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Objective To observe the influences of uncoupling protein 2 (UCP-2) rs660339 variants transfection on cell proliferation and apoptosis of human umbilical vein endothelial cell (HUVEC). Methods Two UCP-2 green fluorescent protein (GFP) lentivirus constructs were created with the rs660339 locus carried C or T (UCP-2C or UCP-2T), respectively. HUVEC were cultured after lentiviral infection of UCP-2C or UCP-2T. The expression of UCP-2C or UCP-2T was detected with real time polymerase chain reaction. Cell proliferation and cell apoptosis were compared among negative control (NC) group, UCP-2T group and UCP-2C group using CCK-8 cell viability and flow cytometry. Western blot and immunostaining were employed to examine the expression of Bcl-2 gene. Results The lentivirus constructs were successfully created.>80%of the transfected cells were found to express GFP under fluorescent microscope. The mRNA levels of UCP-2 gene were significantly increased (F=29.183, P=0.001) in the UCP-2T group and UCP-2C group. The CCK-8 assay revealed that on day two (F=15.970, P=0.004), day three (F=16.738, P=0.004), day four (F=5.414, P=0.045) post-infection, UCP-2T and UCP-2C group showed significantly greater proliferation than the NC cells. The apoptotic rate in the UCP-2T and UCP-2C group was significantly lower than NC group (F=277.138, P=0.000), and the apoptotic rate of UCP-2T was significantly lower than that of UCP-2C (P=0.003). The protein levels of Bcl-2 in the UCP-2T and UCP-2C group were significantly greater than that in the NC group (F=425.679, P=0.000), and the Bcl-2 expression of UCP-2T was greater than that of UCP-2C (P=0.002). The Bcl-2 density in the UCP-2T and UCP-2C group were greater than that in the NC group (F=11.827, P=0.008), while there was no difference between UCP-2T and UCP-2C group (P=0.404). Conclusion The variants of UCP-2 rs660339 may influence HUVEC proliferation and apoptosis, and UCP-2T showed a stronger effect of inhibiting apoptosis than UCP-2C.
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Although there are numerous case resource,there still exist a relatively long distance in clinical research in China in comparison with developing countries because of inadequate recognition and funding,which has greatly affected the development of standardization of clinical medical in China for the past few decades.With the deepening of recognition and increasing of special support to clinical medical research in China,the development based on clinic research will face new opportunities.As a result,it is necessary for ophthalmologists to explore clinical issues,leverage from the mega trend of clinical research and deliver high-quality results.To achieve this goal,ophthalmologists will have to constantly update their knowledge base,standardize clinical practice and improve diagnosis methods.So far,challenges persist in clinical research in ophthalmology as follows:It is difficult to get funding for clinical research project;It is difficult to get ethnics approval for clinical research project;The research design for forward-looking projects is yet to be standardized;Cross-disciplinary collaboration needs to be enhanced.This review was expected to inspire more attention from clinical research specialists and consequently more highquality clinical studies.
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As the precision medicine has been proposed , personalized therapy in cancer steps into a new stage.Since the development of gene molecular biology in lung cancer , numerous molecular targets were discovered.However, selecting patients who will be the most likely to benefit from the therapy is still a great challenge in clinical practice .Companion diagnosis could help evaluate the status of patients , which may provide better suggestions for health care professionals during treatment decision , in order to benefit the patients and decline the side effects risk together .Besides, high-throughput sequencing technics and diverse detection samples has provided a novel direction to understand the tumorigenesis and find more potential gene targets.
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<p><b>BACKGROUND</b>Diabetic macular edema (DME) is one of the major causes of visual impairment in patients with diabetes mellitus. DME shows a variety of clinical characteristics with unpredictable results to treatment. The purpose of this study was to investigate the characteristics of fundus autofluorescence (FAF) in DME, which is a rapid, noninvasive technique for fundus diseases.</p><p><b>METHODS</b>A total of 18 patients (30 eyes) with clinically significant macular edema (CSME) were enrolled. FAF imaging was performed with a confocal scanning laser ophthalmoscope. Other ophthalmic examinations included best corrected visual acuity (BCVA), MP-1 microperimetry, and spectral-domain optical coherence tomography (SD-OCT). Main outcome measurements included BCVA, macular sensitivity (MS), central retinal thickness (CRT), central retinal volume (CRV), the integrity of the inner segment-outer segment junction (IS/OS), and the integrity of the external limiting membrane (ELM).</p><p><b>RESULTS</b>Among the 30 eyes, four eyes (13.3%) had normal foveal FAF and 26 eyes (86.7%) had abnormal FAF. Abnormal FAF was mainly divided into three types: cystoid increased FAF (iFAF) 16 eyes (53.3%), spot iFAF six eyes (20%), irregular decreased FAF (dFAF) four eyes (13.3%). According to the FAF morphology, patients were categorized into four groups: normal, cystoid iFAF, spot iFAF, and irregular dFAF. There was a significant difference in BCVA (P < 0.001) and MS (P < 0.05) among the four groups. The visual function of patients with spot iFAF and irregular dFAF was relatively poor. However, there was no difference in CRT (P = 0.186) and CRV (P = 0.191) among the four groups. In the normal FAF group, the photoreceptor layers were mostly intact. Regarding the cystoid iFAF group, the photoreceptor layers were relatively intact, while in the other two groups, IS/OS and ELM were disrupted in most patients. No one had intact IS/OS or ELM layer.</p><p><b>CONCLUSIONS</b>FAF might reflect the damage of the retina and had a relationship with visual function as well as photoreceptor integrity, which gives new insight into the evaluation of DME. Dynamic FAF monitoring helps to better evaluate the disease progression of DME as well as visual function.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Diabetic Retinopathy , Diagnosis , Fundus Oculi , Macular Edema , Diagnosis , Optical Imaging , Retrospective StudiesABSTRACT
Rapid development of personalized medicine in cancer treatments needs urgent support of proper tumor biomarkers.Comparing to tissue tests,serum/plasma tumor biomarkers show advantages in easy sample collection and flexibility of detecting methods.Nowadays,large scales of investigations have discovered sufficient blood tumor biomarkers for preclinical diagnosis and evaluation of clinical treatment or prognosis,with the development of screening techniques and strategies.Moreover,tremendous promotion would appear in cancer diagnosis and treatment with future profound investigation and rigorous verification from clinical practice.
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Objective To observe the subfoveal choroidal thickness (SFCT) in eyes of patients with diabetic macular edema (DME).Methods Twenty patients (32 eyes) with DME were enrolled in this crosssectional observational study.The patients included 12 males and eight females,with a mean age of (47.3± 10.2) years.All the patients were examined documenting best corrected visual acuity (BCVA),spectraldomain optical coherence tomography (OCT) and ophthalmological examination.According to OCT DME morphology,samples are divided into diffuse macular edema,cystoid macular edema,serous retinal detachment and hard exudate groups.The SFCT was measured by a Cirrus HD-OCT with enhanced depth imaging (EDI) and was compared with the average SFCT (286.84 ± 28.80) μm of same age group.Correlation between SFCT and age,diopter,diabetic duration,fasting blood glucose,BCVA and central retinal thickness were analyzed by Pearson Analysis.SFCT of different DME types were analyzed by ANOVA Analysis.Results The mean SFCT of 32 eyes was (223.81±43.74) μm (ranging from 120.50 to 361.50 μm),which was lower by 63.03 μm (95％ confidence interval,-78.80 to-47.26 μm,P＜0.01)from normal SFCT.SFCT was independent of age (r=0.124),diopter (r=0.277),diabetic duration (r=0.286),fasting blood glucose (r=0.408),BCVA (r=0.087),and central retinal thickness (r=0.036).There was no significant difference of SFCT between different DME types (F =0.042,P ＞ 0.05).Conclusion SFCT is thinner in eyes with DME as compared to normal eyes of the same age.
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Objective To determine the mutant status of BRAF gene in Chinese colorectal cancer (CRC) patients and analyze the association with clinicopathological parameters.Methods 676 CRC samples were collected in Cancer Institute/ Hospital,Chinese Academy of Medical Science from December 2009 to December 2011.The direct sequencing was conducted to detect mutations in the BRAF (exon 15 and exon 11).The correlation between mutant status with clinicopathological parameters were analyzed.Results Beside 10 colorectal cancer samples,among the 666 colorectal cancer patients 4.35％ (29/666) of the tumors harbored a BRAF mutation,of which 1.94％ (13/669) in exon 15 (V600E),2.39％ (16/670) in exon 11.Statistical analysis revealed that BRAF15 mutations appeared to occur more frequently in poor-differentiation tumors than high or moderate-differentiation tumors (5.81％ vs 1.46％,r =0.105,P=0.040).But BRAF15 mutations were not correlated with age,gender,smoking and drinking history,tumor site,tumor type,tumor(T/N) staging,histological type,or distant metastasis [r equals to 0.007,-0.018,-0.049,-0.023,-0.098,-0.038,0.040(0.034/0.059),0.065,0.042,respectively,P ＞ 0.05] ; BRAF11 mutations appeared to occur more frequently in patients with drinking history (6.02％vs 1.81％,r =0.093,P =0.035).However,age,gender,smoking history,tumor site,tumor type,tumor differentiation,tumor(N) staging,histological type,or distant metastasis showed no significant correlation with this mutation [r equals to-0.004,0.047,0.020,0.042,0.029,0.040,0.006 (-0.008),0.008,0.030,respectively,P ＞ 0.05].Conclusion A higher proportion of BRAF15 (V600E) mutations occurred in poor-differentiation tumors among the Chinese patients with CRC; BRAF11 mutations appeared more frequently in patients with drinking history.