[Study on feasibility of enhanced recovery after surgery combined with mobile microendoscopic discectomy-transforaminal lumbar interbody fusion in the treatment of lumbar spondylolisthesis].

Objective: To explore the feasibility of enhanced recovery after surgery (ERAS) combined with mobile microendoscopic discectomy-transforaminal lumbar interbody fusion (MMED-TLIF) in the treatment of lumbar spondylolisthesis and its influence on postoperative rehabilitation. Methods: From October 1 2014 to July 1 2016 , a cohort of 52 patients with lumbar spondylolisthesis who received the program of ERAS-MMED-TLIF were retrospectively reviewed in Department of Minimally Invasive Spine Surgery, Tianjin Hospital.The primary outcomes include the operation time, intraoperative blood loss, length of hospital stay, VAS score (low back pain and leg pain) and Oswestry Disability Index (ODI) at different follow-up time and complication.The height of intervertebral space and fusion rate were also recorded as radiographic indicators. Results: All cases had an average follow-up of 12 months. The mean operative time was (115±30) min with a mean blood loss of (100±35) ml.Compared with preoperative condition, VAS score of low back pain (6.3±3.3 vs 3.5±2.3, P<0.05), VAS score of leg pain (7.1 ± 4.2 vs 3.1 ± 2.6, P<0.05) and the ODI disability index score (43.5±9.6 vs 20.9±7.3, P<0.05) at the postoperative 24 h were decreased and the difference was statistically significant.The mean hospitalized time were (4.9±1.3) days with mean postoperative hospital stay (2.1±1.2) days.Fusion rate was 92.31% (48/52) at the last follow-up time. Conclusion: ERAS combined with MMED-TLIF is feasible in the treatment of lumbar spondylolisthesis, which can significantly reduce intraoperative bleeding, shorten the total length of stay and postoperative hospital stay, improve postoperative pain and promote rapid rehabilitation of patients after operation without increasing the operation time and influencing the long-term effect, it can be applied in clinical practice.

Lung carcinogenesis induced by chronic tuberculosis infection: the experimental model and genetic control.

Coexistence of pulmonary tuberculosis (TB) and lung cancer in clinic poses significant challenges for the diagnostic and treatment of both diseases. Although association of chronic inflammation and cancer is well-documented, causal relationship between TB infection and lung cancer are not understood. We present experimental evidence that chronic TB infection induces cell dysplasia and squamous cell carcinoma (SCC) in a lung-specific manner. First, squamous cell aggregates consistently appeared within the lung tissue associated with chronic TB lesions, and in some cases resembled SCCs. A transplantable tumor was established after the transfer of cells isolated from TB lung lesions into syngeneic recipients. Second, the (Mycobacterium tuberculosis) MTB-infected macrophages play a pivotal role in TB-induced carcinogenesis by inducing DNA damage in their vicinity and by the production of a potent epidermal growth factor epiregulin, which may serve as a paracrine survival and growth factor responsible for squamous metaplasia and tumorigenesis. Third, lung carcinogenesis during the course of chronic TB infection was more pronounced in animals with severe lung tissue damage mediated by TB-susceptibility locus sst1. Together, our experimental findings showed a causal link between pulmonary TB and lung tumorigenesis and established a genetic model for further analysis of carcinogenic mechanisms activated by TB infection.

Localization of isoprenylated antigen of hepatitis delta virus by anti-farnesyl antibodies.

Hepatitis delta virus (HDV) is a subviral pathogen that requires pre-existing or concurrent infection with hepatitis B virus (HBV). HDV expresses two forms of a single protein, the delta antigen (HDAg), which are identical except for an additional 19 residues at the C terminus of the large form. Within this C-terminal extension a cysteine residue is isoprenylated; this isoprenylation is critical for interaction with HBV envelope proteins to enable virus assembly and release into the medium. Therefore, large HDAg must be recruited to an extracellular compartment. However, immuno-staining with HDAg-specific antibodies has localized the large antigen mainly to the nucleus and supports the notion that large HDAg suppresses virus replication in the nucleus. Since isoprenylation would increase the hydrophobicity of the protein and may favour transport towards specific membranes, the question remains whether the large HDAg detected in the nucleus carries an isoprenyl group. To address this issue, antibodies against the farnesyl modification were generated to allow direct visualization of the antigen by immunofluorescence microscopy. The anti-farnesyl antibodies specifically stained large HDAg expressed in Huh-7 cells, and the signal was largely restricted to the nucleus; the staining pattern could be superimposed on those of cells stained for large HDAg. The large HDAg translocated into the nucleus was therefore isoprenylated. In addition, antibodies specific for the farnesyl modification should be applicable to the study of other similarly isoprenylated proteins.

Self-association of the C-terminal domain of the hepatitis-C virus core protein.

The N-terminal region of the hepatitis-C virus (HCV) core protein is rich in basic residues, while the C-terminal end of the protein comprises of a stretch of hydrophobic amino acids. Between these two extremes is an amphipathic region with two predicted alpha-helical segments. This region embodies Leu or hydrophobic residues in positions of heptad repeats and is possibly capable of self-association. To investigate this possibility, the core sequence was divided into two fragments and expressed separately as recombinant proteins. Recombinant proteins with the N-terminal fragment remained as monomers even at high concentrations in SDS/PAGE. Recombinant protein with the C-terminal fragment appeared largely monomeric on denaturing gels but some oligomers were also detected. Furthermore, proline mutations in either one of the predicted alpha helices adversely affected the observed oligomerization. The self-association capacity of the core protein C-terminal region was further supported by results from a yeast two-hybrid system. To affirm our conclusion, a peptide covering the heptad repeats and the predicted alpha helices was synthesized. Data from mass spectrometry and gel-filtration chromatography concluded that this peptide readily self-associated into the homodimer. Therefore, our results suggest that the oligomerization motifs of the HCV core protein may not be limited to the previously suggested N-terminal region.

Truncating the putative membrane association region circumvents the difficulty of expressing hepatitis C virus protein E1 in Escherichia coli.

The putative E1 of hepatitis C virus (HCV) was expressed in Escherichia coli using a glutathione-S-transferase (GST) fusion protein system. The full length E1 protein is difficult to express. A series of E1 DNA fragments was generated and used for expression vector construction. Fusion proteins containing the E1 C-terminal region could not be expressed. When this region was truncated, the fusion proteins were synthesized to high levels. The possibility of this C-terminal region hampering the production of fusion protein was further explored. A construct with this segment directly fused to the C-terminus of GST indeed generated no detectable recombinant protein. According to the predicted structure of E1, this region may have membrane-associating properties. The expression results suggest a general approach to facilitate the production of viral membrane proteins in prokaryotes. Furthermore, these recombinant E1 proteins generated as antigens were used for Western blotting with sera from HCV-infected individuals. It was found that E1 is antigenic during HCV natural infection.