Discovery of species-specific peptide markers and development of quality-evaluation strategies for deer horn gelatin using liquid chromatography-tandem mass spectrometry and a label-free methodology.

Deer horn gelatin (DHG) is a valuable nutritional dietary supplement. Due to the significant variation in the price of DHG from different sources, it is important to assess its quality and to clarify the species of its raw material. However, due to the similarity in appearance and physicochemical properties, as well as the destruction of genetic material during the manufacturing process, it is difficult to distinguish DHG from gelatin derived from other sources. Furthermore, current methods are unable to evaluate the overall quality of DHG. Using Nano LC-Orbitrap MS and data analysis software, DHG samples from five deer species were analyzed to identify peptide markers specific to alpha-2-HS-glycoprotein (AHSG) and collagen. The peptide markers were validated using HPLC-Triple Quadrupole MS, and strategies for assessing the quality of DHG were developed. Eighteen peptide markers were discovered, comprising peptides with differing specificities. Three strategies for the identification, characteristic mapping, and content determination of DHG were developed. These strategies can be used to assess the quality of deer gelatin.

Modified ACDF Technique for the Treatment of Centrum Focal Ossification of the Posterior Longitudinal Ligament: A Case Report.

BACKGROUND: Anterior cervical discectomy fusion (ACDF) is a surgical procedure used to treat cervical spondylosis with anterior spinal cord compression. However, there are limitations to traditional ACDF and posterior indirect decompression when the anterior source lesion is in the center of the cervical vertebra. CASE PRESENTATION: On June 8, 2022, our department treated a patient with cervical spondylotic myelopathy-whose high posterior longitudinal ligament (OPLL) occupied the central position of the vertebral body-with modified ACDF. The preoperative surgical plan was designed based on the relevant imaging data and assay index. Also, the visual analogue scale (VAS), Japanese Orthopaedic Association (JOA) scores, and imaging parameters of neck pain were recorded and compared. Postoperative imaging data showed that cervical curvature was recovered and spinal canal compression was relieved. The VAS score for neck pain decreased from 7 preoperatively to 1.5 at the last follow-up, while the JOA score increased from 10 preoperatively to 29 at the last follow-up. The volume of the spinal canal was restored. Simultaneously, the patient's extremity muscle strength improved and muscle tension decreased. CONCLUSIONS: Modified ACDF may be an effective surgical method for resolving spinal cord compression in a specific location when bone mineral density is good. We can effectively avoid iatrogenic nerve injury and symptom recurrence by removing the vertebral body and the lesion directly.

Peptide Biomarkers Discovery for Seven Species of Deer Antler Using LC-MS/MS and Label-Free Approach.

Deer antler is a globally widely used precious natural medicine and the material of deer horn gelatin. However, identification of deer antler species based on traditional approaches are problematic because of their similarity in appearance and physical-chemical properties. In this study, we performed a comprehensive antler peptidome analysis using a label-free approach: nano LC-Orbitrap MS was applied to discover peptide biomarkers in deer adult beta-globin (HBBA), and HPLC-Triple Quadrupole MS was used to verify their specificity. Nineteen peptide biomarkers were found, on which foundation a strategy for antlers and a strategy for antler mixtures such as flakes or powder are provided to identify seven species of deer antler including Eurasian elk (Alces alces), reindeer (Rangifer tarandus), white-tailed deer (Odocoileus viginianus), white-lipped deer (Przewalskium albirostris), fallow deer (Dama dama), sika deer (Cervus nippon), and red deer (Cervus elaphus) simultaneously. It is worth noting that our search found that the HBBA gene of sika deer, red deer, and North American wapiti (Cervus canadensis) in China may have undergone severe genetic drifts.

Identification of chemosensitizing agents of colorectal cancer in Rauvolfia vomitoria using an NMR-based chemometric approach.

Searching for new adjuvants of conventional chemotherapeutic approaches against colorectal cancer cells is extremely urgent. In current research, a non-targeted analytical approach was established by combining proton nuclear magnetic resonance spectroscopy with a chemometrics data mining tool to identify chemosensitizing agents from Rauvolfia vomitoria. This approach enabled the identification of potential active constituents in the initial fractionation process and provided their structural information. This strategy was validated by its application to Rauvolfia vomitoria extract exhibiting chemosensitizing activity on 5-fluorouracil against colorectal cancer cells. After the workflow, the biochemometrics analysis showed that at least 15 signals (Variable influence on projection (VIP) > 1) could have contributions in the differentiation of various fractions. Through systematic literature and database searches, we found that the most active fraction (fraction 7) exhibited the highest presence of sabazin-type and armaniline-type alkaloids, which were potential chemosensitizers as previously reported. To validate the results of the strategy, the effect of 5-FU and compounds isolated from fraction seven incubation on HCT-8 and LoVo cell vialibilty were evaluated. These results evidenced that compound ß-carboline (3), 1-methyl-ß-carboline (4), and lochnerine (6) could enhance the cytotoxicity of 5-fluorouracil against to Colorectal cancer cells. Besides, 21 compounds including two new compounds were isolated from Rauvolfia vomitoria. The experimental results verify the reliability of the method, and this approach provides a new and efficient tool to overcome some of the bottlenecks in natural products drug discovery.

Transforaminal en bloc resection for the treatment of thoracic ossification of the ligamentum flavum: Retrospective cohort study.

BACKGROUND: Our purpose is to introduce transforaminal en bloc resection for the treatment of thoracic ossification of ligamentum flavum, and report outcomes of the procedure. PATIENTS AND METHODS: The record of patients with symptomatic thoracic ossification of the ligamentum flavum who received transforaminal en block resection from October 2010 to September 2014 were retrospectively reviewed. Outcomes were Japanese Orthopaedic Association (JOA) lower extremity motor dysfunction and sphincter dysfunction scores. RESULTS: Seventeen patients (10 males, 7 females) with a median age of 60 years were included in the analysis. Surgery was performed as planned without complications in all patients. One segment decompression was performed in 1 case, 2 segment in 9 cases, 3 segment in 3 cases, and 4 segment in 4 cases. The median lower extremity motor dysfunction score was significantly higher at 2 weeks, 3 months, and 6 months postoperatively, and the end of follow-up, than before surgery (median score: 2, 2, 3, 3, respective, vs. 1, all, p ≤ 0.008). There were no significant differences in JOA sphincter dysfunction score between the different time points. CONCLUSION: Transforaminal en bloc resection is a safe and effective method for the treatment of thoracic ossification of the ligamentum flavum.

Activation of nuclear factor-κB subunit p50/p65 enhances gefitinib resistance of lung adenocarcinoma H1650 cell line / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the intrinsic connection between activation of classical nuclear factor-κB (NF-κB) pathway and gefitinib resistance in human lung adenocarcinoma H1650 cells.</p><p><b>METHODS</b>Human lung adenocarcinoma H1650 cells were exposed to gefitinib continuously for 60 days to obtain resistant H1650 cells. The expressions of P-IκBα, P-p50 and P-p65 in the cytoplasm or nuclei were detected using Western blotting in human lung adenocarcinoma HCC827 cells, parental H1650 cells and gefitinib-resistant H1650 cells. The effects of gefitinib alone or in combination with PDTC on the survival rate and expressions of NF-κB P-p50 and P-p65 were compared among the 3 cell lines.</p><p><b>RESULTS</b>Gefitinib-resistant H1650 cells showed increased cytoplasmic and nuclear P-IκBα expressions. The expressions of P-p50 and P-p65 differed significantly among the 3 cell line, decreasing in the order of resistant H1650 cells, parental H1650 cells, and gefitinib sensitive HCC827 cell lines (P<0.05 or 0.01). Treatment with gefitinib alone resulted in a significantly lower cell inhibition rate in resistant H1650 cells than in the parental H1650 cells (P<0.05) and HCC827 cells (P<0.01). The resistant H1650 cells had a significantly higher expression of P-p50 and P-p65 than other two cell lines (P<0.05). In both the resistant and parental H1650 cells, gefitinib significantly lowered P-p50 and P-p65 expressions (P<0.05 or 0.01), and the combined treatment with gefitinib and PDTC significantly decreased the cell survival rate and further lowered the cytoplasmic and nuclear expressions of P-p50 and P-p65 (P<0.01 or 0.01).</p><p><b>CONCLUSION</b>The activation of classical NF-κB pathway is a key factor contributing to transformation of the parental H1650 cells into gefitinib-resistant cells. Gefitinib combined with PDTC can inhibit P-IκBα production and NF-κB P-p50 and P-p65 activation to suppress the survival of residual H1650 cells and the generation of gefitinib-resistant cells.</p>

Development of a mouse model of arecoline-induced oral mucosal fibrosis

OBJECTIVE@#To develop a BALB/c mouse model of oral submucous fibrosis (OSF) induced by arecoline and to exhibit an accumulation of collagen and angiogenesis changes.@*METHODS@#BALB/c mice were randomly assigned to either the control (distilled water) or experimental group (arecoline) (n = 40). Eight mice from each group were sacrificed every 4 weeks since 8 weeks post treatment. Changes in histopathologic features, levels of collagen type I and collagen type III, and angiogenesis were measured.@*RESULTS@#In the 8th week, epithelium atrophy, collagen cumulation and micrangium pathologic changes in the lamina propria were observed in the oral mucosa. In the 20th week, hyaline degeneration of the connective tissues was observed on the tongue and palate mucosa. The angiogenesis and collagen type I changed significantly as the diseases advanced (P < 0.05); however, collagen type III was not statistically different.@*CONCLUSIONS@#An OSF model involving mice can be rapidly induced by drinking a high-dose of arecoline. OSF angiogenic changes in mice primarily decrease and collagen accumulation is mainly collagen type I.

Development of a mouse model of arecoline-induced oral mucosal fibrosis

Objective To develop a BALB/c mouse model of oral submucous fibrosis (OSF) induced by arecoline and to exhibit an accumulation of collagen and angiogenesis changes. Methods BALB/c mice were randomly assigned to either the control (distilled water) or experimental group (arecoline) (n = 40). Eight mice from each group were sacrificed every 4 weeks since 8 weeks post treatment. Changes in histopathologic features, levels of collagen type I and collagen type III, and angiogenesis were measured. Results In the 8th week, epithelium atrophy, collagen cumulation and micrangium pathologic changes in the lamina propria were observed in the oral mucosa. In the 20th week, hyaline degeneration of the connective tissues was observed on the tongue and palate mucosa. The angiogenesis and collagen type I changed significantly as the diseases advanced (P < 0.05); however, collagen type III was not statistically different. Conclusions An OSF model involving mice can be rapidly induced by drinking a high-dose of arecoline. OSF angiogenic changes in mice primarily decrease and collagen accumulation is mainly collagen type I.

Comparison of Allograft and Autograft in Lumbar Fusion for Lumbar Degenerative Diseases: A Systematic Review.

Purpose/Aim of the study: The purpose of this analysis was to compare the clinical outcomes of vertebral fusion with allograft versus autograft bone. MATERIALS AND METHODS: Medline, PubMed, Cochrane, EMBASE, and Google Scholar databases were searched until July 27, 2015 using the keywords: lumbar vertebrae, surgery, spondylolisthesis, bone transplantation, allograft, autograft. Inclusion criteria were randomized controlled trials (RCTs), two-arm prospective studies, retrospective studies, and cohort studies comparing the results of autograft and allograft in patients receiving lumbar spinal fusion. The outcomes were changes of Oswestry Disability Index (ODI) and visual analogue scale (VAS) pain scores from baseline at 1, 2, and 3 years after surgery and fusion rates. RESULTS: Of 154 studies identified in the database searches, five were included in the quantitative analysis (one RCT, one prospective, and three retrospective studies). The mean patient age ranged from approximately 40 to 65 years, and approximately half of the patients were males. The total number of patients who received allografts was 333 and the total that received autografts was 175. The analysis revealed the change of ODI and VAS pain score at 1, 2 and 3 years was similar between the allograft and autograft groups (all, p > .05), as were the fusion rates (p > .05). CONCLUSIONS: Both allograft and autograft provide acceptable outcomes for spinal fusion.