Ivermectin induces apoptosis of esophageal squamous cell carcinoma via mitochondrial pathway.

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is the most predominant primary malignant tumor among worldwide, especially in China. To date, the successful treatment remains a mainly clinical challenge, it is imperative to develop successful therapeutic agents. METHODS: The anti-proliferative effect of ivermectin on ESCC is investigated in cell model and in nude mice model. Cell apoptosis was assessed using flow cytometry, TUNEL assay and western blotting. Mitochondrial dysfunction was determined by reactive oxygen species accumulation, mitochondrial membrane potential and ATP levels. RESULTS: Our results determined that ivermectin significantly inhibited the proliferation of ESCC cells in vitro and in vivo. Furthermore, we found that ivermectin markedly mediated mitochondrial dysfunction and induced apoptosis of ESCC cells, which indicated the anti-proliferative effect of ivermectin on ESCC cells was implicated in mitochondrial apoptotic pathway. Mechanistically, ivermectin significantly triggered ROS accumulation and inhibited the activation of NF-κB signaling pathway and increased the ratio of Bax/Bcl-2. CONCLUSIONS: These finding indicated that ivermectin has significant anti-tumour potential for ESSC and may be a potential therapeutic candidate against ESCC.

Cognition in patients with benign epilepsy with centrotemporal spikes: A study with long-term VEEG and RS-fMRI.

OBJECTIVE: The purpose of this study was to investigate the relationship between alterations of functional brain network and cognition in patients with benign epilepsy with centrotemporal spikes (BECTS) as a function of spike-wave index (SWI) during slow wave sleep. METHODS: Resting-state functional magnetic resonance imaging (RS-fMRI) data and Intelligence Quotient (IQ) were collected from two groups of patients with BECTS, including a SWI<50% group (5 cases) and a SWI≥50% group (7 cases). The SWI was calculated from the long-term video-electroencephalogram monitoring (one sleep cycle was included at least). The RS-fMRI data were analyzed by regional homogeneity (ReHo) method. RESULTS: There were three main findings. Firstly, Full Intelligence Quotient (FIQ), Verbal Intelligence Quotient (VIQ), and Performance Intelligence Quotient (PIQ) of the SWI≥50% group were significantly lower than SWI<50% group (p<0.05). Secondly, there was a negative correlation between the FIQ, VIQ, PIQ, and SWI (p<0.05), and the FIQ, VIQ, and PIQ were not dependent on age, age of onset, disease course, years of education, and total number of seizures (p>0.05). Finally, compared with the SWI<50% group, the SWI≥50% group showed increased ReHo in the bilateral precentral gyrus, bilateral premotor area, bilateral subcortical structure, right temporal lobe, and bilateral insular lobe, while they showed decreased ReHo in the posterior cingulate cortex and posterior of right inferior temporal lobe. CONCLUSIONS: The alterations of functional brain network caused by the frequent discharges during slow wave sleep could affect cognition in patients with BECTS.

[Value of electroencephalography/functional magnetic resonance imaging localizing epileptic foci in focal epilepsy].

OBJECTIVE: To explore the diagnostic value of simultaneous electroencephalography/ functional magnetic resonance imaging ( EEG-fMRI) for epileptic foci in focal epilepsy of normal brain structure. METHODS: The normal structural images of EEG-fMRI and resting state-fMRI (RS-fMRI) with 3. OT high-field MR system were performed for 15 focal epileptic patients from April 2013 to July 2014. On EEG-fMRI, epileptiform discharges were regarded as nerve stimulation. And off-line analysis yielded the blood oxygen level-dependent (BOLD) responses related to epileptiform discharges. The relationship was analyzed between BOLD activation reaction and electroencephalography-clinical symptomatology position. And the electroencephalography-clinical symptomatology position was selected as a major indicator of judging the degree of concordance. RESULTS: The involved lobes were temporal (n = 7), frontal (n = 5) and frontotemporal (n = 3). On EEG-fMRI, all patients demonstrated BOLD responses related to epileptiform discharges. Compared with electroencephalography-clinical symptomatology position, 12 cases (12/15) were fully concordant and 3 cases (3/15) partially concordant. Three patients had BOLD responses in accordance with electroencephalography-clinical symptomatology position on RS-fMRI. And the detection rate of BOLD responses was higher for EEG-fMRI than that for RS-fMRI. CONCLUSION: The BOLD responses related to epileptiform discharges may help locate noninvasively the epileptogenic region of focal epilepsy and provide additional useful information for preoperative evaluations of epilepsy.

SpALF4: a newly identified anti-lipopolysaccharide factor from the mud crab Scylla paramamosain with broad spectrum antimicrobial activity.

Anti-lipopolysaccharide factors (ALFs) are antimicrobial peptides with binding and neutralizing activities to lipopolysaccharide (LPS) in crustaceans. This study identified and characterized a novel ALF homolog (SpALF4) from the mud crab Scylla paramamosain. The complete cDNA of SpALF4 had 756 bp with a 381 bp open reading frame encoding a protein with 126 aa. The deduced protein contained a signal peptide and a LPS-binding domain. SpALF4 shared the highest identity with PtALF5 at amino acid level but exhibited low similarity with most of other crustacean ALFs. Furthermore, different from the previously identified three SpALF homologs and most of other ALFs, SpALF4 had a low isoelectric point (pI) for the mature peptide and the LPS-binding domain with the values of 6.93 and 6.74, respectively. These results indicate that SpALF4 may be a unique ALF homolog with special biological function in the mud crab. Similar to the spatial structure of ALFPm3, SpALF4 contains three α-helices packed against a four-strand ß-sheet, and an amphipathic loop formed by a disulphide bond between two conserved cysteine residues in LPS-binding domain. SpALF4, mainly distributed in hemocytes, could be upregulated by Vibrio harveyi, Staphylococcus aureus, or white spot syndrome virus. Recombinant SpALF4 could inhibit the growth of Gram-negative bacteria (V. harveyi, Vibrio anguillarum, Vibrio alginolyticus, Aeromonas hydrophila, Pseudomonas putida), Gram-positive bacteria (S. aureus and Bacillus megaterium), and a fungus Candida albicans to varying degrees. Further study showed that it could also bind to all the aforementioned microorganisms except S. aureus. These results demonstrate that SpALF4 is a unique ALF homolog with potent antimicrobial activity against bacteria and fungi. This characteristic suggests SpALF4 plays an essential function in immune defense against pathogen invasion in mud crab.

[Executive dysfunction in patients with temporal lobe epilepsy and its correlation with P300].

OBJECTIVE: To explore the changes of executive function in patients with temporal lobe epilepsy and analyze its correlation with P300 event-related potentials. METHODS: Fifty patients with temporal lobe epilepsy and 30 age, gender and education-matched healthy control subjects were assessed by neuropsychological tests, including Montreal cognitive assessment (MoCA), working memory, verbal fluency, trail making, digit span, digit symbol and Stroop color-word interference to detect P300 event-related potentials. RESULTS: In temporal lobe epilepsy group, the scores in MoCA, verbal and non-verbal working memory, verbal fluency, digit span, digit symbol and Stroop test were lower than those of the normal control group. And trail making tests A and B became prolonged (P < 0.05). Comparing with normal control group, temporal lobe epilepsy patients had prolonged latency [(332 ± 33)ms] and decreased P300 amplitude [(10 ± 8)µV] (P < 0.05). Comparing epileptogenic focus group on the left and right sides, there was statistically significant difference in verbal working memory (P < 0.05). There was a negative correlation of P300 latency and MoCA, non-verbal working memory, digit span and digit symbol test scores (r = -0.29--0.45, P < 0.05). And a positive correlation of P300 amplitude and MoCA, non-verbal working memory, digit symbol conversion and Stroop scores was also found (r = 0.37-0.47, P < 0.05). P300 amplitude was more relevant to overall cognitive level and executive functions. CONCLUSION: Temporal lobe is involved in the regulation of executive functions. Besides a wide range of cognitive impairment, temporal lobe epilepsy patients have a number of executive dysfunctions, including working memory, cognitive flexibility, attention and inhibitory control ability. And an impairment of verbal working memory is evident in left-sided lesion. Their manifestations include decreased latency and amplitude of P300 on executive function tests. Therefore these two objective parameters may be employed to evaluate the cognitive impairment in patients with temporal lobe epilepsy.

[Values of stereotactic guidance of intraoperative high-field magnetic resonance imaging for multiple intracranial lesions].

OBJECTIVE: To explore the diagnostic and therapeutic values of stereotactic guidance of intraoperative high-field magnetic resonance imaging (iMRI) for multiple intracranial lesions. METHODS: We retrospectively assessed 18 patients with multiple intracranial lesions undergoing stereotactic guidance of high-field iMRI between June 2011 and May 2013. The procedures included biopsy of stereotactic guidance (n = 6), stereotactic aspiration and drainage for brain abscess (n = 4), stereotactic aspiration and intracavitary irradiation with (32)P for mixed solid and cystic craniopharyngiomas (n = 3) and stereotactic hematoma evacuation (n = 5). RESULTS: For 6 cases with high-field iMRI stereotactic guidance, the target lesions were precisely predetermined and pathological specimens confirmed by clinical follow-up results. For 12 cases with multiple cystic lesions and multiple intracranial hematoma, aspiration of hematoma and liquids was satisfactory. And the course of clinical treatment was significantly shortened. And there was a lower incidence of postoperative complications. CONCLUSION: iMRI may guide precisely stereotactic surgery. And it has important clinical significance for confirming the diagnosis of multiple intracranial lesions and shortening their treatment courses.

Mechanism of HDAC1 Regulating Iron Overload-Induced Neuronal Oxidative Damage After Cerebral Hemorrhage.

Iron overload is associated with brain edema in the context of intracerebral hemorrhage (ICH). Here, we investigated the role of histone deacetylase 1 (HDAC1) in mediating oxidative damage induced by iron overload after ICH. Utilizing ICH mouse models and FeCl2-induced HT-22 cell models, we assessed HDAC1 expression and its impact on iron overload and oxidative damage. We examined the levels of Kruppel like factor 4 (KLF4), RAN binding protein 9 (RANBP9), as well as the acetylation levels of HDAC1 and histones H3 and H4 in the KLF4 promoter, and the KLF4 level in the RANBP9 promoter. Additionally, we investigated the binding relationships between KLF4 and the RANBP9 promoter, HDAC1 and miR-129-5p. Our results demonstrated elevated HDAC1 expression in ICH mice and FeCl2-induced HT-22 cells. HDAC1 silencing improved neurological function in mice, reduced brain edema, and alleviated iron overload and oxidative damage in vitro. HDAC1 downregulated KLF4 expression by reducing acetylation levels in the KLF4 promoter, leading to decreased KLF4 enrichment in the RANBP9 promoter and increased RANBP9 expression. Furthermore, upstream miR-129-5p inhibited HDAC1, and the downregulation of miR-129-5p mitigated the protective effect of HDAC1 silencing. Collectively, our findings highlight the significant role of HDAC1 in exacerbating iron overload-induced oxidative damage following ICH and its regulation by miR-129-5p.

Impact of Abl2/Arg deficiency on anxiety and depressive behaviors in mice.

Abl2/Arg (ABL-related gene) is a member of the Abelson family of nonreceptor tyrosine kinases, known for its role in tumor progression, metastasis, tissue injury responses, inflammation, neural degeneration, and other diseases. In this study, we developed Abl2/Arg knockout (abl2-/-) mice to explore its impact on sensory/motor functions and emotion-related behaviors. Our findings show that abl2-/- mice exhibit normal growth and phenotypic characteristics, closely resembling their wild-type (WT) counterparts. Behavioral tests, including the elevated plus maze, marble-burying behavior test, and open field test, indicated pronounced anxiety-like behaviors in abl2-/- mice compared to WT mice. Furthermore, in the tail suspension test, abl2-/- mice showed a significant decrease in mobility time, suggesting depressive-like behavior. Conversely, in the Y-maze and cliff avoidance reaction tests, no notable differences were observed between abl2-/- and WT mice, suggesting the absence of working memory deficits and impulsivity in abl2-/- mice. Proteomic analysis of the hippocampus in abl2-/- mice highlighted significant alterations in proteins related to anxiety and depression, especially those associated with the GABAergic synapse in inhibitory neurotransmission. The expression of Gabbr2 was significantly reduced in the hippocampus of abl2-/- compared to WT mice, and intraperitoneal treatment of GABA receptor agonist Gaboxadol normalized anxiety/depression-related behaviors of abl2-/- mice. These findings underscore the potential role of Abl2/Arg in influencing anxiety and depressive-like behaviors, thereby contributing valuable insights into its broader physiological and pathological functions.

The effect of the whole-process care model of the medical union on the improvement of kinesiophobia and bone mineral density in patients with osteoporosis.

OBJECTIVE: To observe the effect of the whole-process care model of the medical union on the improvement of kinesiophobia and bone mineral density in patients with osteoporosis. METHODS: In this descriptive study, a convenient sampling method was used to select 148 patients with osteoporosis who visited the hospital from January 2020 to December 2021. Patients aged ≥ 18 years and diagnosed with osteoporosis through quantitative computed tomography (QCT) were included in the study. They were able to cooperate during follow-up and had normal cognitive function. Patients with combined spinal curvature, thoracic deformity, and pulmonary dysfunction, accompanied by severe cardiovascular or limb dysfunction, and those who withdrew midway or participated in other clinical studies were excluded. According to whether to use the whole-process care model of the medical union, they were divided into intervention group and control group, with 74 cases each. The control group used conventional care, and the intervention group used the whole-process care model of the medical association. The occurrence of kinesiophobia between the two groups were compared. The dual-energy X-ray absorption detector is used to measure differences in bone density changes. RESULTS: There was no significant difference between the two groups in the TSK scale score and the incidence of kinesiophobia before intervention (P > 0.05). The TSK scale scores of patients in the intervention group were higher than those in the control group at 3 months and 6 months after operation (P < 0.05). The incidence of kinesiophobia in the intervention group for 3 months and 6 months was significantly lower than that in the control group (P < 0.05). There was no significant difference in bone mineral density between the two groups before and 3 months after intervention (P > 0.05). The bone mineral density of lumbar spine, femoral neck and total hip in the intervention group was significantly higher than that in the control group after 6 months of intervention (P < 0.05). CONCLUSION: The whole-process care model of the medical association is used for osteoporosis patients, which might reduce the risk of kinesiophobia and improve the bone density of the lumbar spine and total hip in patients. But further promotion and improvement of relevant support systems are needed to achieve comprehensive promotion and maximize clinical benefits in this field.

Rare oxoisoaporphine alkaloids from Menispermum dauricum with potential anti-inflammatory activity.

Eleven alkaloids including four previously undescribed oxoisoaporphine alkaloids, menisoxoisoaporphines A-D (1-4), four known analogues (5-8), and three aporphine alkaloids (9-11), were isolated and identified from the rhizomes of Menispermum dauricum. Their structures were elucidated by extensive spectroscopic data and single-crystal X-ray diffraction analyses. Among them, compounds 1 and 4 were the first samples of oxoisoaporphine with C-6 isopentylamino moiety, and 2 was a rare C-4 methylation product of oxoisoaporphine alkaloid. The in vitro anti-inflammatory activity of compounds 1-11 was performed by evaluating the inhibition of NO level in LPS-induced RAW264.7 macrophages. Among them, compound 4 exhibited the most potent NO inhibition activity with an IC50 value of 1.95 ± 0.33 µM. The key structure-activity relationships of those oxoisoaporphine alkaloids for anti-inflammatory effects have been summarized.

Successful Treatment of Minocycline-Induced Facial Hyperpigmentation with a Combination of Chemical Peels and Intense Pulsed Light.

Minocycline is a tetracycline derivative antibiotic commonly used to treat acne, rosacea, and other inflammatory skin conditions. Taking minocycline risks inducing skin pigmentation. If minocycline-induced hyperpigmentation is not treated, it may take months to years for the symptoms to subside after discontinuation of the drug, or the hyperpigmentation may never disappear completely, which can lead to cosmetic anxiety and affect people's quality of life. Previous treatment options for hyperpigmentation were mainly q-switched nd: YAG, ruby, and alexandrite lasers. This article reports a case of facial hyperpigmentation caused by minocycline using a combination of chemical peel and intense pulsed light in a patient with eosinophilic cellulitis (Wells syndrome) who was taking oral minocycline. This case suggests combining chemical peel and intense pulsed light is an effective treatment option for minocycline-induced hyperpigmentation.

The Role of Reflectance Confocal Microscopy in the Diagnosis and Therapeutic Evaluation of Rare Disease Eosinophilic Pustular Folliculitis.

Eosinophilic pustular folliculitis (EPF) is a rare skin disease for which the gold standard of diagnosis relies on the invasive examination of pathological tissue sections. However, due to its invasive nature, many patients tend to refuse this diagnostic test. In such situations, reflectance confocal microscopy (RCM) can be a valuable diagnosis tool. Reflectance confocal microscopy (RCM) can accurately identify the specific structures for biopsy and provide objective imaging data to evaluate clinical symptoms following treatment. Therefore, we present a case report demonstrating the utility of RCM in diagnosing and assessing the treatment of the rare disease EPF for reference.

Clinical and Pathological Analysis of 10 Cases of Eosinophilic Pustular Folliculitis.

We conducted a retrospective analysis of clinical and pathologic data from January 2020 to June 2023, focusing on 10 patients diagnosed with eosinophilic pustular folliculitis at our dermatology clinic. Four of the ten patients had the first rash on the face, five on the trunk, and one on the palms and feet, all of which were initially scattered papules that gradually increased and fused into erythematous plaques with a circular distribution. Seven had pustules with small surface desquamation, and three cases had micro swelling on the face. The rash involved only the face in 5 cases, the face and trunk in 5 cases, and the face, trunk, hands, and feet in 1 case. Seven of the ten patients were pruritic, and 3 had no obvious pruritus. The histopathological features were mild epidermal hyperplasia, lymphocytic and eosinophilic infiltration around the superficial middle dermal vessels and appendages, and eosinophilic and neutrophilic abscesses in the local hair follicles. Treatment with oral indomethacin, prednisone, and minocycline was effective.

Brain-derived neurotrophic factor gene polymorphism affects cognitive function and neurofilament light chain level in patients with subcortical ischaemic vascular dementia.

Objective: To investigate the effects of brain-derived neurotrophic factor (BDNF) gene polymorphism on cognitive function, neuroimaging and blood biological markers in patients with subcortical ischaemic vascular dementia (SIVD). Methods: A total of 81 patients with SIVD were included. According to their BDNF gene polymorphism, the participants were divided into the Val/Val (n = 26), Val/Met (n = 35), and Met/Met (n = 20) groups. A comprehensive neuropsychological evaluation and multimodal brain MRI scan were performed. MRI markers for small vessel disease were visually rated or quantitatively analysed. Moreover, 52 patients were further evaluated with blood marker assays, including amyloid beta (Aß), phosphorylated tau at threonine-181 (P-tau181), glial fibrillary acidic protein (GFAP), total tau (T-tau) and neurofilament light chain (NfL). Results: There were no significant differences in demographics, disease duration or MRI markers of small vessel disease between the three groups. Compared with the Val/Val and Val/Met groups, the Met/Met group showed worse performance in the verbal fluency test and higher levels of plasma NfL. Conclusion: The rs6265 polymorphism of the BDNF gene is associated with semantic language fluency in patients with SIVD. The Met genotype may be a risk factor for cognitive impairment and neuronal injury.

Daurisoline alleviated experimental colitis in vivo and in vitro: Involvement of NF-κB and Wnt/ß-Catenin pathway.

Daurisoline (DS) is one of the most abundant alkaloids extracted from the rhizome of Menispermum Dauricum DC, which is traditionally used to treat inflammatory diseases, especially intestinal inflammation. In this study, we established lipopolysaccharide (LPS)-induced RAW 264.7 macrophages in vitro and Dextran sulfate sodium (DSS)-induced colitis mice model in vivo to investigate the anti-inflammatory effect of DS and its underlying mechanisms. Disease activity index (DAI) was detected during drug intervention. The colon length, macroscopic changes and histopathological scores were adopted to observe the physiological status and the colon injury. The apoptosis of intestinal mucosa was detected using TUNEL. In addition, involved molecular indicators were measured by ELISA kits, RT-qPCR, immunofluorescence (IF), immunohistochemistry (IHC) and western blotting. The vitro experiments indicated that DS significantly suppressed the production of Nitric oxide (NO), reactive oxygen species (ROS) and glutathione (GSH), as well as inhibited the expression of NF-κB signaling pathway in RAW 264.7 cells induced by LPS. Consistent with the vitro experimental results, different doses of DS significantly reduced the incidence of diarrhea, DAI, shortening of the colon, visible damage and histological damage in DSS-induced colitis mice. Moreover, DS treatment decreased the levels of pro-inflammatory mediators cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2) and interleukin (IL)-1ß, and increased the anti-inflammatory cytokines IL-4 and IL-10 in colon tissues. RT-qPCR, western blotting and immunofluorescence analyses further demonstrated that DS inhibits the expression of Wnt/ß-Catenin pathway. We reported for the first time that DS may be an active ingredient in treating ulcerative colitis. Its mechanism might be related to the regulation of the NF-κB and Wnt/ß-Catenin signaling pathway.

TFAM downregulation promotes autophagy and ESCC survival through mtDNA stress-mediated STING pathway.

The dynamics of mitochondrial biogenesis regulation is critical in maintaining cellular homeostasis for immune regulation and tumor prevention. Here, we report that mitochondrial biogenesis disruption through TFAM reduction significantly impairs mitochondrial function, induces autophagy, and promotes esophageal squamous cell carcinoma (ESCC) growth. We found that TFAM protein reduction promotes mitochondrial DNA (mtDNA) release into the cytosol, induces cytosolic mtDNA stress, subsequently activates the cGAS-STING signaling pathway, thereby stimulating autophagy and ESCC growth. STING depletion or mtDNA degradation by DNase I abrogates mtDNA stress response, attenuates autophagy, and decreases the growth of TFAM depleted cells. In addition, autophagy inhibitor also ameliorates mitochondrial dysfunction-induced activation of the cGAS-STING signaling pathway and ESCC growth. In conclusion, our results indicate that mtDNA stress induced by mitochondria biogenesis perturbation activates the cGAS-STING pathway and autophagy to promote ESCC growth, revealing an underappreciated therapeutic strategy for ESCC.

Qualitative analysis of a nonautonomous stochastic SIS epidemic model with Le´vy jumps.

In this paper, we study a nonautonomous stochastic SIS epidemic model with Le´vy jumps. We first establish that this model has a unique global positive solution with the positive initial condition. Then, we investigate the condition for extinction of the disease. Moreover, by constructing suitable stochastic Lyapunov function, sufficient conditions for persistence and existence of Nontrivial T-periodic solution of system are obtained. Finally, numerical simulations are also presented to illustrate the main results.

Developmental validation of the Microreader™ Y Prime Plus ID System: An advanced Y-STR 38-plex system for forensic applications.

Y-STR is widely used in sexual assaults and familial searches of suspects. Here, we reported a novel 38-plex STR genotyping system designed for forensic applications. Microreader™ Y Prime Plus ID System (YPP) amplifies 38 loci in one reaction, including 29 loci from commonly used Yfiler® Plus PCR Amplification Kit & PowerPlex® Y23 System (DYS393, DYS570, DYS19, DYS392, DYS549, Y GATA H4, DYS460, DYS458, DYS481, DYS635, DYS448, DYS533, DYS449, DYS456, DYS389I, DYS390, DYS389Ⅱ, DYS438, DYS391, DYS439, DYS437, DYS385a/b, DYS643, DYS518, DYS576, DYF387S1a/b, and DYS627), 6 commonly used loci for the Y-STR database (DYS444, DYS447, DYS596, DYF404a/b, DYS527a/b, DYS557) and one Y-indel specific for the Chinese population. YPP is designed for different types of samples, such as blood card and swabs. In this work, YPP was validated following SWGDAM guidelines (2016) and guidelines from Ministry of Public Security of the People's Republic of China, including PCR-based, sensitivity, accuracy and precision, mixture, stability and inhibitor, and species specificity. The results indicate that the Microreader™ Y Prime Plus ID System is a powerful identification kit designed for forensic databases.

Identification, characterization, and functional analysis of Tube and Pelle homologs in the mud crab Scylla paramamosain.

Tube and Pelle are essential components in Drosophila Toll signaling pathway. In this study, we characterized a pair of crustacean homologs of Tube and Pelle in Scylla paramamosain, namely, SpTube and SpPelle, and analyzed their immune functions. The full-length cDNA of SpTube had 2052 bp with a 1578 bp open reading frame (ORF) encoding a protein with 525 aa. A death domain (DD) and a kinase domain were predicted in the deduced protein. The full-length cDNA of SpPelle had 3825 bp with a 3420 bp ORF encoding a protein with 1140 aa. The protein contained a DD and a kinase domain. Two conserved repeat motifs previously called Tube repeat motifs present only in insect Tube or Tube-like sequences were found between these two domains. Alignments and structure predictions demonstrated that SpTubeDD and SpPelleDD significantly differed in sequence and 3D structure. Similar to TubeDD, SpTubeDD contained three common conserved residues (R, K, and R) on one surface that may mediate SpMyD88 binding and two common residues (A and A) on the other surface that may contribute to Pelle binding. By contrast, SpPelleDD lacked similar conservative residues. SpTube, insect Tube-like kinases, and human IRAK4 were found to be RD kinases with an RD dipeptide in the kinase domain. SpPelle, Pelle, insect Pelle-like kinases, and human IRAK1 were found to be non-RD kinases lacking an RD dipeptide. Both SpTube and SpPelle were highly expressed in hemocytes, gills, and hepatopancreas. Upon challenge, SpTube and SpPele were significantly increased in hemocytes by Gram-negative or Gram-positive bacteria, whereas only SpPelle was elevated by White Spot Syndrome Virus. The pull-down assay showed that SpTube can bind to both SpMyD88 and SpPelle. These results suggest that SpTube, SpPelle, and SpMyD88 may form a trimeric complex involved in the immunity of mud crabs against both Gram-negative and Gram-positive bacteria.