Changes in the centre of rotation and the anterior bone loss of the vertebral body in Mobi-C artificial disc replacement segments after cervical hybrid surgery: a retrospective study.

OBJECTIVE: To examine the short-term efficacy and imaging results of using the Mobi-C in cervical hybrid surgery on 2-level cervical spondylolisthesis. To observe post-operative changes in the flexion-extension centre of rotation (FE-COR) and anterior bone loss (ABL) of the anterior cervical disc replacement (ACDR) segment. METHODS: Forty-two patients (20 males and 22 females, aged 42‒67 years) who underwent cervical hybrid surgery were retrospectively analysed. Their ACDR segment used Mobi-C, and the fusion segment used ROI-C, with a follow-up of 25‒42 months (31.1 ± 4.8 months). The modified Japanese Orthopaedic Association (mJOA) score, Neck Disability Index (NDI), and visual analogue scale (VAS) were used to assess clinical outcomes. Pre-operative, 6-month post-operative, and final follow-up radiographs were collected to compare total cervical spine curvature (C2-C7), curvature of the operated segments, range of motion (ROM) in the total cervical spine, operated segmental ROM, ACDR segmental ROM, and operated adjacent segmental ROM. The height of the superior articular process (HSAP), the orientation of zygapophyseal joint spaces (OZJS), and the length of the superior articular surface (LSAS) were measured. The FE-COR of the ACDR segment was measured using the mid-plumb line method. The translation distance of the Mobi-C was measured. The degree of disc degeneration in the adjacent segment, bony fusion of the ACDF segment, and ABL of the upper and lower vertebra of the ACDR segment were observed. RESULTS: In our group, all patients have shown improvements in their postoperative mJOA, NDI, and VAS scores. Overall cervical ROM and surgical segmental ROM decreased (P < 0.05). However, there was no significant decrease in ACDR segmental ROM and upper or lower adjacent segmental ROM compared with pre-operatively (P > 0.05). For FE-COR-X, only the last follow-up compared with pre-surgery showed statistical significance (46.74 ± 7.71% vs. 50.74 ± 6.92%, P < 0.05). For FE-COR-Y, the change was statistically significant at both 6 months post-operation and the final follow-up compared to pre-operation (45.37% ± 21.11% vs. 33.82% ± 10.87%, 45. 37% ± 21.11% vs. 27.48% ± 13.58%, P < 0.05). No significant difference in the Mobi-C translation distance was observed (P > 0.05). Moreover, the difference in HSAP was not statistically significant at each node (P > 0.05). The OZJS and LSAS were significantly different at the final follow-up compared to the pre-operative period (P < 0.05). All the ACDF segments were observed in a stable condition at the final follow-up. Furthermore, 9 of the adjacent segments showed imaging ASD (9/82, 10.98%), and all were present at the last follow-up, of which 6 were mild, and 3 were moderate. Twenty of the 42 Mobi-C segments had no significant ABL (grade 0) 6 months post-operatively (47.62%). Sixteen cases (38.10%) showed mild ABL (grade 1), and 6 cases (14.28%) showed moderate ABL (grade 2). No severe ABL occurred. CONCLUSION: The cervical hybrid surgery using Mobi-C artificial cervical discs can achieve satisfactory results. The Mobi-C segmental FE-COR-X shows a slow forward shift trend, and FE-COR-Y drops noticeably within 6 months post-surgery before stabilizing. It's common to see mild to moderate ABL after cervical hybrid surgery using Mobi-C, and significant progression is unlikely in the short term. Furthermore, changes in the FE-COR after hybrid surgery in the Mobi-C segment might not affect clinical outcomes.

Effect of use of NSAIDs or steroids during the acute phase of pain on the incidence of chronic pain: a systematic review and meta-analysis of randomised trials.

BACKGROUND: This study is the first to summarize the evidence on how the use of anti-inflammatory drugs during acute pain has an impact on the development of chronic pain. METHODS: Randomized controlled trials retrieved from nine databases included anti-inflammatory drugs (NSAIDs or steroids) versus non-anti-inflammatory drugs in patients with acute pain and reported the incidence of chronic pain. No specified date, age, sex, or language restrictions. Subgroup analyses were performed according to pain classification, follow-up time, and medication. The GRADE method was used to evaluate quality of evidence. RESULTS: A total of 29 trials (5220 patients) were included. Steroids or NSAIDs did not reduce the incidence of chronic nociceptive pain. Steroid use in acute phase significantly reduced the incidence of chronic neuropathic pain. In subgroup analysis, benefits were observed for methylprednisolone and dexamethasone, with some adverse effects. Steroids or NSAIDs were statistically significant in reducing pain intensity over 1 year, but the effect size was too small, and whether the long-term effect is clinically relevant needs to be further studied. CONCLUSION: Quality of the evidence was low to moderate. No drug can be recommended to prevent chronic nociceptive pain. Injections of steroids (methylprednisolone or dexamethasone) during the acute phase reduce the incidence of chronic neuropathic pain, but most included studies also used local anesthetics. The results are indirect and need to be interpreted with caution. The pooled data effect sizes for pain intensity were small, so the clinical relevance was unclear. Study registration PROSPERO (CRD42022367030).

Etiological spectrum and treatment outcome of wound infection in patients with open tibia and fibula fractures.

OBJECTIVE: To explore the etiology of wound infections in patients with open tibia and fibula fractures and the treatment effects. METHODS: In this retrospective study, a total of 76 patients with open tibia and fibula fractures were included in this research. These patients were divided into the control group (n=38) and the observation group (n=38) according to the treatment methods for wound infection. The distribution and drug resistance of pathogenic bacteria in wound infections were analyzed. Clinical effects, time for body temperature returning to normal, time for disappearance of exudates, time for clearance of pathogenic bacteria, recovery effects and patients' satisfaction rate were also compared between two groups. RESULTS: A total of 152 strains of pathogenic bacteria were separated. The main pathogenic bacterium was Acinetobacter baumannii, accounting for 30.92% (47/152). Pathogenic bacteria were demonstrated to be highly sensitive to vancomycin and imipenem. The proportion of wound healing by first intention and the Johner-Wruhs scores in observation group were significantly higher than those in control group, while recurrent infection rate, the time to restore normal body temperature, the time for exudates to disappear, the time to remove pathogenic bacteria, hospital stays and VAS scores in observation group were obviously shorter or lower than those in control group (all P<0.05). Moreover, the satisfaction rate of patients in the observation group was significantly higher than that in the control group (P<0.05). CONCLUSION: Understanding pathogenic characteristics and drug resistance of wound infection in patients with open tibia and fibula fractures is helpful to subsequent treatment. Comprehensive control measures should be taken to decrease incidence of wound infection.

Potential biocontrol efficiency of Trichoderma species against oomycete pathogens.

Plant health is of utmost importance for optimal agricultural production and sustainability. Unfortunately, biotic and abiotic factors put a major constraint on crop safety and productivity. Plant diseases caused by oomycetes inflict serious damage to various crops. Moreover, the injudicious use of chemical pesticides poses threats related to pesticide resistance development in pathogens and environmental pollution. Biocontrol offers an effective solution for disease control; however, research on biocontrol of oomycete-related diseases is scarce. Thus, this study undertakes the screening of biocontrol resources for the effective management of oomycete-related plant diseases. In this regard, 86 isolates of Trichoderma spp. were assessed against Phytophthora nicotianae, P. capsici, Pythium vexans, P. ultimum, and P. dissotocum through dual culture assay. Furthermore, the antagonistic effect of selected isolates was studied against tobacco black shank disease and damping-off of cucumber seedlings in the greenhouse. The relative control effect of the three antagonistic Trichoderma strains AR-4, Tv-1, and ST4-1 on tobacco black shank was more than 60%, which was not significantly different from 6.88 gl-1 fluopicolide-propamocarb. Whereas, the relative control effect of Trichoderma AR-4 and ST4-1 on damping-off of cucumber seedlings was 80.33% and 82.67%, respectively, which were significantly higher than Trichoderma Tv-1 (35.49%) and fluopicolide-propamocarb (47.82%). According to the morphological and molecular characterization, the fungal strains AR-4, Tv-1, and ST4-1 were identified as Trichoderma koningiopsis, T. asperellum, and T. gamsii, respectively. In conclusion, the strains exhibited a strong antagonistic effect against oomycete pathogens and can be integrated into disease management strategies.

Soil mineral nitrogen regulation by a novel porous material in structurally degraded soils.

BACKGROUND: The availability of soil nitrogen (N) decreases as the structure of agricultural soils degrades. Traditional methods focus on organic amendments that indirectly affect the porosity and N content of soil. Due to the low efficiency of such amendments, new materials, particularly highly porous materials, are needed to improve the quality of soil, which has opened new directions. RESULTS: The addition of 2 to 7 mm of porous clay ceramic (PLC) significantly increased the fresh weight of Brassica chinensis. The soil aeration porosity (>50 µm) increased by 0.69% on average in response to 1% PLC application. Soil NO3 - -N, NH4 + -N and mineral N increased by 3.3, 1.3 and 4.6 mg kg-1 on average, respectively, following a 1% PLC application rate. The initial N content of the high PLC treatments was the lowest in the incubation experiment. The parameters of soil N mineralization, i.e. potentially mineralizable N (N0 ), the first-order rate constant (k) and the mineralization composite index (N0  × k), increased obviously as the amount of PLC increased. Porosities larger than 1000 µm were significantly more positively correlated with the parameters of soil N mineralization than those <500 µm. The Pearson correlation coefficients suggested that high porosity, mineral N and N0 values had significant positive relationships with the fresh weights in double seasons. CONCLUSION: The application of PLC increased soil aeration and enhanced the availability of soil N, which yielded large vegetable harvests in clayey soils in the short term. © 2022 Society of Chemical Industry.

Extracellular vesicles-encapsulated microRNA-29b-3p from bone marrow-derived mesenchymal stem cells promotes fracture healing via modulation of the PTEN/PI3K/AKT axis.

Bone marrow-derived mesenchymal stem cells (BM-MSCs) are well-established as vital regulators of fracture healing, whereas angiogenesis is one of the critical processes during the course of bone healing. Accordingly, the current study sought to determine the functions of microRNA (miR)-29b-3p from BM-MSCs-derived extracellular vesicles (EVs) on the angiogenesis of fracture healing via the PTEN/PI3K/AKT axis. Firstly, BM-MSCs-EVs were extracted and identified. The lentiviral protocol was adopted to construct miR-29b-3pKD-BMSCs or miR-negative control-BMSCs, which were then co-cultured with human umbilical vein endothelial cells (HUVECs) in vitro to determine the roles of EVs-encapsulated miR-29b-3p on the proliferation, migration, and angiogenesis of HUVECs in vitro with the help of a CCK-8 assay, scratch test, and tube formation assay. Subsequent database prediction, luciferase activity assay, RT-qPCR, and Western blot assay findings identified the downstream target gene of miR-29b-3p, PTEN, and a signaling pathway, PI3K/AKT. Furthermore, the application of si-PTEN attenuated the effects induced by miR-29b-3pKD-EVs. Finally, a mouse model of femoral fracture was established with a locally instilled injection of equal volumes of BM-MSCs-EVs and miR-29b-3pKD-BM-MSCs-EVs. Notably, the mice treated with BMSC-EVs presented with enhanced neovascularization at the fracture site, in addition to increased bone volume (BV), BV/tissue volume, and mean bone mineral density; whereas miR-29b-3pKD-BMSCs-EVs-treated mice exhibited decreased vessel density with poor fracture healing capacity. Collectively, our findings elicited that BM-MSCs-EVs carrying miR-29b-3p were endocytosed by HUVECs, which consequently suppressed the PTEN expression and activated the PI3K/AKT pathway, thereby promoting HUVEC proliferation, migration, and angiogenesis, and ultimately facilitating fracture healing.

Comparison of 2-year follow-up results of the hybrid surgery using Mobi-C combined with ROI-C and anterior cervical discectomy and fusion for the treatment of contiguous two-level cervical degenerative disc diseases.

Although the Mobi-C artificial disc and the ROI-C cervical cage have been widely used in the treatment of cervical degenerative disc diseases (CDDD), few reports addressed the features of combined application of both devices. This study is aimed at comparing the clinical and radiological outcomes of treating contiguous two-level CDDD using Mobi-C and ROI-C combined in a hybrid surgery (HS) with anterior cervical discectomy and fusion (ACDF) using ROI-C. We reviewed ninety-one patients who underwent HS (n = 48) or ACDF (n = 43) surgery for symptomatic contiguous two-level CDDD. >2 years' clinical and radiological outcomes were reviewed and evaluated retrospectively. At the last follow-up, significant improvement in the mean VAS, JOA, and NDI scores was found both in the HS and ACDF groups (p < 0.05), while the differences between groups were not significant (p > 0.05). The global range of motion (ROM) in the HS group was significantly larger than that in the ACDF group (p < 0.05). The local lordosis improved significantly after surgery in all patients (p < 0.05). Bone resorption and heterotopic ossification (HO) were found after surgery. The result showed that, for the selected patients, HS may provide an alternative approach for the treatment of contiguous two-level CDDD. HS also offers the benefit of both greater global ROM and greater ROM at the Mobi-C index level. Some degree of bone resorption may be an integral component in the early stage of bony fusion in the cage index level. Further studies and long-term follow-up are still needed.

Dynamic Fusion Process in the Anterior Cervical Discectomy and Fusion with Self-Locking Stand-Alone Cages.

BACKGROUND: Self-locking stand-alone cages can achieve satisfactory clinical results and fusion rate. However, there have been no reports on the causes and relationship of different fusion state. This study is to classify the different fusion states of the index level and to explore the potential contributing factors and links of them. METHODS: From June 2008 to October 2011, 42 patients underwent anterior cervical discectomy and fusion with MC+ cages. More than 5 years' follow-up was reviewed. The fusion state and the relevant clinical and radiologic records were reviewed retrospectively. RESULTS: At the last follow-up, the fusion proportion of type I, II, III, and IV was 11.7%, 16.9%, 26.9%, and 42.9%, respectively. The overall fusion rate was 97.4%. For all the fused types, significant improvement for the visual analog scale, Japanese Orthopaedic Association, and Neck Disability Index scores was found at the last follow-up (P < 0.05). However, there were no significant differences between the 4 types (P > 0.05). For sagittal vertical axis, type IV was significantly larger than that of type I, II, and III (P < 0.05), and for range of motion, type III was significantly larger than that of type II and IV (P < 0.05). CONCLUSIONS: For anterior cervical discectomy and fusion with self-locking stand-alone cages, the fusion of the index level seems to be a progressive dynamic process during the mid-term follow-up, which may be influenced by the location of the cage, the aagittal vertical axis of the index level, and the global range of motion of the cervical spine. Satisfactory clinical results could be achieved by all the fused types.

[Application of perfusion bone cement screw in lumbar degenerative disease with osteoporosis].

OBJECTIVE: To observe the early clinical effect of perfusion bone cement screw for lumbar degenerative diseases with osteoporosis. METHODS: The clinical data of 28 patients with lumbar degenerative diseases combined with moderate to severe osteoporosis treated by posterior lateral graft fusion with perfusion of bone cement screws from June 2015 to June 2017 were retrospectively analyzed. There were 9 males and 19 females, aged from 55 to 86 years old with an average of 76 years. Anteroposterior, oblique, and dynamic radiography were performed before operation, and the diagnosis was confirmed by CT, MRI and dual energy X-ray absorptionmetry(DXA). All the patients had moderate to severe lumbar spinal stenosis, including 16 cases with degenerative scoliosis, 12 cases with degenerative lumbar spondylolisthesis, and 16 cases with lumbar disc herniation. According to Jikei grade of osteoporosis, 9 cases were grade II and 19 cases were grade III. Visual analogue scale (VAS) and Japanese Orthopaedic Association (JOA) score were used to assess the improvement of lumbar leg pain, and neurological function. Imaging data were used to observe the circumstance of pedicle loosening, prolapse, breakage and bone cement leakage, and comprehensively evaluate the fusion. RESULTS: The hospital stay was from 10 to 14 days with an average of 12 days; the operative time was 100 to 150 min with an average of 120 min;the blood loss was 200 to 600 ml with an average of 350 ml (for operations more than 3 vertebral segments, blood filtration recovery was intraoperatively used);the postoperative drainage volume was 150 to 600 ml with an average of 300 ml, no allogeneic blood was used in all the patients. Bone cement of 2 to 3 ml were injected into each vertebral body, and bone cement leakage occurred in 2 cases during injection, both of which were paravertebral vessel leakage, and there was no evidence of intravertebral leakage. The injection of bone cement was terminated in a timely manner without serious complications such as nerve injury, bone cement toxicity, and vascular embolization and pulmonary embolism. According to the fusion criteria by X-ray, 18 cases achieved strong bone fusion, and 10 cases were inaccurate fusion, but no pseudarthrosis occurred in the fusion segment. No screw loosening, prolapse or fracture were found, and postoperative VAS, JOA scores were significantly improved. CONCLUSIONS: Perfusion bone cement screw technique can obtain satisfactory effect in treating lumbar degenerative diseases with osteoporosis.

Genotyping-by-sequencing application on diploid rose and a resulting high-density SNP-based consensus map.

Roses, which have been cultivated for at least 5000 years, are one of the most important ornamental crops in the world. Because of the interspecific nature and high heterozygosity in commercial roses, the genetic resources available for rose are limited. To effectively identify markers associated with QTL controlling important traits, such as disease resistance, abundant markers along the genome and careful phenotyping are required. Utilizing genotyping by sequencing technology and the strawberry genome (Fragaria vesca v2.0.a1) as a reference, we generated thousands of informative single nucleotide polymorphism (SNP) markers. These SNPs along with known bridge simple sequence repeat (SSR) markers allowed us to create the first high-density integrated consensus map for diploid roses. Individual maps were first created for populations J06-20-14-3×"Little Chief" (J14-3×LC), J06-20-14-3×"Vineyard Song" (J14-3×VS) and "Old Blush"×"Red Fairy" (OB×RF) and these maps were linked with 824 SNPs and 13 SSR bridge markers. The anchor SSR markers were used to determine the numbering of the rose linkage groups. The diploid consensus map has seven linkage groups (LGs), a total length of 892.2 cM, and an average distance of 0.25 cM between 3527 markers. By combining three individual populations, the marker density and the reliability of the marker order in the consensus map was improved over a single population map. Extensive synteny between the strawberry and diploid rose genomes was observed. This consensus map will serve as the tool for the discovery of marker-trait associations in rose breeding using pedigree-based analysis. The high level of conservation observed between the strawberry and rose genomes will help further comparative studies within the Rosaceae family and may aid in the identification of candidate genes within QTL regions.

Downregulation of miR-10b promotes osteoblast differentiation through targeting Bcl6.

MicroRNAs (miRNAs or miRs) have been shown to play a critical role in osteoblast differentiation. miR-10b has been found to be downregulated during osteoblast differentiation; however, its precise effect on osteoblast differentiation remains unknown. In this study, we aimed to investigate the potential role of miR-10b and the potential underlying mechanism in regulating osteoblast differentiation. We found that miR-10b was downregulated during osteoblast differentiation. Overexpression of miR-10b inhibited osteoblast differentiation, whereas the suppression of miR-10b promoted osteoblast differentiation. Bioinformatics analysis and the dual-luciferase reporter assay demonstrated that miR-10b could target the 3'-untranslated regions of B cell lymphoma 6 (Bcl6) which is an important regulator of osteoblast differentiation. Real­time quantitative polymerase chain reaction and western blot analysis showed that miR-10b directly regulated Bcl6 expression. Further experiments showed that the overexpression of miR-10b increased the expression of signal transducer and activator of transcription 1 (STAT1) and blocked Runt-related transcription factor 2 (Runx2) nuclear translocation, whereas miR-10b suppression showed an opposite effect. Moreover, the miR-10b suppression-induced effects were partially reversed by Bcl6 knockdown. Taken together, our study suggests that miR-10b contributes to osteoblast differentiation through targeting Bcl6, providing a novel insight into understanding the molecular mechanism underlying osteoblast differentiation and suggesting a potential target for inhibiting bone loss.

Exploiting aberrant mRNA expression in autism for gene discovery and diagnosis.

Autism spectrum disorder (ASD) is characterized by substantial phenotypic and genetic heterogeneity, which greatly complicates the identification of genetic factors that contribute to the disease. Study designs have mainly focused on group differences between cases and controls. The problem is that, by their nature, group difference-based methods (e.g., differential expression analysis) blur or collapse the heterogeneity within groups. By ignoring genes with variable within-group expression, an important axis of genetic heterogeneity contributing to expression variability among affected individuals has been overlooked. To this end, we develop a new gene expression analysis method-aberrant gene expression analysis, based on the multivariate distance commonly used for outlier detection. Our method detects the discrepancies in gene expression dispersion between groups and identifies genes with significantly different expression variability. Using this new method, we re-visited RNA sequencing data generated from post-mortem brain tissues of 47 ASD and 57 control samples. We identified 54 functional gene sets whose expression dispersion in ASD samples is more pronounced than that in controls, as well as 76 co-expression modules present in controls but absent in ASD samples due to ASD-specific aberrant gene expression. We also exploited aberrantly expressed genes as biomarkers for ASD diagnosis. With a whole blood expression data set, we identified three aberrantly expressed gene sets whose expression levels serve as discriminating variables achieving >70 % classification accuracy. In summary, our method represents a novel discovery and diagnostic strategy for ASD. Our findings may help open an expression variability-centered research avenue for other genetically heterogeneous disorders.

Epistasis and destabilizing mutations shape gene expression variability in humans via distinct modes of action.

Increasing evidence shows that phenotypic variance is genetically determined, but the underlying mechanisms of genetic control over the variance remain obscure. Here, we conducted variance-association mapping analyses to identify expression variability QTLs (evQTLs), i.e. genomic loci associated with gene expression variance, in humans. We discovered that common genetic variants may contribute to increasing gene expression variance via two distinct modes of action­epistasis and destabilization. Specifically, epistasis explains a quarter of the identified evQTLs, of which the formation is attributed to the presence of 'third-party' eQTLs that influence the gene expression mean in a fraction, rather than the entire set, of sampled individuals. On the other hand, the destabilization model explains the other three-quarters of evQTLs, caused by mutations that disrupt the stability of the transcription process of genes. To show the destabilizing effect, we measured discordant gene expression between monozygotic twins, and estimated the stability of gene expression in single samples using repetitive qRT-PCR assays. The mutations that cause destabilizing evQTLs were found to be associated with more pronounced expression discordance between twin pairs and less stable gene expression in single samples. Together, our results suggest that common genetic variants work either interactively or independently to shape the variability of gene expression in humans. Our findings contribute to the understanding of the mechanisms of genetic control over phenotypic variance and may have implications for the development of variance-centred analytic methods for quantitative trait mapping.

Rapid determination of 88 veterinary drug residues in milk using automated TurborFlow online clean-up mode coupled to liquid chromatography-tandem mass spectrometry.

A novel method based on TurborFlow online solid phase extraction (SPE) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been established for simultaneous screening and confirmation of 88 wide-range veterinary drugs belonging to eight families (20 sulfonamides, 7 macrolides, 15 quinolones, 8 penicillins, 13 benzimidazoles, 4 tetracyclines, 2 sedatives, and 19 hormones) in milk. The preparation method consists of sample dilution and ultrasonic extraction, followed by an automated turbulent flow cyclone chromatography sample clean-up system. The detection was achieved in selected reaction monitoring mode (SRM). The total run time was within 39 min, including automated extraction, analytical chromatography and re-equilibration of the turboflow system. The optimization of different experimental parameters including extraction, purification, separation, and detection were evaluated separately in this study. The developed method was validated and good performing characteristics were obtained. The linear regression coefficients (R(2)) of matrix-match calibration standard curves established for quantification were higher than 0.9930. The limits of detection (LOD) were in the range of 0.2-2.0 µg/kg given by signal-noise ratio ≥3 (S/N) and the limits of quantification (LOQ, S/N≥10) ranged between 0.5 µg/kg and 10 µg/kg. Average recoveries of spiked target compounds with different levels were between 63.1% and 117.4%, with percentage relative standard deviations (RSD) in the range of 3.3-17.6%. The results indicated that the developed method has great potential for the routine laboratory analysis of large numbers of samples on measuring different classes of compounds. In comparison to traditional procedures, the automated sample clean-up ensures rapid, effective, sensitive analyses of veterinary drugs in milk.

[Determination of poppy ingredients in chafing dish materials by isotopic internal standard coupled with multiple reaction monitoring and online full scan mass spectrometry].

A confirmative method was developed for determining five poppy alkaloids including morphine, codeine, papaverine, tibane, noscapine in chafing dish ingredients by high performance liquid chromatography coupled with triple quadrupole linear ion trap mass spectrometry (HPLC-Q Trap MS). The sample was extracted with dilute HCl solution under heating condition. The removal of lipid procedure was performed with hexane. The purification was carried out on a mixed-cation solid-phase extraction column (MCX) and ethyl acetate-methanol containing 5% aqueous ammonia was used for elution. A PAK ST column was used to separate the analytes, and 5 mmol/L ammonium acetate methanol and 10 mmol/L ammonium acetate (pH 3. 6) were used as mobile phases. The five alkaloids was detected in the positive mode simultaneously by multiple reaction monitoring (MRM) and online enhanced product ion full scan (EPI). The LODs were 0.05-0.5 µg/kg and the LOQs were 0. 2-2 µg/kg for the five poppy alkaloids. The overall recoveries of the method varied from 64. 2% to 110. 6%, and the RSD were between 4. 2% and 12. 5%. The EPI mass spectra of positive samples were searched through standard library for qualitative confirmation. The detection of real hot pot material samples showed this method can be used for the simple and accurate determination of the five poppy alkaloid residues in chafing dish.

[The expression of BUB1 in laryngeal squamous cell carcinoma and its clinical significance].

OBJECTIVE: To investigate the expression and clinical significance of BUB1 protein in laryngeal squa mous cell carcinoma (LSCC) tissues. METHOD: The expression of BUB1 protein was measured by immunohistochemistry (EliVision two steps) in 55 cases of LSCC tissues,30 cases of adjacent normal laryngeal mucosa tissues, the relationship of BUB1 expression with clinical pathologic factors in LSCC was also analyzed. RESULT: The positive rate of BUB1 protein in LSCC tissues (50.9%, 28/55) was significantly lower than that in adjacent normal laryngeal mucosa tissues (83.3%, 25/30), P < 0.01. The positive rate of BUB1 protein was correlated with differentiation grading and lymph node metastasis of LSCC (P < 0.05, respectively), but not with patient's sex, age, smoking, tumor site, T-stage and clinical stage(P > 0.05, respectively). CONCLUSION: The low expresssion of BUB1 protein is closely related to the tumorigenesis and development of LSCC, and can be a molecular marker for prognosticating metastasis and prognosis of LSCC.

[Rapid determination of total residues of ribavirin and its metabolites in chicken and chicken products by ultra-performance hydrophilic interaction chromatography-tandem mass spectrometry].

An analytical method was developed for the determination of total residues of ribavirin and its phosphorylated metabolites in chicken and its products by hydrophilic interaction chromatography-tandem mass spectrometry. The samples were extracted with acetonitrile containing 1% (v/v) acetic acid under ultrasonic condition and then enzymatically hydrolysed with acid phosphatase at 37 degrees C. After liposoluble substances being removed with hexane, C18 and PSA dispersion solid phase extraction (DSPE) was introduced to cleanup procedures. The separation was performed on an ultra-performance hydrophilic interaction chromatographic (HILIC) amide column under a gradient elution using acetonitrile and 0.1% formic acid. The analytes were detected in the positive electrospray ionization and multi-reaction monitoring (MRM) mode. In the range of 1-200 microg/kg, the correlation coefficient was greater than 0.999. The limit of detection (LOD, S/N > or = 3) was 1 microg/kg and the limit of quantification (LOQ, S/N > or = 10) was 5 microg/kg. The recoveries of ribavirin spiked at three levels ranged from 67.8% to 112.7% with the relative standard deviations (RSDs) of 6.1%-13.6%. The results of the determination of ribavirin in various real samples showed that the method is simple, rapid, accurate and suitable for the determination of total residues of ribavirin and its metabolites in chicken and its products.

[Simultaneous determination of 11 aminophenols in hair dyes by high performance liquid chromatography].

An analytical method was developed for the simultaneous determination of 11 aminophenols in direct and oxidized hair dyes by high performance liquid chromatography (HPLC). The samples were extracted with methanol using sodium bisulfite for anti-oxidation. The purification was carried out with the high speed frozen centrifugation. The separation was performed on an Agilent Zorbax SB C18 column with the mobile phases of acetonitrile and an ion pair system of sodium heptanesulfonate and phosphate under a gradient elution. The analytes were detected at three different wavelengths of 230, 270 and 400 nm. In the concentration range of 0.05 - 500 mg/L, the correlation coefficients for the 11 aminophenols were not less than 0.9992. The limits of quantification including 4-amino-3-nitrophenol, 2-amino-5-nitrophenol, and 4-(2-hydroxyethyl) amino-3-nitrophenol were 5 mg/kg. Other aminophenols including 4-aminophenol, 4-methyl aminophenol, 3-aminophenol, 2-aminophenol, 5-amino-o-cresol, 4-amino-3-methylphenol, 5-(2-hydroxyethyl) amino-o-cresol and 2-amino-4-nitrophenol were 20 mg/kg. The recoveries of the aminophenols spiked at different levels ranged from 88.5% to 109. 5% with the relative standard deviations (RSDs) within the range of 2.2% - 8.3%. The commercially available hair dye samples were analyzed for the aminophenols and the results showed that the method was suitable for the determination of the 11 aminophenols in direct and oxidized hair dyes.

[Determination of four triorganotin residues in leather products by ultra performance liquid chromatography-tandem mass spectrometry and cation solid-phase extraction].

A confirmative method was developed for the determination of four triorganotins (TOTs) included tributyltin (TBT), triphenyltin (TPhT), di-triphenyltin oxide (DTPhT), and cyhexatin (CYT) in leather products by cation solid-phase extraction (SPE) and ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The organotin residues were extracted with HCl solution. The ionized compounds were purified with SPE. The reconstituted sample solution was analyzed by UPLC-MS/MS in positive ion electrospray and multi-reaction monitor (MRM) mode, with a UPLC C18 column as the separation column. The limits of quantification were 10 microg/kg for TBT, TPhT, DTPhT (calculated as TPhT) and 20 microg/kg for CYT. The method was validated at three levels for each compound. The recoveries were from 54.1% to 101.4%, and the relative standard deviations (RSD) were between 4.3% and 9.8%. The method is simple, reliable and accurate. It can meet the requirements of the domestic and international legislations. The method adapts to simultaneously confirm the residues of the four TOTs in leather products.