Objective To investigate the effect of epidermal growth factor (EGF) on the expression of Matrix metalloproteinase-9 (MMP-9) and the signalling pathways involved in the trophoblast cell line JEG-3. Methods The JEG-3 trophoblast cell line was used in this study. (1) JEG-3 cells were cultured with various concentrations of EGF (0, 1, 10,20 ng/ml) for 24 hours and the expression of MMP-9 was tested by western blotting and reverse transcription PCR (RT-PCR). (2) Western blotting and RT-PCR were also used to investigate the expression of MMP-9 expression after incubation for 0,4,12 and 24 hours with EGF treatment (10 ng/ml) in JEG-3 cells. (3) According to the different added ingredients, JEG-3 cells were divided into some groups: control group (without EGF), EGF group (exposure to l0ng/ml EGF),EGF + inhibitors group (exposure to 10 ng/ml EGF +20 ng/ml SB203580 or exposure to 10 ng/ml EGF + 10ng/ml U0126) inhibitors group (exposure to 20 ng/ml SB203580 or exposure to 10 ng/ml U0126). Western blotting were used to investigate the expression levels of MMP-9, nuclear factor kappa B (NF-kB) ,p38MAPK, phospho-p38MAPK (p-p38MAPK) , extracellular -signal regulated kinase (ERK) and phospho-ERK (p-ERK) protein in JEG-3 cells after incubation for 24 hours. Results (1) The profiles of MMP-9mRNA were increased by various concentrations of EGF (0, 1 , 10, 20 ng/ml) in JEG-3 cells after 24hculture. The expression of MMP-9 mRNA in JEG-3 cells exposure at 1 ng/ml of EGF (0. 567 ±0. 056) , 10ng/ml of EGF (1. 392 ±0. 133) , 20 ng/ml of EGF (1. 971 ±0. 067) were significantly higher respectively (P <0. 05) , compared with 0 ng/ml of EGF treatment (0. 166 ±0. 015). Similarly, MMP-9 mRNAs were also increased with the increasing incubation time. Compared to EGF (10 ng/ml) stimulation for 0 h (0.253 ±0.044), the MMP-9 mRNA profiles were 0. 470 ±0. 026, 1.061 ±0. 115, 1. 453 ±0. 180 for 4,12 and 24 hours, respectively (P < 0. 05). (2) In accordance to the mRNA profiles, the expression of MMP-9 protein was also increased by different concentrations of EGF (0,1, 10, 20 ng/ml) in JEG-3 cells after 24 h-culture. The abundance of MMP-9 protein in the three groups was 0. 043 ±0. 012, 0. 085 ±0. 008, 0. 142 ±0. 015, with a significantly higher expression, compared with 0 ng/ml of EGF treatment (0. 004 ±0.001, P < 0.05) respectively. Similarly, MMP-9 proteins were also increased with the increasing incubation time. Compared to EGF(10 ng/ml) stimulation for 0 h (0. 030 ±0. 009) , the profiles of MMP-9 protein were 0. 137 ± 0. 010, 0. 240 ± 0. 010, 1.240 ±0.061 for 4, 12 and 24 hours, respectively (P < 0.05). (3) Both p38MAPK and ERK signalling pathways were activated by EGF in JEG-3 cells. The expression of p-p38MAPK was significantly higher (without or with 10 ng/ml EGF, 234. 1 ± 4. 1 vs.260. 9 ± 2. 5 , P < 0. 05) , however, the p38 MAPK inhibitor SB203580 markedly suppressed the increase in p-p38MAPK content induced by EGF(227. 9 ±2. 4 vs. 260. 9 ±2. 5, P<0. 05). Similarly, the expression of p-ERK was significantly higher with EGF treatment (812. 2 ±3. 5) vs. without EGF group (453.4±5.8) (P <0. 05) , while the ERK inhibitor U0126 significantly inhibited the increased p-ERK content in response to EGF treatment (71. 0 ± 1. 2 vs. 812. 2 ± 3. 5, P < 0. 05) . (4) The p38MAPK inhibitor SB203580 significantly reduced the expression of EGF-induced MMP-9 (0. 645 ± 0. 270 vs. 1. 476 ± 0. 452, P < 0. 05)and NF-kB (0.530 ± 0.026 vs. 0.959 ± 0. 017, P < 0. 05) . (5) The ERK inhibitor U0126 also significantly reduced the expression of EGF-induced MMP-9 (0. 623 ±0. 030 vs. 2. 112 ±0. 056, P <0. 05)and NF-kB (0. 325 ± 0. 082 vs. 0. 939 ± 0. 153, P < 0. 05). Conclusion EGF induced the expression of MMP-9 in a time and dose-dependant manner in JEG-3 cells. EGF enhanced MMP-9 expression through the activation of p38MAPK and ERK signalling pathways in JEG-3 cells.

Objective To investigate the feasibility of ADC values in the evaluation of normal fetal brain development by measuring ADC values changes in specific regions with advancing gestational age.Methods Forty fetuses(gestational age:24 to 41 weeks) with normal brain underwent DWI(b value were 0 and 600 s/mm2).ADC values of the frontal white matter(WM),occipital WM,thalamus,basal ganglia,and cerebellum were measured by post-processing software.The differences among different regions' ADC values were calculated by repeated measurements of ANOVA,and simple linear regression was used to evaluate the relationship between ADC values and gestational age.Results The mean ADC valuesof 40 fetuses were(1 800±214) ×10-6mm2/s in frontal WM,(1 400±100) ×10-6mm2/s in basal ganglia,(1 300±126) ×10-6mm2/s in thalamus,(1 700±133) ×10-6mm2/s in occipital WM and(1 400± 155) × 10-6mm2/s in cerebellum,respectively.There was significant difference in the ADC values among the five regions(F=80.813,P＜0.01).In pair-wise comparison,ADC values of basal ganglia,thalamus and cerebellum had no significant difference; however,others had significant difference between each other.With the increasing gestational age,ADC values of basal ganglia,thalamus,occipital WM and cerebellum decreased,and had significant negative correlations with gestational age(Pearson correlation coefficient were-0.568,-0.716,-0.830 and-0.700,respectively,all P＜0.01).In terms of ADC value,occipital WM declined fastest with gestational age,followed by cerebellum and thalamus,and the slowest was basal ganglia.Frontal ADC values showed no correlation with gestational age(P＞0.05).Conclusions Specific regions of fetal brain have specific ADC values,and ADC values of the region undergo regular change with advancing gestational age.ADC value is a specific quantitative parameter that could help to evaluate normal brain development and early diagnosis of fetal brain lesions.

Objective To compare the effects of three kinds of fixative solutions on paraffin section of mouse lens tissue and optimize the fixing?method of paraffin section in mouse lens tissue. Methods Three kinds of conventional fixatives were selected for the test ,including the conven?tional Davison’s solution,modified Davison’s solution and 10%neutral buffered formalin. Mice eyeball tissues were fixed with three different fixa?tives,embedded,sliced and then stained with HE method. The paraffin slices were observed under the light microscope. Results The structures of lens and retina fixed in conventional Davison ’s fixative solutions were clear and intact ,and the cells were arranged regularly and compactly. There was no eyeball distortion,contraction and retinal detachment in the eyeballs fixed in modified and conventional Davison’s fixative solution. However,the ones fixed in 10%neutral buffered formalin showed eyeball distortion and contraction,space and spherules. Conclusion The mice lens slides made from tissues fixed by conventional Davison ’s fixative solution are better than fixed by modified Davison ’s fixative solution and the 10%neutral buffered formalin fixed ones.

Objective To demonstrate the value of eombined application of prenatal ultrasonography with fetal magnetic resonance imaging(MRI) in the diagnosis of monochorionic muhifetal realformations. Methods Fourteen cases of muhifetal malformations,detected by prenatal ultrasonography,received MRI within 48 h afterwards.All diagnosis were confirmed after delivery or mid-term termination.All imaging results of the 14 cases were retrospectively reviewed. Results Among the 14 cases,there were 7 acardias,5 Conjoined twins and 2 demise of multifetuses.Comparing ultrasound with MRI,we found that:(1)In cases with acardia and demise of multifetusea,ultrasound could diagnose correctly and be an important tool for follow-up,while MRI could demonstrate organs and structures of the acardiac recipient more clearly and detect the secondary changes of brain in the donor and survived fetus.(2)In Conjoined twins,ultrasound was superior to MRI in demonstrating the structure and function of cardiovascular system : and equivalent to MRI in identifying stomach,kidney,bladder and limbs;but inferior to MRI in identifying esophagus,lung,liver and intestinal,especially in the brain. And MRI could demonstrate two fetuses and the relationship between them in COnjoined twins simultaneously. Conclusions Prenatal ultrasonography and MRI have their own advantages and disadvantages in diagnosing monochorionic multifetal malformations.But the combination of prenatal ultrasonography and fetal MRI may be more valuable.

Objective To investigate the effect of transforming growth factor β1 (TGF-β1) on the expression of matrix metalloproteinase 9 (MMP-9),tissue inhibitor of metalloproteinase 1 (TIMP-1),nuclear factor kappa B(NF-κB) and the possible signalling pathways in human amniotic cells WISH.Methods The WISH cell line was cultured.WISH cells were added with TGF-β1 of different concentrations (0,2,10 and 20 ng/ml,respectively) for 24 hours.Then,reverse transcription (RT) PCR and western blotting were used to analyze the protein and mRNA expression of TIMP-1 and MMP-9; and the expression of NF-κB was analyzed by western blot.Results (1) The profile of TIMP-1 mRNA (0.413 ±0.036,0.623 ±0.058,1.392 ±0.124,1.387 ±0.102) in WISH cells elevated when the concentration of TGF-β1 increased (0,2,10,20 ng/ml).In accordance with TIMP-1 mRNA,the expression of TIMP-1 also elevated with the increase of TGF-β1 (0.357 ± 0.031,0.596 ± 0.048,1.243 ± 0.097 and 1.359 ± 0.121,respectively).And when 2,10 or 20 ng/ml of TGF-β1 was added,the TIMP-1 mRNA and protein were significantly higher than the TIMP-1 mRNA and protein when no TGF-β1 was added(P ＜ 0.05).(2)In contrast with TIMP-1,MMP-9 mRNA (1.325 ±0.056,0.987 ±0.081,0.610 ±0.034,0.347 ±0.023) in WISH cells decreased when the concentration of TGF-β1 increased (0,2,10,20 ng/ml).The MMP-9 protein (1.119 ±0.064,1.008 ±0.052,0.578 ±0.041,0.401 ±0.015) also decreased with the increase of TGF-β1.And when 2,10 or 20 ng/ml of TGF-β1 was added,the MMP-9 mRNA and protein were significantly lower than the MMP-9 mRNA and protein when no TGF-β1 was added (P ＜ 0.05).(3) The NF-κB protein (1.423 ±0.065,1.116 ± 0.045,0.796 ± 0.041,0.359 ± 0.021) was significandy reduced with the increase of TGF-β1 (0,2,10,20 ng/ml; P ＜ 0.05).Conclusions The mRNA and protein expression of TIMP-1 decreased when TGF-β1 was low in WISH cells,whereas those of MMP-9 elevated when TGF-β1 was low.The unbalance of TIMP-1 and MMP-9 was related to the pathology of the premature rupture of membrane.And the NF-κB singalling pathway might be an important mechanism in the regulation of TIMP-1 and MMP-9 system.

Objective To explore expression and clinical value of VEGF-C and p63 in early esophageal carcinoma and intraepithelial neoplasia. Methods 146 cases were randomized into normal esophageal mucosa, low level intraepithelial tumor, high level intraepithelial tumor and early esophageal carcinoma. The expression of VEGF-C and p63 were detected by using the immunohistochemistry dyeing.Results The expression of VEGF-C immunohistochemistry dyeing had statistical differences among different levels(X~2= 47.455, P <0.001). Normal esophageal mucosa v.s. high level intraepithelial tumor (X~2=36.721, P <0.001), Normal esophageal mucosa v.s. early esophageal carcinoma (X~2=26.483, P <0.001), low level intraepithelial tumor v.s. high level intraepithelial tumor(X~2= 10.025, P<0.0083), low level intraepithelial tumor v.s. early esophageal carcinoma(X~2=16.734, P<0.001). There was a significant correlation between pathological classification and the expression amount of VEGF-C (r = 0.462, P <0.001). The expression of p63 had statistical differences among different levels(X~2=28.962, P <0.05). There was a significant difference on normal esophageal mucosa comparing with low level, high level intraepithelial tumor or early esophageal carcinoma (X~2=12.735, P =0.005, X~2=20.421, P<0.001, X~2=20.854, P<0.001). There was a significant correlation between pathological classification and the expression of p63 (r= 0.272, P<0.05). Conclusion There is a significant correlation in the express of either VEGF-C or p63 comparing with either intraepithelial tumor or early esophageal carcinoma. It may be an early warning indicator.

Glucagon-like peptide-1 (GLP-1) is a 30-amino acid peptide hormone that is mainly expressed in the intestine and hypothalamus. In recent years, basic and clinical studies have shown that GLP-1 is closely related to lipid metabolism, and it can participate in lipid metabolism by inhibiting fat synthesis, promoting fat differentiation, enhancing cholesterol metabolism, and promoting adipose browning. GLP-1 plays a key role in the occurrence and development of metabolic diseases such as obesity, nonalcoholic fatty liver disease, and atherosclerosis by regulating lipid metabolism. It is expected to become a new target for the treatment of metabolic disorders. The effects of GLP-1 and dual agonists on lipid metabolism also provide a more complete treatment plan for metabolic diseases. This article reviews the recent research progress of GLP-1 in lipid metabolism.

Objective To explore the real experience during treatment of patients with multiple myeloma, so as to provide evidences for clinical nursing staff to implement targeted nursing measures and to carry out research on nursing staff of high evidence level. Methods Using phenomenological methods, nine cases of patients with multiple myeloma conducted semi-structured and in-depth interviews and then using Colaizzi 7-step for analysis. Results Three major topics were extracted: poor physiological experience (feeling abnormal, numbness, pain, urinary incontinence), positive response (family support, Increased medical standards, self-regulation), self-perceived burden (economic stress, short cycle of relapse, psychological burden). Conclusion Treatment of multiple myeloma have many adverse reactions, and patients bear too much psychological burden. Medical staff need to be evaluated in a timely manner, strengthen symptom management, provide comprehensive information support, analysis of patient self-perception burden causes, propose targeted interventions, and build continuation care system to improve patient treatment experience.

Objective:To explore the micro RNA (miR)-193b-3p expression in hippocampus tissues of neonatal sepsis rats, as well as its role in neuroinflammatory response and possible mechanisms.Methods:Twenty-four 2-d-old SD rats were randomly divided into control group, lipopolysaccharide (LPS) group, negative control group, and miR-193b-3p group ( n=6); except for the negative control group, the rats in the other 3 groups were intraperitoneally injected with LPS to establish sepsis models; in miR-193b-3p group and NC group, 5 μL miR-193b-3p mimics/controls (20 nmol/L)were injected into the lateral ventricle 3 d before LPS injection. In vitro, PC12 cells were divided into control group, LPS group, miR-193b-3p group, and LPS+miR-193b-3p group; miR-193b-3p mimics were transfected into cells of miR-193b-3p group and LPS+miR-193b-3p group; cells in the LPS group and LPS+miR-193b-3p group were exposed to 100 ng/mL LPS. The miR-193b-3p downstream target gene was analyzed by gene microarray and dual luciferase reporter. Neurobehavioral scale used to assess the neurological function at specific time points in each group. The mRNA expression levels of miR-193b-3p and cytokines (interleukin [IL]-1β, IL-6, tumor necrosis factor [TNF]-α) in the brain tissues or PC12 cells were analyzed by real-time fluorescence quantification PCR (RT-qPCR). The protein expression levels of retinoic acid-related orphan receptor α (RORα), neuronal nuclear antigen (NeuN), ionized calcium binding adaptor molecule-1 (IBA-1) and glial fibrillary acidic protein (GFAP) were detected by double-labelling immunofluorescence. The protein expression levels of RORα in PC12 cells were detected by immunofluorescence staining. Results:(1) Gene microarray and dual luciferase reporter analysis confirmed that RORα was the target gene of miR-193b-3p. (2) As compared with those in rats of the negative control group, the neurobehavioral scores in rats of the LPS group were significantly decreased since 6 h of LPS injection and reached to the lowest at 24 h after LPS ( P<0.05). As compared with those in the negative control group, the IL-1β, IL-6, and TNF-α mRNA expression levels in the hippocampus of LPS group were significantly increased, while the miR-193b-3p expression was significantly decreased ( P<0.05). (3) As compared with those in negative control group (3.23±0.92), the neurobehavioral scores in rats of the miR-193b-3p group (7.51±0.84) 48 h after LPS injection were significantly higher ( P<0.05). As compared with those in the negative control group, the IL-1β, IL-6 and TNF-α mRNA expression levels in the hippocampus of the miR-193b-3p group were significantly deceased, while the miR-193b-3p expression was significantly higher ( P<0.05). (4) As compared with that in negative control group, the number of NeuN(+)RORα(+) cells in the hippocampus of the LPS group was significantly reduced ( P<0.05); as compared with negative control group, miR-193b-3p group had significantly increased number of NeuN(+)RORα(+) cells in the hippocampus ( P<0.05). (5) As compared with the control group, the LPS group had significantly decreased RORα expression, and significantly increased TNF-α, IL-1β and IL-6 mRNA expression in PC12 cells ( P<0.05). As compared with those in the LPS group, the RORα expression was significantly increased, and the TNF-α, IL-1β and IL-6 mRNA expression levels in PC12 cells were significantly decreased in LPS+miR-193b-3p group ( P<0.05). Conclusion:The miR-193b-3p inhibits the neuroinflammatory response in neonatal sepsis rats by regulating its target molecule RORα mRNA expression in hippocampal neurocyte.

Objective To investigate whether vascular endothelial cells in choroidal neovascularization whether hypoxia condition can up-regulate SNAI1 and activate matrix metalloproteinase (MMP)2 and MMP9 therefore to participate in choroidal neovascularization(CNV).Methods Sixteen SPF male C57 mice aged 6-8 weeks were divided into control group and model group.CNV models were induced by retinal laser photocoagulation,and flatmount and frozen sections of retinal pigment epithelium (RPE)-choroid-sclera compound were prepared at 7 days after modeling.The CNV in flat-mount was examined by Isolectin B4 staining,and the location of SNAI1,MMP2 and MMP9 in frozen sections was determined by immunofluorescence technology.The expression of SNAI1,MMP2 and MMP9 at mRNA level in CNV was detected by real-time fluorescence quantitative PCR (real-time PCR).The use and care of experimental animals complied with Statement for the Use of Animals in Ophthalmic and Visual Research.The RF/6A cells were divided into normoxia group and hypoxia group and cultured for 24 hours in 5％ CO2condition and mix condition of 94％ N2,5％ CO2 and 1％ O2,respectively.The expression of SNAI1,MMP2 and MMP9 in the cells at mRNA and protein levels was detected by real-time PCR and Western blot assay,respectively.Small interfering RNA of SNAI1 (siSNAI1) was transfected into the cells,and then the expression of MMP2 in the cells at mRNA and protein levels was detected by real-time PCR and Western blot assay,respectively,and the migrating number of the cells was assayed by Transwell chamber assay.Results CD31 and SNAI1 positive-response cells were seen in RPE-choroid-sclera flat-mounts under the laser scanning confocal microscope.The relative expression levels of SNAI1mRNA and MMP2 mRNA in RPE-choroid-sclera tissues were higher in the model group than those in the control group (SNAI1 mRNA:1.291 ±0.060 vs.0.759±0.074,P =0.001;MMP2 mRNA:1.610±0.424 vs.0.772 ±0.080,P =0.044).The expression of MMP9 mRNA was not significantly elevated between model group and control group (P＞0.05).The relative expression level of MMP2 mRNA was higher in comparison with MMP9 mRNA in the model group (P＜0.01).The relative expressions of hypoxic induced factor 1α (HIF-1α),SNAI1 and MMP2 at mRNA level and protein level in RF/6A cells were significantly higher in the hypoxia group than those in the normoxia group (all at P＜0.05) and no considerable difference was seen in MMP9 mRNA expression between the two groups (P＞0.05).The relative expressions of MMP2 mRNA in the cells were 0.217±0.036 and 0.818±0.105,and those of MMP2 protein in the cells were 0.236±0.009 and 1.043±0.120 in the hypoxia+siSNAI1 group and only hypoxia group,respectively,with significant differences between them (P =0.002,0.003).The migrating number of the cells was (254.60 ±71.31)/field in the hypoxia+siSNAI1 group,which was significantly less than (534.10±96.21) /field in the control group (P =0.029).Conclusions The hypoxic environment at CNV can activate MMP2 by up-regulating the expression of SNAI1,which promotes the migration of vascular endothelial cells and therefore participates in CNV formation,and the intervention of SNAI1 activation under the hypoxic condition can inhibit this process.