[Quantification of optical coherence tomographic angiography images in type 2 diabetic patients].

Objective: To quantify optical coherence tomographic angiography (OCTA) images and to explore potential clinical values of each parameter in diabetic patients. Methods: A case-control and cross-sectional study of diabetic (129 eyes) and healthy (32 eyes) subjects was conducted. 3-mm OCTA scan, fundus photography and best corrected visual acuity measurement were performed. Image-pro plus was used to calculate microvascular and foveal avascular indices in nonsegmented and segmented OCTA images. Intraclass correlation coefficient and relative standard deviation were used to examine the reliability, reproducibility and accuracy. Correlation of each parameter was calculated, and so was the area under the receiver operating characteristic curve. Results: Vessel density (VD) of diabetic patients measured in the nonsegmented retinal layer (RL) and superficial retinal layer (SRL) was 49.146%±6.097%, 44.038%±5.641%, respectively, significantly lower than healthy subjects 52.212%±6.250%, 46.698%±5.417% (t=2.534, 2.405; P=0.012, 0.017), and vessel length (VL) was (19.905±2.285)mm, (17.596±2.149)mm, (14.479±2.091)mm in the three layers, which was significantly shorter than that in the healthy control group (21.037±2.185)mm, (18.739±1.994)mm, (15.343±2.266)mm (t=2.529, 2.731, 2.059; P=0.012, 0.007, 0.041). Compared with non-diabetes, foveal avascular zone (FAZ) in diabetes was (0.441±0.167)mm(2), larger compared to health eyes 0.352±0.109mm(2) (t=-2.831, P=0.005). FAZ demonstrated a negative correlation with VD and VL in RL or SRL(r=-0.227, -0.338; P<0.05) (r=-0.241, -0.332; P<0.05), while best corrected visual acuity showed a negative correlation with VL in each layer (r=-0.225, -0.201, -0.250; P=0.01, 0.022, 0.004), and VD in DRL (r=-0.197, P=0.026). All parameters showed high reproducibility between graders(intraclass correlation coefficient>0.965), while relative standard deviation was greater than 3.049%. AR had the highest area under the receiver operating characteristic curve (0.737), compared with AI (0.724). Conclusions: Our study demonstrates good reliability, reproducibility and accuracy of quantification of OCTA images and reveals changes of VD, VL and FAZ in diabetes, which may occur earlier than vision impairment. AR and AL may have great values in prompting early diabetic retinopathy in diabetic patients. (Chin J Ophthalmol, 2019, 55:273-279).

[Clinicopathological features of second primary lung cancer and pulmonary metastasisin patients with breast cancer].

Objective: To compare the clinicopathological characteristics of second primary lung cancer following breast cancer and lung metastases from breast cancer, and then to analyze the risk factors in breast cancer patients with second primary lung tumor. Methods: Clinical data of 55 breast cancer patients with second primary lung tumor and 205 breast cancer patients with solitary pulmonary metastasis in Shandong Cancer Hospital from January 2006 to January 2017 were retrospectively analyzed. The risk factors of primary lung cancer following breast cancer were analyzed using logistic regression model. Results: Second primary lung cancer in patients with first breast cancer accounted for approximately 21.2%(55/260) of pulmonary malignant solitary nodules, and 0.84%(55/6 580) of all breast cancer patients. The median intervals between the diagnosis of second primary lung cancer or lung metastasis and first breast cancer were 52 months and 42 months, respectively. These two groups showed significant difference between age, time interval between diagnoses, breast tumor size, axillary lymph node metastasis, estrogen receptor, molecular subtype (luminal B and triple-negative) and history of radiotherapy (P<0.05 for all). A multivariate logistic regression model confirmed that age (OR=1.088, P<0.001), breast tumor size(OR=0.480, P<0.001), and radiotherapy history (OR=3.460, P=0.004) were all independent factors for second primary lung cancer. Conclusions: For isolated pulmonary nodules in patients with breast cancer, especially for those with elder age, larger tumor size and radiotherapy history, we should distinguish the second primary lung cancer from pulmonary metastasis. The treatment regimen for lung metastasis and primary lung cancer in patients with breast cancer are entirely distinct. The timely histopathology examinations for pulmonary nodes in patients with breast cancer are recommended.

[Effect of autophagy inhibitor combined with EGFR inhibitor on triple-negative breast cancer MDA-MB-468 and MDA-MB-231 cells].

OBJECTIVE: To investigate the effect of combined administration of autophagy inhibitor 3-methyladenine/bafilomycin A1 and EGFR inhibitor gefitinib on triple-negative breast cancer MDA-MB-468, MDA-MB-231 cells and estrogen receptor-positive MCF-7 cells. METHODS: All the cells were treated with 3-methyladenine/bafilomycin A1 and/or gefitinib. The effect of autophagy inhibitor and gefitinib on the cell growth was evaluated by MTT assay. Cell apoptosis was detected by flow cytometry. Western blot analysis was used to determine the alteration of autophagy-related protein (such as LC3) and apoptosis-related proteins (such as caspase-3 and caspase-9). RESULTS: MTT assay showed that the IC50 in the GE+ 3-MA and GE+ BAF groups were (4.1±0.2) µmol/L and (3.8±0.3) µmol/L, significantly lower than that of the gefitinib alone group [(7.0±0.2) µmol/L] in MDA-MB-468 cells (P<0.05). Similarly, the IC50 in the GE+ 3-MA and GE+ BAF groups were (9.7±0.1) µmol/L and (7.7±0.2) µmol/L, significantly lower than that of the gefitinib alone group [(14.7±0.1) µmol/L]in MDA-MB231 cells (P<0.05). The flow cytometry assay revealed that the apoptosis rates of MDA-MB-468 cells in GE, GE+ 3-MA and GE+ BAF groups were (12.43±3.18)%, (23.37±2.71)% and (18.71±2.81)%, respectively. The apoptosis rates of MDA-MB-231 cells of the GE, GE+ 3-MA and GE+ BAF groups were (12.15±1.82)%, (16.94±2.19)% and (33.83±5.92) %, significantly higher than that of the gefitinib alone group (All P<0.05). The apoptosis rates of the MCF-7 cells were not changed significantly among the three groups (P>0.05). Western blot data showed that the expression levels of LC3 and p-Akt were decreased in the combined groups than that of the gefitinib alone group, while the p-PTEN, caspase-3 and caspase-9 were increased. CONCLUSIONS: Autophagy inhibitor may enhance the sensitivity to gefitinib in MDA-MB-468 and MDA-MB-231 cells by activation of the PTEN/P13K/Akt pathway. Apoptosis in MDA-MB-468 and MDA-MB-231 cells might be enhanced by the combination treatment through caspase cascade.

Preparation and characterization of tea polyphenols and vitamin E loaded nanoscale complex liposome.

The purpose of the present work was to prepare nanoscale complex liposome loaded both aqueous soluble drug tea polyphenols (TP) and insoluble drug vitamin E (VE) by reverse-phase evaporation method. The preparation formulation of TP-VE complex liposome was optimized by the orthogonal experiment. The entrapment efficiency of TP was (50.81 +/- 1.91)% while that of VE was (94.05 +/- 3.45)% for the optimal formulation. The results of penetration experiments of TP-VE liposome demonstrated a slight influence of the concentration of TP and the content of cholesterol on the penetration quantity of TP. These results indicate that the complex liposome offers a new approach to entrap aqueous soluble drug and poorly soluble drug, an inner liposome protection, a relatively high drug encapsulation efficiency and a sustained transdermal penetration.

Photoprotection by tocopherol submicron emulsion against UV-mediated damage in HaCaT cells.

alpha-Tocopherol is a lipophilic vitamin E that shows antioxidative, antiaging and antiphotodamage activity. Nanometer biotechnology is more widely used in the entrainment system of drug carriers and the development for new pharmaceutical preparations. Ultraviolet irradiation to human skin in the long term can result in photoaging and photocarcinogenesis. The purpose of this study was to observe the biological features of tocopherol submicron emulsion (vitE SME) and to clarify the roles of vitE SME on UVB-induced photodamage in HaCaT keratinocytes (KC). VitE SME was prepared by high-pressure homogenization and microemulsion technique. HaCaT KC was incubated in the culture medium supplied with 1/200 and 1/400 of VitE SME prior to different dosages of UVB irradiation. The vitamin E amount in the culture medium was measured by high-performance liquid chromatography (HPLC). Cell growth and cellular viability was detected by MTT assay. The amount of vitamin E remaining in the culture medium significantly decreased during the first 8 h, and less than 10% can be detected by the terminal experiment (24 h). No cytotoxicity effect of tocopherol NM on HaCat KC was observed. In contrast to the control group, the cellular viability of VitE SME-treated group increased 44.22% by 24 h. Compared with irradiated groups without VitE SME, cell proliferation decreased by 17.77% and 40.42% when the HaCaT KC was irradiated with 30 mJ/cm(2) and 90 mJ/cm(2) UVB irradiation, respectively. VitE SME has no toxicity to cell culture system and is characterized by stable release and penetration. Pre-incubation with VitE SME can partly reduce UV-induced cell damage, and the photoprotective efficiency to UVB irradiation also shows time dependence.

HUWE1 plays important role in mouse preimplantation embryo development and the dysregulation is associated with poor embryo development in humans.

HUWE1 is a HECT domain containing ubiquitin ligase implicated in neurogenesis, spermatogenesis and cancer development. The purpose of the current study is to investigate the role of HUWE1 in early embryo development. Here we demonstrate that Huwe1 is expressed in both nucleus and cytoplasm of preimplantation mouse embryos as well as gametes. Hypoxia (5% O2) treatment could significantly increase Huwe1 expression during mouse embryo development process. HUWE1 knockdown inhibited normal embryonic development and reduced blastocyst formation, and increased apoptotic cell numbers were observed in the embryos of HUWE1 knockdown group. Human embryo staining result showed that reduced HUWE1 staining was observed in the poor-quality embryos. Furthermore, Western blot result showed that significantly reduced expression of HUWE1 was observed in the villi of miscarriage embryos compared with the normal control, indicating that reduced expression of HUWE1 is related to poor embryo development. Oxidative reagent, H2O2 inhibited HUWE1 expression in human sperm, indicating that HUWE1 expression in sperm is regulated by oxidative stress. In conclusion, these results suggest that HUWE1 protein could contribute to preimplantation embryo development and dysregulated expression of HUWE1 could be related to poor embryo development and miscarriage in IVF clinic.

APP intracellular domain acts as a transcriptional regulator of miR-663 suppressing neuronal differentiation.

Amyloid precursor protein (APP) is best known for its involvement in the pathogenesis of Alzheimer's disease. We have previously demonstrated that APP intracellular domain (AICD) regulates neurogenesis; however, the mechanisms underlying AICD-mediated regulation of neuronal differentiation are not yet fully characterized. Using genome-wide chromatin immunoprecipitation approaches, we found that AICD is specifically recruited to the regulatory regions of several microRNA genes, and acts as a transcriptional regulator for miR-663, miR-3648 and miR-3687 in human neural stem cells. Functional assays show that AICD negatively modulates neuronal differentiation through miR-663, a primate-specific microRNA. Microarray data further demonstrate that miR-663 suppresses the expression of multiple genes implicated in neurogenesis, including FBXL18 and CDK6. Our results indicate that AICD has a novel role in suppression of neuronal differentiation via transcriptional regulation of miR-663 in human neural stem cells.

[Immunological study of inefficiency schizophrenics with deficiency syndrome treated with xin shen ling].

This paper reports 30 cases of chronic schizophrenics with deficiency syndrome who had chronically taken many sorts of neuroleptic medications and other therapies to be ineffective. The 7 immunological functioning markers were detected: phytohemagglutinin (PHA) intradermic test; circulating immune complex (CIC); T, B lymphocytes, null (N), double (D) cell; and large granular lymphocyte (LGL) and to be compared with a control group of 30 healthy individual. The result showed that 6 immunological markers (PHA, CIC and T, B, N and D cell) were significantly different as compared with the control group (P less than 0.01). In order to regulate proportion and function to immune cell, the 30 patients were given to take immuno-modulating herbs (xin shen ling, XSL) during 6 weeks, while their 7 immunological markers were detected before treatment (BT) and after treatment (AT). The results showed that 5 immunological markers (PHA, CIC, T, N, and D cell) of BT were significantly different as compared with that of AT (P less than 0.01). However, the 5 immunological markers (PHA, CIC, and T, N and D cell) of AT were not significantly different as compared with that of the control group (P less than 0.05). The brief psychosis rating scale (BPRS) and nurses observation scale for inpatient evaluation (NOSIE) were used as evaluating changes of clinical symptoms BT and AT. The results showed that BPRS and NOSIE of BT were significantly different as compared with that of AT (P less than 0.05). The clinical efficacious rate was 67%. We have followed up the results for near 3 years which had a relapse of 5 cases of 20 cases be discharged. It seemed that XSL may be one of the preventive relapse agent for these patients.