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Objective To investigate the influence of Nei endonuclease Ⅷ-like protein 1(NEIL1)on H2O2-induced apoptosis and autophagy in lens epithelial cells(LECs).Methods Reverse transcription polymerase chain reaction(RT-PCR)was used to detect the expression of NEIL1 messenger ribonucleic acid(mRNA)in the anterior lens capsule and LEC line SRA01/04 cells from age-related cataract(ARC)patients and macular epiretinal membrane patients who underwent the clear lens extraction.SRA01/04 cells were then divided into the control group,OE-Vector group and OE-NEIL1 group.RT-PCR and Western blot(WB)were used to detect the overexpression efficiency.Subsequently,the cells were divided into the control group,H2O2 group,OE-Vector H2O2 group and OE-NEIL1 H2O2 group;cell viability was detected by Cell Count-ing Kit-8,WB was used to evaluate the expression of autophagy-related proteins(ATG7,P62,Beclinl,LC3-Ⅰ and LC3-Ⅱ)and apoptosis-related proteins(Bax and Bcl-2),and fluorescent staining was adopted to measure cell apoptosis and mito-chondrial membrane potential.Results The expression of NEIL1 mRNA in the anterior lens capsule samples from ARC patients was significantly lower than that of macular epiretinal membrane patients,and the expression of NEIL1 mRNA in the SRA01/04 cells in the H2O2 group was also lower than that in the control group(both P<0.05).The results of RT-PCR and WB revealed that the expressions of NEIL1 mRNA and protein in the SRA01/04 cells of the OE-NEIL1 group were signif-icantly higher than those in the OE-Vector group(both P=0.000).Compared with the control group,the viability of SRA01/04 cells in the H2O2 group decreased,the expression of the Bax protein was up-regulated,the Bcl-2 protein was down-regulated,viable cells labeled with Mito-Tracker decreased,and apoptotic cells labeled with Annexin V-FITC in-creased,with statistical significances(all P<0.05).Compared with the OE-Vector H2O2 group,in the OE-NEIL1 H2O2 group,the viability of SRA01/04 cells significantly improved,the autophagy-related proteins(ATG7,P62,Beclin1,LC3-Ⅰand LC3-Ⅱ)and Bcl-2 protein were significantly up-regulated,the Bax protein was down-regulated,viable cells marked with Mito-Tracker increased,and apoptotic cells labeled with Annexin V-FITC decreased(all P<0.05).Conclusion Under the oxidative stress induced by H2O2,NEIL1 can promote autophagy of LECs,maintain homeostasis of LECs,and then increase cell viability and reduce apoptosis of LECs,participating in the occurrence and development of ARC.
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Objective To evaluated the prevalence of pterygium among people aged ≥40 in China by using Meta analysis.Methods A literature retrieval was performed in China biology medicine disc (CBMdisc),China national knowledge infrastructure (CNKI),VIP database,WanFang database,PubMed,excerpta medica database (EMBASE),Web of Science from 1990 to 2016 according to designed searching strategy.Tailored quality evaluation standard of epidemiological research was used to evaluate the study quality of each included study.Stata 13.0 software was used to estimate the prevalence of pterygium,subgroup analysis of age,sex and region was also carried out.Results Thirty-six studies were involved in this Meta analysis,including 8 English researches and 28 Chinese researches.The random effect model was used to merge 36 research data.The prevalence of pterygium among people aged over 40 in China was 13.4% [95% confidence interval (CI):10.6%-16.5%].Subgroup analysis results showed that the prevalence of pterygium increased with age.The prevalence of pterygium in rural region was 15.3% (95% CI:12.1%-18.8%),which was higher than 4.0% (95% CI:2.4%-6.1%) in urban region.The prevalence of pterygium among rural people aged ≥ 40 was 12.1% (95% CI:8.6%-16.2%),which was lower than 14.7% (95% CI:10.5%-19.5%) among rural people aged ≥50.The highest prevalence of pterygium in different age and sex subgroups was in the west region of China,followed by the east region of China and the lowest was in the central region of China.Conclusion The prevalence of pterygium among people aged ≥ 40 in China was high.Over the past decade,the prevalence of pterygium were different among different ages,and places of residence.
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Objective To investigate the prevalence and associated risk factors for pterygium among people aged 50 years and above in Funing County,Jiangsu Province.Methods A cluster random sampling method was performed,the subjects aged 50 years or above were randomly selected from 30 survey sites in Funing County,Jiangsu Province.Questionnaires,visual acuity tests,the examinations of eye surface,anterior segment,fundus examinations were conducted.Pterygium was diagnosed and graded clinically by slit lamp examination.The risk factors were acquired from questionnaires and analyzed by the multivariate logistic regression analysis.This study protocol was approved by Ethic Committee of Affiliated Hospital of Nantong University (NO.2010-05).Written informed consent was obtained from each subject prior to entering study cohort.Results A total of 6 145 persons aged 50 years and above were enumerated,and actually 5 947 (96.8%) participants were examined.Among them,1 950 cases were diagnosed as pterygium in either eye and 1 228 cases were diagnosed as pterygium in binoculus,which was equivalent to the 32.79% [95% confidence interval(CI):31.60%-33.98%] of pterygium in either eye and 20.65% (95% CI:19.62%-21.68%) in bilateral pterygium.Among 2467 male subjects,838 were diagnosed as pterygium (33.97%,95% CI:32.10%-35.84%).Among 3480 female subjects,1 112 were diagnosed as pterygium (31.95%,95% CI:30.40%-33.51%).There was no significant difference in the prevalence of pterygium between genders (P =0.135).Multivariate Logistic analysis showed that,older age (50 ~ <60 years:odds ratio [OR] =1.00;60 ~ <70 years:OR=1.54,P<0.001;70 ~ <80 years,OR=1.83,P<0.001;≥80 years:OR=1.99,P<0.001),low educational level (no education:OR =1.00;<primary:OR =0.87,P =0.031;primary education:OR =0.72,P =0.002;≥ secondary education:OR =0.63,P =0.002),farmer occupations (OR =1.34,P =0.020),and long outdoor work time (OR =1.13,P =0.026) were independent risk factors for pterygium.Gender,marriage,income,hypertension,diabetes,smoking and alcohol use history were not associated with pterygium (all at P>0.05).Conclusions The prevalence of pterygium in Funing County is 32.79% in people aged 50 years and above.The high prevalence of pterygium may be associated with older age,low education level and long outdoor work time.
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Long non-coding RNA (lncRNA) widely exists in living organisms.It,whose length is more than 200 bp,can regulate the genic expression in the epigenetic,transcriptional and post-transcriptional level.Therefore,it can influence cell proliferation,apoptosis,vitality,immune response and oxidative stress.In recent years,the study of lncRNA is developing rapidly in the ophthalmic research,and this article reviews the latest research progress on mechanism of lncRNA in the ophthalmology disease.
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Background Researches showed that triterpenoids,with a similar structure to lanosterol,has therapeutical effect on many systemic diseases,and lanosterol was determined to have a therapeutical effect on cataract recently.However,how the lanosterol plays effects on other eye diseases is still unelucidated.Understanding the distribution of lanosterol in ocular tissue is helpful for us to elucidate the relationship of lanosterol with eye diseases.Objective This study attempted to investigate the distribution of lanosterol synthase (LSS) and lanosterol in cornea,lens and retina tissue of rats and offer a basis for the targeting treatment of eye diseases.Methods Fifteen SPF male SD rats were sacrificed by excessive anesthesia to obtain the eyeballs.The relative expressions of LSS protein and gene in the cornea,lens and retina tissue of the rats were detected by Western blot and reverse transcription (RT)-PCR,respectively.Immunofluorescence staining technology was used to locate the distribution of LSS in cornea,lens and retina tissue.The contents of lanosterol in the cornea,lens and retina tissue were analyzed by liquid chromatograph mass spectrometer (LC-MS).Results No LSS protein and mRNA was expressed in the retinal tissue in normal rats.The mean relative expression of LSS protein in the lens and cornea was 0.43±0.05 and 0.25±0.03,respectively,showing a significant difference between them (t =-5.35,P< 0.01).The relative expression of LSS mRNA was 0.51 ±0.04 and 0.29 ±0.02 in the lens and cornea,respectively,with a stronger expression in the lens in comparison with the cornea (t =-8.34,P<0.01).Immunofluorescence staining showed that LSS primarily located in corneal epithelial layer,stromal layer and endothelial layer as well as lens epithelial cells and shallow cortex layer and hardly expressed in retina,and no co-expression of LSS with the neuron marked by NeuN and the Müller cell marked by glutamine synthetase (GS) in retinal tissue.LC-MS analysis revealed that the contents of lanosterol in lens and cornea was (24.37 ±2.91) ng/mg and (5.31 ±0.58) ng/mg,respectively,with a significant difference between them (t =-11.13,P<0.01).Conclusions LSS and lanosterol extensively distribute in cornea and lens of normal rats,but not in retina tissue.These results offer new strategies for the target treatment of relevant eye diseases.
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<p><b>OBJECTIVE</b>To explore the effect of 20 (S)-ginsenoside Rg3 [20 (S)-Rg3] on the proliferation inhibition and secretion of vascular endothelial growth factor (VEGF) of multiple myeloma (MM) cell line U266.</p><p><b>METHODS</b>The proliferation inhibition rate of U266 cells after treatment with different doses of 20 (S)-Rg3 was detected by MTT method, the cell cycle and apoptosis by flow cytometry, the expression of apoptosis related proteins of caspase-3, 8 and 9 by Western blot, VEGF concentration in the culture supernatant by ELISA.</p><p><b>RESULTS</b>It showed that 20 (S)-Rg3 could inhibit the proliferation of U266 in a dose-dependent manner (P<0.05) with IC50 of (71.07 ± 2.63)μmol/L and (44.06 ± 3.98) μmol/L at 24 h and 48 h, respectively. VEGF concentration in the culture supernatant showed a dosedependent reduction (P<0.05), decreased from (419.93 ± 36.76) pg/106 cells in the control group to (314.82 ± 27.05) pg/106 cells in 80 μmol/L 20 (S)-Rg3 treated group by ELISA assay. Flow cytometry with Annexin-V/PI double staining revealed that 20(S)-Rg3 may induce U266 cells apoptosis in a concentration-dependent manner from (0.51 ± 0.05)% at control group to (8.32 ± 0.83)%, (10.72 ± 1.29)% and (15.27 ± 2.26)% at 20, 40 and 80 μmol/L treatment groups, respectively (P<0.05). Flow cytometry with PI staining showed that the ratio of cells in G0/G1 phase increased from (49.11 ± 1.71)% to (52.72 ± 7.75)%, (60.29 ± 5.76)% and (61.81 ± 3.46)%, respectively (P<0.05). Western blot analysis indicated that the expression of caspase-3, 8 and 9 declined, and that of cleaved-caspase-3, 8 and 9 significantly increased (P<0.05) with 20 (S)-Rg3 concentration increased.</p><p><b>CONCLUSION</b>20(S)-Rg3 can inhibit the proliferation of U266 cells by cell cycle arrest in G1 phase and induce cell apoptosis by increasing the expressions of cleaved-caspase-3, -8 and -9. It can also inhibit VEGF secretion of U266 cells, which makes it a potential agent for multiple myeloma therapy.</p>
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Humans , Apoptosis , Caspases , Metabolism , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation , Ginsenosides , Pharmacology , Multiple Myeloma , Metabolism , Pathology , Vascular Endothelial Growth Factor A , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To analyze the characteristics of lymph node metastasis and prognosis in patients with T1 breast cancer.</p><p><b>METHODS</b>The clinicopathological data of 354 patients with T1 breast cancer after standard treatment from March 2007 to September 2011 were collected to analyze the relationship between the clinical characteristics of T1 breast cancer, lymph node metastasis and prognostic features.</p><p><b>RESULTS</b>In the 354 patients with T1 breast cancer, 105 patients (29.7%) had lymph node metastasis, among them 73 cases (69.5%) had 1-3 lymph node metastasis, and 32 cases (30.5%) had more than 4 lymph node metastasis. The lymph node metastasis rate was 8.3% in T1a patients, 39.7% in T1b patients, and 30.4% in T1c cases (P = 0.005). Pairwise comparison showed that the difference of lymph node metastasis rate between T1a, T1b and T1c patients was statistically significant (P = 0.001 and P = 0.006, respectively). The difference of lymph node metastasis rates in T1b and T1c patients was statistically insignificant (P = 0.171). In the 354 patients of T1 breast cancer, 92 patients had vascular tumor thrombi and their lymph node metastasis rate was 71.7%, while the lymph node metastasis rate in 262 patients without vascular tumor thrombus was 14.9% (P < 0.001). The median follow-up was 49 months (range 27-81 months). 12 patients developed recurrence, and 3 patients died, one of them died of cerebrovascular accident. The 4-year disease-free survival for all patients was 96.6%, and the 4-year overall survival rate was 99.2%.</p><p><b>CONCLUSIONS</b>There is a correlation between vascular tumor thrombus, tumor size and lymph node metastasis rate. The lymph node metastasis rate is lower in T1a patients and relatively higher in T1b/c patients. Compared with patients without vascular tumor thrombus, the T1 breast cancer patients with vascular tumor thrombi have a higher lymph node metastasis rate. Generally speaking, there is a still good prognosis in patients with T1 breast cancer.</p>
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Adult , Aged , Female , Humans , Middle Aged , Young Adult , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Breast Neoplasms , Drug Therapy , Pathology , General Surgery , Carcinoma, Ductal, Breast , Drug Therapy , Pathology , General Surgery , Carcinoma, Lobular , Drug Therapy , Pathology , General Surgery , Disease-Free Survival , Follow-Up Studies , Lymphatic Metastasis , Mastectomy, Radical , Neoplasm Recurrence, Local , Neoplasm Staging , Neoplastic Cells, Circulating , Prognosis , Survival RateABSTRACT
Objective To evaluate the efficacy and safety of bortezomib-based chemotherapy and MPT regimen in the MM patients who were newly diagnosed or relapsed/refractory. Methods Twenty-seven MM patients were treated with bortezomib-based chemotherapy, median cycles:3 (range 1-5 cycles). Other 30patients received MPT chemotherapy. EBMT and WHO criteria were used to evaluate the therapeutic effects and the adverse effects, respectively. Results Bortezomib group: 21 patients (77.8 %) showed effects after the first cycle chemotherapy and 24 patients (88.8 %) showed effects after the whole therapy. In wich, 15 patients(94.0 %) and 9 patients (82.0 %) were newly diagnosed and relapsed/refractory, respectively. MPT group: 15patients (50.0 %) showed effects after the whole therapy. In wich, 12 patients (44.0 %) were newly diagnosed.And the other 3 were relapsed/refractory patients. The ORR in Bortezomib group was better than MPT group (P <0.05). The incidence of peripheral neuropathy, herpes and Ⅲ - Ⅳ grade thrombocytopenia in the bortezomib group was 10 patients (37.0 %), 7patients (26.0 %), 10 patients (37.0 %) respectively,and they were more common than MPT group, but the incidence of Ⅲ-Ⅳgrade anemia was 21 patients (70.0 %) and more comumom in the MPT group. The theraputic efficacy of bortezomib for renal insufficiency and normal renal function patients was similar, and no significant increase in all kinds of adverse effects. In MPT group,there were 4 patients with renal insufficiency, the serum level of creatinine in the 3 patients returned to normal after 5 cycles therapy. Conclusion Bortezomib-based chemotherapy is more effective than MPT regimen in the treatment of MM. The newly diagnosed, relapsed/ refractory and with renal insufficiency patients all can benefit from it. The adverse effects are mild and with better tolerance.
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Objective To identify the clinical phenotypic diagnosis and gene mutation detection of two kindreds with PS deficiency. MethodsPS: A was measured by chromogenic substrate method;TPS:Ag, FPS: Ag levels were measured by ELISA method; PS gene(PROS1 gene)was detected by amplifying 15 exons and flanking intron sequences from the propositus with PCR method. PCR products were purified and directly sequenced. Results For propositus 1,PS: A was 48.6% ,TPS: Ag was 136 mg/L, FPS : Ag was 41 mg/L, PROSI gene exon 2 was in c. Heterozygous base substitutions was detected in C121T locus, which led to Arg-1Cys (R-1C) heterozygous roissense mutation encoded in PS proteins. For propositus 2, PS: A was 29.2%, TPS: Ag was 83 mg/L, FPS: Ag was 26 mg/L, PROSI gene exon 14 was in c. Heterozygous base substitutions was identified in CI687T locus, in which Gln.522Stop heterozygous nonsense mutation was encoded in PS proteins. Conclusions c. C121T is a novel mutation locus detected in PROS1 gene. This heterozygous mutation could lead to type Ⅱ PS hereditary deficiency, while c. C1687T heterozygous mutation could bring about type Ⅰ PS hereditary deficiency.
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The genetic indentification of 16S rDNA or ITS, capability of phosphate-solubilization and pH of medium, and optimization of medium of some microorganism isolated from mangrove were investigated in this study. The result showed that the fungi normally had much higher capacity to dissolve the inorganic phosphate than the bacteria, the capacity of the fungi was closely correlated to the pH of medium, but the relationship was weak for the bacteria. It was illustrated by single factor experiments that the optimal carbon and nitrogen sources were maltose and urea respectively. The orthogonal design was employed in testing the optimum composition of medium composed of 5 g/L maltose, 0.05 g/L urea, 5 g/L NaCl, pH 5. In this optimal medium, the effectively enrichment of bacteria could reach up to 6.06?109 CFU/mL under 30?C for 48 hours cultivation.
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Objective To set up an effective and low-price way for enriching dendritic cells precursor from chronic myelogeous leukemia by Percoll density gradients.Methods Peripheral blood mononuclear cells(PBMCs) were collected by separation through Ficoll-Hypaque,then PBMCs were separated by 55% Percoll gradients. The cell type and DCs expressing CD1a,CD86 were detected in the high-density group(C),low-density group(B) and non-Percoll separation group(A).Results After separation of 55% Percoll,the percentage of promyelocytes and myelocytes in group B was obviously higher than those in group A and group C(P
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The interaction of Klebsiella mytoca NG13/pMC73A with Eucalyptus were studied. The results showed that the root surface and even the inner cortex of Eucalyptus were colonized by K. oxytoca. The K. oxytoca was reisolated from rhizosphere, root surface and inner root of inoculated Eucalyptus. The inoculation with K. oxytoca stimulated the excretion of Eucalyptus root and affected the contents of amino acids, carbohydrates, phytohormones of root exudates. The seedlings of Eucalyptusto to inoculation with K. oxytoca increased growth, total dry matter and N content by 29.81% - 100.40% after 3 months in comparinson to the uninoculated seedlings.