TANK-binding kinase 1 (TBK1) is an important enzyme in the regulation of cellular antiviral effects. TBK1 regulates the activity of the interferon regulatory factors IRF3 and IRF7, thereby playing a key role in type I interferon (IFN) signaling pathways. The structure of TBK1 consists of an N-terminal kinase domain, a middle ubiquitin-like domain (ULD), and a C-terminal elongated helical domain. It has been reported that the ULD of TBK1 regulates kinase activity, playing an important role in signaling and mediating interactions with other molecules in the IFN pathway. In this study, we present the crystal structure of the ULD of human TBK1 and identify several conserved residues by multiple sequence alignment. We found that a hydrophobic patch in TBK1, containing residues Leu316, Ile353, and Val382, corresponding to the "Ile44 hydrophobic patch" observed in ubiquitin, was conserved in TBK1, IκB kinase epsilon (IKKɛ/IKKi), IκB kinase alpha (IKKα), and IκB kinase beta (IKKβ). In comparison with the structure of the IKKβ ULD domain of Xenopus laevis, we speculate that the Ile44 hydrophobic patch of TBK1 is present in an intramolecular binding surface between ULD and the C-terminal elongated helices. The varying surface charge distributions in the ULD domains of IKK and IKK-related kinases may be relevant to their specificity for specific partners.
Amino Acid Sequence ; Conserved Sequence ; Crystallography, X-Ray ; Humans ; Hydrophobic and Hydrophilic Interactions ; I-kappa B Kinase ; chemistry ; metabolism ; Isoleucine ; chemistry ; Models, Molecular ; Molecular Sequence Data ; Protein Binding ; Protein Multimerization ; Protein Structure, Tertiary ; Protein-Serine-Threonine Kinases ; chemistry ; Sequence Alignment ; Static Electricity ; Ubiquitin ; chemistry

Objective:To investigate effects of short-term cigarette smoke exposure combined with poly(I:C)stimulation on lung immune response and interferon expression in mice.Methods:BALB/c mice were randomly divided into 4 groups:control group,smoke group,poly(I:C)group and smoke combined poly(I:C)group.Total cell number and cell classification count of bronchoalveo-lar lavage fluid(BALF)were detected,and cell morphology was observed under ordinary light.Cytokines,chemokines,interferon and interferon stimulating genes expressions in lung tissues were detected by fluorescence quantitative PCR.Results:Compared with control group,total cell count,macrophage count and neutrophil count in smoke combined poly(I:C)group were significantly increased(P<0.05),and macrophage count was higher than that in poly(I:C)group.Macrophages of airway lavage fluid of mice in smoke combined with poly(I:C)group were larger in size,round or irregular in shape,and had more vacuoles in cytoplasm.Com-pared with control group,mRNA expressions of neutrophil chemokine CXCL1(P<0.05),CXCL2(P<0.01)and lymphocyte chemo-kine CCL2(P<0.01)in lung tissues of mice in smoke combined with poly(I:C)group were increased.IL-1β,IL-6,TNF-α mRNA expressions were significantly increased(P<0.01),IFN-β(P<0.01),IFN-γ(P<0.05),MX2(P<0.01)and IP-10(P<0.01)expre-ssions in lung tissues were significantly increased,and compared with poly(I:C)group,mRNA expressions of CXCL2(P<0.05),TNF-α(P<0.01)and IFN-β(P<0.05)in lung tissues of mice in smoke combined with poly(I:C)group were significantly increased.Conclusion:Cigarette smoke combined with poly(I:C)induces lung inflammation and expressions of interferon and interferon stimu-lating genes in mice.Cigarette exposure also increases poly(I:C)-induced acute lung inflammation and type Ⅰ interferon expression in mice.

Objective To investigate the association between phage-mediated shiga toxin and molecular distribution of CRISPR in Escherichia (E.) coli O26:H11 or NM.Methods A total of 135 E.coli O26:H11 or NM strains were collected from NCBI database.Software CRT and CRISPR Finder were used to extract CRISPR and Excel was used to assign the spacer of unique number and type CRISPR.And the relationship between CRISPR and stx phage was analyzed.Results All the 135 E.coli O26:H11 or NM strains had the CRISPR.For CRISPRI,CRISPR2.1,CRISPR2.2 and CRISPR3-4,19,22,1 and 1 subtypes were found,respectively.According to the four CRISPR sites,the strains could be divided into 40 subtypes.Stx-phage was only observed in the group C of CRISPR.Compared with E.coli of stx-phage negative,E.coli with stx-phage harbored more spacers.Conclusions CRISPR loci was extensively existed in E.coli O26:H11 or NM,and many subtypes were found in these strains.The presence of stx-phage was related to the molecular distribution of CRISPR in E.coli O26:H11 or NM.CRISPR might be a valuable biomarker to identify strains with high virulent potential.

Objective:To verify the efficacy and safety of daily oral minodronate in postmenopausal women with established osteoporosis.Methods:In this randomized, double-blinded, placebo-controlled trial, 262 postmenopausal women were enrolled. Patients were randomized to receive daily oral minodronate 1 mg with supplements of 500 mg calcium and 200 U vitamin D 3 ( n=130) or placebo ( n=132) with daily supplements of 500 mg calcium and 200 U vitamin D 3, for 48 weeks. The primary endpoint was the average bone mineral density (BMD) change in the lumbar vertebrae 48 weeks post-treatment. Secondary outcome measures was the incidence of vertebral fractures. Safety assessments included the rate of adverse events. Results:At the end of 48 weeks treatment, the average BMD change rate from baseline were: full analysis set results: (3.52±4.82)% in the minodronate group and (2.00±5.74)% in the placebo group; per-protocol set results: (3.99±5.05)% in the minodronate group and (2.07±6.20)% in the placebo group; the differences were all significant (all P<0.05). Vertebral fracture occured in 3 patients (2.3%, 3/132) in the placebo group, and 1 case (0.8%, 1/130) in the minodronate group ( P>0.05). The incidence of adverse events was 71.5% (93/130) in the minodronate group and 78.0% (103/132) in the placebo group ( P>0.05). Conclusion:Minodronate is effective and safe in the treatment of postmenopausal osteoporosis without severe side effects.

Objective:To investigate the correlation of isokinetic thigh muscle asymmetry with gait asymmetry at one year after anterior cruciate ligament (ACL) reconstruction.Methods:A retrospective case series analysis was made on 25 patients treated by ACL reconstruction in Third Hospital of Peking University from January 2014 to January 2019. All the subjects were male, aged 17-47 years [(29.4±5.2)years]. Standard rehabilitation treatment procedures were conducted after surgery. The isokinetic muscle strength of the thigh was collected at one year after operation to evaluate knee extensor and flexor concentric strength at 60°/s, 180°/s and 300°/s, and knee extensor and flexor eccentric strength at 60°/s. The maximum concentric and eccentric strength of the knee extensor and flexor were analysed between the injured and intact side. The three-dimensional motion information and ground reaction force was collected during the stance phase of gait, and knee kinematic and kinetic parameters were calculated by inverse dynamics analysis, including the peak flexion moment, peak extension moment, first and second peak adduction moment, peak external and internal rotation moment, peak flexion angle, peak extension angle during the terminal stance phase, flexion angle at heel strike, peak abduction angle, and peak external rotation angle during terminal stance phase. Spearman correlation analysis was used to study the correlation between limb symmetry index (LSI) of isokinetic muscle strength and LSI of gait parameters.Results:One year after ACL reconstruction, the maximum concentric strength of the knee extensor and flexor at 60°/s, 180°/s and 300°/s and maximum eccentric strength of the knee flexor and extensor at 60°/s on the injured side were significantly lower than those on the normal side (all P<0.01). Compared with the intact side, the peak knee extension moment on the injured side was significantly lower during the stance phase of gait ( P<0.01), and the extension angle was insufficient during the terminal stance phase ( P<0.01). There were no significant differences between the injured and intact side in terms of peak flexion moment, first and second peak adduction moment, peak external and internal rotation moment, peak flexion angle, flexion angle at heel strike, peak abduction angle, and peak external rotation angle during terminal stance phase (all P>0.05). One year after ACL reconstruction, Spearman correlation analysis during gait stance phase showed that the LSI of 60°/s concentric strength of the isokinetic knee extensor was significantly related to LSI of peak internal rotation moment ( R=0.42, P<0.05), the LSI of 180°/s concentric strength of the isokinetic knee extensor was significantly related to LSI of peak flexion moment ( R=0.45, P<0.05), the LSI of 180°/s concentric strength of the isokinetic knee flexor was significantly correlated with LSI of peak flexion angle ( R=0.46, P<0.05), the LSI of 300°/s concentric strength of the isokinetic knee extensor was significantly correlated with LSI of peak knee flexion angle ( R=0.42, P<0.05), and the LSI of 60°/s eccentric strength of the isokinetic flexor was significantly correlated with LSI of peak knee flexion angle ( R=0.54, P<0.01). Conclusions:For patients following ACL reconstruction at one year postoperatively, a significant correlation of isokinetic thigh extensor strength asymmetry with peak knee flexion and rotation moment and peak knee flexion angle, and that of isokinetic thigh flexor strength asymmetry with peak knee flexion angle are observed during the stance phase of gait, which suggests that patients with ACL reconstruction still need systematic rehabilitation training one year postoperatively to improve muscle strength and motor function so as to reduce the risk of ACL reinjury and secondary injury.

Objective:To evaluate knee biomechanics of patients about 12 months after anterior cruciate ligament(ACL)reconstruction during cutting and determine the abnormal biomechanical characteristics.Methods:Sixteen males about 12 months after ACL reconstruction were recruited for this study.Three-dimensional kinematic and kinetic data were collected during cutting movement.Knee joint angles and moments were calculated.Paired t-tests were used to compare the differences in knee biomechanics be-tween the surgical leg and nonsurgical leg.Results:The peak posterior ground reaction force(surgical leg:0.380±0.071;nonsurgical leg:0.427±0.069,P=0.003)and vertical ground reaction force(surgical leg:1.996±0.202,nonsurgical leg:2.110±0.182,P=0.001)were significantly smaller in the surgical leg than in the nonsurgical leg.When compared with the uninjured leg,the surgical leg demonstrated a smaller knee flexion angle(surgical leg:38.3°±7.4°;nonsurgical leg:42.8°±7.9°,P＜0.001)and larger external rotation angle(surgical leg:10.3°±2.4°;nonsurgical leg:7.7°±2.1°,P=0.008).The surgical leg also demonstrated a smaller peak knee extension moment(surgical leg:0.092±0.031;nonsurgical leg:0.133±0.024,P＜0.001)and peak knee external rotation moment(surgical leg:0.005±0.004;nonsurgical leg:0.008±0.004,P=0.015)when com-pared with the nonsurgical leg.Conclusion:The individuals with ACL reconstruction mainly showed asymmetrical movements in the sagittal and horizontal planes.The surgical leg demonstrated a smaller peak knee flexion angle,knee extension moment,and knee external rotation moment,with greater knee external rotation angle.

Objective Gait analysis plays an important role in the description of the site and degree of injury in the lower limbs, and symmetry is an important indicator of gait characteristics. The aim of this study is to quantitatively describe gait information and its asymmetry in three-dimensional space, dig deeper into gait information and improve the analysis of abnormal gait characteristics.Methods The information about the change of coronal-sagittal plane angle in different parts was obtained based on the inertial gait sensor, and the gait space vector was inverted to build the movement curve and form a motion symmetry detection method based on three-dimensional space angle vector distance. The characteristics of normal subjects (control group) and the anterior cruciate ligament (ACL) rupture patients (patients group) were analyzed from July, 2017 to January, 2018, and its effectiveness was verified.Results There was a better discrimination effect in the calf, thigh and knee asymmetry, etc., between the control group and the patient group (P<0.05). The characteristics of the calf space motion vector were highest among the control group and the patient group, and the discrimination was relatively stable, and not various with gender, age, height or body mass index.Conclusion The time-coronal-sagittal vector diagrams of left and right limbs were obtained by inertial sensor and the average of the square of the Euclidean distances in space at the selected gait cycle time points was calculated in three-dimensional space, which could be used as a gait asymmetry analysis method.

Previously, we reported that Y, a new epigallocatechin gallate derivative, is efficacious in reversing doxorubicin (DOX)--mediated resistance in hepatocellular carcinoma BEL-7404/DOX cells. In this study, we evaluated the efficacy of Y in reversing drug resistance both and by determining its effect on the adenosine triphosphate-binding cassette protein B1 transporter (ABCB1 or P-glycoprotein, P-gp). Our results showed that Y significantly sensitized cells overexpressing the ABCB1 transporter to anticancer drugs that are ABCB1 substrates. Y significantly stimulated the adenosine triphosphatase activity of ABCB1. Furthermore, Y exhibited a higher docking score as compared with epigallocatechin gallate inside the transmembrane domain of ABCB1. In addition, in the nude mouse tumor xenograft model, Y (110 mg/kg, intragastric administration), in combination with doxorubicin (2 mg/kg, intraperitoneal injection), significantly inhibited the growth of BEL-7404/DOX cell xenograft tumors, compared to equivalent epigallocatechin gallate. In conclusion, Y significantly reversed ABCB1-mediated multidrug resistance and its mechanisms of action may result from its competitive inhibition of the ABCB1 drug efflux function.

Objective To investigate the prevalence of Cryptosporidium infection and the distribution of parasite species and genotypes among HIV-positive individuals in Jiangxi Province. Methods HIV-positive individuals' sociodemographic and clinical data were collected from three AIDS designated hospitals in Jiangxi Province from January 2022 to March 2023. Subjects' stool samples were collected, and genomic DNA was extracted from stool samples. Nested PCR assay was performed based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium, and Cryptosporidium gp60 gene was amplified in stool samples positive for the SSU rRNA gene. The second-round PCR amplification product was checked with 1.5% agarose gel electrophoresis, and the products of suspected positive amplifications were sequenced, followed by sequence alignment. The phylogenetic tree was created using the Neighbor-Joining method with the software MEGA 11.0, to characterize the species, genotypes and sub-genotypes of Cryptosporidium. Results A total of 382 HIV-positive individuals were enrolled, with two cases identified with Cryptosporidium infection (0.52% prevalence), and both cases had no abdominal pain or diarrhea. Following sequencing and sequence alignment, the gene sequences of these two Cryptosporidium isolates shared 99.76% and 99.88% similarity with the gene sequence of C. meleagridis isolates. Phylogenetic analysis based on the Cryptosporidium SSU rRNA gene sequence identified the species of these two Cryptosporidium-positive stool samples as C. meleagridis. Following nested PCR amplification of the Cryptosporidium gp60 gene, sequencing and sequence alignment, the two C. meleagridis isolates were characterized as III eA17G2R1 and III bA25G1R1a sub-genotypes, and the sub-genotype III bA25G1R1a was firstly described in humans. Conclusion The prevalence of Cryptosporidium is low among HIV-positive individuals in Jiangxi Province. The likelihood of Cryptosporidium infection cannot be neglected among HIV-positive individuals without diarrhea.

The complexity of oral ulcerations poses considerable diagnostic and therapeutic challenges to oral specialists. The expert consensus was conducted to summarize the diagnostic work-up for difficult and complicated oral ulcers, based on factors such as detailed clinical medical history inquiry, histopathological examination, and ulceration-related systemic diseases screening. Not only it can provide a standardized procedure of oral ulceration, but also it can improve the diagnostic efficiency, in order to avoid misdiagnosis and missed diagnosis.
Consensus ; Humans ; Oral Ulcer/therapy*