Complete Genome Sequence Analysis of Bacillus subtilis MC4-2 Strain That against Tobacco Black Shank Disease.

The MC4-2 bacterium strain was isolated and purified from the Periplaneta americana intestine as a biocontrol agent with good antagonistic effect against the pathogens of a soil-borne disease called tobacco black shank. The MC4-2 strain was found to have good broad-spectrum inhibition by plate stand-off test. Based on 16S rRNA and gyrB genes, ANI analysis, and other comparative genomics methods, it was determined that the MC4-2 strain was Bacillus subtilis. The complete genome sequence showed that the genome size was 4,076,630 bp, the average GC content was 43.78%, and the total number of CDSs was 4,207. Genomic prediction analysis revealed that a total of 145 genes were annotated by the CAZy, containing mainly GH and CE enzymes that break down carbohydrates such as glucose, chitin, starch, and alginate, and a large number of enzymes involved in glycosylation were present. A total of ten secondary metabolite clusters were predicted, six clusters of which were annotated as surfactin, bacillaene, fengycin, bacillibactin, subtilosin A, and bacilysin. The present investigation found the biological control mechanism of B. subtilis MC4-2, which provides a strong theoretical basis for the best use of this strain in biological control methods and provides a reference for the subsequent development of agents of this bacterium.

Role of prognostic gene DKK1 in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is one of the most common squamous cell carcinomas of the head and neck. The morbidity and mortality rates of OSCC have increased in recent years. However, the pathogenesis of this disease remains unknown. The present study aimed to identify predictive biomarkers and therapeutic targets for OSCC. Bioinformatics screening of differentially expressed genes in OSCC was performed based on data from The Cancer Genome Atlas and Gene Expression Omnibus databases. Dickkopf Wnt signaling pathway inhibitor 1 (DKK1) was identified to be associated with survival, tumor immunity and DNA repair in OSCC. Furthermore, the effects of DKK1 were evaluated by the knockdown of DKK1 in two OSCC cell lines. The proliferation, clonogenicity, migration and invasion of the cells were assessed in vitro using Cell Counting Kit-8, colony formation, wound healing and Transwell assays, respectively, and were found to be inhibited by DKK1 knockdown. The present study suggests that DKK1 may be used in the prognosis of patients with OSCC and that targeting DKK1 is a potential strategy for OSCC therapy.

Risk Factors for Postoperative Knee Stiffness in Patients with Anteromedial Knee Osteoarthritis Undergoing Unicompartmental Knee Arthroplasty with Cemented Prostheses: A Short-Term, Retrospective, Case-Control Study.

BACKGROUND The present study was performed to determine the potential risk factors for postoperative knee stiffness in patients with anteromedial knee osteoarthritis undergoing unicompartmental knee arthroplasty with cemented prostheses. MATERIAL AND METHODS This retrospective cohort study evaluated patients with anteromedial knee osteoarthritis who underwent medial unicompartmental knee arthroplasty at our hospital between May 2017 and May 2020. The patients were divided into 2 groups according to their prognosis: those who experienced knee stiffness after undergoing unicompartmental knee arthroplasty and those who did not. The factors associated with stiffness after UKA were identified using univariate analysis. Frequencies are used to express categorical variables, while mean±SD is used to express continuous variables. The t test and chi-square test were used. A multivariate logistic regression model was built to identify the risk factors for postoperative stiffness. RESULTS We included 590 knees in the study after unicompartmental knee arthroplasty. The overall incidence of postoperative stiffness in unicompartmental knee arthroplasty surgery was 10.17%. In terms of the radiological measurements, varus deformity (70.34% vs 29.66%) and tibial component posterior slope angle (4.8±2.0 vs 4.6±2.0, P<0.001) were significantly differences between the 2 groups. Four independent risk factors for stiffness after unicompartmental knee arthroplasty were identified: age (95% CI, 1.022-1.048), varus deformity (95% CI, 1.186-1.192), tibial component posterior slope angle (95% CI, 0.550-0.870), and preoperative maximum flexion (95% CI, 0.896-0.923). CONCLUSIONS The overall incidence of postoperative knee stiffness in patients with anteromedial knee osteoarthritis undergoing unicompartmental knee arthroplasty with cemented prostheses was 10.17%, which was at a moderate level compared to patients with other diseases undergoing unicompartmental knee arthroplasty. Four independent risk factors were identified: age, varus deformity, preoperative maximum flexion, and tibial component posterior slope angle. Awareness these risk factors might help surgeons prevent the occurrence of postoperative knee stiffness in patients with UKA.

The indications for core decompression surgery in patients with ARCO stage I-II osteonecrosis of the femoral head: a new, comprehensive prediction system.

BACKGROUND: Core decompression (CD) is considered the most popular treatment method for patients with Association Research Circulation Osseous (ARCO) stage I-II osteonecrosis of the femoral head (ONFH). However, the definitive indication for CD is currently not well established. METHODS: This was a retrospective cohort study. Patients who were diagnosed with ARCO stage I-II ONFH and who underwent CD were included. According to the prognosis, the patients were divided into two groups: collapse of the femoral head after CD and noncollapse of the femoral head. Independent risk factors for the failure of CD treatment were identified. Subsequently, a new scoring system that included all these risk factors was built to help estimate the individual risk of CD failure in patients who were planning to undergo CD. RESULTS: The study included 1537 hips after decompression surgery. The overall failure rate of CD surgery was 52.44%. Seven independent prognostic factors for failed CD surgery were identified, such as male sex (HR = 75.449; 95% confidence interval (CI), 42.863-132.807), Aetiology (Idiopathic HR = 2.762; 95% CI, 2.016-3.788, Steroid-induced HR = 2.543; 95% CI, 1.852-3.685), if the patient had a seated occupation (HR = 3.937; 95% CI, 2.712-5.716), age (HR = 1.045; 95% CI, 1.032-1.058), haemoglobin level (HR = 0.909; 95% CI, 0.897-0.922), disease duration (HR = 1.217; 95% CI, 1.169-1.267) and the combined necrosis angle (HR = 1.025; 95% CI, 1.022-1.028). The final scoring system included these seven risk factors, and the area under the curve of this scoring system was 0.935 (95% confidential interval = 0.922-0.948). CONCLUSION: This new scoring system might provide evidence-based medical proof for determining whether a patient with ARCO stage I - II ONFH might benefit from CD surgery. This scoring system is crucial for making clinical decisions. Consequently, this scoring system is recommended before CD surgery, which could help determine the potential prognosis of patients.

Comparison of Outcomes Following Total Hip Arthroplasty Between Patients Diagnosed with Association Research Circulation Osseous (ARCO) Stage III and Stage IV Osteonecrosis of the Femoral Head: A Retrospective Study of 302 Patients.

BACKGROUND This retrospective study from a single center aimed to compare outcomes following total hip arthroplasty (THA) in 302 patients diagnosed with Association Research Circulation Osseous (ARCO) stage III and stage IV osteonecrosis of the femoral head (ONFH). MATERIAL AND METHODS The study included 302 patients who underwent THA for ONFH between January 2018 and September 2021. Patient groups included ARCO stage III (n=145) and ARCO stage IV (n=157). Outcomes measured included duration of disease, operative time, intraoperative blood volume, postoperative length of hospital stay, pain measured by the visual analog scale (VAS) score, Harris hip score (HHS), and forgotten joint score (FJS). RESULTS Patients with ARCO stage III ONFH had a significantly shorter operative time (P=0.009), shorter length of hospital stay (P=0.021), and reduced volume of intraoperative bleeding (P=0.021) compared with patients with ARCO stage IV ONFH. There were no significant differences in disease duration (P=0.310), postoperative complications (P=0.07), preoperative (P=0.086, P=0.156) and postoperative (P=0.062, P=0.173) HHS and VAS scores, respectively. Patients with stage III ONFH reported significantly higher FJS scores at 3 months, 6 months, and 1 year after THA. CONCLUSIONS Patients who underwent hip arthroplasty for ARCO stage III femoral head necrosis experienced shorter operative time and hospital stay, less intraoperative bleeding, and better restoration of hip function than those with ARCO stage IV. Moreover, stage III patients were more prone to "forget" their hip arthroplasty experience within 1 year of surgery.

Does drainage affect development of heterotopic ossification after total hip arthroplasty?

OBJECTIVE: To investigate the effect of drainage on heterotopic ossification (HO) after total hip arthroplasty (THA) and to evaluate other postoperative complications and joint dysfunction between patients with and without drainage. METHODS: In this retrospective cohort study, the medical records of patients who underwent THA from 2017 to 2019 were reviewed. The patients were divided into a drainage group and non-drainage group. Standard preoperative anteroposterior and lateral radiographs were assessed by senior radiologists for HO analysis. Clinical indicators included the hemoglobin concentration, superficial infection, blood transfusion, hematoma formation, hip range of motion (ROM), erythrocyte sedimentation rate, C-reactive protein concentration, dressing changes, visual analogue scale score, and Harris Hip Score (HHS). RESULTS: The incidence of HO was significantly higher in the drainage than non-drainage group (32.0% vs. 16.3%). The presence of severe HO (Brooker grade III or IV) was also different between the groups. Patients in the non-drainage group had smaller ROM early after surgery, but the final ROM and HHS did not differ significantly between the groups. CONCLUSIONS: The rate and degree of HO after THA were significantly different between patients with and without drainage. There is no added advantage of closed suction drainage over no drainage in primary THA.

Two-Stage Total Hip Arthroplasty for Primary Advanced Septic Arthritis of the Hip in Adults.

Background: Comparing the outcomes of debridement and total hip arthroplasty (THA) with antibiotic-loaded spacer implantation and subsequent THA for the treatment of patients affected by primary advanced septic arthritis (SA) of the hip in adults. Methods: All of the 20 patients (20 hips) underwent two-stage surgery. Nine patients were submitted to surgical debridement first and then THA (group 1), while 11 patients were treated with antibiotic-loaded spacer and subsequent THA (group 2). Patients were evaluated based on the recurrence of infection, Harris hip score, visual analogue scale (VAS) pain score, and leg length discrepancy. Results: No cases of infection, deep vein thrombosis, death, and loosening of the hip prosthesis were observed during follow-up. The mean follow-up time was 29.09 ± 10.80 months in group 1 and 28.22 ± 14.80 months in group 2. Before the THA surgery, the mean leg length discrepancy was 2.80 ± 2.03 cm in group 1 and 0.50 ± 0.23 cm in group 2 (P < 0.05). In the latest follow-up, the Harris hip scores of patients were 90.33 ± 4.85 in group 1 and 94.36 ± 2.34 in group 2 (P < 0.05), respectively. There was no statistically significant difference in the VAS pain score of the hip between the two groups (P > 0.05). Conclusions: Debridement and antibiotic-loaded spacer and subsequent THA were effective in eradicating the infection for advanced SA. However, antibiotic-loaded spacer and subsequent THA was superior for effectively maintaining the length of the lower limb and function of the hip.

The negative feedback regulation of microRNA-146a in human periodontal ligament cells after Porphyromonas gingivalis lipopolysaccharide stimulation.

OBJECTIVE: Toll-like receptors (TLRs) pathway has been demonstrated to play an important role in periodontitis. However, the regulatory mechanism of microRNAs (miRNAs) on TLRs pathway is still unclear. Hence, this study is to explore the function of miRNA-146a in inflammatory reaction induced by Porphyromonas gingivalis lipopolysaccharide (LPS) in human periodontal ligament cells (hPDLCs). METHODS: Cells were treated with 1 or 10 µg/ml P. gingivalis LPS. The expression of TLR2, TLR4 and miRNA-146a were measured by real-time polymerase chain reaction (PCR). Enzyme-linked immunosorbent assay (ELISA) was applied to detect nuclear factor (NF)-κ B p65 nuclear activity, interleukin-1ß (IL-1ß), IL-6, IL-8 and tumor necrosis factor-α (TNF-α). To examine the underlying mechanisms, cells were exposed to anti-TLR2/4 mAb or miRNA-146a inhibitor/mimic and evaluated by real-time PCR and ELISA. RESULTS: 10 µg/ml P. gingivalis LPS increased the expressions of TLR2 (3.79 ± 0.31), TLR4 (2.21 ± 0.31), and miRNA-146a (4.91 ± 0.87), NF-κ B p65 nuclear activity (6.51 ± 0.77 fold) (p < 0.05). 1 µg/ml P. gingivalis LPS induced TLR2 (3.05 ± 0.23), miRNA-146a (3.66 ± 0.83) and NF-κ B p65 nuclear activity (4.06 ± 0.78 fold) (p < 0.05), except TLR4 (1.11 ± 0.30, p > 0.05). Also, cytokines production increased (p < 0.05). The up-regulation of miRNA-146a could be blocked by anti-TLR2/4 mAb (p < 0.05). After the blockage of miRNA-146a, TLR2, TLR4, NF-κ B p65 nuclear activity and proinflammatory cytokines increased. However, after application of miRNA-146a mimic, the levels of these indexes decreased obviously (p < 0.05). CONCLUSION: MiRNA-146a functions as a negative feedback regulator via down-regulating proinflammatory cytokine secretion and blocking TLRs signaling pathway in hPDLCs after P. gingivalis LPS stimulation.

[Effect of high glucose on the secretion of cytokines induced by Porphyromonas gingivalis lipopolysaccharide].

OBJECTIVE: To investigate the influence of high glucose on Porphyromonasgingivalis (Pg) lipopolysaccharide (LPS) stimulating human gingival fibroblasts (HGF) to secret the cytokines. METHODS: HGF were obtained from the primary culture of the tissue explants. Cells were divided into four groups, low glucose (5.5 mmol/L) + 1 mg/L Pg LPS (group A);low glucose + 10 mg/L Pg LPS (group B); high glucose (25 mmol/L) +1 mg/L Pg LPS(group C); high glucose+10 mg/L Pg LPS (group D). The levels of tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) in cell supernatants were detected by enzyme- linked immunosorbent assay at 6 h and 12 h. The expressions of toll-like receptor 2, 4 (TLR-2, 4) were examined by real-time polymerase chain reaction. After pretreatment with anti-TLR2 and anti-TLR4 monoclonal antibody in HGF, TNF-α and L-1ß levels were detected. RESULTS: TNF-α concentration increased obviously in high glucose 6 h and 12 h after Pg LPS stimulation (P < 0.01). IL-1ß secretion increased (P < 0.01). Meanwhile, TLR2, 4 mRNA expression increased, especially in high glucose+10 mg/L Pg LPS (P < 0.01). After inhibition of the TLR2, 4 in high glucose + 10 mg/L Pg LPS respectively, TNF-α level [(297.16±11.49), (390.01±12.81) ng/L] decreased (F = 166.02, P < 0.01), and IL-1ß level [(49.90±4.08), (99.35±5.01) ng/L] also decreased (F = 153.51, P < 0.01). CONCLUSIONS: High glucose may promote Pg LPS to stimulate the secretion of TNF-α and IL-1ß through regulating TLR2, 4 expression, which suggests that the elevating blood glucose precipitate in aggravating the process of periodontal disease.

High glucose induces inflammatory cytokine through protein kinase C-induced toll-like receptor 2 pathway in gingival fibroblasts.

Toll-like receptors (TLRs) play a key role in innate immune response and inflammation, especially in periodontitis. Meanwhile, hyperglycemia can induce inflammation in diabetes complications. However, the activity of TLRs in periodontitis complicated with hyperglycemia is still unclear. In the present study, high glucose (25 mmol/l) significantly induced TLR2 expression in gingival fibroblasts (p<0.05). Also, high glucose increased nuclear factor kappa B (NF-κB) p65 nuclear activity, tumor necrosis factor-α (TNF-α) and interleukin-lß (IL-1ß) levels. Protein kinase C (PKC)-α and δ knockdown with siRNA significantly decreased TLR2 and NF-κB p65 expression (p<0.05), whereas inhibition of PKC-ß had no effect on TLR2 and NF-κB p65 under high glucose (p<0.05). Additional studies revealed that TLR2 knockdown significantly abrogated high-glucose-induced NF-κB expression and inflammatory cytokine secretion. Collectively, these data suggest that high glucose stimulates TNF-α and IL-1ß secretion via inducing TLR2 through PKC-α and PKC-δ in human gingival fibroblasts.