ObjectiveTo develop a national standard for genomic titer determination of recombinant type 5 adeno-associated virus(rAAV5).MethodsThe rAAV5-GFP stock solution prepared by the three-plasmid system was identified and verified for the appearance,pH,sterility,genomic titer,purity and infection titer according to the relevant requirements of Chinese Pharmacopoeia(Volume Ⅲ,2020 edition),which was diluted and subpackaged to prepare candidate standards according to the results;The stability of candidate standards was investigated by thermal acceleration test;Three laboratories were organized to collaboratively calibrate the candidate standards using droplet digital PCR(ddPCR).ResultsAll the detection indexes of the candidate standard and the stock solution met the relevant requirements;The genomic titer showed no significant decrease at 25,4,-20,-40,-80 ℃ for 1,3,4,6 months;Through collaborative calibration by three laboratories,the candidate standard was assigned a value of 2. 56 × 10(12)copies/mL,and the 95% confidence interval was 2. 48 ×10(12)copies/mL,and the 95% confidence interval was 2. 48 ×10(12)copies/mL ～ 2. 64 × 10(12)copies/mL ～ 2. 64 × 10(12)copies/mL.ConclusionThe developed national standard for the determination of rAAV5 genomic titer had good stability and might be used for the quality evaluation of rAAV5 related products.

Pertussis is an severe infectious respiratory disease that continues to be a serious public health problem,which is caused by Bordetella Pertussis.Because it's highly infectivity,it is still an important etiological factor associated with persistent cough in adults and adolescents.With the introduction of mass vaccination in 1974,there has been a dramatic decrease in the morbidity and mortality.However,in recent years,increasing numbers of pertussis are reported in adolescents and adults in many countries where with high vaccination coverage.The thesis summarize and discuss the progress of pertussis cpidemiology and clinical characteristics.

Objective To observe the effect of combination therapy with methotrexate,sulfasalazine and hydroxychloroquine on serum lipids in elderly patients with rheumatoid arthritis (RA).Methods 136 patients with RA were randomly divided into group A and group B (n=68,each).Patients in group A were treated with methotrexate,sulfasalazine and hydroxychloroquine sulfate plus life style intervention.Patients in group B were treated with the therapy as same as that of group A and received xuezhikang additionally.The levels of cholesterol (TC),low density lipoprotein cholesterol (LDL-C),triglyceride (TG),high density lipoprotein cholesterol (HDL-C),C-reactive protein (CRP),erythrocyte sedimentation rate (ESR) were determined before and 6 months after treatment.Disease activity was assessed by DAS28,swelling index,joint pain index,duration of morning stiffness.Results The levels of TC,TG and LDL-C were lower and HDL-C level was higher in the two groups after 6 months of the treatment than before treatment (all P＜ 0.05).DAS28,ESR,CRP,joint tenderness,joint swelling and duration of morning stiffness were significantly reduced in two groups after 6 months of the treatment as compared with pretreatment (all P＜0.05).There was no significant difference in levels of TC,TG,LDL-C,HDL-C,DAS28,ESR,CRP,joint tenderness,joint swelling and duration of morning stiffness between the two groups after treatment.There were no obvious differences in liver and kidney function between the two groups before and after treatment.Conclusions Blood lipids levels are abnormal in RA patients.The slow acting antirheumatic drugs have certain effect on blood lipids when alleviating disease symptoms.

Objective To explore the effects of psychological intervention on negative emotions and quality of life in patients with rheumatoid arthritis (RA).Methods A total of 112 patients with rheumatoid arthritis were divided into two groups:the intervention group and the control group.All patients were given drugs for treatment of RA.Additionally,patients in the intervcntion group were received psychological intervention.Zung's self-rating depression scale (SDS),Zung's self-rating anxiety scale (SAS),RA disease activity score (DAS28),visual analog scale (VAS),disability index of health assessment questionnaire (HAQ-DI) were assessed,and the related laboratory parameters such as erythrocyte sedimentation rate (ESR),C reactive protein (CRP),rheumatoid factor (RF)were determined.Results 6 months after the treatment,there were significant differences in SDS and SAS between the treatment group and the control group [SDS:(31.7±4.7)scores vs.(29.6±4.4)scores; SAS:(34.7±5.3)scoresvs.(32.6±5.1)scores; t=2.441,-2.119,both P＜0.05].The patients with tender joints,joint swelling and morning stiffness time were less in the treatment group than in control group [(5±3) vs.(7±4),(4±3) vs.(5±3),(36±16) min vs.(42±19)min.respectively,t=2.054,2.393,1.995,all P＜0.05].The scores of HAQ-DI,VAS and DAS28were lower in the treatment group than in control group [(0.89±0.73) vs.(1.20±0.77),(6.1±1.6) vs.(6.8±1.8),(3.2±1.7) vs.(3.9±1.9),respectively,t=2.136,2.406,2.135,all P＜0.05].There was a significant difference in CRP between the treatment group and the control group [(34±14) mg/L vs.(37± 17)mg/L,t=2.406,P＜0.05].Conclusions Psychological intervention can improve the negative emotions and quality of life in patients with rheumatoid arthritis.

Objective To observe the effect of combination therapies with methotrexate, sulfasalazine and hydroxychloroquine on hyperlipemia of rheumatoid arthritis.Method From 2009 November to 2010 November, 68 RA inpatients and outpatients in the department of rheumatism of our hospital were randomly divided into two groups: A group and B group, 34 cases in each group. The patients in A group were given combination of methotrexate, sulfasalazine, hydroxychloroquine sulfate for the treatment of RA,patients in B group, based on A group's treatment, were treated by xuezhikang. Some indexes were observed in two group before treatment and 6 months after treatment, including cholesterolTC,low density lipoprotein cholesterol LDL-C , triglyceride TG , high density lipoprotein cholesterol HDL-C , swelling index, joint pain index, time of morning stiffness, C-reactive protein CRP , erythrocyte sedimentation rate ESR and DAS28. Results After 6 months’ treatment, the serum levels of TC, TG,, LDL-C, both in two groups of patients, were lower than those before treatment, and HDL-C was higher than that before treatment, there were significant differences (P<0.05 or P<0.01) . After 6 months of treatment,DAS28,ESR,CRP,joint tenderness,joint swelling and morning stiffness time, both in two groups RA patients, significantly reduced compared with before treatment, there were a very significant difference (P<0.05 or P<0.01) .After treatment, the above indexes had no statistically significant difference between two groups (P>0.05) . Conclusion RA patients have abnormal blood lipid levels, disease-modifying anti-rheumatic drugs have effect on blood lipid during remission joint disease in RA patients.

Objective To assess the efficacy of clinical intervention of early swallowing syndrome on neonates. Methods Eighty-six cases of normal newborns were enrolled in the study, including 44 cases of natural delivery,32 cases of cesarean section,8 cases of premature infant. The cases started vomitting before feeding the milk, became more severe after feeding the milk. once the pathological factors were excluded, the cases were treated with breastfeeding, warm water enema, massaging the lower abdomen, 1% sodium bicarbonate solution to the gastric lavage. Results A variety of clinical interventions on the control of neonatal swallowing syndrome results are obvious. Conclusions Early detection early intervention can prevent complications occurring.

Objective To investigate the incidence of neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD)in neonates with idiopathic neonatal cholestasis (INC)in Nanjing,China,SLC25A13 gene mutations in these neonates,and clinical features.Methods A total of 152 neonates with INC,who were admitted to the Affiliated Nanjing Children's Hospital of Nanjing Medical University from Sep-tember 2009 to August 2013,underwent gene analysis for detecting SLC25A13 gene mutations.The neonates were divided into NICCD group,who had been diagnosed definitely,and INC group at a ratio of 1∶2,considering the age and gender.Several biochemical indices were compared between the two groups.Comparison of continuous data between the two groups was made by Mann-Whitney U test after Bonferroni correction.Results There were 21 confirmed cases of NICCD (21/152,13.82%)among the 152 neonates with INC;five types of SLC25A13 mutations were identified in the 21 neonates with NICCD,including 851_854del (27/42,64.29%),IVS6+5 G→A (7/42, 16.67%),1638ins23 (5/42,11.90%),IVS11 +1 G→A (2/42,4.76%),and Q259X (1/42,2.38%).The alanine aminotransferase (ALT)level,aspartate aminotransferase (AST)level,bile acid concentration,albumin level,fasting blood glucose,blood ammonia,and prothrombin time for the NICCD group were 39.42 ±23.40 U/L,124.85 ±92.65 U/L,142.43 ±24.34μmol/L,30.66 ±2.70 g/L,2.79 ± 0.54 mmol/L,117.57 ±27.88 μmol/L,and 14.03 ±2.79 s,respectively,versus 136.02 ±113.67 U/L,226.12 ±129.26 U/L,80.47 ± 31.53 μmol/L,36.87 ±4.96 g/L,3.14 ±0.45 mmol/L,76.43 ±20.80 μmol/L,and 11.40 ±1.55 s for the INC group.The NICCD group had significantly lower ALT and AST levels than the INC group (Z=-5.02,P=0.000;Z=-3.66,P=0.000);the NICCD group had a significantly higher bile acid concentration than the INC group (Z=-5.58,P=0.000);the NICCD group had significantly lower albumin level and fasting blood glucose than the INC group (Z=-4.52,P=0.000;Z=-2.56,P=0.010);the NICCD group had a significantly higher blood ammonia level than the INC group (Z=-4.75,P=0.000);the NICCD group had a significantly longer prothrombin time than the INC group (Z=-4.10,P=0.000).Conclusion Citrin deficiency due to SLC25A13 gene mutations is an im-portant cause of INC in Nanjing.The three most common mutations are 851_854del,IVS6+5 G>A,and 1638_1660dup23,which account for 92.86% of the SLC25A13 gene mutations.More attention should be paid to clinical analysis and detection of SLC25A13 gene mutations to confirm the diagnosis of NICCD.

Objective To carry out the detection of JAGI gene in children with chronic cholestasis and to im-prove the diagnostic level and understanding of Alagille syndrome. Methods Two cases of chronic cholestasis with multiple organ involvement were selected as the research subjects and their clinical data,laboratory test results were col-lected. Two milliliter peripheral intravenous heparin anticoagulan blood was drawn from each patient. All fragments of 26 exons of the JAGI gene were amplified by polymerase chain reaction - sequence based on typing method. Results One patient with chronic cholestasis,heart murmur and dysmorphic face showed bile duct paucity in liver biopsy and a novel heterozygous mutation c. 809 809delG(p. G270Dfs*142)in 6 exon. Abnormal amino acid replaced JAG1 protein and resulted in truncation of the JAG1 protein. The part of epidermal growth factor(EGF)like repeats region loss and the cysteine rich region completely lost. One case with typical chronic cholestasis and dysmorphic face showed a known IVS20 - 2 5delTAAG heterozygous mutation which resulted in splice site changes. Conclusion A novel JAGI gene mutation c. 809 809delG(p. G270Dfs*142)is helpful to screen JAGI gene of Notch signal transduction pathway for chronic cholestasis with multiple organs involvement in children.

Objective To investigate the relationship between acupuncture analgesia and cellular immunity, the acupuncture effect on expression of c fos mRNA, ppENKmRNA,iNOSmRNA and iNOS activity in the mouse peritoneal macrophage were studied. Methods Twenty BALB/c mice were randomly divided into 2 groups:a,the acupuncture group treated with 5Hz electroacupuncture (EA); b, the control group treated with no EA. The pain threshold was detected by K + ionophoresis method before and after EA or restraining. The experiment was performed by using in situ hybridization, NADPH NBT histochemistry, RNA dot blot and protein dot blot techniques. All dot blot signals were scanned for statistical analysis. Results The hybridization signals and iNOS activity appeared as granules in bluish violet color, localized in the cytoplasm of the macrophage. All dot blot signals were increased in the acupuncture group, when compared with those in the control group( P

Objective To observe the manifestation of fundus angiographs of polypoidal choroidal vasculopathy (PCV). Methods Twelve PCV patients involved 7 males and 5 females aging from 40 to 69 year old (average 56.4). Fundus examination, fundus fluorescein angiography (FFA), and indocyanine green angiography (ICGA) were performed on 12 patients (12 eyes) with PCV, out of whom 5 underwent optical coherenece tomography (OCT). Results In 12 eyes, deep and (or) superficial hemorrhage and yellow hard exudations were found, including orange-red lesions in 6 and pre-retinal hemorrhage in 2. The results of FFA discovered orange-red spotty fluorescence in 6 eyes and choroidal vascular network in 4 eyes. At the late phase, leakage of polypoidal hyperfluorescence spot in all of the eyes except 2 without leakage were found. The images of ICGA showed typical dotted or clustered polypoidal hyperfluorescence in 12 eyes at the late phase. OCT disclosed protrusion of the retinal pigment epitelium (RPE) with a bumpy surface at polypoidal structure in 4 eyes and no change in 1 eye. Conclusions PCV mainly affects the elderly persons and mostly on unilateral eyes. Macular hemorrhage, serous RPED, and (or) neuroepithelial detachment with yellow hard exudations are the main manifestations. Branching choroidal vascular net with ployplike terminal anourysmal dilations can be discovered in FFA and ICGA.