Targeting ZIP8 mediated ferroptosis as a novel strategy to protect against the retinal pigment epithelial degeneration.

The degeneration of retinal pigment epithelium (RPE) plays an important role in the development of age-related macular degeneration (AMD). However, the underlying mechanism remains elusive. In this study, we identified that ZIP8, a metal-ion transporter, plays a crucial role in the degeneration of RPE cells mediated by ferroptosis. ZIP8 was found to be upregulated in patients with AMD through transcriptome analysis. Upregulated ZIP8 was also observed in both oxidative-stressed RPE cells and AMD mouse model. Importantly, knockdown of ZIP8 significantly inhibited ferroptosis in RPE cells induced by sodium iodate-induced oxidative stress. Blocking ZIP8 with specific antibodies reversed RPE degeneration and restored retinal function, improving visual loss in a mouse model of NaIO3-induced. Interestingly, the modification of the N-glycosylation sites N40, N72 and N88, but not N273, was essential for the intracellular iron accumulation mediated by ZIP8, which further led to increased lipid peroxidation and RPE death. These findings highlight the critical role of ZIP8 in RPE ferroptosis and provide a potential target for the treatment of diseases associated with retinal degeneration, including AMD.

LY2874455 and Abemaciclib Reverse FGF3/4/19/CCND1 Amplification Mediated Gefitinib Resistance in NSCLC.

Non-small cell lung carcinoma (NSCLC) patients who initially received tyrosine kinase inhibitor (TKI) therapy often acquired resistance via multiple complex mechanisms. The amplification of FGF3/4/19/CCND1 on chromosome 11q13 was found in many cancers with TKI resistance. However, the role of these amplifications in TKI-resistant NSCLC remains uncovered. Here, we generated the FGF3/4/19/CCND1 amplification model in the NSCLC cell lines PC-9 and HCC827. Upregulation of FGF3/4/19/CCND1 strongly promoted cell proliferation and gefitinib resistance in NSCLC cells. To find out the potential therapeutic strategies, we screened the combination of inhibitors against the FGF/FGFR signaling pathway and the CCND1/CDK4 complex and revealed that gefitinib combined with LY2874455 and abemaciclib exhibited the most effective inhibition of resistance in vitro and in vivo. Mechanistically, FGFs/CCND1 activated the MAPK pathway, which was abolished by the combination drugs. Our study provides a rationale for clinical testing of dual targeting FGFR and CCND1 with LY2874455 and abemaciclib in NSCLC patients who harbored FGF3/4/19/CCND1 amplification.

Methyltransferase like 7B is a potential therapeutic target for reversing EGFR-TKIs resistance in lung adenocarcinoma.

BACKGROUND: Identification of potential novel targets for reversing resistance to Epidermal Growth Factor Receptor (EGFR)-tyrosine kinase inhibitors (EGFR-TKIs) holds great promise for the treatment of relapsed lung adenocarcinoma (LUAD). In the present study, we aim to investigate the role of methyltransferase-like 7B (METTL7B) in inducing EGFR-TKIs resistance in LUAD and whether it could be a therapeutic target for reversing the resistance. METHODS: METTL7B-overexpressed LUAD cell lines, gefitinib and osimertinib-resistant Cell-Derived tumor Xenograft (CDX) and Patient-Derived tumor Xenograft (PDX) mouse models were employed to evaluate the role of METTL7B in TKIs resistance. Ultraperformance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS) was used to identify the metabolites regulated by METTL7B. Methylated RNA immunoprecipitation (MeRIP)-qPCR analysis was performed to measure the N6-methyladenosine (m6A) status of mRNA of METTL7B targeted genes. Gold nanocluster-assisted delivery of siRNA targeting METTL7B (GNC-siMETTL7B) was applied to evaluate the effect of METTL7B in TKIs resistance. RESULTS: Increased expression of METTL7B was found in TKIs-resistant LUAD cells and overexpression of METTL7B in LUAD cells induced TKIs resistance both in vitro and in vivo. Activated ROS-metabolism was identified in METTL7B-overexpressed LUAD cells, accompanied with upregulated protein level of GPX4, HMOX1 and SOD1 and their enzymatic activities. Globally elevated m6A levels were found in METTL7B-overexpressed LUAD cells, which was reduced by knock-down of METTL7B. METTL7B induced m6A modification of GPX4, HMOX1 and SOD1 mRNA. Knock-down of METTL7B by siRNA re-sensitized LUAD cells to gefitinib and osimertinib both in vitro and in vivo. CONCLUSIONS: This study uncovered a new critical link in METTL7B, glutathione metabolism and drug resistance. Our findings demonstrated that METTL7B inhibitors could be used for reversing TKIs resistance in LUAD patients.

A Novel Nonsense Mutation in the EXT2 Gene Identified in a Family with Hereditary Multiple Osteochondromas.

Aims: Identification of genetic mutations linked to hereditary multiple osteochondromas (HMO) is crucial for understanding the molecular mechanisms leading to disease pathogenesis. In this study, we investigated four patients and eight healthy individuals from a family with HMO. Methods: Clinical HMO data and Sanger sequences of the coding regions of the exostosin glycosyltransferase 1 (EXT1) gene (18q24.11) and the EXT2 gene (11p12) of all 12 members of the family were analyzed. Results: A novel nonsense mutation in the EXT2 gene (c.526C>T; p.Gln176*) was detected, which was present in all four patients but absent in their healthy relatives. This mutation encodes a stop codon that results in a truncated EXT2 protein that consists of only 176 amino acids and lacks the remaining 522 amino acids at its C-terminus, missing the entire glycosyltransferase domain. Conclusions: Association of a truncated EXT2 protein with HMO provides new insights into exostosis pathogenesis, highlighting potential roles of the EXT2 gene and its glycosyltransferase domain. Further research is required to understand the mechanisms underlying the development of exostosis.

METTL7B Is Required for Cancer Cell Proliferation and Tumorigenesis in Non-Small Cell Lung Cancer.

Lung cancer remains a leading cause of cancer-associated mortality worldwide, however, molecular mechanisms underlying lung cancer tumorigenesis and progression remain unknown. Here, we report evidence showing that one member of the mammalian methyltransferase-like family (METTL), METTL7B, is a potential molecular target for treatment of non-small cell lung cancer (NSCLC). METTL7B expression was elevated in the majority of NSCLC comparing to normal tissues. Increased expression of METTL7B contributed to advanced stages of tumor development and poor survival in NSCLC patients. Lentivirus-mediated shRNA silencing of METTL7B suppressed proliferation and tumorigenesis of cancer cells in vitro and in vivo. Investigation on gene expression profiles of NSCLC cells revealed that abundant cell cycle related genes were downregulated in the absence of METTL7B. Pathway enrichment analysis indicated that METTL7B participated in cell cycle regulation. Notably, CCND1, a key regulator for G1/S transition, was significantly decreased with the depletion of METTL7B, resulting in G0/G1 arrest, indicating that METTL7B is critical for cell cycle progression. Taken together, our findings implicate that METTL7B is essential for NSCLC development and progression. METTL7B might serve as a potential therapeutic target for NSCLC.

The influences of anesthesia methods on some complications after orthopedic surgery: a Bayesian network meta-analysis.

BACKGROUND: Although several anesthesia procedures have been explored for orthopedic surgery, the complications of anesthesia remain not well resolved. This study aimed to explore the influence of different anesthesia methods on the complications after orthopedic surgery. METHODS: According to the searching strategy, anesthesia associated studies in orthopedic surgery were screened from Pubmed, Embase, and the Cochrane Library up to Mar. 10th, 2018. Then, complications and demographic data were extracted and quality of studies was assessed using Cochrane Collaboration recommendations. ADDIS software was used to perform the network meta-analysis. Pooled effect size was calculated using random effective model or consistency model, and presented with odds ratio (OR) and 95% confidence interval (CI). RESULTS: According to the selective criteria, a total of 23 studies with 2393 patients were enrolled in this study. Quality assessment revealed all studies had an ordinary quality. Network meta-analyses revealed that nerve block analgesia (NBA) presented a lower effect on the occurrence of post-operative nausea or vomiting (PONV; OR = 0.17, 95% CI: 0.06-0.39) and urine retention (OR = 0.07, 95% CI: 0.01-0.37) compared with epidural anesthesia (EA). Interscalene block (ISB) and local infiltration analgesia (LIA) could significantly reduce the occurrence of back pain compared with EA (OR = 0.00, 95% CI = 0.00-0.30; OR = 0.00, 95% CI = 0.00-0.25). CONCLUSION: NBA presented an effective role in reliving the occurrence of PONV and urine retention, and ISB and LIA relieved the back pain compared with EA after orthopedic surgery.

Cervical disc herniation causing Brown-Sequard syndrome: Case report and review of literature (CARE-compliant).

RATIONALE: Brown-Sequard syndrome (BSS) is manifested as ipsilateral motor deficit and contralateral sensory loss. BSS caused by herniated cervical disc is extremely rare and easily be misdiagnosed, and clinical features of this problem were not fully understood. PATIENT CONCERNS: A 57-year-old man presented with a 3-month history of weakness in his right arm, and he experienced progressive right hemiparesis at 2 days before admission, along with contralateral deficit in sensation of pain and temperature below T2. DIAGNOSES: Magnetic Resonance Imaging (MRI) showed severe cord compression due to a large paracentral extradural C4-C5 cervical disc herniation (CDH). INTERVENTIONS: Subtotal cervical corpectomy, decompression, and fusion through anterior approach were performed. The patient recovered rapidly after surgery. OUTCOMES: Complete recovery of sensory and motor functions was obtained at a 4-months follow-up after surgery. LESSONS: Our case, along with a review of the literature, highlights that careful medical history inquiries, detailed neurologic examinations, and cervical spinal MRI scans are essential for diagnosis of CDH caused BSS. Prompt surgical decompression according to individual condition is commonly warranted. Early diagnosis with prompt surgical decompression could lead to favorable recovery.

[Current advances on surgical treatment for knee articular cartilage injuries].

Chondral injuries are short of self-healing ability and need to surgical repair after articular cartilage injury. Conventional treatment includes debridement and drainage under arthroscope, micro-fracture, osteochondral autograft transplantation (OATS), mosaiplasty and osteochondral allografts (OCA), autologous chondrocyte implantation (ACI). Debridement and drainage could remove pain factor, and has advantages of simple operation, wide clinical application and early clinical effect. Micro-fracture and osteochondral autograft transplantation is suitable for small area of cartilage repair, while the further effect showed that fibrous cartilage permeated by drill could decrease postoperative clinical effect. Osteochondral autograft transplantation has better advantages for reconstruction complete of wear-bearing joint. Autologous chondrocyte implantation and allogeneic cartilage transplantation are suitable for large area of cartilage defect, postoperative survival of allogeneic cartilage transplantation is effected by local rejection reaction and decrease further clinical effect. Cartilage tissue engineering technology could improve repair quality of autologous chondrocyte implantation, and make repair tissue close to transparent cartilage, but has limit to combined subchondral bone plate, reactive bone edema, bone loss and bad axis of lower limb. New technology is applied to cartilage injury, and has advantages of less trauma, simple operation, rapid recover, good clinical effect and less cost;and could be main method for treat cartilage injury with surgical repair technology. How to improve repair quality with compression resistance and abrasive resistance are expected to be solved.

Genetic variation of SMAD3 is associated with hip osteoarthritis in a Chinese Han population.

Objective This study was performed to investigate the association between genetic variation in SMAD3 and hip osteoarthritis (OA) in a Chinese Han population. Methods The frequency of two single nucleotide polymorphisms of SMAD3, rs1470002 and rs12901499, was examined in 500 patients with hip OA and 1080 healthy controls in a Chinese Han population. Further analysis was performed according to sex and age. Results We detected statistically significant differences in the allele frequency and genotype between the hip OA and healthy control groups. The frequency of the GA+GG and GA genotypes of rs12901499 and the G variant were much higher in patients with hip OA than in healthy controls. This association was also present when the participants were stratified by sex and age. However, there was no significant association between the risk of hip OA and the presence of rs1470002 GA, AA, or GA+AA genotypes, even after sex- and age-stratified analysis. Conclusions The SMAD3 SNP rs12901499 GA genotype and G variant may increase the risk of hip OA in Chinese Han patients.

siRNA-mediated c-Rel knockdown ameliorates collagen-induced arthritis in mice.

Previous studies have shown that inflammatory mediators involved in the development of rheumatoid arthritis (RA) are regulated by the Rel/nuclear factor-κB (Rel/NF-κB) transcription factor family. c-Rel, a member of the Rel/NF-κB family that is preferentially expressed by immune cells, is a risk factor for several inflammatory diseases including RA. In the current study, we investigated whether targeting c-Rel can be used to treat collagen-induced arthritis, an animal model for RA. c-Rel specific siRNA (siRel) delivered by nanoparticles was used to knockdown the expression of c-Rel. Our results showed that siRel treatment significantly ameliorated collagen-induced arthritis. Further study revealed that c-Rel expression in the dendritic cells and macrophages from mice treated with siRel was significantly down-regulated. Consistent with the phenotypical result, the expression of inflammatory cytokines TNF-α, IL-1ß, IL-6, IL-12 and IL-23 by peritoneal macrophages and splenocytes were significantly decreased. In addition, attenuated systemic and collagen-specific Th1 and Th17 immune responses were observed. Furthermore, we found that the expression of inflammatory cytokines was significantly down-regulated and the infiltration of CD3+ T cells and F4/80+ macrophages was markedly reduced in hind paws of mice treated with siRel. Collectively, our study provides strong evidence that siRNA-mediated c-Rel knockdown can suppress the development of collagen-induced arthritis in mice. Therefore, blocking c-Rel may represent an attracting strategy for the treatment of human rheumatoid arthritis.

[Clinical study of medicinal-cake-separated moxibustion for senile osteoporosis].

OBJECTIVE: To explore the clinical efficacy and partial mechanism of medicinal-cake-separated moxibustion for senile osteoporosis. METHODS: Sixty cases of senile osteoporosis were randomly divided into an observation group and a control group according to the random digits table, 30 cases in each one. The two groups were both treated with basic treatment of western medicine. The acupoints included four groups:① Dazhui (GV 14), Dazhu (BL 11) and Ganshu (BL 18); ② Zhongwan (CV 12), Danzhong (CV 17) and Zusanli (ST 36); ③ Pishu (BL 20), Shenshu (BL 23) and Mingmen (GV 4); ④ Shenque (CV 8) and Guanyuan (CV 4). Each group of acupoints was selected for one treatment. The observation group was treated with medicinal-cake-separated moxibustion, and the medicinal cake was consisted of fructus psoraleae (30 g), prepared rehmannia root (30 g), atractylodes (30 g), codonopsis pilosula (30 g), epimedium herb (20 g), rhizoma curculiginis (20 g), syzygium aromaticum (5 g) and cinnamon (5 g). The control group was treated with wheat-flour-cake moxibustion. Each acupoint was treated with 5 moxa cones in the two groups. The treatment was given once every other day for six months. The symptom score, lumbar and hip bone mineral density (BMD), serum type Ⅰ procollagen amino-terminal propeptide (PINP) and serum ß-type Ⅰ collagen carboxy-terminal peptide (ß-CTX) were observed before and after treatment. RESULTS: After treatment, the symptom score and serum ß-CTX were significantly lowered (all P<0.05), while the lumbar and hip BMD and serum PINP were significantly increased (all P<0.05) of the two groups. After treatment, the symptom score and serum ß-CTX in the observation group were significantly lower than those in the control group (both P<0.05), while the lumbar and hip BMD and serum PINP in the observation group were significantly higher than those in the control group (all P<0.05). CONCLUSIONS: The medicinal-cake-separated moxibustion has significant efficacy for senile osteoporosis, which is superior to wheat-cake-se-parated moxibustion.

Association between transforming growth factor-beta 1 gene single nucleotide polymorphisms and knee osteoarthritis susceptibility in a Chinese Han population.

Objective To investigate associations between single polymorphisms (SNPs) rs1800469 and rs1982073 in the transforming growth factor-ß1 gene ( TGF-ß1) and knee osteoarthritis (OA) susceptibility in a Chinese Han population. Methods TGF-ß1 rs1800469 and rs1982073 were genotyped in patients with knee OA and age- and sex-matched OA-free controls from a Chinese Han population. The association was further analyzed according to gender and age. Results A total of 765 patients with knee OA and 780 controls were included. CT and CT + CC genotypes of rs1982073, and variant C, were associated with a significantly increased risk of knee OA. Stratification analysis showed that the association between the OA risk and rs1982073 CT heterozygotes compared with TT homozygotes was stronger in females and those aged >65 years. In contrast, CT, TT, and CT + TT genotypes of rs1800469 were not significantly associated with the risk of knee OA, even after further stratification analysis for gender and age. Conclusions The TGF-ß1 rs1982073 T to C change and the variant C genotype may contribute to knee OA risk in the Chinese Han population.

[Current knowledge in the treatment of anterolateral ligament injury of knees].

Although the function of the anterolateral stabilizing structures of the knee in the anterior cruciate ligament (ACL) injuries has been recognized since many years, most of orthopedic surgeons do not take the anterolateral structure into consideration when performing an ACL reconstruction. Anatomic ACL reconstruction will improve knee stability, but a small subset of patients may experience some residual anteroposterior and rotational instability. For this reason, some researchers have paid attention to the anterolateral aspects of the knee, especially the anterolateral ligament. We don't know the best time to perform ACL and ALL reconstruction. And we lack the evidence to prove which technique is the best one. So we look forward to more random controlled trial.

Minimal invasive annulotomy for induction of disc degeneration and implantation of poly (lactic-co-glycolic acid) (PLGA) plugs for annular repair in a rabbit model.

BACKGROUND: The rabbit disc model is useful for the study of intervertebral disc (IVD) degeneration and experimental therapeutic interventions. The annulotomy-induced disc models present several drawbacks, particularly an excessive disruption of disc integrity and a rapidly disc degeneration; therefore, this study sought to establish a minimal invasive annulotomy for induction of disc degeneration model, combined to annulus repair using implantation of a PLGA (poly lactic-co-glycolic acid) plug. METHODS: New Zealand white rabbits (n = 24) received annular injuries in three discs (L3/4, L4/5 and L5/6). The experimental discs were randomly assigned to four groups: (a) annular defect with a 1.8 mm diameter mini-trephine; (b) annular puncture by 16G needle; (c) annular defect with a PLGA plug implanted by press-fit fashion; (d) uninjured L2/3 disc served as control. Postsurgical x-ray, MRI examination, and real-time PCR analysis were performed at 1, 3 and 6 months. Gross morphology and histology were evaluated at postoperative 6 months. RESULTS: Radiographic examinations showed a slow, progressive disc space narrowing and a significant degree of disc degeneration on MRI grade in the injured discs at 6 months in all rabbits. Histological examinations and aggrecan, Col1A1, Col2A1 and matrix metalloprotease (MMP)-3 mRNA expression confirmed the disc degeneration, supporting the imaging results. The PLGA implantation reduced the marked loss of T2-weighted signal intensity seen at MRI in the injured discs and slowly decreased the disc height index (DHI) over the follow-up period. HE/Safranin O staining showed that annular defect was replaced by regenerated connective tissue with significant loss of proteoglycan content. CONCLUSIONS: The minimally invasive approach for the creation of annular defects by an appropriately sized mini-Trephine is a suitable option for the study of disc degeneration in a rabbit model. Implantation of a suitable PLGA plug induced a successful repair of the annulus fibrosus within the degenerated disc, and retarded the degenerative process in the annular injury model.

[Case-control studies on therapeutic effects for the treatments of tibial plateau fractures between arthroscopic technique in minimally invasion surgery and minimally invasive internal fixation with plates and screws].

OBJECTIVE: To investigate therapeutic effects of minimally invasive arthroscopic internal fixation with plates and screws in treating tibial plateau fractures. METHODS: A retrospective study from December 2006 to June 2010 was done on 69 patients with tibial plateau fractures. According to Schatzker classification, 5 patients were type I, 5 patients were type II, 25 patients were type III, 20 patients were type IV, 9 patients were type V and 5 patients were type VI. Thirty-six patients were treated with arthroscopy-assisted reduction and internal fixation, including 21 males and 15 females, ranging in age from 17 to 59 years (averaged, 34.2 years); another 33 patients were treated with small incision and fixed with plates and screws,including 19 males and 14 females, ranging in age from 18 to 62 years (averaged, 35.4 years). The operation time, blood loss during operation,drainage volume of blood, healing time, weight-bearing time and function of effected knee were compared between the two groups. RESULTS: All the patients were followed up,and the duration ranged from 6 to 12 months (averaged, 10.3 months). All the patients had no complications such as infection, articular collapse, re-fracture and joint stiffness. There were no significant differences in weight-bearing time, complications and Rasmussen scores between two groups (P>0.05); but there were significant differences in the operative time, blood loss, drainage volume of blood, hospital stay time, the healing time between two groups (P<0.05). The results showed that arthroscopy-assisted technique was better than minimally invasive internal fixation in operation duration, blood loss during operation and the healing time. CONCLUSION: Different types of fracture of tibial plateau should be treated with different surgical treatments. Arthroscopic technique for reduction of fractures, which has less influence on bony union and minimally invasive, and can provide a good clinical outcome.