Background The retina ischemia reperfusion injury (RIRI) can lead to apoptosis,which is associated with many genes.It is very significant to explore the protection of drugs on RIRI-induced apoptosis.Objective This study was to explore the relationship between the expression of Nogo-A and the cell apoptosis as well as the therapeutic effect of NEP1-40 in RIRI retina.Methods The device of raising intraocular pressure (IOP)was used to elevate the IOP for 60 minutes and then restore the IOP to normal for the establishment of RIRI models.Seventy-eight SD rats were randomized to the normal group,RIRI group and NEP1-40 group.PBS of 5 ml/(kg · d)was injected intraperitoneally in the rats of the RIRI group,and NEP1-40 of 8 mg/(L · kg) was used in the same way in the NEP1-40 group.The rats were sacrificed in overdose anesthesia at 6 hours,12 hours and 1 day,2,3,7 days after RIRI to prepare the retinal specimens.The ultrastructure of rat retinas was examined under the transmission electron microscope.Cell apoptosis was assessed by the TUNEL method,and the apoptotic index (AI) was calculated.The expressions of Nogo-A protein and mRNA in rat retinas were detected by immunohistochemistry and semiquantitative reverse transcription PCR (RT-PCR).Results The ultrastructure of retinal cells were normal in the normal group.Retinal cell organelle dissolution,mitochondria swelling,cavitation,chromatin edge heterochromatin and apoptotic body were seen in the rats of the RIRI group from 12 hours through 7 days after injection.However,only the slight loose of outer membranous disk,inner and outer nuclear layer and retinal ganglion cells,the nucleus gap broadening,shortness of some mitochondrial cristae in the NEP1-40 group.TUNEL-positive cells appeared 6 hours after RIRI and reached peak in 1 day,and gradually declined after that in the RIRI group.A similar pattern was seen in the rats of the NEP1-40 group with a more mild manifestation.Significant differences were seen in the AI values among the different groups at various time points (Fgroup =100.850,P =0.000 ; Ftime =34.309,P =0.000),and the AI values were significantly higher in the RIRI group and NEP1-40 group compared with normal group;while the AI values in the NEP1-40 group was lower than those of the RIRI group (all at P＜0.05).The expressions of Nogo-A protein and mRNA showed a coincident pattern with apoptosis procedure,with a gradually elevated level from 6 hours through 7 days after RIRI and a peak in 2 days,and those in the NEP1-40 group were weaker in comparison with the RIRI group in various time points.Significant differences were detected in the expression of Nogo-A protein and the expression of Nogo-A mRNA among different groups and various time points(protein:Fgroup =164.139,P =0.000;Ftime =21.772,P =0.000.mRNA:Fgroup =93.889,P =0.000 ; Ftime =6.349,P =0.000).Conclusions Nogo-A probably plays an important role in RIRI.NEP1-40 can protect retinal cells from apoptosis following RIRI through down-regulating the expression of Nogo-A.

Objective: To investigate the value of lipid accumulation product (LAP) and visceral fat index (VAI) for prediction of metabolic syndrome (MS). Methods: Based on the 2018 Survey on Chronic Diseases and Risk Factors in Yantai City of Shandong Province, residents at ages of 45 years and older were sampled, and subjects' age, disease history, waist circumstance (WC), body mass index (BMI), blood pressure and blood lipid were collected to calculate LAP and VAI. MS was diagnosed with the a joint interim statement of the International Diabetes Federation Task Force on Epidemiology and Prevention; National Heart, Lung, and Blood Institute; American Heart Association; World Heart Federation; International Atherosclerosis Society; and International Association for the Study of Obesity (JIS definition) and the recommended criteria proposed by the Chinese Diabetes Society (CDS) of Chinese Medical Association (CDS criteria), and the values of LAP and VAI for MS screening were evaluated using the receiver operating characteristic (ROC) curve analysis. Results: Totally 9 366 subjects were enrolled, including 4 340 men (46.34%) and 5 026 women (53.66%), and had a mean age of (54.49±9.73) years. According to the CDS criteria, the prevalence of MS was 24.58%, and LAP and VAI showed areas under the ROC curve (AUC) of 0.837 (95%CI: 0.828-0.846) and 0.751 (95%CI: 0.739-0.762), sensitivities of 78.82% and 63.31% and optimal cut-off values of 44.64 and 1.86 for screening of MS. According to the JIS definition, the prevalence of MS was 35.26%, and LAP and VAI showed AUC values of 0.842 (95%CI: 0.834-0.850) and 0.790 (95%CI: 0.780-0.800), sensitivities of 75.73% and 68.42% and optimal cut-off values of 42.01 and 1.67 for screening of MS. Conclusions Both LAP and VAI are effective for screening MS among middle-aged and elderly residents, and LAP presents a higher accuracy than VAI.

BACKGROUND: Studies of biological characteristics of mesenchymal stem cells (MSCs) and regulatory factors that influenced the differentiation of MSCs have shown that the proportion of the natural differentiation from in vitro primarily cultured MSCs into hepatocytes was low, and to select a suitable inductor is important to enhance the differentiation of MSCs into hepatocytes.OBJECTIVE: To verify the feasibility of induced differentiation of rat bone marrow MSCs (BMSCs) into hepatocytes using the combination of hepatocyte growth factor (HGF), epidermal growth factor (EGF) and fibroblast growth factor (FGF-4).DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Experimental Center, Weifang Medical College in August 2007.MATERIALS: Totally 40 Sprague-Dawley rats were supplied by the Experimental Animal Center, Weifang Medical College.METHODS: Rat BMSCs were incubated by adherent method. BMSCs at passage 3 were assigned to 2 groups. BMSCs in the blank control group were treated with L-DMEM containing 10% fetal bovine serum. BMSCs in the combination group were treated with 10 μg/L FGF, 8 μg/L HGF and 8 μg/L EGF following above-mentioned procedures.MAIN OUTCOME MEASURES: Inverted microscope was used to observe the morphological changes in cells.Immunofluorescence method was used to observe the expression of alpha-fetoprotein (AFP) and albumin (ALB). PAS was employed to detect the expression of glycogen. Fox green intake experiment was conducted. Enzymology was utilized to test the contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP).RESULTS: BMSCs in the combination group presented polygonal, orbicular or round shape. BMSCs in the blank control group remained spindle. BMSCs in the combination group were positive for AFP and ALB at day 14 following culture, and a few PAS-positive and fox green-positive cells were found at day 7. Positive cells became more over time. Synthesis of ALT, AST and ALP was detected at day 14, reached a peak at 21 days, and then decreased. Above-described indexes were negative in the blank control group.CONCLUSION: After induced by the FGF, HGF and EGF, BMSCs have the ability to differentiate into hepatocytes in vitro.

Objective To explore the possibility that rat bone mesenchymal cells (BMSCs) can differentiate into hepatocytes under the affection of fetal liver filtrate. Methods PAS and green indigo dye were used to detect glycogen and differential level of hepatocytes, respectively. The concentration of ALT, AST, ALP in the culture supernatant were served as markers of hepaocyte function. Results Fourteen days after induced by the fetal liver filtrate, BMSCs changed their shapes into polygon, oval or round. Some of BMSCs were positive for AFP and ALB at 7 days after induction, then the number of positive cells increased, and most of BMSCs expressed AFP and ALB till 21days. The PAS reaction and indocyanine green(ICG) intaking also appeared at 7days. Enzyme in supernatant such as ALT, AST, ALP were fristly detected at 7days and peaked at 14days,then the level declined. Conclusion The fetal rat liver filtrate was able to induce BMSCs into cells with function and characteristics of hepatocytes.

Objective To study the putative mechanisms underlying fetal alcohol syndrome by comparative protein-profile analysis between normal and ethanol-treated zebrafish embryo with twodimensional electrophoresis (2-DE).Methods Zebrafish embryos were exposed in 400 mmol/L ethanol at dome stage for 3 hours,and then ethanol-induced abnormalities were observed.Proteomes of zebrafish embryos at early stages including zygote stage,dome stage,shield stage and 5-somite stage,were separated by 2-DE.The subtraction analysis method was applied to eliminate the interference from maternal derived proteins.The ethanol-treated embryos at 5-somite stage was analyzed by 2-DE,and the protein profile was compared with that generated from control embryos at the same stage.The data obtained from 2-DE analysis were verified by in-situ hybridization.Results 400 mmol/L ethanol treatment caused axial malformation (62%) and cyclopia (60%) in zebrafish embryos.The 2-DE analysis showed that the expression of Collagen2al (Col2a1) and TAR DNA binding protein (TDP) was decreased in 12 hours post fertilization (12 hpf) ethanol-treated embryos by 81% and 73%,respectively.The in-situ hybridization also demonstrated that the expression of Col2al in axial mesoderm was reduced by ethanol treatment at the same stage.But for 24 hpf ethanoltreated embryos,the expression of Col2al in axis recovered to a comparable level to that in control embryos,while the structure of neural tube was disrupted severely by ethanol exposure.Conclusions It is suggested that the expressions of Col2al and TDP were disrupted by ethanol during early stage,which might induce the zebrafish developmental abnormalities.The ethanol interference on early expression of Col2al is supposed to be one of the major reasons leading to later abnormalities of axis and neutral tube.

OBJECTIVE: To observe the therapeutic effect of radiotherapy combined with TPF in patients with nasopharyngeal carcinoma (NPC) and to explore the influencing factors on prognosis of NPC. METHOD: A retrospective analysis of 144 patients with nasopharyngeal carcinoma receiving radiotherapy combined with chemotherapy, from January 2006 to December 2011, was conducted. The survival analysis in patients with nasopharyngeal carcinoma were performed. The impact of age, sex, clinical stage, alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), lactate dehydrogenase (LDH), homocysteine (Hcy), uric acid, NK cell proportion, T cell proportion, Carbohydrate antigen on survival time was observed with univariate and multivariate methods. RESULT: By the last follow-up, 9 cases were lost . The follow-up rate was 93.75 percent and the average follow-up time were 54.67 ±17.17 (37-88) months. The age, clinical stage, LDH, proportion of T cells and NK cells were independent factor in patients with NPC with univariate analysis (P < 0.01). COX regression multivariate analysis revealed that age, clinical stage and LDH were strong risk factor affecting prognosis (P < 0.01) and the proportion of NK cells and T cells were protective factors (P < 0.01). CONCLUSION Radiotherapy combined with TPF is more effective than radiotherapy alone in patients with nasopharyngeal carcinoma . Age, clinical stage and LDH may adversely influence the disease prognosis, and NK cells and T cell proportion may act as protective factors.
Age Factors ; Biomarkers, Tumor ; Carcinoma ; Combined Modality Therapy ; Follow-Up Studies ; Humans ; Induction Chemotherapy ; Multivariate Analysis ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; diagnosis ; therapy ; Neoplasm Staging ; Prognosis ; Radiotherapy ; Retrospective Studies ; Risk Factors ; Survival Analysis

Objective To study the clinical characteristics, diagnosis, treatment and prognosis of primary pulmonary peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS). Methods Two cases of primary pulmonary PTCL-NOS were studied and relevant literature were reviewed. Results Case 1 diagnosed as primary pulmonary PTCL-NOS was a 44 years old woman and disease progressed after GLD (Gem+L-OHP+DXM) chemotherapy regimen. At last, the patient died of respiratory failure after one month. Case 2 diagnosed as primary pulmonary PTCL-NOS was a 46 years old man and reach partial response after CHOP regimen, and still alive now.Conclusion Primary pulmonary PTCL-NOS is very rare. It is easy to be misdiagnosed due to non-specific clinical and imaging manifestations. Acquiring enough tissue specimens for pathologic examination is the key to a definitive diagnosis. At present, there is no standard chemotherapy regimen for these patients, the prognosis is relatively poor.

Objective To study pulmonary wedge angiography ( PWA ) with hemodynamic the evaluation of children with congenital heart disease and pulmonary artery hypertension ( PAH) . Methods Hemodynamic measurement and pulmonary wedge angiography were performed in 50 children with congenital heart disease. Comparison and analysis were made from the data obtained from PWA and catheterization. Results After PWA, the patients were categorized into 3 groups according to the measured hemodynamics parameters:group A [ n=15, patients with normal mean pulmonary artery pressure ( mPAP≤25 mmHg) and normal pulmonary vessel resistance (PVR﹤300 dyne?s?cm5)], group B [n=24, patients with PAH (mPAP﹥25 mmHg) but normal PVR] and group C (n=11, patients with PAH and elevated PVR (PVR≥300 dyne?s?cm5). Rote of tapering (ROT) was significant lower in group C than in group A and B (F=42. 559,P﹤0. 05). Pulmonary circulation time (PCT) was higher in group C than in group A and B (F=6. 037,P﹤0. 05). ROT correlated negatively with PVR (r = -0. 606, P ﹤0. 05). PCT index correlated positively with PVR (r=0. 783,P=0. 01). There was no significant correlation between PCT and mean pulmonary artery hypertension (mPAP). Conclusions PWA may help to make quantitative analysis of the pulmonary vascular status in patients with congenital heart disease.

Objective To investigate the difference of expression profiling of plasma miRNAs (microRNA) between the patients with small artery occlusive stroke (SAO) and the healthy subjects.Methods Eight patients with SAO classified by TOAST were selected and 8 healthy subjects were used as a control group.High-throughput sequencing technology was used to detect the expression profiling of plasma miRNAs.The differentially expressed miRNAs were screened.Real-time fluorescent quantitative polymerase chain reaction was used to validate the results,and the target gene prediction and bioinformatics analysis were performed.Results The miRNA difference analysis showed that the expression profilings of miRNA-127,miRNA-99b-5p,miRNA-320,and other 19 miRNAs in the SAO group were significantly upregulated compared with those in the control group (all P＜0.01),while miRNA-451a and other 5 miRNAs in the SAO group were significantly downregulated compared with those in the control group (all P ＜0.01).The validated results of miRNA-127,miRNA-99b-5p,miRNA-320,and miRNA-451a with real-time fluorescent quantitative polymerase chain reaction were consistent with those of the high-throughput sequencing.Bioinformatics analysis showed that the miRNA-regulated target genes expressed differentially were mainly correlated with cell proliferation,adhesion,phylogenetic development,macromolecule metabolism,and other biological functions.Conclusions There are significant differences in the expression profiling of plasma miRNAs between the patients with SAO and the healthy subjects,suggesting that miRNAs may play a regulatory role via target genes in pathogenesis of SAO.

Objective To investigate the changes of circulating miRNAs expression profiles in different subtypes of ischemic stroke according to the Trial of Org 10 172 in Acute Stroke Treatment.Methods We selected 16 patients diagnosed as acute ischemic stroke at first time in the Department of Neurology,the Affiliated Hospital of Medical College Qingdao University from November to December in 2012.They were divided into large artery atherosclerosis (LAA) stroke group (n =8) and small artery occlusive (SAO) stroke group (n =8).At the same time,8 healthy checkup subjects were selected as control.High-throughput sequencing was used to detect the expression profiles of miRNAs in the plasma,and the high-throughput sequencing results were validated by quantitative real-time polymerase chain reaction.We performed the miRNAs variance analysis and associated bioinformatics analysis.Results Thirty hundred and sixty-nine miRNAs were detected in LAA group,SAO group and the control group.We found remarkable differences (fold change ＞2,P ＜0.01) in the expression of 12 miRNAs,including let-7a-5p,miR-744-5p,etc.,between any two groups of the three groups.Thirty-four miRNAs,containing miR-126-5p,miR-23a-3p,miR-143-3p,etc.,had a lower expression (fold change ＞2,P ＜0.01)in LAA group than that in the control group.In comparison with the control group,miR-1304-3p and miR-451a were significantly down-regulated (fold change ＞ 2,P ＜ O.01),while 27 miRNAs were significantly up-regulated (fold change ＞2,P ＜0.01) in SAO group.The expression levels of miR-146b-5p,miR-23a-3p and miR-451 a were validated in accordance with the results of real-time PCR.Target gene prediction and functional analysis revealed that target genes regulated by differential expression of miRNAs were mainly associated with cell proliferation,adhesion,metabolism and other biological functions.Conclusion miRNAs are differently expressed in the plasma of LAA group,SAO group and healthy control group,which suggest that miRNAs might play different roles in the pathogenesis of LAA stroke and SAO stroke by regulating downstream target genes.