N6-methyladenosine methyltransferase METTL3 modulates the cell cycle of granulosa cells via CCND1 and AURKB in Haimen goats.

N6-methyladenosine (m6A) plays a crucial role in many bioprocesses across species, but its function in granulosa cells during oocyte maturation is not well understood in animals, especially domestic animals. We observed an increase in m6A methyltransferase-like 3 (METTL3) in granulosa cells during oocyte maturation in Haimen goats. Our results showed that knockdown of METTL3 disrupted the cell cycle in goat granulosa cells, leading to aggravated cell apoptosis and inhibition of cell proliferation and hormone secretion. Mechanistically, METTL3 may regulate the cell cycle in goat granulosa cells by mediating Aurora kinase B (AURKB) mRNA degradation in an m6A-YTH N6-methyladenosine RNA binding protein 2 (YTHDF2) manner and participating in AURKB transcription via the Cyclin D1 (CCND1)-Retinoblastoma protein (RB)-E2F transcription factor 1 (E2F1) pathway. Overall, our study highlights the essential role of METTL3 in granulosa cells during oocyte maturation in Haimen goats. These findings provide a theoretical basis and technical means for understanding how RNA methylation participates in oocyte maturation through granulosa cells.

Percutaneous Endoscopic Lumbar Discectomy for Recurrent Lumbar Disc Herniation: An Updated Systematic Review and Meta-Analysis.

Objective: This meta-analysis evaluated surgical outcomes following endoscopic or conventional discectomy for recurrent lumbar disc herniation. Methods: Medline, Cochrane, EMBASE, and Google Scholar were search until October 16, 2016 using these terms: recurrent lumbar disc herniation, endoscopic surgery, and discectomy. Randomized controlled trials (RCTs), prospective, retrospective, and cohort studies were eligible for inclusion. Pooled difference in mean (PDM) with 95% confidence interval (CIs) or relative risks (RRs) were calculated using fixed-effects methods. Results: One RCT and 15 studies were included with a total of 820 patients. Patients received endoscopic surgery experienced shorter operation time than those received conventional surgery (PDM: -52.01, 95% CI: -76.84 to -27.18, P < 0.001). A significantly lower risk in complication was displayed in patients received endoscopic surgery compared to those received conventional surgery (RR: 0.209, 95% CI: 0.076-0.581, P = 0.003). No significant difference in the improvement in VAS (PDM: -2.19, 95% CI: -5.78 to 1.39, P = 0.231), length of stay (PDM: -6.44, 95% CI: -13.76 to 0.89, P = 0.085) and re-recurrence rate (PDM: 0.88, 95% CI: 0.22-3.50, P = 0.861) between groups. Conclusions: Endoscopic and conventional discectomy reduced patient pain comparably, but endoscopic discectomy had significantly lower operation time and lower risk in complications, which may impact other outcomes such as recovery and healthcare costs. More studies are needed to confirm our findings. Supplementary Information: The online version contains supplementary material available at 10.1007/s43465-022-00636-1.

The PI3K/Akt/FOXO3a pathway regulates regeneration following spinal cord injury in adult rats through TNF-α and p27kip1 expression.

The aim of the present study was to elucidate the expression and role of the phosphatidylinositol 3­kinase (PI3K)/Akt/forkhead box O3 (FOXO3a) pathway in the regeneration of the spinal cord following spinal cord injury (SCI), and its regulatory effect on tumor necrosis factor (TNF)-α and cyclin-dependent kinase inhibitor 1B (p27kip1) expression. Firstly, in a Sprague-Dawley rat model of SCI, western blot analysis revealed that the protein levels of PI3K, phosphorylated Akt and FOXO3a were markedly inhibited compared with those in the sham control group. In vitro experiments were also conducted, in which primary dissociated cultures of rat dorsal spinal cord cells were induced with lipopolysaccharide (LPS; 4 µg/ml). The downregulation of PI3K using LY294002 markedly suppressed cell viability, reduced the protein levels of FOXO3a and p27kip1, and increased TNF-α protein production in the LPS-induced spinal cord cells. In addition, when the LPS-induced spinal cord cells were infected with FOXO3a adenoviral vectors, the overexpression of FOXO3 markedly promoted cell proliferation, activated p27kip1 protein levels and inhibited TNF-α protein production in the spinal cord cells. These results suggest that the PI3K/Akt/FOXO3a pathway regulates regeneration following SCI in adult rats via its modulatory effects on TNF-α and p27kip1 expression.

Tetrahydrocurcumin­induced autophagy via suppression of PI3K/Akt/mTOR in non­small cell lung carcinoma cells.

Lung carcinoma is the leading cause of mortality due to cancer worldwide. Autophagy has a significant role in the development and progression of non­small cell lung carcinoma (NSCLC). A previous study has revealed that tetrahydrocurcumin (THC), a traditional Chinese medicine isolated from Curcuma wenyujin (Chen & Ling, 1981), induces autophagy in human A549 NSCLC cells. The present study evaluated THC­induced autophagy in A549 cells using various assays, including the Cell Counting Kit­8, acridine orange staining, flow cytometry, reverse transcription­quantitative polymerase chain reaction (RT­qPCR), and western blot analysis of the markers of autophagy. THC inhibited the growth and proliferation of A549 cells (P<0.05). Acridine orange staining and flow cytometry revealed that THC treatment significantly enhanced autophagic cell proliferation inhibition (P<0.05). The RT­qPCR analysis revealed that THC treatment increased Beclin­1 expression level and compared with the control group (P<0.05). The light chain 3 (LC3)­II/LC3­I ratio was reduced in THC­treated cells when compared with the control group (P<0.05). Protein expression of various markers of autophagy, including p62, phosphorylated (p)­mechanistic target of rapamycin (mTOR), phosphoinositide 3­kinase (PI3K), p­PI3K, protein kinase B (Akt), and p­Akt was significantly reduced in THC­treated cells (P<0.05). In conclusion, the present study revealed the underlying mechanisms associated with THC­induced autophagy. A promising method of enhancing the therapeutic efficacy of THC against NSCLC cells may include inducing autophagy via inhibition of the PI3K/Akt/mTOR signaling pathway.

Lessons from Anti-Amyloid-ß Immunotherapies in Alzheimer Disease: Aiming at a Moving Target.

BACKGROUND: Available drugs for the global Alzheimer disease (AD) epidemic only treat the symptoms without modifying disease progression. Accumulating evidence supports amyloid-ß42 (Aß42)as the key triggering agent in AD, making it the ideal target for disease-modifying therapies. Preclinical studies provided extensive support for passive Aß42 immunotherapy, leading to human clinical trials with different antibodies. OBJECTIVE: Examine the status of clinical trials for passive immunotherapy against Aß42. METHODS: We performed a thorough literature review of passive Aß42 immunotherapy. RESULTS: Ten anti-Aß42 antibodies targeting lineal or conformational epitopes have been tested in clinical trials. Antibody engineering and appropriate dosing have overcome undesired side effects, leading to increased safety profiles. Unfortunately, few trials have shown cognitive protection, leading to legitimate questions about the utility of Aß42 as an AD target. There is still hope that solanezumab, aducanumab, and other ongoing trials will identify antibodies, patient subpopulations, and administration protocols, with consistent clinical benefits. CONCLUSIONS: Despite the overall disappointing results, there is still hope that Aß immunotherapy in presymptomatic patients will prevent neuronal loss and provide significant clinical benefits that can be applied to larger populations as preventive therapies. Advances with other targets may soon provide additional therapeutic options for AD with increased efficacy.

Value of a new pathological classification of lumbar intervertebral disc herniation based on transforaminal endoscopic observations.

Removal of herniated disc materials based on an imaging only method may not relieve symptoms in many patients. Therefore, the aim of the present study was to develop a transforaminal endoscopic method of classifying the pathological type of lumber intervertebral disc herniation and to compare the outcomes of surgery based on the pathological type with those of conventional endoscopic disc removal. The records of patients who received endoscopic transforaminal nucleotomy with foraminoplasty for symptomatic lumbar disc herniation between 2009 and 2013 were retrospectively reviewed. Patients were then divided into two groups: Group A, which consisted of 275 patients who received conventional endoscopic transforaminal nucleotomy with foraminoplasty between 2009 and 2011 and group B, which consisted of 316 patients who received 'targeted' endoscopic transforaminal nucleotomy with foraminoplasty between 2011 and 2013 (based on the pathological type of disc herniation identified at surgery, including fresh, calcified and scar type based on intraoperative observations). The results showed that there were no significant differences in age, gender, body mass index, symptom duration, operated segments or previous invasive therapies between the two groups. Moreover, evaluation of visual analogue scale pain scores and Oswestry disability index scores revealed that the patients in group B had a greater improvement in symptoms than those in group A (P<0.05). In addition, an age >40 years and a longer symptom duration were associated with the calcified type, and previous invasive therapy was associated with the scar type. Therefore, specific surgical treatment based on the transforaminal endoscopic pathological type can result in better outcomes for patients with lumbar disc herniation.

Isolation and purification of psoralen and isopsoralen and their efficacy and safety in the treatment of osteosarcoma in nude rats.

BACKGROUND: Modern studies have shown that psoralen has a significant inhibitory effect on tumor growth in a variety of animals and humans. OBJECTIVE: To obtain coumarin compounds - psoralen and isopsoralen - from traditional Chinese medicine Psoralea corylifolia L. using chromatographic techniques and isolation and purification methods, and to observe the transplanted tumor growth inhibitory effects and adverse reactions of psoralen and isopsoralen in nude rats with osteosarcoma. METHODS: Dried ripe fruits of Psoralea corylifolia L. were taken as the raw material to prepare crude extract of Psoralea corylifolia L. by ethanol reflux method. Column chromatography was used to isolate the crude extract; compounds were structurally identified based on (1)H-NMR, (13)C-NMR spectra, the two compounds were identified as psoralen andisopsoralen, and their contents were 99.7% and 99.6, respectively. Nude rat model of osteosarcoma was established; the rats were randomized into: normal saline group, psoralen low- and high-dose groups, isopsoralen low- and high-dose groups, and cisplatin group. Osteosarcoma volume and weight inhibition rates in nude rats in each group were observed; radioimmunoassay was used to determine the serum alkaline phosphatase activity; peripheral blood cell and bone marrow nucleated cell counts were determined; light microscopy was used to observe heart, liver, spleen, lung, kidney, and tumor histopathology; and electron microscopy was used to observe the fine structure of tumor cells. RESULTS: Tumor volume inhibition rates were 43.75% and 40.18%, respectively, in the psoralen and isopsoralen low-dose groups, and tumor weight inhibition rates were 38.83% and 37.77%. Tumor volume inhibition rates were 67.86% and 66.96%, respectively, in the psoralen and isopsoralen high-dose groups, and tumor weight inhibition rates were 49.47% and 47.87%. Psoralen and ispsoralen markedly lowered serum AKP level. Psoralen and isopsoralen induced apoptosis or necrosis of osteosarcoma. After administration of high doses of psoralen and isopsoralen, toxic reactions such as writhing, lassitude, and hypoactivity were seen. Kidney histopathology showed tubulointerstitial dilatation and congestion, and inflammatory cell aggregation in the renal intercellular space. Psoralen and isopsoralen did not cause any significant toxic side effects to the bone marrow, or other organs such as heart, lung, liver, and spleen. CONCLUSION: Psoralen and isopsoralen have growth inhibitory effects on transplanted tumor in nude rats with osteosarcoma, and can induce tumor cell apoptosis or necrosis, without significant toxic effects.

Use of the remnant ear for reconstruction in lobule-type microtia.

OBJECTIVE: To present in detail a reconstruction technique using the remnant ear in lobule-type microtia. METHODS: By comparing the location of residual ear and the contralateral normal ear, we classified 3 different types of the remnant ear. Three techniques of transposition were applied to adjust the location of the reconstructed ear. RESULTS: The reconstructed earlobes not only looked real but also were symmetrical with the contralateral normal ears. CONCLUSION: The remnant ear is a key factor-in addition to the framework and covering skin-in ear reconstruction in microtia.