated with that of Shh, Smo only in carcinoma tissue. P16^INK4a and the Hh-Gli signaling pathways were extensively activated in the development and evolution of cervical cancer, and the overexpression of P16^INK4a was correlated with Hh-signaling pathways. The abnormal Hh-signaling pathways maybe much associated with Smo protein overexpression induced by Shh, which can upregulate the expression of Gli protein.

Abstract:As DNA repair associated gene, NBS1 plays a key role in the repair of DNA double strand breaks and the maintenance of genomic stability. It has been shown in recent studies some common NBS1 variants maybe associated with genetic susceptibility of tumors. In the present study, rare allele frequency of single nucleotide polymorphisms (SNPs) of NBS1 gene in primary liver cancer were detected by the method of high resolution Single Strand Conformation Polymorphism (SSCP) analysis, with the aims to analyze the correlation between NBS1 SNPs and primary liver cancer, and evaluate the applicability of the high resolution SSCP technique in the genotyping of SNPs. The rare allele frequency of NBS1 SNPs were detected in 327 cases of primary liver cancer and 295 negative controls from Han people of China by the method of high resolution SSCP analysis. The correlation was analyzed between NBS1 SNPs and primary liver cancer. Genotyping of six common NBS1 SNPs in part samples was carried out by both SSCP analysis and direct sequencing simultaneously to compare the difference between them and evaluate the accuracy and applicability of SSCP analysis in genotyping of SNPs. The results from the tissue samples showed, among six NBS1 SNPs(102G>A, 320+208G/A, 553G>C, 1197T>C, 2016A>G and 2071-30A>T), the rare allele frequency of NBS1 SNP 1197T>C was significantly higher in the 119 cases of primary liver cancer (68.1%) than that in the 95 controls of cirrhosis/chronic hepatitis B (57.9%)(P=0.0298). Similar results was obtained by detection with blood samples of 208 cases of hepatocellular carcinoma(66.8%) and 200 cases of health controls(58.8%)(P=0.0170). There was no any significant difference of the rare allele frequency of the other five NBS1 SNPs. The same result was observed for genotyping of NBS1 SNPs either by the high resolution SSCP analysis or by the direct sequencing. However, higher quality of PCR products is requested for the direct sequencing compared with SSCP analysis. These results suggest that the NBS1 SNP 1197T>C may be associated with the risk of primary liver cancer in the Chinese Han population. High resolution SSCP analysis, with the same accuracy as direct sequencing in the genotyping of NBS1 SNPs and easier to handle, is suitable for the genotyping of multiple known SNPs in large scale cohort study.