Coexistence of Multiple Functional Variants and Genes Underlies Genetic Risk Locus 11p11.2 of Alzheimer's Disease.

BACKGROUND: Genome-wide association studies have identified dozens of genetic risk loci for Alzheimer's disease (AD), yet the underlying causal variants and biological mechanisms remain elusive, especially for loci with complex linkage disequilibrium and regulation. METHODS: To fully untangle the causal signal at a single locus, we performed a functional genomic study of 11p11.2 (the CELF1/SPI1 locus). Genome-wide association study signals at 11p11.2 were integrated with datasets of histone modification, open chromatin, and transcription factor binding to distill potentially functional variants (fVars). Their allelic regulatory activities were confirmed by allele imbalance, reporter assays, and base editing. Expressional quantitative trait loci and chromatin interaction data were incorporated to assign target genes to fVars. The relevance of these genes to AD was assessed by convergent functional genomics using bulk brain and single-cell transcriptomic, epigenomic, and proteomic datasets of patients with AD and control individuals, followed by cellular assays. RESULTS: We found that 24 potential fVars, rather than a single variant, were responsible for the risk of 11p11.2. These fVars modulated transcription factor binding and regulated multiple genes by long-range chromatin interactions. Besides SPI1, convergent evidence indicated that 6 target genes (MTCH2, ACP2, NDUFS3, PSMC3, C1QTNF4, and MADD) of fVars were likely to be involved in AD development. Disruption of each gene led to cellular amyloid-ß and phosphorylated tau changes, supporting the existence of multiple likely causal genes at 11p11.2. CONCLUSIONS: Multiple variants and genes at 11p11.2 may contribute to AD risk. This finding provides new insights into the mechanistic and therapeutic challenges of AD.

Influence of volar margin of the lunate fossa fragment fixation on distal radius fracture outcomes: A retrospective series.

BACKGROUND: Distal radius fractures accompanied by the volar margin of the lunate fossa (VMLF) lesions are often overlooked or inadequately reduced in clinical practice. AIM: To investigate the impact of VMLF fragment in distal radius fractures on the stability and function of the wrist joint. METHODS: This was a retrospective study of patients with distal radius fractures who underwent surgical treatment between January 2013 and December 2017. The patients were divided into two groups according to whether the VMLF fragments were fixed or not. X-rays and computed tomography were performed before surgery, immediately postoperatively, and at 1, 3, and 6 mo to measure the scapholunate angle, radiolunate angle, capitolunate angle, and effective radiolunate flexion (ERLF). The Mayo wrist score and disabilities of the arm, shoulder, and hand (DASH) score were determined at 1 year. RESULTS: Thirty-five patients were included. There were 15 males and 20 females. Their mean age was 52.5 ± 14.3 (range: 19-70) years. There were 38 wrists (17 on the left side, 15 on the right, and three bilateral; 16 in the fixed group, and 22 in the unfixed group). The interval between trauma and surgery was from 1 h to 1 mo. The incidence of postoperative wrist instability in the unfixed group (86.4%) was higher than in the fixed group (25.0%) (P ≤ 0.001). Ten patients had ERLF > 25° in the unfixed group and none in the fixed group (P = 0.019). The Mayo wrist score was 94 ± 5.7 in the fixed group and 68 ± 15.1 in the unfixed group (P < 0.001). The DASH score was 4.6 ± 2.5 in the fixed group and 28.5 ± 19.5 in the unfixed group (P < 0.001). CONCLUSION: Injuries of VMLF, even small fractures, might damage the radial-lunar ligament, leading to postoperative wrist instability, sagittal force line imbalance, and poor recovery of wrist joint function.

Folic acid contributes to peripheral nerve injury repair by promoting Schwann cell proliferation, migration, and secretion of nerve growth factor.

After peripheral nerve injury, intraperitoneal injection of folic acid improves axon quantity, increases axon density and improves electromyography results. However, the mechanisms for this remain unclear. This study explored whether folic acid promotes peripheral nerve injury repair by affecting Schwann cell function. Primary Schwann cells were obtained from rats by in vitro separation and culture. Cell proliferation, assayed using the Cell Counting Kit-8 assay, was higher in cells cultured for 72 hours with 100 mg/L folic acid compared with the control group. Cell proliferation was also higher in the 50, 100, 150, and 200 mg/L folic acid groups compared with the control group after culture for 96 hours. Proliferation was markedly higher in the 100 mg/L folic acid group compared with the 50 mg/L folic acid group and the 40 ng/L nerve growth factor group. In Transwell assays, the number of migrated Schwann cells dramatically increased after culture with 100 and 150 mg/L folic acid compared with the control group. In nerve growth factor enzyme-linked immunosorbent assays, treatment of Schwann cell cultures with 50, 100, and 150 mg/L folic acid increased levels of nerve growth factor in the culture medium compared with the control group at 3 days. The nerve growth factor concentration of Schwann cell cultures treated with 100 mg/L folic acid group was remarkably higher than that in the 50 and 150 mg/L folic acid groups at 3 days. Nerve growth factor concentration in the 10, 50, and 100 mg/L folic acid groups was higher than that in the control group at 7 days. The nerve growth factor concentration in the 50 mg/L folic acid group was remarkably higher than that in the 10 and 100 mg/L folic acid groups at 7 days. In vivo, 80 µg/kg folic acid was intraperitoneally administrated for 7 consecutive days after sciatic nerve injury. Immunohistochemical staining showed that the number of Schwann cells in the folic acid group was greater than that in the control group. We suggest that folic acid may play a role in improving the repair of peripheral nerve injury by promoting the proliferation and migration of Schwann cells and the secretion of nerve growth factors.

Non-invasive quantification of age-related changes in the vertebral endplate in rats using in vivo DCE-MRI.

BACKGROUND: Small animal models that can mimic degenerative disc disease (DDD) are commonly used to examine DDD progression. However, assessments such as histological studies and macroscopic measurements do not allow for longitudinal studies because they can only be completed after the animal is sacrificed. Dynamic contrast-enhanced MRI (DCE-MRI) may provide a reliable, non-invasive in vivo method for detecting the progression. METHODS: The present study investigated the progression of changes in lumbar discs and the effect of endplate conditions on diffusion into the lumbar discs of aging sand rats after intravenous administration of gadolinium-containing contrast medium through the tail vein. Contrast enhancement was measured in the lumbar intervertebral discs on each image. The results were compared with those from conventional histological characterizations. RESULTS: T2-weighted images revealed that with aging, the shape of L3-L4, L4-L5, L5-L6, and L6-S1 nucleus pulposus (NP) became irregular, while the mean areas, signal intensities, and T2 values of the NP were significantly decreased. Each of the observed disc changes demonstrated a progressive increase in phase during 2-min scout scans. Post-contrast MRI showed impaired endplate nutritional diffusion to the disc with aging, enhancement was significantly greater in young animals than in old animals. Endplate calcification or sclerosis was histologically confirmed; histologic score was correlated with the age. We found the histological score of the endplate negatively corresponded to the DCE-MRI results. CONCLUSIONS: DCE-MRI studies offer a non-invasive in vivo method for investigating the progress of diffusion into the discs and the functional conditions of the endplate. We conclude that quantitative DCE-MRI can identify the severity of disc degeneration and efficiently reflect the progression of vertebral endplate changes in the aging sand rat lumbar spine via the NP contrast enhancement patterns.

[Case-control study on the relationship of chronic low back pain of facet joint origin with the distribution of nerve endings and neuropeptide: a quantitative histological analysis].

OBJECTIVE: To investigate the density and distribution of nerve endings and neuropeptide Y (NPY) in lumbar facet joints of patients with low back pain. METHODS: Fifteen patients without low back pain were selected as control group (group A). Facet joint samples in group A were obtained during the operation or lumbar spinal canal tumor they suffered from. Those patients with low back pain were divided into three groups according to their different origins of pain, such as not from facet joint (group B, 15 patients) ,from facet joint only (group C, 20 patients), or from facet joint partially (group D, 20 patients). Different origins were determined by VAS after facet joint block. The density and distribution of nerve ending and neuropeptide in the capsular tissues were analyzed by a modified gold chloride staining and immunochemistry respectively. RESULTS: Compared with the ones in group A and B, the fact joints in group C and D were more inclined to be degenerated and got more nerve endings. NPY was expressed mainly in the facet joint of patients with low back pain in group C and D. In addition, there was a significant relationship between the distribution of nerve endings and NPY expression,while none of them were related with MRI Fujiwara grade of facet joint. CONCLUSION: These results suggest that the number of mechanoreceptors, neural sprouting and secreted peptides in the facet joint capsules vary with the change of mechanical or nociceptive stimulation, which may promote the development of low back pain in return.