[Comparison of early clinical effects between Activ C cervical disc replacement and anterior cervical discectomy and fusion for single-level cervical spondylosis].

OBJECTIVE: To compare the early clinical effects of Activ C cervical disc replacement (ACDR) and anterior cervical discectomy and fusion (ACDF) in treating single-level cervical spondylosis. METHODS: The clinical data of 76 patients with single-level cervical spondylosis underwent surgery from July 2009 to September 2012 were retrospectively analyzed. Among them, 28 patients were treated with ACDR (ACDR group), including 18 males and 10 females, aged from 32 to 62 years old with an average of (45.2±6.2) years; and 48 patients were treated with ACDF (ACDF group), including 28 males and 20 females, aged from 33 to 60 years old with an average of (45.8±6.4) years. Visual analogue scale (VAS), Japanese Orthopedics Association (JOA) score, Short Form-36 (SF-36), imaging data were used to assess the clinical effects after operation. RESULTS: A total of 76 patients were followed up from 6 to 24 months with an average of 13.2 months. VAS of neck pain and brachialgia were improved in all patients after operation (P<0.05), there was no significant difference between two group (P>0.05). Somato-score and psycho-score of SF-36 of two groups were obviously increased (P<0.05), ACDR group was better than that of ACDF group (P<0.05). In ACDR group, there was no significant difference in the range of motion of surgical segments and adjacent segments between preoperative and postoperative (P>0.05); heterotopic ossification around the edge of vertebral body occurred in 1 case on the 6th month after operation, no fusion was found on the 1st year after operation. In ACDF group, the adjacent vertebral disease occurred in 1 case and the patient underwent the reoperation. CONCLUSION: Activ C cervical disc replacement can reduce the degeneration of adjacent segments and its early outcomes for the treatment of single-level cervical spondylosis are satisfactory, but the long-term effects still need study.

Cloning the sterol carrier protein 2 genes of Japanese toad (Bufo japonicus formosus) and Chinese toad (Bufo gargarizans) and its tissue expression analysis.

In this study, to clarify the bioactive polypeptides included in the skins and secretions of Bufo, we screened the Japanese toad (Bufo japonicus formosus) skin cDNA liary by colony polymerase chain reaction (PCR), and obtained a transcript of 1 075 bp consisting of 1 37 bp 5' untranslated region (UTR), 515 bp 3' UTR and a 423 bp open reading frame (ORF) encoding a polypeptide of 140 amino acid residues (GenBank accession number: KF359945). Homolog analysis showed a 70%-96% homology with sterol carrier protein-2 (SCP-2) present in other animals, which is implicated in lipid metabolism of other organisms. The gene SCP-2 of Chinese toad (B. gargarizans) was cloned from a first strand cDNA of Bufo skin (GenBank accession number: KF381341) via PCR, whose encoding polypeptide has only one amino acid difference from that of Japanese toad. Tissue distribution analysis showed that SCP-2 expressed in all organs tested, though in the liver and spleen it manifested lower expression than in other organs. These findings might indicate SCP-2 being one of the active ingredients in toad skin. These findings may in turn have implications for further drug development from traditional Chinese medicine sources.

[Cloning and bioinformatic analysis of TAGLN2 cDNA of Bufo japonicus formosus].

To study the bioactive polypeptides included in Bufo skin and its secretions the plasmid skin cDNA library of adult Japanese toad Bufo japonicus formosus was prepared. The pSD64TR has been used as the vector and the cloning sites are Xho I and EcoR I. To screen cDNAs encoding bioactive components, the plasmid cDNA library was transformed into E. coli DH5 competent cells, and positive colonies were screened by colony PCR (polymerase chain reaction). The suspension of a single colony in LB medium was used as the template, SP6 (the upstream primer of the plasmid cDNA library) and a primer with Xho I site and polyT were used as the primers. As the result, 465 positive colonies out of 1 344 were obtained and their plasmid were collected and sequenced. By homologous analysis, it was found that one of the cDNAs encoding a peptide with high homolog with transgelin-2, which was registered in GenBank (accession number: JX197456), and it was indicated as a partial cDNA sequence with a deletion at the 5' end. The transcript is 997 bp consisting of 31 bp 5', 618 bp 3' untranslated region (UTR) and an open reading frame (ORF) of 348 bp encoding a polypeptide of 115 amino acids. In the putative protein product, there is a calponin homology domain, two cysteine residues for a disulfide bond and three a-helix domains, and five potential phosphorylation sites. The homologous analysis indicates 90% similarity with Xenopus (Silurana) tropicalis and 89% with Xenopus laevis, and 71%-85% with other species.

[Bufo gargarizans mcl-1 cloning and its prokaryotic recombinant protein expression].

MCL-1 is encoded by myeloid cell leukemia-1 gene (mcl-1), which is one of the anti-apoptotic members of bcl-2 cell apoptotic gene superfamily. ChanSu is made of dorsal secretions of several Bufo species, commonly used in the prescriptions of traditional Chinese medicine for treating many diseases including cancer. To clarify if mcl-1 is expressed in the dorsal skin of B. gargarizans, the PCR (polymerase chain reaction) was performed with its dorsal skin first strand cDNA as the template and a pair of specific primers of mcl-1, and PCR products were cloned into the pGM-T vector. DNA sequencing indicated that the ORF length was 639 bp encoding 212 amino acid residues, and the homology of 44%-95% with the MCL-1 of several other animals. For the further studies on MCL-1 biological functions during the oncogenesis and preparation of its antibody, the prokaryotic expression construct of pET-28b-mcl-1 was prepared which was confirmed by DNA sequencing, and its recombinant protein expression (0.02% wet weight) in E. coli BL21 (DE3) strain was confirmed by SDS-PAGE and Western blotting.

[Analysis of volatile oil in Fructus Aurantii processed with Zhangband method by GC-MS].

OBJECTIVE: To analyze the chemical components of the volatile oil in processed pieces of Fructus Aurantii (PPFA) with JiangXi Zhangband methods by GC-MS. METHODS: The volatile oil was obtained from PPFA by steam distillation. The amount of the components from the volatile oil were determinated by area normalization method. The separated components were identified by GC-MS. RESULTS: Composed of the total volatile oil over 90% , 70 components were separated and identified. All of the processed Fructus Aurantii produced new chemical composition, in the meantime,the contents were changed in some chemical composition after processed. CONCLUSION: The method is reliable, stable and has good repeatability. This method can be applied to the analysis of volatile oil components in PPFA. It will provide the certain scientific methods for further evaluating of PPFA quality.