Pleiotropic immunoregulation by growth-blocking peptide in Ostrinia furnacalis.

Insects rely on their innate immune system to eliminate pathogenic microbes. As a system component, cytokines transmit intercellular signals to control immune responses. Growth-blocking peptide (GBP) is a member of the stress-responsive peptide family of cytokines found in several orders of insects, including Drosophila. However, the physiological role of GBP in defence against pathogens is not thoroughly understood. In this study, we explored the functions of GBP in a lepidopteran pest, Ostrinia furnacalis. Injection of recombinant O. furnacalis GBP (OfGBP) precursor (proGBP) and chemically synthesised GBP significantly induced the transcription of antimicrobial peptides (AMPs) and other immunity-related genes including immune deficiency (IMD) and Dorsal. The level of OfGBP mRNA was upregulated after bacterial infection. Knockdown of OfGBP expression led to a decrease in IMD, Relish, MyD88 and Dorsal mRNA levels. OfGBP induced phenoloxidase activity and affected hemocyte behaviours in O. furnacalis larvae. In summary, GBP is a potent cytokine, effectively regulating AMP synthesis, melanization response and cellular immunity to eliminate invading pathogens.

Identification and characteristic analysis of an extracellular signal-regulated kinase from Ostrinia furnacalis Guenée.

The extracellular signal-regulated kinase (ERK) pathway, a critical genetic determinant, controls diverse physiological functions, including innate immunity, development, and stress response. In the current study, a full-length cDNA (1592bp) encoding the ERK gene (OfERK) was cloned from Ostrinia furnacalis Guenée (GenBank accession number: MF797866). The open reading frame of the OfERK gene encoded 364 amino acids and shared 96.43%-98.08% amino acid identities with other insect mitogen-activated protein kinases. For spatiotemporal analysis of the expression pattern, OfERK exhibited a significant peak expression on the 3rd day of the pupa stage and showed the highest expression in hemocytes specifically. Indirect immunofluorescence assays and immuno-electron microscopy revealed a wide distribution of the OfERK protein in hemocytes and epidermis. Moreover, the results demonstrated that the Bt Cry1Ab-activated toxin significantly induces the expression of OfERK. Other genes related to immune response, development, and stress response exhibited dynamic changes in expression after Cry1Ab oral treatment. The expression of OfERK was downregulated through RNA interference, and the correlation of its expression with other related genes was verified using quantitative real-time polymerase chain reaction. Our study provides valuable insights into the regulatory mechanism of ERK in insects for future studies.

Transdural circumferential decompression for thoracic spinal stenosis caused by beak-type ossification of the posterior longitudinal ligament: a technical note.

PURPOSE: Thoracic myelopathy caused by ossification of the posterior longitudinal ligament (OPLL) in the thoracic spine is usually progressive and responds poorly to conservative therapy, making surgery the only effective treatment option. A variety of surgical procedures have been developed to treat thoracic OPLL. However, the optimal surgical approach for removal of thoracic OPLL remains unclear. In the present study, we described a newly modified posterior approach for the removal of OPLL: circular decompression via dural approach, and complete removal of OPLL can be achieved under direct vision and without neurological deficit. MATERIALS AND METHODS: Three patients with beak-type thoracic OPLL presented with progressive thoracic myelopathy and leg weakness. Magnetic resonance imaging showed the spinal cord severely compressed. The surgical management of the three patients involved the 'cave-in' circular decompression and transdural resection of OPLL. RESULTS: Transdural circumferential decompression was successfully performed in all three patients. Clinical outcome measures, including pre- and postoperative radiographic parameters, were assessed. All of the patients were followed up for an average of 12 months (ranging from 10 to 15 months), and no surgery-related complications occurred. Weakness relief and neural function recovery were satisfactorily achieved in all patients by the final follow-up. CONCLUSIONS: Transdural circumferential decompression was an effective method for thoracic spinal stenosis caused by concurrent beak-type OPLL, by which OPLL could be safely removed. It is especially useful when there is a severe adhesion between the dura OPLL.

Compressed Imaging Reconstruction Based on Block Compressed Sensing with Conjugate Gradient Smoothed l0 Norm.

Compressed imaging reconstruction technology can reconstruct high-resolution images with a small number of observations by applying the theory of block compressed sensing to traditional optical imaging systems, and the reconstruction algorithm mainly determines its reconstruction accuracy. In this work, we design a reconstruction algorithm based on block compressed sensing with a conjugate gradient smoothed l0 norm termed BCS-CGSL0. The algorithm is divided into two parts. The first part, CGSL0, optimizes the SL0 algorithm by constructing a new inverse triangular fraction function to approximate the l0 norm and uses the modified conjugate gradient method to solve the optimization problem. The second part combines the BCS-SPL method under the framework of block compressed sensing to remove the block effect. Research shows that the algorithm can reduce the block effect while improving the accuracy and efficiency of reconstruction. Simulation results also verify that the BCS-CGSL0 algorithm has significant advantages in reconstruction accuracy and efficiency.

Application of open-door laminoplasty with ARCH plate fixation in cervical intraspinal tumors.

BACKGROUND: The open-door laminoplasty is an effective procedure for the treatment of cervical spondylotic myelopathy. However, little information is available about the surgical results of open-door laminoplasty in the treatment of intraspinal tumors. In the present study, we aimed to investigate the clinical effect of open-door laminoplasty with ARCH plate fixation in the treatment of cervical intraspinal tumors. METHODS: This was a retrospective study. From January 2013 to May 2018, 38 patients (13 males and 25 females, the average age of 44 ± 17 years) with cervical intraspinal tumors underwent open-door laminoplasty with ARCH plate fixation in our hospital. The operation time, blood loss, pre- and postoperative visual analog scale (VAS), and Japanese Orthopedic Association (JOA) scores were determined. To determine the radiographic outcomes, cervical X-ray film and magnetic resonance imaging (MRI) were performed before and after the operation, and cervical X-ray sagittal film was used to measure Cobb angle. The clinical data before and after the operation were compared by t-test. RESULTS: A total of 38 patients underwent a successful operation and demonstrated primary healing. The average operation time was 113 ± 12 min. The average blood loss was 120 ± 19 mL. All patients were followed up for 26.1 ± 2.8 months, and the final follow-up time was more than 24 months. VAS scores were much better at 24 months after operation compared with those before the operation, which were decreased from 6.1 ± 1.1 to 1.4 ± 0.7 (t = 32.63, P < 0.01). The JOA score was improved from 9.9 ± 1.5 to 15.5 ± 0.6 (t = - 18.36, P < 0.01), and the mean JOA recovery rate was 79% ± 11% at 24 months after the operation. There was no significant difference in Cobb angle between pre-operation and 24 months after the operation, which was 9.8 ± 2.6 and 10.3 ± 3.1 respectively (t = - 0.61, P > 0.05). Neither spinal malalignment on the coronal plane nor displacement of the laminoplasty flap was observed on postoperative cervical X-ray and MRI examinations at the final follow-up. CONCLUSIONS: Open-door laminoplasty with ARCH plate fixation was a safe and effective surgical approach for the treatment of cervical intraspinal tumors.

Culture-negative versus culture-positive in pyogenic spondylitis and analysis of risk factors for relapse.

OBJECTIVES: This study aims to compare and analyze the clinical features, diagnosis, treatment and prognosis of culture-negative and culture-positive primary pyogenic spondylitis. METHODS: In a retrospective analysis, 202 cases of adult primary pyogenic spondylitis with complete clinical data in our hospital from January 2013 to January 2020 were divided into two groups according to bacterial culture results: culture negative (n = 126) and culture positive (n = 76). We compare the clinical characteristics, diagnosis, treatment and prognosis of patients with different culture results. RESULTS: The culture positive rate was 37.62% (76/202). There were no significant differences in age, gender, affected segment, spinal abscess, diabetes mellitus, course of disease, surgery, recurrence, and follow-up time between the two groups (p>.05). There were statistically significant differences in hospital admission erythrocyte sedimentation rate (ESR), admission C-reactive protein (CRP), admission white blood cell (WBC) count, discharge ESR, discharge CRP, ESR decline rate, CRP (p<.05). There were statistically significant differences in the rate of decline, hospitalization days, and body temperature ≥38 °C (p<.05). Higher CRP levels on admission, antibiotic treatment time <6 weeks, and body temperature ≥ 38 °C are independent risk factors for infection recurrence. CONCLUSIONS: The culture-negative group's admission WBC, admission ESR, admission CRP, discharge ESR, discharge CRP, ESR decline rate, CRP decline rate, and hospital stay were lower than the culture positive group, the difference was statistically significant (p<.05). The independent risk factors for infection recurrence are higher CRP levels in hospital admission, antibiotic treatment time <6 weeks, and body temperature ≥ 38 °C.

Aspergillus spondylitis: case series and literature review.

BACKGROUND: Spinal fungal infections, especially spinal Aspergillus infections, are rare in the clinic. Here, we introduce the clinical features, diagnosis, treatment, and prognoses of 6 cases of Aspergillus spondylitis. METHODS: We retrospectively analysed the complete clinical data of patients with Aspergillus spondylitis treated in our hospital from January 2013 to January 2020. RESULTS: Aspergillus fumigatus was isolated in 4 cases, and Aspergillus spp. and Aspergillus niger were isolated in 1 case each. All six patients reported varying degrees of focal spinal pain; one patient reported radiating pain, one patient experienced bowel dysfunction and numbness in both lower limbs, and three patients had fever symptoms. One case involved the thoracic spine, one case involved the thoracolumbar junction, and 4 cases involved the lumbar spine. Three patients were already in an immunosuppressed state, and three patients entered an immunosuppressed state after spinal surgery. All six patients were successfully cured, and five required surgery. Of the 5 patients who underwent surgical treatment, 2 had spinal cord compression symptoms, and 3 had spinal instability. At the end of follow-up, 1 patient reported left back pain and 1 patient reported left limb numbness. CONCLUSION: The clinical manifestations of Aspergillus spondylitis are non-specific, and the diagnosis depends on typical imaging findings and microbiological and histopathological examination results. When there is no spinal instability, spinal nerve compression symptoms, or progressive deterioration, antifungal therapy alone may be considered. If spinal instability, spinal nerve compression, or epidural abscess formation is present, surgery combined with antifungal therapy is recommended.

Thoracic psammomatous meningioma with osseous metaplasia: a controversial diagnosis of a case report and literature review.

BACKGROUND: Spinal psammomatous meningioma with calcification is commonly observed, but distinctive osseous differentiation rarely occurs. CASE PRESENTATION: Here, we described a 52-year-old female complaining of chronic back pain for 5 years. CT and MRI examinations revealed an intradural extramedullary mass at the T4 level. The tumor was meticulously excised en bloc. Under the microscope, the tumor was found to be composed of conspicuous calcified psammoma bodies with remarkable immature bone formation. A primary diagnosis of psammomatous meningioma was made based on the recent WHO classification of tumors of the CNS, whereas other pathologists focused on the osseous components and preferred metaplastic meningioma as the proper subtype. A literature review was conducted, and only five cases have been reported with the same histopathological condition. Experts finally reached a consensus based on the acknowledged notion of the preferential diagnosis of psammomatous meningioma, as well as the current evidence and popular opinion that ossification is generated from osteogenic differentiation of pluripotent cells rather than the accumulation of psammoma bodies. CONCLUSIONS: A final diagnosis of psammomatous meningioma with osseous metaplasia was made. The rigid and adherent features complicate total resection of the tumor and increase the risk of neurologic deficits.

Dura sac compression due to spinal epidural gas pseudocyst after lumbar decompression surgery: a case report.

BACKGROUND: Intraspinal gas pseudocyst is rare, especially following spinal surgery. Here we present a case of spinal epidural gas pseudocyst following lumbar decompression surgery, which caused dura sac compression. CASE PRESENTATION: A 52-years-old woman with chronic lumbar pain and radiating numbness of left leg was admitted to our hospital and underwent a posterior lumbar decompression surgery. 10 days later, the patient began to have dysfunction of excretion. CT and MRI were taken and epidural gas was detected, which compressed the dura sac. A huge pseudocyst encapsulated with high-tension air was found during debridement with no evidence of infection. RESULTS: Debridement surgery was taken to remove the encapsulated gas and cyst wall and her symptoms disappeared soon after the surgery. 2 weeks later, routine X-ray was repeated and gas pseudocyst disappeared with no signs of infection. CONCLUSION: Gas pseudocyst in the spinal canal is rare, especially after lumbar surgery and causing spinal cord compression. CT and MRI can be used to detect the spinal gas. Once gas pseudocyst causes dura sac compression, proper methods should be chosen to treat this kind of intraspinal gas pseudocyst.

Correlation between Polymorphism of TRAIL Gene and Condition of Intervertebral Disc Degeneration.

BACKGROUND: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been suggested to be related with the pathogenesis and progression of osteoarticular degenerations. This study therefore aimed to investigate the relationship between the polymorphism of the TRAIL gene and the pathogenesis and severity of intervertebral disc degeneration (IDD) via detection of serum TRAIL expression levels. MATERIAL AND METHODS: A total of 100 IDD patients in our hospital were recruited in the experimental group, while another cohort of 100 healthy individuals was employed as the control group. Blood samples collected from all people were quantified for TRAIL level using enzyme-linked immunosorbent assay (ELISA), in addition to allele and genotype frequency analysis via fluorescent PCR for TRAIL gene. RESULTS: At loci 1525 and 1529 in 3'-untranslated region (UTR) of 5th exon of TRAIL gene, 3 different genotypes were identified: experimental group had higher frequency of 1525CG/1595CC, 1525G and 1595C alleles, compared to the control group (p<0.05). Patients under Schneiderman grade IV had significantly higher allele frequency compared to those at grade II or III. Serum TRAIL level was also higher in the experimental group compared to the control group, and in grade IV patients compared to grade II or III patients (p<0.05). CONCLUSIONS: The G/C mutation at loci 1525/1595 of TRAIL gene may induce the progression of IDD, as the down-regulation of TRAIL can aggravate the severity of the disease.

MiR-2b-3p Downregulated PxTrypsin-9 Expression in the Larval Midgut to Decrease Cry1Ac Susceptibility of the Diamondback Moth, Plutella xylostella (L.).

Crystal (Cry) toxins, produced by Bacillus thuringiensis, are widely used as effective biological pesticides in agricultural production. However, insects always quickly evolve adaptations against Cry toxins within a few generations. In this study, we focused on the Cry1Ac protoxin activated by protease. Our results identified PxTrypsin-9 as a trypsin gene that plays a key role in Cry1Ac virulence in Plutella xylostella larvae. In addition, P. xylostella miR-2b-3p, a member of the micoRNA-2 (miR-2) family, was significantly upregulated by Cry1Ac protoxin and targeted to PxTrypsin-9 downregulated its expression. The mRNA level of PxTrypsin-9, regulated by miR-2b-3p, revealed an increased tolerance of P. xylostella larvae to Cry1Ac at the post-transcriptional level. Considering that miR-2b and trypsin genes are widely distributed in various pest species, our study provides the basis for further investigation of the roles of miRNAs in the regulation of the resistance to Cry1Ac and other insecticides.

Characterization and functional analysis of a myeloid differentiation factor 88 in Ostrinia furnacalis Guenée larvae infected by Bacillus thuringiensis.

Myeloid differentiation factor 88 (MyD88) is a pivotal adapter protein involved in activating nuclear factor NF-κB of the Toll pathway in insect innate immunity. MyD88 has been extensively studied in vertebrates and Drosophila. However, the information ascribed to MyD88 in Lepidoptera is scarce. In the present study, an Ostrinia furnacalis MyD88 (OfMyD88) cDNA was cloned and functionally characterized (GenBank accession no. MN906311). The complete cDNA sequence of OfMyD88 is 804 bp, and contains a 630 bp open reading frame encoding 209 amino acid residues. OfMyD88 has the death domain (DD), an intermediate domain, and the Toll/interleukin 1 receptor (TIR) domain. OfMyD88 was widely expressed in immune-related tissues such as hemocytes, fat body, midgut, and integument, with the highest expression level in hemocytes, and the lowest expression level in integument. To clarify the immune function of MyD88, O. furnacalis larvae were challenged with Bacillus thuringiensis (Bt) through feeding. Bt oral infection had significantly up-regulated the expression of OfMyD88 and immune genes, including PPO2 (prophenoloxidase 2), Attacin, Gloverin, Cecropin, Moricin, GRP3 (ß-1, 3-Glucan recognition protein 3), and Lysozyme, and increased the activities of PO and lysozyme in hemolymph of O. furnacalis larvae. Knockdown of OfMyD88 by RNA interference suppressed the expression levels of immune related genes, but not PPO2 in the larvae orally infected with Bt, suggesting that OfMyD88 is involved in defending against Bt invasion through the Toll signaling pathway, but does not affect the PPO expression in O. furnacalis larvae.

Gut microbiota mediate Plutella xylostella susceptibility to Bt Cry1Ac protoxin is associated with host immune response.

Insect gut microbiotas have a variety of physiological functions for host growth, development, and immunity. Bacillus thuringiensis (Bt) is known to kill insect pests by releasing insecticidal protoxins, which are activated in the insect midgut. However, the interplay among Bt infection, host immunity, and gut microbiota are still unclear. Here we show that Bt Cry1Ac protoxin interacts with the gut microbiota to accelerate the mortality of P. xylostella larvae. Cry1Ac protoxin was found to cause a dynamic change in the midgut and hemocoel microbiota of P. xylostella, with a significant increase in bacterial load and a significant reduction in bacterial diversity. In turn, loss of gut microbiota significantly decreased the Bt susceptibility of P. xylostella larvae. The introduction of three gut bacterial isolates Enterococcus mundtii (PxG1), Carnobacterium maltaromaticum (PxCG2), and Acinetobacter guillouiae (PxCG3) restored sensitivity to Bt Cry1Ac protoxin. We also found that Cry1Ac protoxin and native gut microbiota can trigger host midgut immune response, which involves the up-regulation of expression of Toll and IMD pathway genes and most antimicrobial peptide genes, respectively. Our findings further shed light on the interplay between insect gut microbiota and host immunity under the Bt toxin killing pressure, and this may provide insights for improving the management of Bt resistance and lead to new strategies for biological control of insect pests.

The Tripartite Interaction of Host Immunity-Bacillus thuringiensis Infection-Gut Microbiota.

Bacillus thuringiensis (Bt) is an important cosmopolitan bacterial entomopathogen, which produces various protein toxins that have been expressed in transgenic crops. The evolved molecular interaction between the insect immune system and gut microbiota is changed during the Bt infection process. The host immune response, such as the expression of induced antimicrobial peptides (AMPs), the melanization response, and the production of reactive oxygen species (ROS), varies with different doses of Bt infection. Moreover, B. thuringiensis infection changes the abundance and structural composition of the intestinal bacteria community. The activated immune response, together with dysbiosis of the gut microbiota, also has an important effect on Bt pathogenicity and insect resistance to Bt. In this review, we attempt to clarify this tripartite interaction of host immunity, Bt infection, and gut microbiota, especially the important role of key immune regulators and symbiotic bacteria in the Bt killing activity. Increasing the effectiveness of biocontrol agents by interfering with insect resistance and controlling symbiotic bacteria can be important steps for the successful application of microbial biopesticides.

Studies on 11 Cases of Spinal Epidural Abscess and Literature Review.

OBJECTIVE: In the present study, we aimed to describe the clinical features, diagnosis, treatment, and prognosis of spinal epidural abscess (SEA). METHODS: The complete clinical data of 11 SEA patients who were treated in our hospital system from January 2015 to June 2018 were retrospectively analyzed. Moreover, the clinical features, diagnosis, treatment, and prognosis of 642 SEA cases collected from the foreign literature from 2010 to 2019 were also investigated. RESULTS: Among our 11 SEA patients, nine cases had purulent inflammation, two cases had tuberculosis, two cases had infection caused by Staphylococcus aureus, one case had infection caused by Streptococcus constellatus, one case had infection caused by Klebsiella pneumoniae, five cases showed negative bacterial culture, and two cases had Mycobacterium tuberculosis. All 11 cases showed focal spinal pain, eight cases exhibited neurological deficits, and six cases experienced fever. Nine of the 11 cases involved the lumbosacral spine, one case involved the thoracic spine, and one case involved the cervical spine. Eight patients had a longer course of disease (>2 weeks), all 11 patients had vertebral osteomyelitis, and nine patients had intervertebral discitis. One patient had motor dysfunction of arms and legs, one patient had lower limb motor dysfunction, one patient had limb numbness, one patient experienced relapse after the conservative treatment, and one patient experienced relapse after the surgical treatment. The follow-up time was 15-24 months. CONCLUSION: The classic diagnosis of triads (focal spine pain, neurological deficit, and fever) was less specific for SEA. MRI examination, blood culture, tissue culture, and biopsy could be used for the diagnosis for SEA. Suppuritis was a common cause of SEA. Early detection, early diagnosis and early treatment, as well as the selection of the most suitable treatment regimen based on comprehensive evaluation, played crucial roles in a better prognosis of SEA. There was no statistically significant difference in terms of the general condition, diagnosis, treatment and prognosis between the patients with negative and positive culture results (P>0.05). For SEA patient with negative culture, antibiotic treatment should be used empirically.

Pathogenicity and Transgenerational Effects of Metarhizium anisopliae on the Demographic Parameters of Aedes albopictus (Culicidae: Diptera).

Dengue fever is a vector-borne infectious disease that spreads swiftly and threatens human lives in several tropical countries. Most of the strategies employed for the control of Aedes albopictus (Skuse) involve synthetic chemicals. The indiscriminate use of synthetic chemicals has led to the development of resistance and is unsafe for human and environmental health. Therefore, there is a need to develop ecologically safe tactics, such as the use of the entomopathogenic fungus (EPF) Metarhizium anisopliae (Metchnikoff 1879) (Met-11.1). The following study investigated the effectiveness of EPF-Met-11.1 on different demographic parameters of Ae. albopictus. Mortality bioassays showed 92.5% mortality when adult Ae. albopictus were treated with M. anisopliae. Metarhizium anisopliae absorbs the hemolymph sugar which results in retarded development. Metarhizium anisopliae LC50 not only affected the parental generation (F0) but also affected the demographic parameters of the offspring (F1). Transgenerational results (F1) with Met-11.1 showed decreased net reproductive rates (Ro), intrinsic rates of increase (r), and mean generation times (T) compared to those of uninfected controls. The larval developmental duration in the treatment group was 8.22 d, compared to 8.00 d in the control. There was a significant decrease in mean fecundity in the treated group (208.87 eggs) compared to that of the control group (360.27 eggs), and adult longevity was also significantly reduced in the treated group. Therefore, it is concluded that M. anisopliae can have lasting effects on the developmental parameters of Ae. albopictus, indicating that it can be an integral part of mosquito control strategies.

An immune-responsive PGRP-S1 regulates the expression of antibacterial peptide genes in diamondback moth, Plutella xylostella (L.).

Peptidoglycan recognition proteins (PGRPs) are family of pattern recognition receptors (PRRs) and triggers the innate immune system (IIS) against the microbial infection. Although PGRPs have been intensively studied in model insects, they remain uncharacterized in most of the non-model insects. Here, we cloned and characterized a full-length cDNA of PGRP, from P. xylostella (PxPGRP-S1), which encodes a protein of 239 amino acids with PGRP domain, Ami2 domain and transmembrane region. The phylogenetic analysis revealed that the PxPGRP-S1 was closely related to the unigene of Plutella xylostella. Quantitative real-time PCR and immunohistochemistry revealed that PxPGRP-S1 is mainly expressed in the fat body of the healthy larva. The expression of PxPGRP-S1 was significantly upregulated in the midgut at 24 h postinfection by Bacillus thuringiensis. Silencing of the PxPGRP-S1 expression by RNAi, significantly decrease the expression of the antimicrobial peptides (AMPs) in the 4th instar larvae of P. xylostella. Similarly injection of an anti-PxPGRP-S1 serum caused the low expression of the AMPs in P. xylostella. Additionally, PxPGRP-S1 depleted P. xylostella by oral administration of bacterial expressed dsRNA decreased the resistance against B. thuringiensis challenge, leads to high mortality. Together, our result indicates that PxPGRP-S1, served as a bacterial pattern recognition receptor (PRR) and triggers the expression of AMPs in P. xylostella.

Application of Laminoplasty Combined with ARCH Plate in the Treatment of Lumbar Intraspinal Tumors.

OBJECTIVE: To investigate the bone fusion and clinical effect of laminoplasty combined with ARCH plate fixation in the treatment of lumbar intraspinal tumors. METHODS: This was a clinical study. From June 2017 to January 2019, 24 patients (seven males and 17 females, average age 40 ± 16 years) with lumbar intraspinal tumors underwent laminoplasty combined with ARCH plate fixation in our hospital. The bone fusion was evaluated by X-ray and computed tomography (CT) scans that were taken 15.2 ± 2.17 months postoperatively. Each segment showed a bone bridge on one side, which was classified as "segmental partial fusion." Each segment showed bilateral bone bridges, which were classified as "segmental complete fusion". When all segments of the patient showed bilateral bone bridging so that the replanted lamina and the host lamina became a unit on the CT scan, it was defined as "complete fusion". In addition, the operation time and blood loss were recorded. Fisher's exact test was used to analyze the potential influencing factors of bone healing, including age (≤40 years vs >40 years), gender, number of operated levels (single vs two). Paired t-test was used to analyze pre- and postoperative Oswestry Disability Index (ODI) scale and low back and leg pain visual analog scale (VAS). RESULTS: A total of 33 segments of laminoplasty were successfully performed in 24 patients. The average operation time was 128 ± 18 minutes. The average blood loss was 110 ± 19 mL. All patients were followed up at least 12 months after operation (average, 15.2 ± 2.17 months). At the final follow-up, according to the definition of this study, the proportion of "segmental partial fusion" and "segmental complete fusion" were 30.3% (10/33) and 69.7% (23/33), respectively. And the proportion of patients with "complete fusion" was 70.8% (17/24). Age, gender, and number of operated levels were not associated with the fusion (P = 1.0, 0.37, and 0.06, respectively). ODI and VAS were much better at 1 month after operation and the final follow-up than those before the operation (P < 0.01). At 6 months after operation, the results of magnetic resonance imaging (MRI) showed that the supraspinous ligament was repaired, and there were no complications, such as spinal epidural scar recompression. CONCLUSIONS: Laminoplasty combined with ARCH plate was a better surgical method, and 70.8% of the patients showed complete bone fusion and there was no case of bilateral nonunion.

MicroRNA expression profiling of Plutella xylostella after challenge with B. thuringiensis.

The diamondback moth, Plutella xylostella, the main pest of brassica crops, has developed resistance to almost all major classes of insecticides as the farmers rely on insecticides to control this pest. An extensive use of broad-spectrum insecticides against P. xylostella promotes the selection of insecticide resistance, destroy natural enemies, and pollute the environment. In this scenario, it is imperative to use genetic methods such as gene silencing technology as an alternate approach against this pest. Evidence shows that microRNAs play pivotal roles in the regulation of target genes at the post-transcription level and show differential expression under various biological processes. However, the knowledge of their role in insect immunity is still in its infancy. In the present study, we aimed at exploring the response of P. xylostella miRNAs against B. thuringiensis at different time courses (6, 12, 18, 24, and 36 h) by using small RNA sequencing. After data filtration, a combined set of 149 miRNAs was identified from all the libraries. Interestingly, a couple of conserved miRNAs such as miR-1, Let-7, miR-275, miR-184, and miR-10 were listed as abundantly expressed miRNAs after exposure to B. thuringiensis. It is worth mentioning that the differential expression analysis revealed that miR-2, a conserved miRNA, was up-regulated following infection. Furthermore, we experimentally validated the involvement of miR-2b-3p in the regulation of corresponding target trypsin. Our luciferase assay results revealed that miR-2b-3p mimic significantly down-regulated the target gene trypsin indicating that it might play a crucial role in the defense mechanism of P. xylostella against B. thuringiensis infection. On the whole, our findings provide insights into the possible regulatory role of miRNAs in insect immunity in response to microorganisms.

Bacillus thuringiensis Suppresses the Humoral Immune System to Overcome Defense Mechanism of Plutella xylostella.

Background: Plutella xylostella has become a notorious pest of cruciferous crops all over the world. Delta-endotoxins of Bacillus thuringiensis are widely used insecticidal proteins for controlling P. xylostella. However, the interaction mechanism of B. thuringiensis with the immune system of P. xylostella, at the genomic level, is still unclear. This study explored the immune response of P. xylostella to B. thuringiensis, at different time intervals, 6 h, 12 h, 18 h, 24 h, and 36 h, by using RNA-Sequencing (RNA-Seq) and RT-qPCR. Results: In total, 167 immunity-related genes were identified and placed into different families, including pattern recognition receptors (PRRs), signal modulators, immune pathways (Toll, IMD, and JAK/STAT), and immune effectors. It is worth mentioning that the analyses of the differentially expressed immunity-related genes revealed that most of the differentially expressed genes (DEGs) (87, 56, 76, 67, and 73 genes) were downregulated in P. xylostella following B. thuringiensis oral infection at 6 h, 12 h, 18 h, 24 h, and 36 h. Interestingly, our RNA-Seq analysis also revealed reduced expression of antimicrobial peptides, that play a vital role in the humoral immune system of P. xylostella. Conclusion: This study demonstrates that B. thuringiensis plays a novel role in controlling P. xylostella, by suppressing the immune system.