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"Wang, Song-Ying"
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Functional Characterizations of Chemosensory Proteins of the Alfalfa Plant Bug Adelphocoris lineolatus Indicate Their Involvement in Host Recognition
Insect chemosensory proteins (CSPs) have been proposed to capture and transport hydrophobic chemicals from air to olfactory receptors in the lymph of antennal chemosensilla. They may represent a new class of soluble carrier protein involved in insect chemoreception. However, their specific functional roles in insect chemoreception have not been fully elucidated. In this study, we report for the first time three novel CSP genes (AlinCSP1-3) of the alfalfa plant bug Adelphocoris lineolatus (Goeze) by screening the antennal cDNA library. The qRT-PCR examinations of the transcript levels revealed that all three genes (AlinCSP1-3) are mainly expressed in the antennae. Interestingly, these CSP genes AlinCSP1-3 are also highly expressed in the 5(th) instar nymphs, suggesting a proposed function of these CSP proteins (AlinCSP1-3) in the olfactory reception and in maintaining particular life activities into the adult stage. Using bacterial expression system, the three CSP proteins were expressed and purified. For the first time we characterized the types of sensilla in the antennae of the plant bug using scanning electron microscopy (SEM). Immunocytochemistry analysis indicated that the CSP proteins were expressed in the pheromone-sensitive sensilla trichodea and general odorant-sensitive sensilla basiconica, providing further evidence of their involvement in chemoreception. The antennal activity of 55 host-related semiochemicals and sex pheromone compounds in the host location and mate selection behavior of A. lineolatus was investigated using electroantennogram (EAG), and the binding affinities of these chemicals to the three CSPs (AlinCSP1-3) were measured using fluorescent binding assays. The results showed several host-related semiochemicals, (Z)-3-hexen-1-ol, (E)-2-hexen-1-al and valeraldehyde, have a high binding affinity with AlinCSP1-3 and can elicit significant high EAG responses of A. lineolatus antennae. Our studies indicate the three antennae-biased CSPs may mediate host recognition in the alfalfa plant bug A. lineolatus.
Journal Article
Cervical Abnormalities in Severe Spinal Deformity: A 10‐year MRI Review
Objective To investigate the incidence of cervical anomalies (CA), including cervical intraspinal neural axis abnormalities (CIINAA) and/or cervical osseous abnormalities (COA), and the clinical relevance in severe spinal deformities (SSD) at a single center. Methods A retrospective study of SSD admitted for spinal surgery from January 2003 to January 2015 was conducted at a single center. Inclusion criteria: patients who present with coronal Cobb over 90° (and/or sagittal cobb ≥90°); and patients with complete imaging and clinical data preoperatively. Exclusion criteria: ankylosing spondylitis, adult onset scoliosis, scoliosis secondary to bone destruction. There were 108 SSD patients who fulfilled the criteria in this research (41 males and 67 females). The mean age of the patients was 18.1 ± 2.7 years (range, 10–45 years). The clinical and radiological data of these patients were reviewed to identify CA and to analyze the relationship between clinical and radiographic characteristics in the population of SSD. Results The major curves of scoliosis and segmental kyphosis were 109.1° ± 24.7° and 91.2° ± 29.1°. Cervical abnormalities were detected in 56 patients (51.85%) with 9 different CA, including 28 patients (25.9%) with 6 different COA, 21 patients (19.4%) with 3 different CIINAA, and 7 patients (6.5%) with a combination of COA and cervical intraspinal neural axis abnormalities (CINAA). Basilar invagination and Klippel–Feil syndrome were the most frequent COA. Syringomyelia was the most frequent CINAA. SSD with COA in upper vertebral levels (UVL) had a higher incidence of CINAA than those in subaxial vertebral levels (SVL) (P = 0.024) and SSD with multiple COA (mCOA) in UVL had a higher incidence of CINAA than those with single COA (sCOA) (P = 0.029). In the present study, 83.9% of the SSD with CA were asymptomatic. Conclusion The incidence of CA in SSD was 51.85%, with most presenting with intact neurologic status. As the diversity of COA increased, we found a higher incidence of CINAA, especially in UVL.
Journal Article
The risk factors of neurologic deficits of one-stage posterior vertebral column resection for patients with severe and rigid spinal deformities
Purpose
To determine the risk factors of neurologic deficits during PVCR correction, so as to help improve safety during and after surgery.
Methods
A consecutive series of 76 patients with severe and rigid spinal deformities who were treated with PVCR at a single institution between October 2004 and July 2011 were included in our study. Of the 76 patients, 37 were male and 39 female, with an average age of 17.5 years (range 10–48 years). There were 52 adolescent patients (with an age <18 years) and 24 adult patients (with an age ≥18 years). Preoperatively, postoperatively and 6 months after surgery, we performed systemically neurologic function evaluations of each patients through meticulous physical examination. Any new abnormality or deterioration in evaluation of neurologic function than preoperative is reckoned postoperative neurologic deficits. Ten variables that might affect the safety of neurologic deficits during PVCR procedures, including imaging factors, clinical factors and operational factors, were analyzed using univariate analysis. Then the variables with statistical difference were analyzed by using multi-factor unconditional logistic regression analysis.
Results
No patient in this series had permanent paraplegia and nerve root injury due to operation. Change of neurologic status was found in six patients after surgery. Results of single-factor comparison demonstrated that the following seven variables were statistically different (
P
< 0.05): location of apex at main curve (
X
3
), Cobb angle at the main curve at the coronal plane (
X
4
), scoliosis associated with thoracic hyperkyphosis (
X
5
), level of vertebral column resected (
X
6
), number of segmental vessels ligated (
X
7
), preexisting neurologic dysfunction (
X
8
), and associated with intraspinal and brain stem anomalies (
X
9
). The multi-factor unconditional logistic regression analysis revealed that
X
8
(OR = 49.322),
X
9
(OR = 18.423),
X
5
(OR = 11.883), and
X
6
(OR = 8.769) were independent and positively correlated with the neurologic deficit.
Conclusions
Preexisting neurologic dysfunction, associated with intraspinal and brain stem anomalies, scoliosis associated with thoracic hyperkyphosis and level of vertebral column resected are independent risk factors for neurologic deficits during PVCR procedure.
Journal Article
Expert consensus for PVCR in severe, rigid and angular spinal deformity treatment: The Kunming consensus
In the past decades, an increasing number of surgeons started using posterior vertebral column resection (PVCR) to treat severe, rigid and angular spinal deformities. Little high-level evidence is available to guide surgical treatment. The aims of our study were to identify important surgical strategies and key technical points of Chinese experts who have extensive experience in the management of severe, rigid and angular spinal deformities using PVCR, and to standardize and unify the current core concepts. Workgroups of consensus were formed by selecting nationwide representing experts and comprehensive consultations. Eight task forces for major issues were established, then retrieval of literature, collection of expert opinions and writing of review articles were carried out. A modified Delphi process was chosen in round-table forum with three face-to-face meetings. Consensus was reached with items graded more than seven points including: indications and contraindications of PVCR; review PVCR in the evolution of spinal osteotomies; The corrective mechanism and safety of spinal cord; monitoring and responses of spinal cord crisis; characteristics and therapeutic outcome of pulmonary function; management of bleeding during PVCR; relationship of pedicle screw insertion and spinal cord safety; and analysis of non-neurologic complications and prevention strategies. In conclusion, The essential properties regarding PVCR procedure are tightly linked with various factors such as medical and surgical indication, range and level of vertebral column resection, strategies of correction, corrective efficiency and control of neurological risk. PVCR is used mainly for severe, rigid spinal deformity that is not manageable by other osteotomy techniques.
Journal Article
A five-step remedial screw placement method to treat severe spinal deformity with free-hand transpedicular screw placement
Purpose
Severe spinal deformity is a complex morphological deformation that occurs and develops in three-dimensional space combined with abnormal development and morphology of anatomical structures, which presents great difficulties in the process of transpedicular screw placement. This study tried to explore the methods of transpedicular screw placement in surgical correction of severe spinal deformities.
Methods
Surgical corrections through posterior approach were performed in all the 76 cases (mean age 20.4 years). The averaging preoperative Cobb’s angle of scoliosis was 108.2° ± 33.6° (range 100°–170°). Among these patients, 34 cases were combined with kyphosis; the average Cobb’s angle of kyphosis was 77.3° (range 63°–160°). During operation, the screw tract was first established with the regular free-hand pedicle screw placement method. When this failed, in order to adjust the screw trajectory, a five-step remedial method was performed in the following order: (1) the“funnel” method; (2) exploring the pedicle exterior edge through the costotransverse joint; (3) exploring the superior and inferior edges of pedicle through the nerve root canal; (4) the vertebral plate fenestration; and (5) hemilaminectomy.
Results
Among all 1,472 screws planned to be placed for the patients, 1,210 (82.2 %) were successfully placed after using the regular method, and 262 (17.8 %) failed in this stage. After applying the five-step remedial method, 256 of the failed 262 screws were successfully placed. Among them, 176 screws (68.8 %) were successfully placed after Step 1, 44 (17.2 %) after Step 2, 21 (8.2 %) after Step 3, 12 (4.7 %) after Step 4, and 3 (1.2 %) after Step 5. In only six, pedicles screws could not be placed eventually. No nerve or blood vessel damages occurred in all cases. All final screw positions were validated by CT.
Conclusion
The five-step remedial method proved to be an effective supplementary method for transpedicular screw placement to treat patients with severe spinal deformities. The key points include a detailed preoperative plan, a meticulous hand drilling sensation, and an experienced probing technique for screw tract.
Journal Article
Improvement of chloroperoxidase stability by covalent immobilization on chitosan membranes
Chloroperoxidase (CPO) from Caldariomyces fumago was optimally covalently immobilized on chitosan membranes pretreated with 0.8 M glutaraldehyde at pH 3.5 to give 3.18 mg CPO g⁻¹ support. Using monochlorodimedone (MCD) as assay substrate, the immobilized-CPO retained 40% activity at 50°C after 40 min whereas free CPO retained only 0.02%. The residual activity for immobilized-CPO was 99 and 58% compared with 68 and 43% for free CPO in the presence of 1.5 M urea and 300 μM H₂O₂, respectively, after 20 h.
Journal Article
Development and initial validation of classification for severe spinal deformity based on X-ray features
PurposeTo develop a clinically feasible classification for severe spinal deformity based on X-ray features.MethodsA total of 223 consecutive severe spinal deformity cases who underwent corrective operation were enrolled from 2004 to 2015 retrospectively. Based on X-ray features, a novel classification was developed containing three components: curve types, curve angle and apex location. There were five curve types as follows: single scoliosis (SS), kyphoscoliosis (KS), angular deformity (AD), long curve (LC), and double curves (DC). Curve angle subsection on coronal and sagittal planes including A:90–109, B:110–129, C:130–149, D: > 150. Apex location means the exact level of apex located. Reliability of the classification was tested.ResultsThe kappa values for inter-observer and intra-observer reliability of the curve types, curve angle, and apex level were larger than 0.80. X-ray classification for overall patients with severe spinal deformity showed that there were 101 SS cases, 47 KS, 46 AD, 19 LC and 10 DC. For the curve angle, there were grade A 123 cases, B 43, C 18, D 15 on coronal plane and grade A 38, B 17, C 16, and D 19 on sagittal plane. Apex location showed there were 27 patients at T7 or upper levels, 31 on T8, 58 on T9, 45 on T10, 18 on T11, and 44 at T12 or lower levels.ConclusionA novel classification for severe spinal deformity was described based on X-ray morphology. A high value for inter-observer and intra-observer reliability was shown. Each subgroup has its particular influence on decision-making and prognostic prediction.
Journal Article
A new coumarin derivative, IMM-HOO4, attenuates okadaic acid-induced spatial memory impairment in rats
Aim： A novel coumarin derivative 7-hydroxy-5-methoxy-4-methyl-3-（4-methylpiperazin-l-yl）-coumarin （IMM-HO04） has shown antiapoptotic, anti-inflammatory and neuroprotective activities. In this study we investigated the effects of IMM-HOO4 on spatial memory in rats treated with okadaic acid （OKA）, which was used to imitate Alzheimer＇s disease （AD）-Iike symptoms. Methods： SD rats were administered IMM-HOO4 （8 mg.kg-1d-1, ig） or donepezil （positive control, 1mg.kg-1.d-1, ig） for 25 d. On d 8 and 9, OKA （200 ng） was microinjected into the right ventricle. Morris water maze test was used to evaluate the spatial memory impairments. Tau and 13-amyloid （Aβ） pathology in the hippocampus was detected using Western blot and immunohistochemistry. TUNEL staining was used to detect cell apoptosis. Results： OKA-treated rats showed significant impairments of spatial memory in Morris water maze test, which were largely reversed by administration of IMM-HO04 or donepezil. Furthermore, OKA-treated rats exhibited significantly increased phosphorylation of tau, deposits of Aβ protein and cell apoptosis in the hippocampus, which were also reversed by administration of IMM-HOO4 or donepezil. Conclusion： Administration of IMM-HO04 or donepezil protects rats against OKA-induced spatial memory impairments via attenuating tau or Aβ pathology. Thus, IMM-HOO4 may be developed as a therapeutic agent for the treatment of AD.
Journal Article
Parkin represses 6-hydroxydopamine-induced apoptosis via stabilizing scaffold protein p62 in PC12 cells
Aim： Parkin has been shown to exert protective effects against 6-hydroxydopamine （6-OHDA）-induced neurotoxicity in different models of Parkinson disease. In the present study we investigated the molecular mechanisms underlying the neuroprotective action of parkin in vitro. Methods. HEK293, HeLa and PC12 cells were transfected with parkin, parkin mutants, p62 or si-p62. Protein expression and ubiquitination were assessed using immunoblot analysis. Immunoprecipitation assay was performed to identify the interaction between parkin and scaffold protein p62. PC12 and SH-SY5Y cells were treated with 6-OHDA （200 pmol/L）, and cell apoptosis was detected using PI and Hoechst staining. Results： In HEK293 cells co-transfected with parkin and p62, parkin was co-immunoprecipitated with p62, and parkin overexpression increased p62 protein levels. In parkin-deficient HeLa cells, transfection with wild-type pakin, but not with ligase activity-deficient pakin mutants, significantly increased p62 levels, suggesting that parkin stabilized p62 through its E3 ligase activity. Transfection with parkin or p62 significantly repressed ERK1/2 phosphorylation in HeLa cells, but transfection with parkin did not repress ERK1/2 phosphorylation in p62-knockdown HeLa cells, suggesting that p62 was involved in parkin-induced inhibition on ERK1/2 phosphorylation. Overexpression of parkin or p62 significantly repressed 6-OHDA-induced ERK1/2 phosphorylation in PC12 cells, and parkin overexpression inhibited 6-OHDA-induced apoptosis in PC12 and SH-SY5Y cells. Conclusion： Parkin protects PC12 cells against 6-OHDA-induced apoptosis via ubiquitinating and stabilizing scaffold protein p62, and repressing ERK1/2 activation.
Journal Article