Objective To construct the recombinant plasmid containing human filaggrin gene,purify and identify the immunoreactivity of the recombinant protein,and establish the indirect ELISA to detect AFA for diagnosis of RA.Methods The constructed plasmids were transformed into E. Coli Rosettagami(DE3).This fusion protein was purified by NAT chromatography.ELISA coated with the fusion protein Was established to detect the AFA in serum of patients,which included 114 cases of RA,56 cases of SLE,32 cases of OA and 40 cases of normal controls. The correlation between the results of AFA and anti-CCP in RA group were compared. Results 321 bp fragment of filaggrin gene was amplified and the recombinant expression vector pET-28a( + )-filaggrin was constructed. The sequence of filaggrin gene was the same as the sequence reported in the literatures. The Rosetta-gami (DE3) strains of E. Coli with recombinant vector showed high level of filaggrin protein after induction. The SDS-PAGE showed that the plasmid expressed the filaggrin fusion protein with molecule weight of 14 000 Da. The expression protein could be purified by Ni-NAT with activity. The absorbance value of AFA in RA group was 0.473 ±0. 248 while they were 0. 160 0. 088, 0. 121±0. 070, 0.050 0. ±018 in SLE, OA and normal groups respectively. There were significant differences of absorbance values of AFA between RA and SLE, OA, control group (t = 12.004, 14. 464, 18.078, P<0. 01, respectively). The positivities of anti-filaggrin in RA, SLE and OA were 48.2%, 5.4% and 3. 1% respectively. The positivities of AFA were significantly different between RA, OA and normal control groups (x~2 = 67. 088, P < 0. 01). There was positive correlation of results between AFA and anti-CCP antibody (r = 0.42, P < 0. 05 ) . The consistency rate of results between AFA and anti-CCP was 70. 1%. Anti-CCP was negative in 10 out of 114 patients with AFA positive. AFA can be used to diagnose RA with sensitivity of 48. 2% , specificity of 96.9% , positive predictive value of 93. 2% and negative predictive values of 67. 9% . Conclusions The purified human filaggrin fusion protein is successfully purified. The indirect ELISA method based on the recombinant protein shows good sensitivity and specificity. Joint detection with AFA and anti-CCP can improve the positive rate of detection.

To discuss clinicopathological features and molecular genetic change of congenital mesoblastic nephroma (CMN).Methods Nine cases diagnosed as CMN were analyzed retrospectively in this study.Histological features,immunohistochemical profiles and ETV6 gene rearrangement status were assessed.Results All patients were within two years of age and eight of them were within one year.The average diameter of tumors was 9.5 cm (3.2-15.0 cm).These series cases included 3 classic CMN,5 cellular CMN and 1 mixed CMN.Cystic degeneration was found in 5 cases,and cartilage islands were observed in 2 cases.Compared with classic CMN,tumor size was bigger,and hemorrhage,necrosis and mitotic figures were easily to see in cellular CMN.All the tumor cells were positive for vimentin and negative for WT-1 by immunohistochemistry.ETV6 gene rearrangement was detected in 5 cases (including 4 cellular CMN and 1 classic CMN).Three cellular CMN harbored ETV6 gene translocation,1 mixed CMN and 1 cellular CMN were negative for ETV6 gene translocation by FISH analysis.The follow up data were obtained in 7 cases and 2 cases were lost.All the 7 patients were alive without evidence of recurrence and metastasis from 5 to 46 months.Conclusion CMN is a rare infant renal tumor with unique clinicopathological characteristics.Most of cellular CMNs harbor ETV6 gene translocation.The prognosis of CMN is relative good and needs to be differentiated from other malignant renal tumors.

Objective To compare the features of OCT angiography (OCTA) between neovascular agerelated macular degeneration (nAMD) and myopic ehoroidal neovascularization (mCNV) patients before and after intravitreal anti-VEGF treatment.Methods A prospective cohort study.Twenty-nine patients (37 eyes) with nAMD (19 males and 10 females,aged 68.20±8.76) and 31 patients (34 eyes) with mCNV (9 males and 22 females,aged 43.10± 11.80,with the mean diopter of-9.71 ± 1.20 D) from Department of Ophthalmology,West China Hospital of Sichuan University during May and December 2017 were included in this study.Ranibizumab or Conbercept (0.5 mg/0.05 ml) was intravitreally injected in all eyes.The patients were follow-up for 3-6 months.The OCTA was conducted before treatment and 1 day,1 week,1 month and 3-6 months after treatment.In order to ensure that the scanning position was the same,the tracking mode was adopted for each scanning.According to the OCTA images,the lesion area,parafoveal superficial vessel density and perfusion area were measured and analyzed contrastively between nAMD and mCNV patients.Results The mean lesion area before and 1 month after treatment in nAMD patients were 0.38± 1.87 mum2 and 0.06±0.12 mm2,while in mCNV patients,those were 0.26± 1.06 mm2 and 0.03 ± 0.05 mm2,respectively.There were statistically significant differences (Z=4.181,4.475;P＜0.001) in CNV lesion area before and 1 month after treatment between nAMD and mCNV patients.Compared with those before treatment,the absolute change (Z=1.853,P=0.064) and the percentage changes (t=2.685,P=0.010) of CNV lesion area l month after treatment in nAMD and mCNV patients show a statistical meaning.There were significantly decreases in both parafoveal superficial vessel density (F=8.997,P=0.003) and perfusion area (F=7.887,P=0.015) 3 months after treatment in nAMD patients,while decreases in parafoveal superficial vessel density (F=11.142,P=0.004) and perfusion area (F=7.662,P=0.013) could be detected 1 day after treatment in mCNV patients,before rising 1 month after treatment.Conclusions There are significantly differences in lesion area before and after the treatment of intravitreal anti-VEGF between nAMD and mCNV patients by OCTA examination.Moreover,the changes of both parafoveal superficial vessel density and perfusion area after anti-VEGF treatment are statistically different in two groups.

Objective : To evaluate the immune effects of inactivated H7N9 influenza vaccine. Methods : We searched several common databases（The Cochrane Library,PubMed,China Biology Medicine disc,China National Knowledge Infrastructure,etc.）for research articles about immune effects of H7N9 influenza vaccine published from the time the database built to July 10th of 2018,using H7N9 and vaccine as keywords. After screening the articles according to the inclusion and exclusion criteria,we assessed the quality of the studies and then employed seroconversion rate（SCR）as an outcome indicator to analyze the immune effects of different doses and adjuvants. Results : We recruited 5 articles on inactivated H7N9 influenza vaccine from 1 679 articles. The sample size was 2 579. The results of the meta-analysis showed that the rate difference（RD）values of SCR in each dose group after the first dose ranged from 1% to 10%,which indicated a poor protective effect;after the second dose of immunization,the RD values of SCR in the vaccines without adjuvants ranged from 13% to 19%,which was not effective enough;the RD values of SCR in the vaccines with adjuvants ranged from 62% to 69%,which met the licensing criteria for influenza vaccine;better results could be achieved when immunized with two doses of vaccines with adjuvants（ RR=1.19,95%CI：1.02-1.39）;vaccines with AS03 or MF59 at the lowest dose of 3.75 μg had the same immune effects as ones at a dose of 15 μg;vaccines with AS03（RD=89%,95%CI：85%-93%）were superior to those with MF59（RD=42%,95%CI：9%- 75%）. Conclusion Inactivated H7N9 influenza vaccines could achieve good immune effects when inoculated two doses with adjuvants,and the minimum effective dose was 3.75 μg.

Objective To observe the clinical characteristics ofretinoblastoma (RB) in Southwest China.Methods A retrospective clinical study.From January 2010 to December 2017,66 RB patients diagnosed in Ophthalmology Department of West China Hospital of Sichuan University were included in the study.All the patients underwent ocular B-ultrasound,orbital CT or MRI examination.Ten patients underwent RetCam examination at the same time.Twenty-nine patients were diagnosed by histopathological examination,and 37 patients were diagnosed by clinical symptoms and imaging examination.According to whether the tumor invaded the orbit and optic nerve,it could be divided into extraocular stage and intraocular stage.Intraocular tumors were divided into A-E stages according to the international intraocular RB classification.Treatments were performed according to different stages.The general information,age at diagnosis,course of diseases (the time between onset symptoms and diagnosis),causes of visiting a doctor,classification,treatment methods and eyeball preservation rate were retrospectively analyzed.Results Patients all came from Southwest China (56 patients from Sichuan Province,2 patients from Yunnan Province,2 patients from Guizhou Province,and 6 patients from Tibet).The permanent residence were identified in 43 patients,including 27 patients (62.8％) from rural areas.There were 38 males (57.6％);50 unilateral tumors (75.8％) and 16 bilateral tumors (24.2％);51 firstvisiting patients (77.3％) and 15 re-visiting patients (22.7％).The average diagnostic age of first-visiting patients was 20.9 ± 14.4 months,with 23.2 ± 14.7 and 11.2 ± 7.6 months for unilateral and bilateral tumors,respectively.There were 41 patients had definite course and causes,of whom the average course was 90.6± 115.2 days.The most common cause was leucocoria in 32 patients (62.7％),followed by redness and swelling in 4 patients (9.8％),and other causes in 5 patients (12.2％).Among the 15 re-visiting patients,the average diagnostic age was 63.6± 46.8 months,the average course was 32.8 ± 45.5 months.Recurrence was occurred in 5 patients (33.3％),leucocoria in 4 patients (26.7％),postoperative complication in 3 patients (20.0％),protrusion in 2 patients (13.3％) and redness in 1 (6.7％) patient,respectively.Fifty out of 82 eyes were admitted to hospital,including 37 eyes of first-visiting patients and 13 eyes of re-visiting patients.Among 37 first-visiting eyes,there were 5 eyes (13.5％) in stage A-C,26 eyes (70.3％) in stage D-E,6 eyes (16.2％) in extraocular stage.Five eyes in stage A-C were treated with laser photocoagulation and (or) cryotherapy combined with systemic chemotherapy.Four eyes in stage D were treated with intraocular arterial chemotherapy.Nineteen eyes (51.3％) were performed with enucleation,2 eyes (5.4％) with evisceration and 7 eyes (18.9％) abandoned treatment.Among 13 re-visiting eyes,6 eyes (46.2％,with 5 eyes of recurrence) had been enucleated before,4 eyes (30.8％) were in extraocular stage and 3 eyes (23.1％) in stage D-E.Five eyes (38.5％) were treated with evisceration,4 eyes (30.8％) with enucleation,1 eye with oculoplastic surgery and 3 eyes (23.1％) abandoned treatment.The rate of eye preservation was 18.0％,29.0％ for intraocular stage and 0％ for extraocular stage,respectively.Conclusion RB patients in Southwest China have a longer course between onset symptoms and diagnosis,more advanced classification and lower rate of eye preservation.

Objective To investigate the effect of miRNA-199a-3p overexpression on the expression of MAP3K4 protein in gastric cancer. Methods 35 gastric cancers and the matched adjacent tissue specimens were collected. Expression of miRNA-199a-3p and MAP3K4 were detected by stem-loop real-time reverse transcription polymerase chain reaction and western blot. Cell transfection was employed to explore the regulation of miRNA-199a-3p on MAP3K4 gene. Luciferase reporter assay was performed to identify target MAP3K4 gene. Results Com-pared with the adjacent tissue specimens ,miRNA-199a-3p was upregulated in the gastric cancers ,and MAP3K4 protein was down-regulated in the gastric cancers. Cells transfected with miR-199a-3p mimics showed lower MAP3K4 protein. MAP3K4 was identified as target gene of miR-199a-3p. Conclusions miRNA-199a-3p acts as an oncogene in gastric cancer and functions by targeting MAP3K4.

Objective To investigate the prevalence of urine abnormalities for school children in Chengdu city and to evaluate the significance of urinary screening.Methods During January to December 2013,morning urine of 6 615 students were collected and screened by urine reagent paper.Two weeks later,the repeated screening was conducted in the children whose urine samples were positive for the first screening.Urine samples with positive testing results for twice were submitted to urine routine tests at local hospital,and the children with the urine positive results were defined as urine abnormalities.The children with urine abnormalities were transferred to a tertiary hospital and given treatment and follow-up.Results There were 6 615 cases receiving urine screening,including 2 624 cases (39.67 ％) of the grade I,and 3 991 cases(60.33％) at junior middle school.During the first screening,323 cases (4.83％) children had urinary occult blood positive,43 cases (0.65％) had urinary protein,20 cases (0.30％) had occult blood positive and proteinuria,and 103 cases (1.56％) had white cells in urine.During the second urine screening,62 cases (0.94％) had occult blood positive,6 cases (0.09％) had urinary protein,2 cases (0.03％) had proteinuria and occult blood positive,46 cases (0.70％) had white cells in urine.The incidence of urine abnormalities with occult blood positive,proteinuria,occult blood positive and proteinuria,and white cells in urine of children at junior middle school [1.38％ (55/3 991 cases),0.13％ (5/3 991 cases),0.05％ (2/3 991 cases),0.70％ (28/3 991 cases)] were significantly higher than those of children at primary school [0.27％ (7/2 624 cases),0.04％ (1/2 624 cases),0 (0/ 2 624 cases),0.69％ (18/2 624 cases)],and all the differences were statisticallysignificant (x2 =64.16,168.53,178.09,98.16,all P ＜ 0.05).In children transferred to a tertiary hospital for treatment,there were 4 cases with IgA nephropathy,1 case with minor glomerular abnormalities,and 12 cases with urinary tract infection.Conclusion Urinary screening is an effective way to find out kidney disease and urinary tract infection in children.Follow-ups should be strengthened.