Effect of bacillus subtilis strain Z15 secondary metabolites on immune function in mice.

BACKGROUND: Previous studies have shown that secondary metabolites of Bacillus subtilis strain Z15 (BS-Z15) are effective in treating fungal infections in mice. To evaluate whether it also modulates immune function in mice to exert antifungal effects, we investigated the effect of BS-Z15 secondary metabolites on both the innate and adaptive immune functions of mice, and explored its molecular mechanism through blood transcriptome analysis. RESULTS: The study showed that BS-Z15 secondary metabolites increased the number of monocytes and platelets in the blood, improved natural killer (NK) cell activity and phagocytosis of monocytes-macrophages, increased the conversion rate of lymphocytes in the spleen, the number of T lymphocytes and the antibody production capacity of mice, and increased the levels of Interferon gamma (IFN-γ), Interleukin-6 (IL-6), Immunoglobulin G (IgG) and Immunoglobulin M (IgM) in plasma. The blood transcriptome analysis revealed 608 differentially expressed genes following treatment with BS-Z15 secondary metabolites, all of which were significantly enriched in the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) terms for immune-related entries and pathways such as Tumor Necrosis Factor (TNF) and Toll-like receptor (TLR) signaling pathways, and upregulated expression levels of immune-related genes such as Complement 1q B chain (C1qb), Complement 4B (C4b), Tetracyclin Resistant (TCR) and Regulatory Factor X, 5 (RFX5). CONCLUSIONS: BS-Z15 secondary metabolites were shown to enhance innate and adaptive immune function in mice, laying a theoretical foundation for its development and application in the field of immunity.

LncRNA BC promotes lung adenocarcinoma progression by modulating IMPAD1 alternative splicing.

BACKGROUND: The therapeutic value of targeted therapies in patients with lung cancer is reduced when tumours acquire secondary resistance after an initial period of successful treatment. However, the molecular events behind the resistance to targeted therapies in lung cancer remain largely unknown. AIMS: To discover the important role and mechanism of lncRNA BC in promoting tumor metastasis and influencing clinical prognosis of LUAD. MATERIALS & METHODS: Microarrays were used to screen a comprehensive set of lncRNAs with differential expression profiles in lung cancer cells. The functional role and mechanism of lncRNA were further investigated by gain- and loss-of-function assays. RNA pull-down, protein assays, and mass spectrometry were used to identify proteins that interacted with lncRNA. TaqMan PCR was used to measure lncRNA in lung adenocarcinoma and adjacent nontumor tissues from 428 patients. The clinical significance of lncRNA identified was statistically confirmed in this cohort of patients. RESULTS: In this study, we show that the long non-coding RNA BC009639 (BC) is involved in acquired resistance to EGFR-targeted therapies. Among the 235 long non-coding RNAs that were differentially expressed in lung cancer cell lines, with different metastatic potentials, BC promoted growth, invasion, metastasis, and resistance to EGFR-tyrosine kinase inhibitors (EGFR-TKIs), both in vitro and in vivo. BC was highly expressed in 428 patients with lung adenocarcinoma (LUAD) and high BC expression correlated with reduced efficacy of EGFR-TKI therapy. To uncover the molecular mechanism of BC-mediated EGFR-TKI resistance in lung cancer, we screened and identified nucleolin and hnRNPK that interact with BC. BC formed the splicing complex with nucleolin and hnRNPK to facilitate the production of a non-protein-coding inositol monophosphatase domain containing 1 (IMPAD1) splice variant, instead of the protein-coding variant. The BC-mediated alternative splicing (AS) of IMPAD1 resulted in the induction of the epithelial-mesenchymal transition and resistance to EGFR-TKI in lung cancer. High BC expression correlated with clinical progress and poor survival among 402 patients with LUAD. DISSCUSSION: Through alternative splicing, BC boosted the non-coding IMPAD1-203 transcript variant while suppressing the IMPAD1-201 variant. In order to control the processing of pre-mRNA, BC not only attracted RNA binding proteins (NCL, IGF2BP1) or splicing factors (hnRNPK), but also controlled the formation of the splicing-regulator complex by creating RNA-RNA-duplexes. CONCLUSION: Our results reveal an important role for BC in mediating resistance to EGFR-targeted therapy in LUAD through IMPAD1 AS and in implication for the targeted therapy resistance.

The remarkable enhancement of photo-stability and antioxidant protection of lutein coupled with carbon-dot.

Lutein is a carotenoid that is beneficial to human health. However, its low stability and bioavailability have limited its application in the pharmaceutical and food industries. Herein, lutein has been successfully modified with carbon-dots (CDs) via a simple and mild solvent-ultrasonic method at room temperature. The synthesized lutein/CDs composites (LCs) showed 3.4 times higher photostability than the pristine lutein under UV irradiation, and 3.5 times higher than that under visible light, and the retention rate of lutein in the air rose from 3.6 % to 68 % over 25 days. Meanwhile, the antioxidant capacity of lutein has been improved by 6.4 times. Based on the photoluminescence (PL) measurements, a possible mechanism for the enhanced photostability and antioxidant ability of LCs has been proposed. The work provides a simple but effective approach to enhancing the stability and antioxidant capacity of lutein, and is expected to extend its application in food and biomedical fields.

Genome sequencing and functional annotation of Bacillus sp. strain BS-Z15 isolated from cotton rhizosphere soil having antagonistic activity against Verticillium dahliae.

In the present study, antagonistic activity of bacterial strain BS-Z15, was evaluated against Verticillium dahlia. The fermented broth of BS-Z15 inhibited the growth of Verticillium dahliae. The genome of strain BS-Z15 had a total size of 4,068,702 base pairs and contained 4318 genes, of which 4196 are coding sequences and 122 are non-coding RNA. Among these genes, nine genomic islands, 86 tRNAs, 13 sRNAs, and one prophage was determined. With the help of annotation databases, most unigene functions were identified. At the same time, genomic comparison between BS-Z15 and 12 Bacillus members showed that the genes of BS-Z15 were closely related to the Bacillus group, and were conserved between the two groups, including most of the genes associated with fungal antagonism. BS-Z15 contains genes involved in a variety of antagonistic mechanisms, including genes encoding or synthesizing mycosubtilin, chitinases (but not CHIA and CHIB), glycoside hydrolases, iron nutrients, and antibiosis. However, it only contained the complete mycosubtilin- and bacilibactin-related operators in the reported main antifungal gene cluster of B. subtilis. Mycosubtilin and bacilibactin may be the main active antifungal substance. Besides, some genes could encode products related to biofilm production, which may be related to the colonization ability of the strain in plant rhizospheres. The complete genome of B. subtilis BS-Z15 provided new insights into the potential metabolites it produces related to its biocontrol activity.

Hydrothermally activated TiO2 nanoparticles with a C-dot/g-C3N4 heterostructure for photocatalytic enhancement.

Dye degradation via photocatalysis technology has been investigated intensively to tackle environmental issues and energy crisis concerns. In this study, a newly designed ternary photocatalyst was facilely prepared by a simple one-pot hydrothermal process by directly mixing TiO2 nanoparticles with carbon dots (C-dots) and graphitic carbon nitride (g-C3N4). The optimized precursor treatments and heterostructure components show significantly enhanced photodegradation activity towards organic dyes Rhodamine B (RhB) and methylene blue (MB). Excellent photocatalytic activities were achieved owing to the better attachment of anatase-type TiO2 nanoparticle-aggregations to the C-dots/g-C3N4 (CC) nanocomposite, which impressively displays superhydrophilicity by employing the hydrothermal activation process. FT-IR spectra revealed that the hydrothermal treatment could remarkably increase the coupling interactions between TiO2 nanoparticles and the CC nanosheets within the ternary catalyst, enhancing the photocatalytic activity. Thus, it was concluded that this ternary photocatalyst is highly suitable for the remediation of dye-contaminated wastewater.

[Short and medium term follow-up study on the changes of spine pelvic parameters in patients with hip-spine syndrome after total hip arthroplasty].

OBJECTIVE: To investigate the influence of total hip arthroplasty on the changes of spine pelvic parameters in patients with hip spine syndrome. METHODS: From January 2013 to October 2014, 22 patients (26 hips) with hip spine syndrome accompanied by necrosis of femoral head, hip osteoarthritis and congenital dysplasia of hip were treated with total hip arthroplasty. There were 12 males and 10 females with an average age of 58.4 years (range, 45 to 76 years). The course of disease was 1.5 to 25 years with an average of 12.8 years. Before and after the operation, the anteroposterior, full length radiographs of both lower limbs, thoracolumbar spine and pelvis in standing position were routinely taken. The balance parameters of spine pelvis coronal plane and sagittal plane before and after the replacement were measured. Visual analogue scale (VAS), Oswestry Disability Index (ODI) and Harris score were performed before and after the operation. RESULTS: All cases were followed up for 21 to 52 (32±8) months. No infection, prosthesis subsidence, loosening, prosthesis dislocation were found in the last follow up. After total hip arthroplasty, sagittal vertical axis(SVA), thoracic kyphosis(TK), lumbar lordosis(LL), pelvic tilt (PT) were significantly reduced(P<0.05), sacral slope(SS) was significantly increased(P<0.05), there was no significant difference in pelvic incidence(PI)(P>0.05); PT and scoliosis (coronal position) were significantly decreased after operation(P<0.05); VAS score of waist, VAS score of hip joint and ODI score of waist were significantly decreased before and after operation (P<0.05). Harris score of hip joint increased significantly after operation, and thedifference was statistically significant (P<0.05). CONCLUSION: After total hip arthroplasty, the coronal and historical balance parameters of spine and pelvis are significantly improved, and the short term and medium-term effects are satisfactory.

[Strategy of acetabular anteversion in total hip arthroplasty with lumbar degenerative kyphosis].

OBJECTIVE: To investigate how to place the anteversion of acetabular prosthesis more reasonably in patients with lumbar degenerative kyphosis. METHODS: A total of 122 patients with degenerative kyphosis of lumbar spine who underwent total hip arthroplasty from December 2017 to October 2019 were included and divided into experimental group and control group, 61 cases in each group. In experimental group, there were 25 males and 36 females, with a median age of 67.0 years;the median course of disease was 46.0 months;the functional pelvic plane with acetabular anteversion was set according to different types of pelvic anterior plane bracket. In control group, there were 27 males and 34 females, with a median age of 67.0 years;the median course was 42.0 months;in control group, the anteversion was set by the traditional method. The patients were followed up for 3 months. The operation time and blood loss were recorded. The incidence of infection and dislocation within 3 months was counted. Harris score before and 3 months after operation was recorded. Functional anteversion angle of standing position was measured 3 months after operation. RESULTS: Compared with control group, there was no difference in operation time and blood loss between the two groups (P=0.918, 0.381);there was no infection between two groups within 3 months after operation;there was 1 case of hip joint dislocation in the control group and no dislocation in experimental group. There was no significant difference in Harris score before and after operation. Three months later, reexamination of pelvic standing radiographs showed that the number of patients with functional anteversion of acetabular prosthesis outside the safe area was less in experimental group thanin control group (P=0.048), and the functional anteversion angle of acetabular prosthesis was more concentrated in the range of 15 ° to 20 ° in experimental group (P<0.001). CONCLUSION: According to the preoperative evaluation and classification of patients, better functional anteversion of acetabular prosthesis can be obtained with the help of pelvic anterior plane reference bracket in hip arthroplasty with lumbar degenerative kyphosis.

BRD1-Mediated Acetylation Promotes Integrin αV Gene Expression Via Interaction with Sulfatide.

Integrin αV gene expression is often dysregulated in cancers especially in hepatocellular carcinoma (HCC); however, the mechanism of regulation is poorly understood. Here, it is demonstrated that sulfatide activated integrin αV gene transcription, through histone H3K9/14 acetylation at the promoter, and high integrin αV expression are closely associated with poor prognosis. To elucidate the mechanism of regulation of acetylation, sulfatide-bound proteins were screened by mass spectrometry (MS), and bromodomain containing protein 1 (BRD1) was identified as an interacting protein that also colocalized with sulfatide in HCC cells. BRD1 was also formed a complex with Sp1, which was recruited to the integrin αV gene promoter. Sulfatide was also found to induce BRD1, monocytic leukemia zinc finger (MOZ) and histone acetyltransferase binding to ORC1 (HBO1) acetyltransferase multiprotein complex recruitment to the integrin αV promoter, which is responsible for histone H3K9/14 acetylation. Finally, knockdown of BRD1 limited sulfatide-induced H3K9/14 acetylation and occupancy of MOZ or HBO1 on integrin αV gene promoter.Implications: This study demonstrates that sulfatide interaction with BRD1 mediates acetylation and is important for regulation of integrin αV gene expression. Mol Cancer Res; 16(4); 610-22. ©2018 AACR.

[Comparison of therapeutic effects of three repair methods for rotator cuff tears under arthroscopy].

OBJECTIVE: To retrospectively study curative effects of three repair methods for rotator cuff tears under arthroscopy, and to explore relationships between different repair methods and cuff lesions. METHODS: From January 2009 to Jaunary 2014, a total of 353 patients with rotator cuff tears treated with surgical repair under arthroscopy were included in this study. All the patients were divided into three groups according to time of visitiny hospital and it was divided into three periods. The patients on the first period were treated with single row rivet fixation(115 cases), including 51 males and 64 females, with an average age of (57.46±9.08) years old. The patients on the second period were treated with double row rivet fixation(163 cases), including 76 males and 87 females, with an average age of (56.93±9.92) years old. The patients in the third period were treated with suture bridge fixation(75 cases), including 32 males and 43 females, with an average age of (55.90±9.15) years old. There were 29 patients with huge rotator cuff injuries, who were treated with single-row suture. The shoulders were protected by a brace for 6 weeks after operations were permit ted to perform passive movement within 6 weeks, and then perform active shoulder exercise 6 to 10 weeks after operation. Constant-Murley score, UCLA score and VAS score were recorded preoperatively and postoperatively. RESULTS: All the patients were followed up, and the duration ranged from 12 to 62 months, with a mean of 30 months. There was no infection or nerve injury. UCLA score was improved from preoperative 10.71±2.45 to postoperative 32.07±3.16; Constant-Murley score was improved from preoperative 43.33±11.55 to postoperative 78.15±12.64; VAS score was improved from preoperative 5.81±1.27 to postoperative 0.52±0.71. There were no statistical differences among three groups in UCLA score, Constant-Murley score and VAS score. Total 337 cases were satisfied with treatment results and 16 cases were not satisfied with the results. Among the 16 cases, 3 cases had huge rotator cuff surgery, and 13 cases had no joint stiffness before operation. The main complaints that resulted in dissatisfaction were weakness of the postoperative muscles and failure to restore the labour capacity(11 cases). CONCLUSIONS: Rotator cuff repair under arthroscopy has a reliable clinical effect for the patients with rotator cuff tears. Stable and reliable clinical results can be obtained regardless different repair methods or different rotator cuff tears. The following factors such as no stiffness before operation, too early active exercise and preoperative rotator cuff atrophy may be the risk factors for postoperative dissatisfaction of patients.

Vimentin is important in the neural differentiation of PC12 cells promoted by sialylation.

Sialic acid modification is a kind of post-translational modification. To investigate the regulation effect of sialic acid on neural differentiation, we used CycloManN propanyl perac (CycloManN pro), a metabolic precursor of sialic acid, to treat PC12 cells. We noted that CycloManN pro indeed robustly promoted global sialylation detected by MAL II lectin blot in PC12 cells. Simultaneously, we interestingly found that the neurite outgrowth of PC12 cells was significantly promoted by the CycloManN pro treatment. The profile analysis of sialylated proteins showed that a protein band at 55KD was greatly enhanced especially in PC12L cells after CycloManN pro treatment. After enrichment with lectin MAL II, the proteins in this band were analyzed by mass spectrometry. The results showed that 23 proteins were in the band, but the score of vimentin was the highest among them. To investigate further the role of vimentin in the process of neurite differentiation, vimentin construct was transfected into PC12 cells. We interestingly observed that ectopic expression of vimentin significantly enhanced the neurite outgrowth induced by CycloManN pro. However, after three potential glycosylation sites (Ser-7, Thr-33, Ser-34:) of vimentin were mutated to alanine, overexpression of the mutated vimentin completely lost the enhancement activity for the neural differentiation even in the presence of CycloManN pro. Taken together, our study demonstrated that vimentin was important in the induction of neural differentiation by CycloManN pro.

Posttranslational modification of E-cadherin by core fucosylation regulates Src activation and induces epithelial-mesenchymal transition-like process in lung cancer cells.

E-cadherin is often dysregulated in aggressive lung cancer, the mechanism of which cannot always be explained at the level of transcription. In 66 patients with lung cancer, immunohistochemical staining demonstrated that co-localization of E-cadherin and core fucose by Lens culinaris agglutinin was significantly less extensive in tumor than in nontumor tissue. Through gain and loss of fucosylation experiments in the giant lung carcinoma cell lines 95C and 95D, our results revealed that E-cadherin core fucosylation in 95C cells overexpressing α-1, 6-fucosyltransferase (Fut8) inhibited Fut8-95C cell migration, whereas knockdown of Fut8 in 95D cells enhanced migration of short-interfering RNA-targeting Fut8 (siFut8)-95D cells. The level of active Src (phosphorylated Src [Y416]) was significantly reduced in Fut8-95C cells, but elevated in siFut8-95D cells. In protein complexes immunoprecipitated from Fut8-95C cell lysates with anti-E-cadherin, less phosphorylated Src (Y416) and more ß-catenin were observed, but immunoprecipitates from siFut8-95D cells, containing less core fucosylated E-cadherin, contained an elevated level of phospho-Src Y416. In Fut8-95C cells, phosphorylation of Akt (Y315, Y326) and GSK-3ß (S9) was significantly reduced, but ß-catenin (S37) phosphorylation was enhanced. Expression of N-cadherin and Snail1 was also reduced in Fut8-95C cells, but significantly increased in siFut8-95D cells. Intriguingly, when Src kinase activity was inhibited by treatment of cells with PP2 and SU6656, regulation of N-cadherin, Snail1 and cell migration by E-cadherin core fucosylation was abrogated in both Fut8-95C and siFut8-95D cells. Therefore, posttranslational modification of E-cadherin by less core fucosylation recruited and activated Src, and induced an epithelial-mesenchymal transition-like process in lung cancer cells.

Circulating MicroRNA Let-7d in Attention-Deficit/Hyperactivity Disorder.

Up to date, there has been no molecular signature available in the clinical practice for attention-deficit/hyperactivity disorder (ADHD). To investigate circulating miRNA let-7d significance in ADHD, we investigated serum miRNA let-7d in 35 newly diagnosed ADHD subjects who were randomly selected from 406 patients out of 7450 children, paired with gender- and age-matched control through case-control study. We observed that circulating miRNA let-7d was significantly higher in ADHD subjects than in control (p < 0.05). Higher circulation level of miRNA let-7d was significantly associated with ADHD (odds ratio 16.7; 95% confidence, p < 0.05). Meanwhile, serum galectin-3 level was down-regulated in ADHD subjects and the subjects with low galectin-3 expression accounted for 66% in ADHD. The difference of the serum galectin-3 levels between ADHD and non-ADHD groups reached significance (p < 0.05). In 1-year follow-up, a significantly higher rate of clinical improvement was noted in subjects with low level of circulating miRNA let-7d (p < 0.05) than those with high level of circulating miRNA let-7d. Our data demonstrated that miRNA let-7d was elevated in the serum of ADHD subjects, which might be a novel, useful molecule signature for ADHD.

Association between Ras association domain family 1A promoter methylation and esophageal squamous cell carcinoma: a meta-analysis.

RASSF1A has been reported to be a candidate tumor suppressor in esophageal squamous cell carcinoma (ESCC). However, the association between RASSF1A promoter methylation and ESCC remains unclear. Eligible studies were identified through searching PubMed, Medline, Web of Science, and the China National Knowledge Infrastucture database. Studies were pooled and odds ratios (ORs) with corresponding confidence intervals (CIs) were calculated. Funnel plots were also performed to evaluate publication bias. Twelve studies involving 859 cases and 675 controls were included in this meta-analysis. A significant association was observed between RASSF1A methylation and ESCC overall (OR = 11.7, 95% CI: 6.59-20.9, z=8.36, P<0.00001). Subgroup analysis showed that the OR for heterogeneous tissues was 5.35 (95% CI = 2.95-9.71) while for autologous tissues it was 16.0 (8.31-30.96). For patient sample size, the OR for the <50 subgroup was 9.92 (95% CI = 2.88-34.2) and for the 50 case group was 13.1 (95% CI = 6.59-25.91). The OR for a relationship between RASSF1A methylation and TNM stages was 0.27 (95% CI=0.10-0.77), whereas there were no significant differences in RASSF1A methylation in relation to gender and differentiation among ESCC cases. This meta-analysis suggests a significant association between RASSF1A methylation and ESCC.

[Analysis of delayed diagnosis of the vertical unstable pelvic fractures].

OBJECTIVE: To explore the causes of delayed diagnosis of the vertical unstable pelvic fractures in patients with multiple trauma so in order to minimize these incidence. METHODS: A retrospective study was conducted in 176 patients with the vertical unstable pelvic fractures from January 2003 to December 2008. The classification of vertical unstable pelvic fractures was recorded. Medical data were compared and statistically analyzed to identify the incidence, and the contributing factors of the delayed diagnosis of the vertical unstable pelvic fractures. Of the 176 patients, 46 (26.1%) had delayed diagnosis of the vertical unstable pelvic fractures. There were 29 males and 17 females with an average age of 34 years old. RESULTS: Of the factors contributing to 46 cases of delayed diagnosis, 41.3% (19/46) closely related to the evaluation by the doctors and were potentially avoidable, and 58.7% (27/46) were associated with the severity of the injuries. CONCLUSION: The patients with multiple trauma had high incidence of delayed diagnosis of the vertical unstable pelvic fractures. The recognition of pelvic fractures, careful clinical assessments, and the awareness of the orthopedic doctors, emergency doctors and ICU doctors. Most of the delayed diagnosis of the vertical unstable pelvic fractures in patients with multiple trauma are potentially avoidable.

[Reconstruction of posterolateral structure in knee with Müller method].

OBJECTIVE: To study the clinical effects of Müller method for reconstruction of posterolateral corner (PLS) of knee joint. METHODS: From June 2005 to June 2007, 13 patients with PLS injured were treated with Müller method. Four patients complicated with injuries in anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL), 7 patients complicated with injuries in PCL, 4 patient also had injuries in ACL, 1 patient complicated with ACL injury, and 1 patient complicated with avulsions fractures of the tibial ACL insertion. Among the patients, 5 patients had chronic injuries with a duration of 2 to 7 months. After the cruciate ligament was reconstructed, the popliteus tendon was reconstructed with iliotibial band and lateral collateral ligament (LCL) reconstructed with biceps femoris tendon. All the ligaments were fixed with interface screws. The patients were encouraged to do strength and motion exercise on bed immediately after operation and permit toe-touch weight-bearing till two months after operation. The brace was used till six months after operation. RESULTS: The average follow-up period was 13 months (ranged from 6 to 27 months). There was no restriction of range of motion. External rotation range were good compared to the normal side. One year after operation, one-grade varus instability at 30 degrees flexion was found only in 2 patients. The postoperative Lysholm scores were from 77 to 94 (average 86). CONCLUSION: The method is easier to perform, easier to get the materiar for reconstruction and promised satisfactory clinical results.

[Ligaments reconstruction for the treatment of lateral ankle instability].

OBJECTIVE: To study therapeutic effects of lateral ankle ligaments reconstruction for the treatment of chronic lateral instability of the ankle joint. METHODS: From July 2005 to January 2008, among 13 patients with chronic lateral instability of the ankle joint, 10 patients were male and 3 patients were female, ranging in age from 24 to 45 years,with an average of 33 years. Anterior talo-fibular ligament (ATFL) and calcanea-fibular ligament (CFL) were anatomy reconstructed with a split peroneus brevis tendon graft for all patients. The ankle scoring system was used to evaluate ankle joint function before and after operation, which including stability, pain, locomotor activity and X-ray films. RESULTS: All the patients were followed up ranged from 6 to 32 months, averaged 16.4 months. The postoperative scores of the ankles increased in respect to stability, pain and locomotor activity. The total average score increased from preoperative (43.54+/-7.04) to postoperative (73.38+/-4.17). There was significant difference between preoperative scores and postoperative scores (P<0.01). All the patients were satisfied with the results. CONCLUSION: Anatomy reconstruct of the ATFL and CFL with a split peroneus brevis tendon graft (Sammarco method) is a practical method for lateral ankle instability and promise good results especially for patients complained of instability.