Application of combined anterior and posterior approaches for the treatment of cervical tuberculosis with anterior cervical abscess formation and kyphosis using a Jackson operating table: a case report and literature review.

BACKGROUND: There have been insufficient reports to date regarding the treatment of cervical spinal tuberculosis, and the optimal surgical approaches to treating this condition have yet to be established. CASE REPORT: This report describes the treatment of a case of tuberculosis associated with a large abscess and pronounced kyphosis through the use of a combined anterior and posterior approach with the aid of the Jackson operating table. This patient did not exhibit any sensorimotor abnormalities of the upper extremities, lower extremities, or trunk, and presented with symmetrical bilateral hyperreflexia of the knee tendons, while being negative for Hoffmann's sign and Babinski's sign. Laboratory test results revealed an erythrocyte sedimentation rate (ESR) of 42.0 mm/h and a C-reactive protein (CRP) of 47.09 mg/L. Acid-fast staining was negative, and spine magnetic resonance imaging revealed the destruction of the C3-C4 vertebral body and a posterior convex deformity of the cervical spine. The patient reported a visual analog pain score (VAS) of 6, and exhibited an Oswestry disability index (ODI) score of 65. Jackson table-assisted anterior and posterior cervical resection decompression was performed to treat this patient, and at 3 months post-surgery the patient's VAS and ODI scores were respectively reduced to 2 and 17. Computed tomography analyses of the cervical spine at this follow-up time point revealed good structural fusion of the autologous iliac bone graft with internal fixation and improvement of the originally observed cervical kyphosis. CONCLUSIONS: This case suggests that Jackson table-assisted anterior-posterior lesion removal and bone graft fusion can safely and effectively treat cervical tuberculosis with a large anterior cervical abscess combined with cervical kyphosis, providing a foundation for future efforts to treat spinal tuberculosis.

Upregulation of miR-802 suppresses gastric cancer oncogenicity via targeting RAB23 expression.

OBJECTIVE: The aberrant expression of microRNAs (miRNAs) has been observed in various types of cancer. Recently, miR-802 was found to play important role in tumor progression. However, the function of miR-802 in gastric cancer (GC) remains unknown. The aim of the present study was to investigate biological effects and underlying mechanisms of miR-802 in GC. PATIENTS AND METHODS: Quantitative RT-PCR was performed to evaluate the expression level of miR-802 in GC tissues and cell lines. The in vitro cell proliferation was measured using the MTT method. Cell invasion and migration assays were performed using the transwell assay. The effects of miR-802 on tumor growth were examined using a GC xenograft model. Flow cytometry method was used to detect the effect of miR-802 in apoptosis of GC cells. Targets of miR-802 were predicted using bioinformatics and validated using luciferase reporter and Western blot analyses. RESULTS: The results showed that miR-802 was significantly down-regulated in GC tissues and cell lines. The enforced expression of miR-802 induced growth suppression and apoptosis of GC cells. Moreover, miR-802 overexpression suppressed the migration and invasion of GC cells. Bioinformatics analysis predicted that the RAB23 was a potential target gene of miR-802. The results of luciferase reporter assay demonstrated that miR-802 could directly target RAB23. Additionally, in vivo analysis, the xenograft mouse model also confirmed the suppressive effects of miR-802 on tumor growth. CONCLUSIONS: Our results are the first to demonstrate the tumor-suppressive role of miR-802 in GC. The identification of miR-802 and its novel target RAB23 will be valuable in developing therapeutic applications for GC.

Analysis of beta-tubulin cDNAs from taxol-resistant Pestalotiopsis microspora and taxol-sensitive Pythium ultimum and comparison of the taxol-binding properties of their products.

The anti-cancer drug taxol binds to beta-tubulin in assembled microtubules and causes cell cycle arrest in animal cells; in contrast, in fungi, the effect of taxol varies. For instance, the taxol-producer Pestalotiopsis microspora Ne32, an ascomycete, is resistant to taxol (IC50 greater than 11.7 microM), whereas Pythium ultimum, an oomycete, is sensitive to taxol (IC50 0.1 microM). In order to understand the differential fungal response to taxol, we isolated cDNAs encoding beta-tubulin from both P. microspora and P. ultimum. The deduced amino acid sequence of beta-tubulin from P. microspora is very similar to those from other Ascomycetes, many of which are resistant to taxol. The sequence of beta-tubulin from P. ultimum is very similar to those from Oomycetes and non-fungal organisms, many of which are sensitive to taxol. To examine the interaction between taxol and fungal microtubules, binding studies were performed with fungal cells, using [3H]taxol. The labeled taxol was found to bind specifically to P. ultimum, but not to P. microspora. In addition, the amount of [3H]taxol specifically bound to P. ultimum was reduced by the microtubule-depolymerizing drug thiabendazole, in a dose-dependent manner. These results suggest efficient binding of taxol to microtubules in P. ultimum, but not in P. microspora, and are consistent with the differential taxol sensitivity of these two organisms. Finally a comparison of previously characterized taxol binding sites in various beta-tubulin sequences showed that beta-tubulins of taxol-sensitive organisms, including P. ultimum, contain Thr219, but beta-tubulins of resistant organisms, including P. microspora, contain Asn or Gln at this position, suggesting an important role for residue 219 in the interaction between taxol and beta-tubulin.

Identification by PCR of receptor-like protein kinases from Arabidopsis flowers.

We have isolated three receptor-like kinase cDNAs from an Arabidopsis flower cDNA library by PCR using degenerate oligonucleotide primers for conserved domains of protein kinases. Cloning and sequencing of the full-length cDNAs, designated RKF1 to 3 (receptor-like kinase in flowers), showed that the putative extracellular domain of the RKF1 protein contains 13 tandem repeats of leucine-rich sequences and those of RKF2 and RKF3 have no significant homology with other plant sequences. RNA blot analysis revealed that the RKF1 mRNA is highly expressed in stamens while RKF2 and RKF3 mRNAs are present at low levels in all organs examined. In situ localization experiments indicated that the RKF1 mRNA is detectable in early flower primordia and during stamen development. In addition, when fused to a GUS reporter gene, the RKF1 promoter directed high GUS expression in pollen grains. Recombinant RKF1, produced in Escherichia coli, was found to have kinase activity with serine/threonine specificity in vitro.

Expression of human muscarinic cholinergic receptors in tobacco.

We expressed human m1, m2 and chimeric muscarinic cholinergic receptors (MAChR) in tobacco plants and in cultured BY2 tobacco cells using Agrobacterium-mediated transformation. The membranes of most transgenic plants and calli bound muscarinic ligands with appropriate affinities, kinetics and pharmacologic specificity, as determined by direct and competitive binding measurements using the muscarinic ligand [3H]quinuclidinyl benzylate (QNB). Membranes of untransformed plants and calli or those transformed with vector alone did not bind [3H]QNB. Preliminary experiments did not suggest regulation of endogenous plant G protein signalling pathways by the recombinant receptors. Membranes from one callus clone expressed m1 MAChR at the level of 2.0-2.5 pmol [3H]QNB bound per mg membrane protein, more than the number of m1 MAChR in mammalian brain and comparable to that expressed in Sf9 insect cells using baculovirus vectors. This work demonstrates high level expression of active G protein-coupled receptors in plants, such that signaling might be genetically reconstituted by co-expression of appropriate G proteins and effectors.

Characterization of a pollen-expressed gene encoding a putative pectin esterase of Petunia inflata.

From a pollen tube cDNA library of Petunia inflata, we isolated cDNA clones encoding a protein, PPE1, which exhibits sequence similarity with plant, bacterial, and fungal pectin esterases. Genomic clones containing the PPE1 gene were isolated using cDNA for PPE1 as a probe, and comparison of the cDNA and genomic sequences revealed the presence of a single intron in the PPE1 gene. During pollen development, PPE1 mRNA was first detected in anthers containing uninucleate microspores; it reached the highest level in mature pollen and persisted at a high level in in vitro germinated pollen tubes. The observed expression pattern of the PPE1 gene suggests that its product may play a role in pollen germination and/or tube growth.

Characterization of a pollen-expressed receptor-like kinase gene of Petunia inflata and the activity of its encoded kinase.

From a pollen tube cDNA library of Petunia inflata, we isolated clones encoding a protein with structural features and biochemical properties characteristic of receptor-like kinases. It was designated PRK1 for pollen receptor-like kinase 1. The cytoplasmic domain of PRK1 is highly similar to the kinase domains of other plant receptor-like kinases and contains nearly all of the conserved amino acids for serine/threonine kinases. The extracellular domain of PRK1 contains leucine-rich repeats as found in some other plant receptor-like kinases, but overall its sequence in this region does not share significant similarity. Characterization of a gene encoding PRK1 revealed the presence of two introns. During pollen development, PRK1 mRNA was first detected in anthers containing mostly binucleate microspores; it reached the highest level of mature pollen and remained at a high level in in vitro-germinated pollen tubes. The recombinant cytoplasmic domain of PRK1 autophosphorylated on serine and tyrosine, suggesting that PRK1 may be a dual-specificity kinase. Monospecific immune serum to the recombinant extracellular domain of PRK1 detected a 69-kD protein in microsomal membranes of pollen and pollen tubes. The characteristics of PRK1 suggest that it may play a role in signal transduction events during pollen development and/or pollination.

Cloning and sequencing of cDNAs encoding two self-incompatibility associated proteins in Solanum chacoense.

We have isolated and sequenced cDNAs for S2- and S3-alleles of the self-incompatibility locus (S-locus) in Solanum chacoense Bitt., a wild potato species displaying gametophytic self-incompatibility. The S2- and S3-alleles encode pistil-specific proteins of 30 kDa and 31 kDa, respectively, which were previously identified based on cosegregation with their respective alleles in genetic crosses. The amino acid sequence homology between the S2- and S3-proteins is 41.5%. This high degree of sequence variability between alleles is a distinctive feature of the S-gene system. Of the 31 amino acid residues which were previously found to be conserved among three Nicotiana alata S-proteins (S2, S3, and S6) and two fungal ribonucleases (RNase T2 and RNase Rh), 27 are also conserved in the S2- and S3-proteins of S. chacoense. These residues include two histidines implicated in the active site of the RNase T2, six cysteines, four of which form disulfide bonds in RNase T2, and hydrophobic residues which might form the core structure of the protein. The finding that these residues are conserved among S-proteins with very divergent sequences suggests a functional role for the ribonuclease activity of the S-protein in gametophytic self-incompatibility.