LncRNA H19 inhibits high glucose-induced inflammatory responses of human retinal epithelial cells by targeting miR-19b to increase SIRT1 expression.

Diabetic retinopathy (DR) is a common complication of diabetes mellitus. Inflammatory responses play crucial roles in the progress of DR. Long noncoding RNA (lncRNAs) and microRNAs (miRNAs) are key signal transduction molecules in retina, and tightly connected with DR occurrence and development. Our study aimed to explore the functions of lncRNA H19, miR-19b and silence information regulator factor related enzymes 1 (SIRT1) in the progress of DR. Retinal pigment epithelial cells (ARPE-19) were used to build high-glucose (HG) model. Quantitative real-time PCR (qPCR) was performed to detect expression of H19, miR-19b and SIRT1 at normal glucose (NG) and HG conditions. And western blotting was performed to test protein level of SIRT1. QPCR and enzyme-linked immunosorbent assay were performed to detect expression of inflammatory cytokines. Finally, the interactions among H19, miR-19b and SIRT1 were determined by dual-luciferase reporter assay. Our results showed that lncRNA H19 and SIRT1 were reduced, while miR-19b was increased in ARPE-19 cells with HG condition. MiR-19b positively regulated the expression of inflammatory cytokines, including TNF-α, IL-1ß and IL-6. Inhibition of miR-19b and overexpression of H19 inhibited the expression of inflammatory cytokines, such as TNF-α, IL-1ß and IL-6, while knockdown of SIRT1 reversed their effects on inflammatory cytokines. Furthermore, overexpression of miR-19b reversed the inhibitory effects of overexpression of H19 on inflammatory cytokines. Importantly, H19 targeted miR-19b to downregulate miR-19b expression. Furthermore, miR-19b bound to SIRT1 and declined SIRT1 expression. H19/miR-19b/SIRT1 axis plays a key role of HG-induced inflammatory response in ARPE-19 cells, which provides new targets for DR treatment.

Long Noncoding RNA MEG3 Inhibits Apoptosis of Retinal Pigment Epithelium Cells Induced by High Glucose via the miR-93/Nrf2 Axis.

Diabetic retinopathy (DR) is the leading cause of visual impairment in developed nations. Though plasma microRNA-93 (miR-93) is associated with the risk of DR, the function and regulatory mechanism of miR-93 during DR remains unclear. Blood samples were collected from 12 DR patients and 12 healthy controls. Primary human retinal pigment epithelium (RPE) cells and ARPE-19 cells were cultured in 5 mmol/L or 33 mmol/L d-glucose medium. Long noncoding (lnc) RNA MEG3 and miR-93 expression was detected by real-time quantitative PCR. The effect of MEG3 and miR-93 on high glucose (HG)-induced apoptosis was detected by MTT and flow cytometry. IL-6 and tumor necrosis factor-α levels were detected by enzyme-linked immunosorbent assay. The relationships among MEG3, miR-93, and Nrf2 (also known as NFE2L2) were explored via dual-luciferase reporter assay. lncRNA MEG3 and Nrf2 were decreased and miR-93 was increased in blood samples of DR patients and HG-treated human RPE and ARPE-19 cells. Overexpression of miR-93 inhibited cell proliferation and promoted apoptosis, whereas overexpression of Nrf2 or MEG3 promoted proliferation and suppressed apoptosis and inflammation. In addition, MEG3 targeted miR-93 and down-regulated miR-93. Moreover, miR-93 directly targeted Nrf2 and negatively regulated Nrf2. This study suggests that lncRNA MEG3 depresses HG-induced apoptosis and inflammation of RPE via miR-93/Nrf2 axis, providing a novel perspective on the genesis and development of DR.

Regulation of the miR-19b-mediated SOCS6-JAK2/STAT3 pathway by lncRNA MEG3 is involved in high glucose-induced apoptosis in hRMECs.

OBJECTIVE: Diabetic retinopathy (DR) is one of the most severe and common complications of diabetes mellitus. The present study aimed to investigate the molecular mechanism of MEG3, miR-19b and SOCS6 in human retinal microvascular endothelial cells (hRMECs) under high glucose conditions. METHODS: HRMECs were cultured in 5 or 30 mM D-glucose medium. qRT-PCR and Western blotting were used to determine the mRNA expression and protein levels. MTT assay and flow cytometry analysis were performed to detect the viability and apoptosis of hRMECs, respectively. TNF-α, IL-6 and IL-1ß levels in cell supernatants were detected by ELISA. The activity of caspase-3/7 was also determined. A luciferase reporter assay was performed to confirm the targeting relationship between miR-19b and SOCS6, as well as MEG3 and miR-19b. RESULTS: Our study demonstrated that miR-19b was increased and SOCS6 was decreased in HG-induced hRMECs. Knockdown of SOCS6 inhibited cell viability and reversed the promotion of cell viability induced by knockdown of miR-19b. Additionally, miR-19b directly targeted and negatively regulated SOCS6. Moreover, miR-19b promoted the cell apoptosis rate and caspase-3/7 activity and increased inflammatory factors through the SOCS6-mediated JAK2/STAT3 signalling pathway. In addition, MEG3 attenuated HG-induced apoptosis of hRMECs by targeting the miR-19b/SOCS6 axis. CONCLUSION: These findings indicate that MEG3 inhibited HG-induced apoptosis and inflammation by regulating the miR-19b/SOCS6 axis through the JAK2/STAT3 signalling pathway in hRMECs. Thus, these findings might provide a new target for the treatment of DR.

lncRNA H19 sponging miR-93 to regulate inflammation in retinal epithelial cells under hyperglycemia via XBP1s.

OBJECTIVE: Diabetic retinopathy (DR) is a major complication of both type 1 and type 2 diabetes. Recently, inflammation was found to play an important role in DR pathogenesis. But the mechanism has not been fully understood. METHODS: ARPE-19 cells were cultured under normal condition and high-glucose condition, then the expressions of miR-93, XBP1s and lncRNA H19 were measured using RT-qPCR or western blots. Besides, the mRNA level of eIF2α and GRP78 and protein level of p-eIF2α and GRP78 were measured by RT-qPCR or western blots. In addition, RT-qPCR and ELISA were adopted to examine the expression and secretion of cytokine factors in these conditions. Dual-luciferase reporter gene assay was used to elucidate the binding and regulation among XBP1s, miR-93 and H19. RNA immunoprecipitation was also performed to verify the interaction between H19 and miR-93. The expressions of DNAJC3 and DNAJB9, the downstream targets of XBP1s, were detected by RT-qPCR. RESULTS: We identified that H19 and XBP1s were down-regulated in ARPE-19 cells under high-glucose condition, while miR-93 was up-regulated. ER stress inducers TM and IRE1 inhibitor STF-083010 were adopted and data suggest that ER stress could be induced during high-glucose treatment. In addition, the altered expressions of miR-93, XBP1s and H19 might mediate the increased level of pro-inflammatory cytokines. Furthermore, miR-93 interacted with either lncRNA H19 or XBP1s then modulating the inflammatory processes. CONCLUSIONS: H19 played an important role in regulating inflammatory processes in retinal endothelial cells under high-glucose condition through modulating miR-93/XBP1s axis.

[Research Advances in Ocular Myasthenia Gravis].

Ocular myasthenia gravis(OMG)is an autoimmune disease caused by neuromuscular junction transmission disorders and manifested mainly as fluctuating blepharoptosis and diplopia,with the extraocular muscles as the main involveed sites.While the pathogenesis of OMG remains unclear,some antibodies,complements,and cytokines may be the contributing factors.The diagnosis and treatment of OMG have been defined in recent years.This article reviews the pathogenesis,diagnosis,and treatment of OMG.

[Advances in Retinopathy of Prematurity].

Retinopathy of prematurity(ROP)is a pathological neovascularization with fibrotic changes in the fundus of premature infants.It is a major cause of preventable blindness in children in both developing and developed countries.Treatment of ROP has long been a hot research topic in ophthalmology and pediatrics.With a clearer knowledge of the pathogenesis of ROP,more basic and clinical studies have been carried out.The anti-vascular endothelial growth factor therapy and surgical treatment have become mature strategies,and a variety of therapeutic drugs including insulin-like growth factor-1,transforming growth factor-ß,polyunsaturated fatty acids,and ß-adrenergic receptor blockers have been developed.This article reviews the recent advances in ROP.

[Mechanism of Th1/Th2 Cytokines in HLA-DQ8 Transgenic Mice Model of Ocular Experimental Autoimmune Myasthenia Gravis].

Objective To analyze the levels of cytokines (IL-2,IFN-γ,IL-6,IL-10) associated with Th1 and Th2 cells in HLA-DQ8 transgenic mice model of ocular experimental autoimmune myasthenia gravis (oEAMG) induced by recombinant H-AChR γ subunit immunization.Methods DQ8 mice were immunized with 20 µg of AChR γ subunit,20 µg of crude E. coli extract (E. coli group),or complete Freund's adjuvant (CFA) only (CFA group). All mice were immunized on days 0,30,and 60. Mice were euthanized 28 days after the third immunization,and draining lymph node cells (LNC) and spleen lymphocytes were cultured in vitro. The supernatant was collected to observe the interleukin(IL)-2,interferon(IFN)-γ,IL-6,IL-10 production by ELISA.Results LNCs and spleen lymphocytes of H-AChR γ subunit-immunized mice exhibited significantly enhanced IFN-γ (F=76.332,P<0.001;F=34.865,P<0.001) and IL-2 (F=42.835,P<0.001;F=38.030,P<0.001),which associated with Th1 cells,as compared to E. coli group and CFA group. There were no significant differences in IL-6 (F=1.325,P=0.284;F=1.935,P=0.166) and IL-10 (F=0.908,P=0.417;F=1.189,P=0.322) levels,which secreted by Th2 cells,among these three groups.Conclusion Th1 cytokines play key roles in the pathogenesis of oEAMG,while the mechanism of Th2 cytokines for oEAMG remains unclear.

Abnormal resting-state functional network centrality in patients with high myopia: evidence from a voxel-wise degree centrality analysis.

AIM: To investigate the functional networks underlying the brain-activity changes of patients with high myopia using the voxel-wise degree centrality (DC) method. METHODS: In total, 38 patients with high myopia (HM) (17 males and 21 females), whose binocular refractive diopter were -6.00 to -7.00 D, and 38 healthy controls (17 males and 21 females), closely matched in age, sex, and education levels, participated in the study. Spontaneous brain activities were evaluated using the voxel-wise DC method. The receiver operating characteristic curve was measured to distinguish patients with HM from healthy controls. Correlation analysis was used to explore the relationship between the observed mean DC values of the different brain areas and the behavioral performance. RESULTS: Compared with healthy controls, HM patients had significantly decreased DC values in the right inferior frontal gyrus/insula, right middle frontal gyrus, and right supramarginal/inferior parietal lobule (P<0.05). In contrast, HM patients had significantly increased DC values in the right cerebellum posterior lobe, left precentral gyrus/postcentral gyrus, and right middle cingulate gyrus (P<0.05). However, no relationship was found between the observed mean DC values of the different brain areas and the behavioral performance (P>0.05). CONCLUSION: HM is associated with abnormalities in many brain regions, which may indicate the neural mechanisms of HM. The altered DC values may be used as a useful biomarker for the brain activity changes in HM patients.

The prognostic value of lymphocyte-to-monocyte ratio in retinopathy of prematurity.

AIM: To evaluate the associations between development of retinopathy of prematurity (ROP) and serum lymphocyte-to-monocyte ratio (LMR), neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR). METHODS: A retrospective cohort study was performed, involving infants who were screened for ROP from January 2015 to December 2015. Preterm newborns of ≤32 gestational weeks with ROP were enrolled as the observation group, and non-ROP infants were enrolled as the control group, whose complete blood cell were measured within the first 24h of life. The levels of NLR, LMR and PLR were determined in all groups. The data obtained were analyzed using univariate and multivariate logistic regression analysis. RESULTS: In this study, 40 cases of ROP were enrolled and 40 cases of non-ROP as controls. The LMR levels were significantly higher (P<0.001) in ROP group (3.96±1.16) compared to non-ROP group (2.85±0.79). The NLR levels were significantly lower (P=0.035) in ROP group {median [interquartile range (IQR)], 0.88 (0.67-1.46)} compared to non-ROP group [median (IQR), 1.20 (0.85-1.89)]. The median PLR values were 61.99 (IQR, 50.23-75.98) in ROP group and 69.24 (IQR, 55.52-88.12) in non-ROP group (P=0.104). Logistic regression analysis suggested that LMR was an independent risk factor for ROP (OR: 0.275; 95% CI: 0.134-0.564; P=0.001). CONCLUSION: The findings demonstrate that higher LMR is independently and significantly associated with the development of ROP, and the LMR may be invoked as a predictive tool for identifying risk for ROP.

Altered spontaneous brain activity pattern in patients with high myopia using amplitude of low-frequency fluctuation: a resting-state fMRI study.

OBJECTIVE: Many previous reports have demonstrated significant neural anatomy changes in the brain of high myopic (HM) patients, whereas the spontaneous brain activity changes in the HM patients at rest are not well studied. Our objective was to use amplitude of low-frequency fluctuation (ALFF) method to investigate the changes in spontaneous brain activity in HM patients and their relationships with clinical features. METHODS: A total of 38 patients with HM (17 males and 21 females) and 38 healthy controls (HCs) (17 males and 21 females) closely matched in age, sex, and education underwent resting-state functional magnetic resonance imaging scans. The ALFF method was used to assess local features of spontaneous brain activity. The relationship between the mean ALFF signal values in many brain regions and the clinical features in HM patients was calculated by correlation analysis. RESULTS: Compared with HCs, the HM patients had significantly lower ALFF in the right inferior and middle temporal gyrus, left middle temporal gyrus, left inferior frontal gyrus/putamen, right inferior frontal gyrus/putamen/insula, right middle frontal gyrus, and right inferior parietal lobule and higher ALFF values in the bilateral midcingulate cortex, left postcentral gyrus, and left precuneus/inferior parietal lobule. However, no relationship was found between the mean ALFF signal values of the different areas and the clinical manifestations in HM. CONCLUSION: The HM patients were affected with brain dysfunction in many regions, which may indicate the presence of neurobiological changes involving deficits in language understanding and attentional control in HM patients.

Reconstruction of degloved thumb with prefabricated flap.

The wrap around the toe flap from the great toe is considered to be a good reconstructive proce-dure for degloved injuries of the thumb. In this study, we used prefabricated flaps of the medial plantar skin to cover a degloved injury of the thumb of a patient unsuitable for application of this method and obtained satisfactory clinical results.

[High tibial osteotomy for the treatment of osteoarthritis of the knee].

OBJECTIVE: To summarize clinical application of the high tibial osteatomy (HTO) with lateral closing-wedge for the treatment of pain of unicompartmental osteoarthritis of the knee. METHODS: From February 2000 to February 2004,9 patients (3 males and 6 females, ranging in age from 52 to 58 years, with an average of 56 years) with unicompartmental osteoarthritis of the knee treated by HTO with lateral closing-wedge. The proximal tibiofibular joint was exposed, the anterior part of the capsule was incised, and to remove a laterally based wedge with saw cuts and osteotomes. Stepped staples were used for the fixation of tibial osteotomies after closing the defect of osteotomy. RESULTS: The operative course was uneventful. There were no complications after operation. The postoperative follow-up period ranged from 2 to 5.5 years (mean, 3.5 years). The results were evaluated with functional assessment criterion of the osteoarthritis of the knee, among the 9 cases, excellent was in 5 cases, good in 3 cases, fair in 1 case. CONCLUSION: HTO with lateral closing-wedge is an effective operative method for the treatment of pain of unicompartmental osteoarthritis of the knee, but except for older patients over 60 years.

[Treatment of occluded iliofemoral veins with great saphenous vein cross-over bypasses].

OBJECTIVE: To summarize clinical application experience of treatment of occluded iliofemoral veins with great saphenous vein cross-over bypasses. METHODS: From September 1998 to December 2005,6 patients(5 men, 1 woman) with occluded iliofemoral veins underwent great saphenous vein cross-over bypasses. They ranged in age from 36 to 52 years (mean, 41 years old). All patients had unilateral thigh and leg edema and swelling with accompanying pain. The normal contralateral great saphenous vein was dissected and it was tunneled across the super-pubic fat pad and anastomosed to the femoral vein or the proximal portion of great saphenous vein in distal to occluded iliofemoral veins. RESULTS: The postoperative course was uneventful. One case sustained superficial infection postoperatively at donor site and the gradual wound healed by daily wound dressings. At 1 to 3.5 years (mean 1.8 years) followed up, all patients had pain disappearance and obvious relief of the edema but some swelling. CONCLUSION: Great saphenous vein cross-over bypasses is effective operative method of treatment of occluded iliofemoral veins. The operation is relatively simple and safe.

[Reconstruction of distal phalanx defect with microsurgical fascia latae flaps combined with rib and skin graft].

OBJECTIVE: To evaluate the clinical application of microsurgical fascia latae flaps combined with rib and skin graft for reconstruction of the distal phalanx defect. METHODS: The phalanx wounds were sutured together like syndactyly. The autologous rib was revised to repair the bone defect of distal phalanx. The fascia latae flap was used to cover the bone exposure with microvascular anastomoses and resurfaced by a meshed split-thickness skin graft. RESULTS: 5 cases were treated successfully. The fascia latae flaps were all survived with only skin graft necrosis at the distal end in one case. It was healed spontaneously. CONCLUSIONS: The fascia latae flap is nourished by the descending branch of the lateral circumflex femoral artery. The flaps has a good blood supply and can be easily obtained with a long vascular pedicle. The flap is ideal for the reconstruction of distal phalanx defect when combined with autologous rib implant and skin graft.

[Repair of soft-tissue defect of amputation stumps of the forearm with free flap from the traumatic amputated extremity].

OBJECTIVE: To report the clinical results of the repair of soft-tissue defect of amputation stumps of the forearm with free flap from the traumatic amputated extremity. METHODS: Five patients, 4 males and 1 female, aged 32 (22 - 43), with soft-tissue defect of the remaining stump of the traumatic amputation of the forearm, 3 cases in the right forearm and 2 cases in the left forearm, underwent repair of the defect by free flap from the traumatic amputated extremity with the size of the flaps ranging from 8 cm x 9 cm to 9 cm x 12 cm. The patients were followed up for 2.6 years (1.5 - 3.5 years). RESULTS: Superficial infection occurred in one patient postoperatively and the wound was gradually healed by daily wound dressings. All the flaps survived completely with satisfactory clinical results. The cosmetic appearance on the recipient area was good and the function of the elbow recovered satisfactorily. CONCLUSION: The flap from the traumatic amputated extremity has a constant vascular anatomy and a long vascular pedicle, so that dissection of the flap can be accomplished easily. The surgery allows to preserve the functional length or the elbow function of the remaining stump of the traumatic amputation.