SP1 transcriptionally activates HTR2B to aggravate traumatic spinal cord injury by shifting microglial M1/M2 polarization.

BACKGROUND: Spinal cord injury (SCI) can result in structural and functional damage to the spinal cord, which may lead to loss of limb movement and sensation, loss of bowel and bladder control, and other complications. Previous studies have revealed the critical influence of trans-acting transcription factor 1 (SP1) in neurological pathologies, however, its role and mechanism in SCI have not been fully studied. METHODS: The study was performed using mouse microglia BV2 stimulated using lipopolysaccharide (LPS) and male adult mice subjected to spinal hitting. Western blotting was performed to detect protein expression of SP1, 5-hydroxytryptamine (serotonin) receptor 2B (HTR2B), BCL2-associated x protein (Bax), B-cell lymphoma-2 (Bcl-2), inducible nitric oxide synthase (iNOS), clusters of differentiation 86 (CD86), Arginase 1 (Arg-1) and clusters of differentiation 206 (CD206). Cell viability and apoptosis were analyzed by MTT assay and TUNEL assay. mRNA levels of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-4 (IL-4) and tumor necrosis factor-ß (TNF-ß) were quantified by quantitative real-time polymerase chain reaction. The association of SP1 and HTR2B was identified by chromatin immunoprecipitation assay and dual-luciferase reporter assay. HE staining assay was performed to analyze the pathological conditions of spinal cord tissues. RESULTS: LPS treatment induced cell apoptosis and inhibited microglia polarization from M1 to M2 phenotype, accompanied by an increase of Bax protein expression and a decrease of Bcl-2 protein expression, however, these effects were relieved after SP1 silencing. Mechanism assays revealed that SP1 transcriptionally activated HTR2B in BV2 cells, and HTR2B knockdown rescued LPS-induced effects on BV2 cell apoptosis and microglial M1/M2 polarization. Moreover, SP1 absence inhibited BV2 cell apoptosis and promoted microglia polarization from M1 to M2 phenotype by decreasing HTR2B expression. SCI mouse model assay further showed that SP1 downregulation could attenuate spinal hitting-induced promoting effects on cell apoptosis of spinal cord tissues and microglial M1 polarization. CONCLUSION: SP1 transcriptionally activated HTR2B to aggravate traumatic SCI by shifting microglial M1/M2 polarization.

[Comparison of unilateral biportal endoscopic transforaminal lumbar interbody fusion versus minimally invasive tubular transforaminal lumbar interbody fusion for lumbar degenerative disease].

Objective: To compare the clinical and radiological outcomes of unilateral biportal endoscopic transforaminal lumbar interbody fusion (UBE-TLIF) and minimally invasive tubular TLIF (MT-TLIF) in treatment of lumbar degenerative diseases. Methods: A clinical data of 75 patients with lumbar degenerative diseases, who met the selection criteria between August 2019 and August 2020, was retrospectively analyzed, including 35 patients in the UBE- TLIF group and 40 patients in the MT-TLIF group. There was no significant difference in general data such as gender, age, body mass index, disease type and duration, and surgical segment between the two groups ( P>0.05), which was comparable. The operation time, intraoperative blood loss, hemoglobin (Hb) before operation and at 1 day after operation, the length of hospital stay, incidence of complications, and visual analogue scale (VAS) score of low back and leg pain, Oswestry Disability Index (ODI), Short-Form 36 Health Survey Scale (SF-36 scale), intervertebral disc height (IDH), sagittal Cobb angle, lumbar lordosis (LL), and the intervertebral fusion were compared between the two groups. Results: Compared with MT-TLIF group, UBE-TLIF group had significantly longer operation time but less intraoperative blood loss and shorter length of hospital stay ( P<0.05). The Hb levels in both groups decreased at 1 day after operation, but there was no significant difference in the difference before and after operation between the two groups ( P>0.05). All patients were followed up, and the follow-up time was (14.7±2.5) months in the UBE-TLIF group and (15.0±3.4) months in the MT-TLIF group, with no significant difference ( t=0.406, P=0.686). In both groups, the VAS score of low back pain, VAS score of leg pain, SF-36 scale, and ODI after operation significantly improved when compared with those before operation ( P<0.05). There was no significant difference between 1 month after operation and last follow-up ( P>0.05). There was no significant difference in the VAS score of low back pain, VAS score of leg pain, and SF-36 scale between the two groups before and after operation ( P>0.05). At 1 month after operation, the ODI in the UBE-TLIF group was significantly better than that in the MT-TLIF group ( P<0.05). At 1 month after operation, IDH, Cobb angle, and LL in both groups recovered when compared with those before operation ( P<0.05), and were maintained until last follow-up ( P>0.05). There was no significant difference in the IDH, Cobb angle, and LL between the two groups at each time point ( P>0.05). Thirty-three cases (89.2%) in the UBE-TLIF group and 35 cases (87.5%) in the MT-TLIF group achieved fusion, and the difference was not significant ( χ 2=0.015, P=0.901). In the UBE-TLIF group, 1 case of intraoperative dural tear and 1 case of postoperative epidural hematoma occurred, with an incidence of 5.7%. In the MT-TLIF group, 1 case of intraoperative dural tear, 1 case of postoperative epidural hematoma, and 1 case of superficial infection of the surgical incision occurred, with an incidence of 7.5%. There was no significant difference in the incidence of complications between the two groups ( χ 2=1.234, P=1.000). Conclusion: Compared with MT-TLIF, UBE-TILF can achieve similar interbody fusion in the treatment of lumbar degenerative diseases, and has the advantages of smaller incision, less bleeding, and shorter length of hospital stay.

Development and Validation of Novel Prognostic Models for Immune-Related Genes in Osteosarcoma.

Immunotherapy has shown excellent therapeutic effects on various malignant tumors; however, to date, immunotherapy for osteosarcoma is still suboptimal. In this study, we performed comprehensive bioinformatic analysis of immune-related genes (IRGs) and tumor-infiltrating immune cells (TIICs). Datasets of differentially expressed IRGs were extracted from the GEO database (GSE16088). The functions and prognostic values of these differentially expressed IRGs were systematically investigated using a series of bioinformatics methods. In addition, CCK8 and plate clone formation assays were used to explore the effect of PGF on osteosarcoma cells, and twenty-nine differentially expressed IRGs were identified, of which 95 were upregulated and 34 were downregulated. Next, PPI was established for Identifying Hub genes and biology networks by Cytoscape. Six IRGs (APLNR, TPM2, PGF, CD86, PROCR, and SEMA4D) were used to develop an overall survival (OS) prediction model, and two IRGs (HLA-B and PGF) were used to develop a relapse-free survival (RFS) prediction model. Compared with the low-risk patients in the training cohort (GSE39058) and TARGET validation cohorts, high-risk patients had poorer OS and RFS. Using these identified IRGs, we used OS and RFS prediction nomograms to generate a clinical utility model. The risk scores of the two prediction models were associated with the infiltration proportions of some TIICs, and the activation of memory CD4 T-cells was associated with OS and RFS. CD86 was associated with CTLA4 and CD28 and influenced the infiltration of different TIICs. In vitro experiments showed that the knockdown of PGF inhibited the proliferation and viability of osteosarcoma cells. In conclusion, these findings help us better understand the prognostic roles of IRGs and TIICs in osteosarcoma, and CD86 and PGF may serve as specific immune targets.

Knockdown of NOLC1 Inhibits PI3K-AKT Pathway to Improve the Poor Prognosis of Esophageal Carcinoma.

OBJECTIVE: Esophageal carcinoma (ESCA) is a common malignant gastrointestinal tumor. The abnormal expression of NOLC1 is involved in the tumorigenesis of various human tumors, whereas the function and mechanism of NOLC1 in ESCA remain unclear. In this study, we explored the relationship between NOLC1 and poor prognosis of ESCA, and its role and mechanism in the occurrence of ESCA. METHODS: The NOLC1 expression in ESCA tissues and cell lines was determined by qRT-PCR, immunohistochemistry, or western blot. The Kaplan-Meier method was conducted to estimate the overall survival. Cox regression analysis was carried out to examine the association between patient characteristics and prognosis. A recombined lentiviral vector containing NOLC1 was applied for transfecting ESCA cells (Eca109 and TE-13) and established a stable cell line with low NOLC1 expression or high NOLC1 expression, in the absence or presence of PI3K inhibitor (LY294002) treatment. Cell proliferation, apoptosis rate, invasion ability, migration ability, and PI3K/AKT pathway were detected by CCK8 assay, flow cytometry, Transwell assay, wound-healing assay, and western blot. RESULTS: NOLC1 overexpression was observed in ESCA tissues and ESCA cell lines (EC9706, Eca109, TE-13, Kyse170, T.TN) compared with adjacent normal tissues and normal esophageal cell line HEEC. NOLC1 overexpression was markedly associated with bigger tumor size, lymph node metastasis, and advanced TNM stage. Patients with NOLC1 overexpression have shorter overall survival than that of those with low NOLC1 expression. NOLC1 overexpression was considered to be an independent poor prognostic factor affecting overall survival. NOLC1 knockdown inhibited proliferation, migration, invasion, and cyclin B1 expression and promoted the apoptosis and cleaved-caspase-3 expression of Eca109 and TE-13 cells. NOLC1 overexpression accelerated proliferation, migration, invasion, and cyclin B1 expression and inhibited the apoptosis and cleaved-caspase-3 expression of ESCA cells via activating PI3K/AKT pathway. Rescue experiments showed that PI3K inhibitor (LY294002) could reverse the phenomenon caused by NOLC1 overexpression. CONCLUSION: NOLC1 may be a marker for poor prognosis. It can participate in the occurrence and development of ESCA via the PI3K/AKT pathway.

Novel combinatorial transdermal drug delivery of alendronate with risedronate for the treatment of osteoporosis.

The presented investigation explores the efficiency of novel transdermal drug delivery system of combination of two drugs i.e risedronate and alendronate in the treatment of osteoporosis. The nanoparticulate transdermal patch was prepared using PLGA as principle polymer which has been found to be suitable for drug delivery. The novel formulation system was found to be more efficient in lowering and maintaining the plasma calcium at a normal level when compared to a pure drug in a study carried out on an excised rat skin.