Cell-free DNA methylation reveals cell-specific tissue injury and correlates with disease severity and patient outcomes in COVID-19.

BACKGROUND: The recently identified methylation patterns specific to cell type allows the tracing of cell death dynamics at the cellular level in health and diseases. This study used COVID-19 as a disease model to investigate the efficacy of cell-specific cell-free DNA (cfDNA) methylation markers in reflecting or predicting disease severity or outcome. METHODS: Whole genome methylation sequencing of cfDNA was performed for 20 healthy individuals, 20 cases with non-hospitalized COVID-19 and 12 cases with severe COVID-19 admitted to intensive care unit (ICU). Differentially methylated regions (DMRs) and gene ontology pathway enrichment analyses were performed to explore the locus-specific methylation difference between cohorts. The proportion of cfDNA derived from lung and immune cells to a given sample (i.e. tissue fraction) at cell-type resolution was estimated using a novel algorithm, which reflects lung injuries and immune response in COVID-19 patients and was further used to evaluate clinical severity and patient outcome. RESULTS: COVID­19 patients had globally reduced cfDNA methylation level compared with healthy controls. Compared with non-hospitalized COVID-19 patients, the cfDNA methylation pattern was significantly altered in severe patients with the identification of 11,156 DMRs, which were mainly enriched in pathways related to immune response. Markedly elevated levels of cfDNA derived from lung and more specifically alveolar epithelial cells, bronchial epithelial cells, and lung endothelial cells were observed in COVID-19 patients compared with healthy controls. Compared with non-hospitalized patients or healthy controls, severe COVID-19 had significantly higher cfDNA derived from B cells, T cells and granulocytes and lower cfDNA from natural killer cells. Moreover, cfDNA derived from alveolar epithelial cells had the optimal performance to differentiate COVID-19 with different severities, lung injury levels, SOFA scores and in-hospital deaths, with the area under the receiver operating characteristic curve of 0.958, 0.941, 0.919 and 0.955, respectively. CONCLUSION: Severe COVID-19 has a distinct cfDNA methylation signature compared with non-hospitalized COVID-19 and healthy controls. Cell type-specific cfDNA methylation signature enables the tracing of COVID-19 related cell deaths in lung and immune cells at cell-type resolution, which is correlated with clinical severities and outcomes, and has extensive application prospects to evaluate tissue injuries in diseases with multi-organ dysfunction.

Effects of Music, Massage, Exercise, or Acupuncture in the Treatment of Depression Among College Students: A Network Meta-Analysis.

Objective: To assess the therapeutic impacts of exercise, massage, and music interventions on college students experiencing depression by employing a mesh meta-analysis approach. This research intends to offer valuable insights to aid in the development of non-pharmaceutical treatment strategies for depression. Methods: We conducted a thorough search across various databases including Cochrane, PubMed, Embase, Web of Science, CNKI, and Wanfang to explore the effects of music, massage, aerobic exercise, fitness Qigong, yoga, tai chi, ball games, strength training, dance, whole body vibration training, and high-intensity interval training on the treatment of depression in college students. The search period was from January 1, 2023, which marks the establishment of each database. Subsequently, a mesh meta-analysis was performed using the "Stata 15.1" software, incorporating outcome indicators from 24 included literature comprising a total of 1458 patients. Results: Based on the ranking of the optimal intervention effects of various non-pharmaceutical methods, the order, from highest to lowest probability, was as follows: high-intensity interval training (96%), yoga (94.90%), dance (78.30%), music (73.30%), ball games (62.50%), strength training (51.70%), aerobic training (45.30%), tai chi (35.40%), vibration training (27.30%), massage (20.10%), qigong (14.30%), and no intervention (1.00%). This ranking aligns closely with the findings obtained from pairwise comparisons between different interventions. Conclusion: High-intensity interval training is likely to yield the most effective therapeutic results for college students with depression. In the pairwise comparison of different interventions, High-intensity interval training is also better than most interventions. However, to establish its intervention effect more conclusively, further validation through additional high-quality randomized controlled trials is necessary.

Short-term outcomes of near-infrared imaging using indocyanine green in laparoscopic lateral pelvic lymph node dissection for middle-lower rectal cancer: A propensity score-matched cohort analysis.

Laparoscopic lateral pelvic lymph node dissection (LPND) is limited by complex neurovascular bundles in the narrow pelvic sidewall and various post-operative complications. Indocyanine green (ICG) has been applied to increase the number of harvested lymph nodes and reduce the injury of irrelevant vessels in patients with rectal cancer. However, few studies on the recurrence rate of ICG fluorescence imaging-guided laparoscopic LPND were reported. This retrospective study enrolled 50 middle- low rectal cancer patients who were treated by LPND. After propensity score matching, 20 patients were matched in each of the indocyanine green (ICG) guided imaging group (ICG group) and non-ICG guided imaging group (non-ICG group). The average follow-up time was 13.5 months (12-15 months). Our results showed that the total number of harvested lymph nodes in the ICG group was significantly higher than that in the non-ICG group (P < 0.05), and intraoperative blood loss and post-operative hospital stay times in the ICG group were less than those in the non-ICG group (P < 0.05). After 12 months of follow-up, no residual lymph node and local tumor recurrence were found for patients in the ICG group. Four patients in the non-ICG group detected residual lymph nodes at the 3-month visit. Our findings highlighted the importance of ICG fluorescence-guided imaging in LPND because it has unique advantages in improving the number of lymph node dissections, surgical accuracy, and decreasing the residual lymph nodes and local tumor recurrence. In addition, ICG fluorescence guidance technology can effectively shorten the operation time, and it is simple to operate, which is worth popularizing.

[Efficacies of bone-marrow-derived mononuclear cells with a hydroxylapatite composite in the treatment of osteonecrosis of the femoral head].

OBJECTIVE: To evaluate the efficacies of core decompression and implantation of concentrated autologous bone marrow containing mononuclear cells (BMMCs) with porous hydroxylapatite composite in the treatment of osteonecrosis of the femoral head. METHODS: A total of 35 patients with 57 osteonecrosis hips with ARCO stage I, stage II and stage IIIA disease were treated by BMMCs with a porous hydroxylapatite composite. The mean age at surgery was 39.4 (26-58) years and the mean period of follow-up 28 (12-40) months. In the control group, cell-free porous hydroxylapatite composite was implanted into 17 patients (27 hips) with osteonecrosis of the femoral head and the outcomes were compared. RESULTS: At the last follow-up, postoperative Harris hip scores significantly increased in both groups (P < 0.0001). The magnitude of increase was significantly greater in the BMMCs group compared with the control group (28.3% ± 0.9% vs 18.4% ± 1.7%, P < 0.01). Postoperative visual analog scale (VAS) scores significantly decreased in both groups (P < 0.01). The magnitude of decrease was significantly greater in the BMMCs group compared with the control group (-70.2% ± 2.1% vs -51.7% ± 2.9%, P < 0.001). The clinical success rate was significantly higher in the BMMCs group compared with the control group (75.4% vs 37.0%, P < 0.01). The radiological success rates were similar between the BMMCs and control groups (59.6% vs 40.7%, P = 0.1046). CONCLUSION: The combined regimen of core decompression and implantation of concentrated autologous BMMCs with porous hydroxylapatite composite appears to confer benefits in the treatment of in stages I-IIIA osteonecrosis of the femoral head.

[Single factor analysis of motor dysfunction and imaging and clinical features in metastatic epidural spinal cord compression].

OBJECTIVE: To explore the relationship of motor dysfunction of the lower extremities with the imaging appearances and clinical features of metastatic epidural spinal cord compression (MESCCs). METHODS: From July 2006 to December 2007, 26 successive patients with metastases of the thoracic, lumbar and the cervical spine were treated in our department. Forty-three main involved vertebra in all 26 patients were evaluated by magnetic resonance imaging and computed tomography, and were scored according motor dysfunction in this study. Fourteen patients (25 vertebrae) had motor dysfunction. RESULTS: Among 26 patients, 12 cases with visceral metastasis,in which had motor dysfunction in 10 cases; 14 cases without visceral metastasis, in which had motor dysfunction in 4 cases; comparison between two groups, P=0.0079. Among vertebral presence of continuity of 43 main involved vertebrae, 16 vertebrae had motor dysfunction;among vertebral absence of continuity, motor dysfunction occurred in 9 vertebrae, comparison between two groups, P=0.1034. Among vertebral presence of lamina involvement of 43 main involved vertebrae, 11 vertebrae had motor dysfunction; among vertebral absence of lamina involvement, motor dysfunction occurred in 14 vertebrae, comparison between two groups, P=0.020 5. Among vertebral presence of protruding of vertebral posterior wall of 43 main involved vertebrae, 12 vertebrae had motor dysfunction; among vertebral absence of protruding of vertebral posterior wall, 13 vertebrae had motor dysfunction, comparison between two groups, P=0.0334. Among vertebral presence of involvement epidural space of 43 main involved vertebrae, 11 vertebrae had motor dysfunction; among vertebral absence of involvement epidural space, 14 vertebrae had motor dysfunction, comparison between two groups, P=0.003 6. Such factors as age, gender, whether or not received regular chem before admission, back pain degree of metastasis, received regular chem before admission, therapeutic efficacy of primary tumor, number of bony metastases outside spine, number of the main involved vertebrae, level of vertebral metastases location, level of continuous involved vertebrae, vertebral-body involvement, fracture of anterior column, fracture of posterior wall, and pedicle involvement had no effects on incidence of motor dysfunction due to MESCC (P>0.05). CONCLUSION: MESCC with visceral metastases, lamina involvement, presence of outstanding buttocks sign of posterior wall,involvement epidural space tended to cause symptomatic MESCC. Incidence of continuity of main involved vertebrae occurred more frequently in the CUTS compared with other levels of spine.

Panax notoginseng reduces atherosclerotic lesions in ApoE-deficient mice and inhibits TNF-alpha-induced endothelial adhesion molecule expression and monocyte adhesion.

It is widely recognized that atherogenesis is associated with vascular inflammation. Panax notoginseng , a commonly used herb in China, has been shown to possess anti-inflammatory activity. In the present study, the antiatherogenic effect of P. notoginseng saponins (PNS) was examined in apolipoprotein E (apoE)-deficient mice. The molecular mechanisms responsible for the antivascular inflammatory effect of PNS on human coronary artery endothelial cells (HCAECs) were also investigated in vitro. PNS, dissolved in drinking water, was administered orally to two treatment groups at dosages of 4.0 and 12.0 mg/day/mouse, respectively. After 8 weeks, atherosclerosis in the entire aortic area was assessed using an en face method. Compared with the control group, both low- and high-dose PNS-treated groups showed a significant decrease in extent of atherosclerotic lesions by 61.4 and 66.2%, respectively (P < 0.01). PNS also notably reduced serum lipid levels. Serum levels of IL-6 and TNF-alpha in all groups of apoE-deficient mice were below the detection limit. In vitro studies showed that PNS dose-dependently inhibited monocyte adhesion on activated endothelium, as well as the expression of TNF-alpha-induced endothelial adhesion molecules, such as ICAM-1 and VCAM-1. In conclusion, PNS has antiatherogenic activity through, at least in part, its lipid-lowering and antivascular inflammatory mechanisms.

Docosahexaenoic acid suppresses arachidonic acid-induced proliferation of LS-174T human colon carcinoma cells.

AIM: To investigate the impact of arachidonic acid (AA) and docosahexaenoic acid (DHA) and their combination on colon cancer cell growth. METHODS: The LS-174T colon cancer cell line was used to study the role of the prostaglandin precursor AA and the omega-3 polyunsaturated fatty acid DHA on cell growth. Cell viability was assessed in XTT assays. For analysis of cell cycle and cell death, flow cytometry and DAPI staining were applied. Expression of cyclooxygenase-2 (COX-2), p21 and bcl-2 in cells incubated with AA or DHA was examined by real-time RT-PCR. Prostaglandin E(2) (PGE(2)) generation in the presence of AA and DHA was measured using a PGE(2)-ELISA. RESULTS: AA increased cell growth, whereas DHA reduced viability of LS 174T cells in a time- and dose-dependent manner. Furthermore, DHA down- regulated mRNA of bcl-2 and up-regulated p21. Interestingly, DHA was able to suppress AA-induced cell proliferation and significantly lowered AA-derived PGE(2) formation. DHA also down-regulated COX-2 expression. In addition to the effect on PGE(2) formation, DHA directly reduced PGE(2)-induced cell proliferation in a dose-dependent manner. CONCLUSION: These results suggest that DHA can inhibit the pro-proliferative effect of abundant AA or PGE(2).

Altered spinal arachidonic acid turnover after peripheral nerve injury regulates regional glutamate concentration and neuropathic pain behaviors in rats.

Spinal glutamate transporters (GT) have been implicated in the mechanisms of neuropathic pain; however, how spinal GT uptake activity is regulated remains unclear. Here we show that alteration of spinal arachidonic acid (AA) turnover after peripheral nerve injury regulated regional GT uptake activity and glutamate homeostasis. Chronic constriction nerve injury (CCI) in rats significantly reduced spinal GT uptake activity ((3)H-glutamate uptake) with an associated increase in extracellular AA and glutamate concentration from spinal microdialysates on postoperative day 8. AACOCF3 (a cytosolic phospholipase A2 inhibitor, 30mug) given intrathecally twice a day for postoperative day 1-7 reversed this CCI-induced spinal AA production, prevented the reduced spinal GT uptake activity and increased extracellular glutamate concentration. Conversely, alteration of spinal AA metabolism by diclofenac (a cyclooxygenase 1/2 inhibitor, 200mug) further reduced spinal GT uptake activity and increased extracellular glutamate concentration in CCI rats. GT uptake activity was also attenuated when AA (10 or 100nM) was directly added into spinal samples of naïve rats in an in vitro(3)H-glutamate uptake assay, indicating a direct inhibitory effect of AA on GT uptake activity. Consistent with these findings, AACOCF3 reduced the development of both thermal hyperalgesia and mechanical allodynia, whereas diclofenac exacerbated thermal hyperalgesia, in CCI rats. Thus, spinal AA turnover may serve as a regulator in CCI-induced changes in regional GT uptake activity, glutamate homeostasis, and neuropathic pain behaviors. These data suggest that regulating spinal AA turnover may be a useful approach to improving the clinical management of neuropathic pain.