In vivo quantification of human aqueous veins by enhanced depth imaging optical coherence tomography and optical coherence tomography angiography images.

AIM: To investigate the aqueous vein in vivo by using enhanced depth imaging optical coherence tomography (EDI-OCT) and optical coherence tomography angiography (OCTA). METHODS: In this cross-sectional comparative study, 30 healthy participants were enrolled. Images of the aqueous and conjunctival veins were captured by EDI-OCT and OCTA before and after water loading. The area, height, width, location depth and blood flow of the aqueous vein and conjunctival vein were measured by Image J software. RESULTS: In the static state, the area of the aqueous vein was 8166.7±3272.7 µm2, which was smaller than that of the conjunctival vein (13 690±7457 µm2, P<0.001). The mean blood flow density of the aqueous vein was 35.3%±12.6%, which was significantly less than that of the conjunctival vein (51.5%±10.6%, P<0.001). After water loading, the area of the aqueous vein decreased significantly from 8725.8±779.4 µm2 (baseline) to 7005.2±566.2 µm2 at 45min but rose to 7863.0±703.2 µm2 at 60min (P=0.032). The blood flow density of the aqueous vein decreased significantly from 41.2%±4.5% (baseline) to 35.4%±3.2% at 30min but returned to 45.6%±3.6% at 60min (P=0.021). CONCLUSION: The structure and blood flow density of the aqueous vein can be effectively evaluated by OCT and OCTA. These may become biological indicators to evaluate aqueous vein changes and aqueous outflow resistance under different interventions in glaucoma patients.

Dauer larva-derived extracellular vesicles extend the life of Caenorhabditis elegans.

There is growing evidence that extracellular vesicles (EVs) play a functional role in tissue repair and anti-aging by transferring the contents of donor cells to recipient cells. We hypothesized that Dauer (C. elegans), known as "ageless" nematodes, can also secrete extracellular vesicles and influence the lifespan of C. elegans. Here, we isolated EVs of dauer larvae (dauer EVs). Dauer EVs were characterized using transmission electron microscopy, nanoparticle tracking analysis (NTA), and Western blot analysis. Wild-type C. elegans were fed in the presence or absence of dauer EVs and tested for a range of phenotypes, including longevity, mobility and reproductive capacity. Results showed that dauer EVs increased the average lifespan of nematodes by 15.74%, improved mobility, slowed age-related pigmentation as well as body length, and reduced the accumulation of reactive oxygen species and lipids, while not impairing nematode reproductive capacity. These findings suggest that dauer EVs can extend the lifespan of C. elegans as well as the healthy lifespan by reducing ROS accumulation, with potential anti-aging capacity.

The association between parenteral nutrition and pancreatic injury in adult patients: a retrospective observational study.

BACKGROUND AND OBJECTIVE: Patients on parenteral nutrition (PN) are at high risk of both liver and pancreatic injury. More efforts were focused on liver, however, limited data is available to evaluate the effects of PN on pancreas. Thus, we performed a retrospective observational study to evaluate the association between PN and pancreatic injury in Chinese adult patients. METHODS: Adult patients (18-80 years), who received PN for a week or longer, and with repeated measurements of pancreatic enzymes, were included in the analysis. Pancreatic injury was confirmed by serum level of pancreatic amylase (P-AMYwas 53 U/L or higher) or lipase (LP was 63 U/L or higher), which were evaluated at baseline and following every week during PN duration. Age, sex, body weight, height, diagnosis of diseases, history of diseases, surgery, white blood cell, c-reactive protein, liver and renal function, fasting blood glucose, lipid profile, and daily energy supplied by PN and enteral nutrition were abstracted from medical records. RESULTS: A total number of 190 adult patients (125 men, 65 women) were included in the study. The average age and BMI were 61.8 ± 13.0 years and 21.7±3.3 kg/m2, while medium serum level of P-AMY and LP were 29.0 U/L (quartile range: 18.0, 47.0) and 33.0 U/L (quartile range: 19.0, 58.0), respectively at baseline. The median duration of PN was 15 days (quartile range: 11.0, 21.0). The prevalence of pancreatic injury was 42.1% (80/190) while it was 28.4% (54/190), 43.3% (77/178), 47.8% (44/92) after one-, two-, and three-week or longer PN adminstration. The proportion of daily energy supplement by PN (OR = 3.77, 95%CI: 1.87, 7.61) and history of infection were positively (OR = 3.00, 95%CI: 1.23, 7.36), while disease history for diabetes mellitus (OR = 0.38, 95%CI: 0.15, 0.98) and cancer (OR = 0.46, 95%CI: 0.23, 0.95), were negetively associated with pancreatic injury. Total bile acids were associated with the increment of P-AMY (beta = 0.98, 95%CI: 0.39, 1.56) and LP (beta = 2.55, 95%CI: 0.98, 4.12) by multi-variate linear regression. CONCLUSION: PN was associated with pancreatic injury, as demonstrated by the increase of both serum P-AMY and LP.

Diagnostic accuracy of mac-2-binding protein glycosylation isomer for diagnosing hepatitis B-related fibrosis: A meta-analysis.

OBJECTIVES: Mac-2-binding protein glycosylation isomer (M2BPGi) is a novel biomarker for liver fibrosis. The aim of this meta-analysis was to evaluate the accuracy of M2BPGi for predicting hepatitis B virus (HBV)-related liver fibrosis. METHODS: EMBASE, PubMed, Web of Science, China National Knowledge Infrastructure, SinoMED, VIP Database for Chinese Technical Periodicals databases were searched comprehensively for articles published up to March 2022. Quality assessment was carried out in accordance with the Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2). The pooled diagnostic estimates including sensitivity, specificity, the area under the summary receiver operating characteristic curve (AUROC) were calculated. Before pooling the estimates, the threshold effect was assessed. Subgroup analysis was performed as well. RESULTS: In all, 11 studies including 1836 patients were included. None of the 11 studies met all the criteria of QUADAS-2. The threshold effect was found (r = 0.757, P = 0.011) for predicting HBV-related severe fibrosis. The sensitivity, specificity and AUROC of M2BPGi for the prediction of significant fibrosis were 0.68 (0.65-0.71), 0.67 (0.64-0.70) and 0.741, respectively, while those for predicting cirrhosis were 0.65 (0.57-0.72), 0.79 (0.77-0.81) and 0.792. Additionally, the AUROC of M2BPGi for predicting severe fibrosis reached 0.766. No publication bias was observed. The results of subgroup analyses were similar to the overall results. CONCLUSIONS: M2BPGi has moderate diagnostic accuracy for predicting HBV-related significant fibrosis, severe fibrosis, and cirrhosis. Further studies stratified by etiology, liver inflammation, treatment, etc, are urgently needed.

The prevalence of hypophosphatemia and refeeding-related hypophosphatemia in hospitalized patients requiring parental nutrition: a retrospective study.

OBJECTIVE: Hypophosphatemia might cause respiratory and heart failure and even death. We aimed to evaluate risk factors for hypophosphatemia and refeeding-related hypophosphatemia in patients requiring parental nutrition (PN). METHODS: This was a single-center, retrospective study. Clinical parameters were obtained from medical records. Serum phosphate (inorganic phosphorus) was measured by photometric analysis. Hypophosphatemia was confirmed when serum phosphate level was less than 0.8 mmol/L (≈2.5 mg/dl). Refeeding related hypophosphatemia was confirmed if serum phosphate level had a decrease of 0.16 mmol/L or more from baseline and if the final assessment was below 0.65 mmol/L. RESULTS: A total number of 655 (426 men and 229 women, aged 62.8 ± 14.8 years) hospitalized patients requiring PN were included in the study, and 60.6% of them were patients with cancer. The average body mass index (BMI) was 21.1 ± 4.1 kg/m2 and the median of serum phosphate was 0.9 mmol/L (quartile range: 0.68 mmol/L, 1.11 mmol/L). The prevalence of hypophosphatemia was 37.6% (246/655). Older age (≥ 65 years vs. < 65 years), lower serum level of pre-albumin (< 160 mg/L vs. ≥ 160 mg/L), calcium (< 2.11 mmol/L vs. ≥ 2.11 mmol/L), and magnesium (< 0.75 mmol/L vs. ≥ 0.75 mmol/L) were associated with high risk of hypophosphatemia by multivariate logistic regression (OR ranged from 1.43 to 3.06, all p < 0.05). Refeeding related hypophosphatemia was 9.5% (16/168). Serum level of calcium at baseline was significantly lower in participants with refeeding related hypophosphatemia than those without it. Total calorie and nitrogen delivered during first week of PN period showed no obvious difference between patients with and without refeeding related hypophosphatemia. CONCLUSIONS: Hypophosphatemia is common (37.6%) in hospitalized patients requiring PN. Monitoring of serum level of phosphorus is necessary to facilitate early treatment of hypophosphatemia.

Changes in Schlemm's canal, trabecular meshwork, and relevant parameters in the early stage after SMILE of myopia patients.

AIM: To observe the changes in Schlemm's canal (SC), trabecular meshwork (TM), and anterior chamber relevant parameters after small incision lenticule extraction (SMILE) of myopia patients. METHODS: A total of 58 eyes from 30 patients who underwent SMILE were divided into a low and moderate myopia group (group A, 32 eyes) and a high myopia group (group B, 26 eyes). The diameter and area of the SC, the width and thickness of TM obtained by CIRRUS HD-OCT5000, and the related anterior chamber parameters obtained by Pentacam anterior segment analysis system, accommodation amplitude (AMP) were observed pre- and postoperatively. The preoperative intraocular pressure (IOP) and postoperative correction of intraocular pressure (IOPcc) were measured. RESULTS: The diameter and area of the SC in the two groups were significantly increased postoperatively (all P<0.01). The TM width of the patients in the two groups were increased at 1mo after surgery (both P<0.01), but the TM thickness did not change (P>0.05). The corneal curvature, central anterior chamber depth, and anterior chamber volume decreased after SMILE surgery (all P<0.01). There was a weak negative correlation between the SC area change and AMP change in group A (r=-0.362, P<0.01). The postoperative IOP decreased after correction by Shah formula (P<0.05). CONCLUSION: SC and TM in myopia patients change in the early postoperative stage of SMILE and the IOP is decline.

Analogs of microgravity: the function of Schlemm's canal, intraocular pressure and autonomic nervous during the head-down tilt test in healthy subjects.

AIM: To evaluate the ocular outcomes and to elucidate possible mechanisms underlying intraocular pressure (IOP) change following the head-down tilt (HDT) test. METHODS: The study included 21 participants at the Department of Ophthalmology of Tongji Hospital. Subjects received the test of I-care tonometry, enhanced depth imaging optical coherence tomography and heart rate variability (HRV) analysis before and after 15min HDT test. The lumen area of Schlemm's canal (SCAR), IOP, HRV were calculated. RESULTS: IOP increased significantly after 20° head down position from 14.0±3.0 to 17.0±3.3 mm Hg (P<0.001). SCAR decreased from 13449.0±5454.9 µm2 at sitting condition to 9576.6±4130.9 µm2 post 15min HDT test. High frequency (HF) indices increased significantly from 1462±865 Hz at baseline to 2128±824 Hz. Heart rate (HR) decreased significantly from 76±11.48 to 70±11.52 bpm after the HDT. The linear regression analysis showed that the difference of HF and SCAR significantly correlated with each other during the HDT (R2 =20%, P=0.04). CONCLUSION: These outcomes perform the first evidence of the activation of autonomic nervous system of HDT may cause the collapse of Schlemm's canal lumen, which in turn leading to the increased IOP.

Numerical simulation of the flow of aqueous humor in the Schlemm's canal.

Schlemm's canal (SC) is a ring structure responsible for maintaining fluid homeostasis in the anterior chamber of the eye by draining aqueous humor (AH) from the trabecular meshwork (TM) into the collecting channel (CC). Obstruction of the AH flow in this conventional pathway leads to an elevation of intraocular pressure in glaucoma, which is internationally recognized as the most common cause of blindness. Although there are obvious morphological differences between patients and healthy individuals, the AH drainage capacity of SC with specific parameters is difficult to quantitatively evaluate due to the limitations of analyses or experimental technologies. In this paper, the AH flow dynamic features under various SC morphological parameters are studied by numerical simulations of 3D models. The axis length of the SC cross-section and the level of openness are chosen as the main morphological factors according to the clinical data. The flow features, including the velocity, streamline and wall shear stress, are analyzed. This study aimed not only to explore the feasibility of 3D numerical modeling in evaluating the SC drainage capacity, but also to reveal the relationship between SC morphological parameters and the dynamic features of AH drainage.

Correlates of Physical Activity in Colorectal Cancer Patients Based on Health Promotion Model.

BACKGROUND: Increasing attention is being given to physical activity in colorectal cancer patients. Some studies have explored a few correlates of physical activity separately. A contemporary study based on the Health Promotion Model may systematically broaden the understanding of physical activity in colorectal cancer patients. OBJECTIVE: To understand the status of physical activity in Chinese colorectal cancer patients and to explore the correlated factors. METHODS: A total of 168 adults with colorectal cancer were recruited at 3 tertiary hospitals in China. Participant data were collected on demographics, physical activity, biological factors, anxiety and depression, benefits/barriers to physical activity, self-efficacy, and social support. SAS 8.2 was used for statistical analysis, including descriptive analysis, correlation analysis, single factor analysis, and multiple stepwise regression analysis. RESULTS: Only 25.60% of colorectal cancer survivors reached the requirements of the Colorectal Cancer Survivorship Care Guidelines. Employment states, number of complications, fatigue, body image, depression, perception of benefits/barriers, and self-efficacy were closely correlated with physical activity in Chinese colorectal cancer patients. CONCLUSIONS: The physical activity status of patients with colorectal cancer is not optimal. The correlated factors that nurses can take measures to improve are fatigue, body image, depression, perception of benefits/barriers, and self-efficacy, which may improve physical activity in colorectal cancer patients in China. Additional research is needed to determine if improving factors correlated with physical activity will assist with directly increasing physical activity. IMPLICATIONS FOR PRACTICE: Nurses should evaluate physical activity of colorectal cancer patients timely and play an active role in health promotion programs to improve colorectal cancer patients' physical activity.

[Diagnostic Value of MPV, PDW, PAIg and Their Combination for Megakaryocyte Dysmaturity in Children with Acute Immune Thrombocytopenic Purpura].

OBJECTIVE: To investigate the changes of mean platelet volume (MPV), platelet distribution width (PDW) and platelet associated antibodies (PAIg) in children with acute immune thrombocytopenic purpura (aITP), and to explore the diagnostic value of MPV, PDW, PAIg and their combination for megakaryocyte dysmaturity in aITP children. METHODS: Plt count, MPV and PDW of 36 aITP children were measured by using Sysmex XN automatic blood cell analyzer, and 33 children with acquired thrombocytopenic purpura (ATP) without megakaryocyte dysmaturity. The expression of PAIg was detected by flow cytometry, and the number and classification of megakaryocytes in the bone marrow were performed by marrow cytology. The diagnostic significances of MPV, PDW, PAIg and their combination as well as the sensitivity and specificity for megakaryocytes dysmaturity in aITP were assessed through calculating the area under ROC curve (AUC), after determining the influence of each parameters on the megakaryocyte dysmaturity by Logistic regression. RESULTS: MPV, PDW and PAIg of aITP children were significantly higher than those of the ATP children (P＜0.05), while the Plt count and number of thromocytogenic megakaryocytes per area (1.5 cm×3 cm) were less than those of the controls (P＜0.05). Count of RBC and WBC, percentages of neutrophil granulocytes and lymphocydes in aITP were similar to those in the controls（P＞0.05）. The results of Logistic regression showed that Plt count, MPV, PDW and PAIg were the factors influencing megakaryocyte dysmaturity in aITP children, and the regression model has a high statistical significance (χ2=65.491，P=0.001) and r square (R2=0.713). The AUC of the combined detection of Plt count, MPV, PDW and PAIg was 0.863, which was much higher than that of Plt count, MPV, PDW, PAIg individually or in pairs. The sensitivity and specificity of the combined detection were 79.167% and 89.697%, which were higher than those of Plt count, MPV, PDW, PAIg individually or in pairs. CONCLUSION: The diagnostic significance of MPV and PDW for megakaryocyte dysmaturity in aITP are insufficient, but the diagnostic efficacy can be improved by combined examination with PAIg.

NaBr-Induced Production of Brominated Azaphilones and Related Tricyclic Polyketides by the Marine-Derived Fungus Penicillium janthinellum HK1-6.

Cultivation of the mangrove rhizosphere soil-derived fungus Penicillium janthinellum HK1-6 with NaBr led to the isolation of two new brominated azaphilones, penicilones G and H (5, 6), two new tricyclic polyketides, penijanthinones A and B (7, 8), and two known azaphilones, penicilones A and B (1, 2). The planar structures and relative configurations of the new compounds were elucidated using comprehensive spectroscopic methods including 1D and 2D NOE spectra. Their absolute configurations were determined by chemical conversions, TDDFT ECD calculations, and comparisons of their ECD spectra. Interestingly, the NaBr-induced brominated azaphilones (5, 6) had the opposite configuration at C-7 compared to the chloro analogues (3, 4) produced by this fungus cultivated with sea salt. Ester hydrolysis of penicilone B (2) afforded the carboxylic acid side chain 2,4-dimethyldec-2-enoic acid (9), with a 4 S configuration assigned by its specific rotation. Penicilone H (6) showed antibacterial activity with MIC values ranging from 3.13 to 12.5 µg/mL.

Penicilones A-D, Anti-MRSA Azaphilones from the Marine-Derived Fungus Penicillium janthinellum HK1-6.

Four new azaphilones, penicilones A-D (1-4), were isolated from the mangrove rhizosphere soil-derived fungus Penicillium janthinellum HK1-6. Their planar structures and absolute configurations were determined by extensive analysis of NMR spectroscopic data, ECD spectra, the modified Mosher's method, and chemical conversions. Interestingly, 1 and 2 had the opposite configuration at C-7 compared to the closely related chloro analogues 3 and 4. Ester hydrolysis of 2 and 4 afforded their parental azaphilones, named penicilones E (5) and F (6). Compounds 1-6 were evaluated for their antibacterial activities in vitro. Penicilones B-D (2-4) showed potent anti-MRSA (Staphylococcus aureus ATCC 43300, ATCC 33591) activities with MIC values ranging from 3.13 to 6.25 µg/mL.

Abnormal expression of p190RhoGAP in colorectal cancer patients with poor survival.

PURPOSE: This study aims to investigate the expression and clinical significance of p190RhoGAP, a member of the RhoGAP family, in colorectal cancer (CRC). METHODS: The expression p190RhoGAP was detected by RT-PCR, western blot (WB) and immunohistochemistry (IHC) in 14 paired CRCs and matched non-cancerous mucosal tissues. The protein content of p190RhoGAP was identified in 114 CRCs by IHC. In addition, the association of the expression of p190RhoGAP with carcinogenesis, distant metastasis and prognosis was further evaluated. RESULTS: In 14 paired fresh tissues, the mRNA (P<0.0001) and protein (P = 0.003) expression levels of p190RhoGAP were significantly higher in primary CRCs than in paired non-cancerous mucosal tissues; and was consistent with WB results. The expression of p190RhoGAP increased from normal mucosa to adenoma, and became even greater in primary carcinoma (P = 0.001). The expression level of p190RhoGAP was highest in liver metastasis compared to primary carcinoma (P = 0.028). The incidence of p190RhoGAP expression-positive cases was 58.77% in 114 CRC tissues. Furthermore, the enhanced expression of p190RhoGAP was significantly associated with shorter disease-specific survival (P<0.001) and shorter disease-free survival (P<0.001). Cox regression analysis indicated that p190RhoGAP was an independent prognostic parameter for CRC. CONCLUSION: p190RhoGAP may be an independent predictive factor for the prognosis of CRC, and the abnormal expression of p190RhoGAP may play a crucial role in colorectal carcinogenesis and distant metastasis.

Ocular biometry in the adult population in rural central China: a population-based, cross-sectional study.

AIM: To describe the distribution and determinants of ocular biometric parameters and to ascertain the relative importance of these determinants in a large population of adults in rural central China. METHODS: A population-based, cross-sectional study performed in rural central China included 1721 participants aged 40 or more years. Ocular biometrical parameters including axial length (AL), anterior chamber depth (ACD), radius of corneal curvature (K) and horizontal corneal diameter [white-to-white (WTW) distance] were measured using non-contact partial coherence interferometry [intraocular lens (IOL)-Master]. RESULTS: Ocular biometric data on 1721 participants with a average age of 57.0±8.7y were analyzed at last. The general mean AL, ACD, mean corneal curvature radius (MCR), WTW were 22.80±1.12, 2.96±0.36, 7.56±0.26 and 11.75±0.40 mm, respectively. The mean values of each parameter in 40 to 49, 50 to 59, 60 to 69, and 70 to 91 years age groups were as follows: AL, 22.77±0.87, 22.76±1.06, 22.89±1.41, 22.92±0.80 mm; ACD, 3.10±0.32, 2.98±0.34, 2.86±0.36, 2.77±0.35 mm; MCR, 7.58±0.25, 7.54±0.26, 7.55±0.26, 7.49±0.28 mm; WTW, 11.79±0.38, 11.75±0.40, 11.72±0.41, 11.67±0.41 mm. The AL, ACD, MCR and WTW were correlated with age and the AL was correlated with height and weight. CONCLUSION: Our findings can serve as an important normative reference for multiple purposes and may help to improve the quality of rural eye care.

Glutamine-supplemented parenteral nutrition and probiotics in four adult autoimmune enteropathy patients.

To evaluate the effects of glutamine-supplemented parenteral nutrition (PN) and probiotics in adult autoimmune enteropathy (AIE) patients. Four adult AIE patients were identified from April 2006 to January 2012. Clinical and nutritional data were obtained from the patients' medical records. Glutamine-supplemented PN started immediately when the AIE diagnosis was confirmed. The total PN duration was 351 days. According to the PN prescription, the average caloric intake ranged from 20 to 25 kcal/kg/day, and the protein intake ranged from 1.2 to 1.5 g/kg/day. Alanyl-glutamine (20 g/day) was administered to AIE patients for 4 weeks followed by a 2-week break, and this treatment schedule was repeated when PN lasted for more than 6 weeks. Body weight gain and an increased serum albumin level were achieved after PN, and defecation frequency and quality also improved. Each patient received oral supplements, 250 mL of Ensure and two probiotics capsules (each capsule containing 0.5×108 colonies) three times a day when enteral nutrition started. Three AIE patients were successfully weaned off PN, and one patient died of pneumonia. Glutamine-supplemented PN and probiotics show promise in managing patients with AIE and related malnutrition.

Expression of endoplasmic reticulum stress-related factors in the retinas of diabetic rats.

Recent reports show that ER stress plays an important role in diabetic retinopathy (DR), but ER stress is a complicated process involving a network of signaling pathways and hundreds of factors, What factors involved in DR are not yet understood. We selected 89 ER stress factors from more than 200, A rat diabetes model was established by intraperitoneal injection of streptozotocin (STZ). The expression of 89 ER stress-related factors was found in the retinas of diabetic rats, at both 1- and 3-months after development of diabetes, by quantitative real-time polymerase chain reaction arrays. There were significant changes in expression levels of 13 and 12 ER stress-related factors in the diabetic rat retinas in the first and third month after the development of diabetes, Based on the array results, homocysteine- inducible, endoplasmic reticulum stress-inducible, ubiquitin-like domain member 1(HERP), and synoviolin(HRD1) were studied further by immunofluorescence and Western blot. Immunofluorescence and Western blot analyses showed that the expression of HERP was reduced in the retinas of diabetic rats in first and third month. The expression of Hrd1 did not change significantly in the retinas of diabetic rats in the first month but was reduced in the third month.

[Functional magnetic resonance imaging in glaucoma].

Neurological damage in glaucoma does not only limit to the eye, but also affects the central nervous system of the visual pathway. Functional magnetic resonance imaging provides a non-invasive method that can help to assess anatomical and functional changes of the central nervous system. This technology has been used in the investigations of nerve fiber damage, metabolic changes in visual cortex, and dysfunction of the visual system in glaucoma. Functional magnetic resonance imaging is introduced, and the progress in its application and prospect in glaucoma diagnosis were summarized in this paper.

Functional characterization of 58-kilodalton inhibitor of protein kinase in protecting against diabetic retinopathy via the endoplasmic reticulum stress pathway.

OBJECTIVE: 58-kilodalton inhibitor of protein kinase (P58(IPK)) plays an important role in preventing endoplasmic reticulum (ER) stress. It is an interferon-induced kinase that targets the eukaryotic translation initiation factor eukaryotic initiation factor 2 alpha. The aim of this study was to determine the roles of P58(IPK) in protecting against diabetic retinopathy (DR) by inhibiting ER stress-signaling mediators. METHODS: A rat diabetic model was established by intraperitoneal injection of streptozotocin. Overexpression of P58(IPK) was achieved by intravitreal injection of purified recombinant adeno-associated virus vector (rAAV2)-P58(IPK) or transfection into rat retinal capillary endothelial cells. Retinal vascular permeability was determined by assessing the Evans Blue retinal leakage. To downregulate the P58(IPK) level in cultured rat retinal capillary endothelial cells, pGIPZ-P58(IPK) RNA interference (P58(IPK)RNAi) was introduced in these cells. Real time reverse transcription (RT)-PCR and western blot analyses were performed to evaluate the mRNA and protein levels of Core/emopamil binding protein (C/EBP) homologous protein (CHOP), vascular endothelial growth factor (VEGF), and tumor necrosis factor-α (TNF-α). RESULTS: Retinal blood vessel leakage was significantly decreased in rAAV2-P58(IPK)-transfected diabetic rats compared with the control diabetic rats. Both mRNA and protein levels of CHOP, TNF-α, and VEGF in the retina of diabetic rats were remarkably reduced in P58(IPK)-transfected rats. In vitro study further demonstrated that overexpression of P58(IPK) downregulated the expression of CHOP, TNF-α, and VEGF under high glucose conditions, whereas introduction of P58(IPK)RNAi enhanced the expression of CHOP, TNF-α, and VEGF. CONCLUSIONS: These results revealed the protecting role of P58(IPK) against ER stress-mediated DR in diabetic rats, suggesting that P58(IPK) may act as a DR-resistant gene during diabetes.

Effects of endothelin-1 on the cytoskeleton protein F-actin of human trabecular meshwork cells in vitro.

AIM: To observe the effect of endothelin-1 (ET-1) on the cytoskeleton protein F-actin of cultured human trabecular meshwork (HTM) cells. METHODS: CULTURED HTM CELLS WERE RANDOMLY DIVIDED INTO FOUR GROUPS: control group, low-dose ET-1 (10(-9) mol/L) treatment group, middle-dose ET-1 (10(-8) mol/L) treatment group, and high-dose ET-1(10(-7) mol/L) treatment group. After treated with ET-1, the expression of cytoskeleton protein F-actin in trabecular meshwork was analyzed with Western-blot and the distribution of F-actin was detected with FITC-Phalloidin probe. RESULTS: ET-1 dose-dependently and significantly increased F-actin in trabecular meshwork cells (P<0.05). The F-actin stress fiber and periphery actin fiber highly increased and manifested mild reorganization after treated with ET-1; and there were much more cell-to-cell and cell-to-extracellular matrix attachments formation in ET-1 treated HTM cells than that in the untreated HTM cells. CONCLUSION: ET-1 promotes the expression of cytoske- leton protein F-actin and induced the trabecular meshwork actin cytoskeleton reorganization.

The influence of He-Ne laser on scar formation after trabeculectomy in rabbits.

AIM: To investigate the influence of He-Ne lasers on scar formation in the filtration canal after trabeculectomy in a rabbit model, as well as to explore the mechanisms for preventing scar formation when using He-Ne lasers in vivo. METHODS: Experiment 1: Four groups were established (four eyes in each group). In 12 eyes, the upper nasal limbus area next to the upper rectus muscle received 10 minutes of He-Ne laser irradiation (100, 150, 200mW/cm(2); 60, 90, 120J/cm(2)) every day for three days. Four eyes served as controls. Twenty-four hours after the final irradiation, the rabbits were sacrificed and the irradiated tissue was excised, fixed with paraformaldehyde and tested for proliferating cell nuclear antigen (PCNA), connective tissue growth factor (CTGF) and apoptosis (TUNEL). Experiment 2: Forty-two rabbits were randomly divided into two groups and standard trabeculectomy was performed in the right eyes either after 200mW/cm(2) He-Ne laser irradiation or not in the filtration area. The expression of PCNA and CTGF, apoptosis and collagen density in the filtration area were tested on the 7(th), 14(th) and 28(th) day after surgery. RESULTS: Experiment 1: There were no more PCNA and CTGF positive cells in the He-Ne irradiation group than in the control group. No apoptotic cells were found in either group. Experiment 2: The expression of PCNA and CTGF was lower in the He-Ne irradiation group than in the control group on the 7(th) and 14(th) day after trabeculectomy surgery (P<0.05); no apoptotic cells were detected in either group. Collagen density was significantly lower in the He-Ne irradiation group than in the control group on the 14(th) and 28(th) day after surgery (P<0.05). CONCLUSION: Pretreating the filtration area with 200mW/cm(2) (120J/cm(2)) of He-Ne laser irradiation may be helpful in preventing scar formation after trabeculectomy, possibly due to the downregulation of the expression of PCNA, CTGF and collagen synthesis in fibroblasts.