Objective Choose an easy method to screen the “leaky" phenotype SCID mice before experimental use. Methods IgG in SCID mice serum was assayed by three kinds of ELISA(sandwich ELISA, sandwich indirect ELISA and Avidin\|biotin sandwich ELISA). Results Avidin\|biotin sandwich ELISA is the most sensitive method of the three, and 41 SCID mice of 6～8 weeks old were assayed with this method. The average IgG of 40 SCID mice was 0 24?0 16mg/L, only one was 2\^3mg/L. The “Leaky" probability was 2\^4%.Conclusion\ The Avidin\|Biotin sandwich ELISA is an easy, sensitive and reliable method to screen“leaky" SCID mice.\;[

Objective To observe the curative effect and adverse reactions of intercostals nerve block combined with fentanyl transdermal system in treatment of post-thoracotomy pain syndrome (PTPS). Methods Intercostals nerve block combined with fentanyl transdermal system was used for treatment of 141 patients with PIPS. The degree of pain relief was evaluated by numerical rating scale (NRS),the quality of life of patients was evaluated by the quality of life score standard,and the adverse reactions were also observed. Results The overall remission rate was 97.85%,there were 51 cases (36.43%)with complete remission, 80 cases (57.14%)with significant remission,and 6 cases (42.85%) with moderate remission. The adverse reactions included nausea,vomiting, constipation,dizziness,skin pruritus and dysuria,no serious liver and kidney function damage and respiratory inhibition. The quality of life of all patients was improved. Conclusion Multimodal analgesia has better curative effect and less adverse reactions in treatment of PTPS,so it deserves clinical promotion.

Objective Fecal biomarkers have emerged as an important tool for assessing and monitoring disease activity in patients with inflammatory bowel disease ( IBD) .We aimed to investigate the diagnostic value of fecal neopterin and calprotectin in pa-tients with active inflammatory bowel disease and made comparison with that of serum C-reactive protein ( CRP) . Methods A total of 151 consecutive patients with IBD (84 CD and 67 UC) provided 2 gram fecal samples for the measurement of fecal neopterin( FNP) and calprotectin( FCP) concentrations and 2 milliliter blood samples for the serum C-reactive protein measurement before undergoing a colonoscopy.ELISA was applied in the measurement.Clinical disease activities were scored independently according to the Best Crohn′s Disease Activity Index(CDAI) in patients with CD, while the Modi-fied Mayo Scores in patients with UC.Comaprison was made in the relativity of each fecal marker and IBD activity score, the optimum value of diagnosing IBD acitivity as to each fecal marker, as well as sensitivity, specificity, moreover, receiver operating characteristic curve ( ROC) was drawn.50 healthy volunteers who received a normal colonoscopy were also enrolled as the control group and asked to give a 2 gram fresh stool sample. Results The FNP and FCP concentrations in patients with IBD were significantly higher than those in healthy control group(P<0.05).Both FNP and FCP concentrations differed significantly in clinically active IBD when compared with those in patients with inactive disease( P<0.001) .In CD patients, the correlation coefficients of FNP and FCP with CDAI were 0.55 and 0.59, respectively(P<0.001).In UC patients, the correlation coefficients of FNP and FCP with Mayo scores were 0.74 and 0.77, respectively( P<0.001) .The correlation coefficients of serum CRP in CD and UC patients with clinical scores were 0.49 and 0.60, respectively(P<0.001).The area under the ROC curve(AUC) of FNP and FCP for the diagnosis of clinical activity in pa-tients with CD were 0.75 and 0.80, respectively.The AUC of FNP and FCP in UC patients were 0.85 and 0.90, respectively.The AUC of serum CRP in patients with CD and UC were 0.65 and 0.74, respectively.When combined FNP with FCP, the AUC in pa-tients with CD and UC were 0.85 and 0.92, respectively. Conclusion FNP is a novel reliable and non-invasive biomarker to evalu-ate clinical disease activity in patients with IBD as accurate as FCP, It is advisable to combine FNP with FCP to evaluate disease activi-ty in patients with IBD.

Background:Peroral endoscopic myotomy(POEM)is a novel minimally invasive endoscopic technique for treatment of esophageal achalasia(EA). It has been demonstrated to be safe and effective in several short-term clinical studies, however,studies focusing on its mid- and long-term outcome are rare. Aims:To investigate the mid-and-long term efficacy of POEM in EA patients. Methods:A retrospective analysis was performed in 41 EA patients who underwent POEM in the Department of Gastroenterology and Hepatology of Nanjing General Hospital of Nanjing Military Region from Jul. 2012 to Dec. 2013. All these patients were followed up periodically for clinical symptoms(Eckardt score),maximum esophageal diameter and lower esophageal sphincter pressure(LESP),and the intra- and post-operative complications and recurrence were recorded. Results:POEM was successfully completed in all 41 EA patients,the mean operative time was(69. 5 ± 20. 6)min(range 35-125 min),and the mean length of hospital stay was 3. 5 days. Compared with the pre-operative data,the post-operative Eckardt score,maximum esophageal diameter and LESP during follow-up(mean 26. 3 months, range 19-36 months)were significantly decreased(P all < 0. 05). Procedure-related complications were seen in 6 patients (14. 6% );the clinical success rate was 95. 1%(39 / 41)and the recurrence rate was 2. 4%(1 / 41). Conclusions:POEM is safe and effective in mid-and-long term follow-up with low recurrence rate. It might become the first-line therapy for EA,and the long-term efficacy and complications need further follow-up study.

Objective To study the effect of autoantibody test which includes antinuclear antibodies(ANA),antinuclear antibody fluorescence patterns,anti-extractable nuclear antigen(ENA) antibodies and anti-double strands DNA(ds-DNA) antibodies in the diagnosis of pediactic autoimmune diseases.Methods Of all the inpatients which had positive results of autoantibody tests,135 cases were reviewed.The autoantibody assay,positive value(PV) analysis were performed respectively.Results PV of ANA test to autoimmune diseases was 0.36 which was proportional to the intensity of fluorescence;Of all the fluorescence patterns,speckled(fine) had a relatively high PV;Anti-ENA and anti-dsDNA antibody tests had higher PV than ANA test.Conclusion Fluorescence intensity,(anti-ENA) antibody test and anti-dsDNA antibody test may be useful in identifying autoimmune diseases in clinic.

OBJECTIVETo study the effect of bifidobacterium genomic DNA on umbilical cord blood mononuclear cell (CBMC), and investigate the immunoregulation of bifidobacterium DNA and explore possible mechanisms by which bifidobacterium acts against allergic reaction.

METHODSBifidobacterium genomic DNA (bDNA) and human DNA (hDNA) were extracted with phenol/chloroform/isoamyl alcohol and stored at -20 degrees C for later use. Parts of bDNA were completely digested with DNaseI (d-bDNA) at 37 degrees C. CBMCs were isolated with Ficoll from umbilical cord blood and incubated at 37 degrees C in a 5% CO2 humidified incubator. These cells were divided into four groups, control group: without any stimulant; bDNA group: stimulated with 25 microg/ml bDNA; d-bDNA group: stimulated with 25 microg/ml d-bDNA; hDNA group: stimulated with 25 microg/ml hDNA. The cells were stimulated with different stimulants in vitro, at the end of incubation culture supernatant and cells were collected. IL-12 and IL-10 levels in the culture supernatant were measured by enzyme linked immuno sorbent assay (ELISA); cells secreting IL-4 and IFN-gamma were counted by enzyme linked immunospot (ELISPOT) assay; and total RNA was isolated from the cells to assay T-bet and GATA3 mRNA expression levels by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSSix hours after stimulation there was no significant difference in IL-12 level in supernatant among the four groups; 12 hours after stimulation, IL-12 level in supernatant of bDNA treated group was significantly higher than that of each of the other groups, so were the results obtained at 24 hours and 48 hours after stimulation (P < 0.05). No significant difference could be detected in IL-12 level in supernatant among the other 3 groups. On the other hand, 6 hours after stimulation there was no significant difference in IL-10 level in supernatant among the four groups. But 12 and 24 hours after stimulation IL-10 level in supernatant of bDNA treated group was lower than that of each of the other groups, but the difference was not statistically significant. The count of IFN-gamma secreting cells of bDNA treated group was higher than that of the other groups, while IL-4 secteting cells of bDNA treated group were lower than that of the other groups. After bDNA stimulation, nuclear factor T-box expressed in T cells (T-bet) mRNA expression was conspicuously enhanced as compared to the other three groups (P < 0.05). GATA3 mRNA transcription in CBMC had no significant change after bDNA stimulation.

CONCLUSIONbDNA could promote secretion of Th1 type cytokine IL-12, while Th2 type cytokine IL-10 level of cell supernatant was decreased. bDNA could stimulate secretion of IFN-gamma by CBMC and inhibit secretion of IL-4. T-bet mRNA expression was highly enhanced after bDNA stimulation. bDNA could upregulate Th1 type response, which may be one of important mechanisms by which bifidobacterium inhibit allergic response.

Bifidobacterium ; cytology ; genetics ; Cell Culture Techniques ; DNA, Bacterial ; biosynthesis ; metabolism ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Fetal Blood ; cytology ; immunology ; GATA3 Transcription Factor ; genetics ; Humans ; Infant, Newborn ; Interferon-gamma ; immunology ; secretion ; Interleukin-10 ; immunology ; secretion ; Interleukin-12 ; immunology ; secretion ; Interleukin-4 ; immunology ; secretion ; Leukocytes, Mononuclear ; immunology ; secretion ; RNA, Messenger ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; T-Box Domain Proteins ; genetics ; Th1 Cells ; drug effects ; immunology ; secretion

OBJECTIVEX-linked agammaglobulinemia (XLA) is the most common disorder among primary immunodeficiency diseases, which is caused by mutations in the cytoplasmic Bruton's tyrosine kinase (BTK) gene, characterized by lack of mature, circulating B lymphocytes, hypogammaglobulinemia, and recurrent bacterial infections. Mutations in BTK are highly diverse. In this study, genetic analysis was performed on BTK to realize the feature of gene mutation of XLA in Mainland of China.

METHODSSeven patients from 7 different families were enrolled in the analysis. RT-PCR was employed to reverse transcript total RNA and 8 couples of primers were designed for PCR. PCR products were sequenced and the mutation sites were identified.

RESULTSSeven completely different mutations were identified in the 7 patients. All the 7 mutations located at BTK coding region. Three of the 7 mutations were located in pleckstrin homology functional area, 2 mutations located in BTK area, and in other 2 cases at Src homology 2 and Src homology 3 regions, respectively. The mutations in 2 of 7 cases were in exon 18, and the others were in exon 2, 5, 6, 8 and 10, respectively. The types of mutation included 3 missense (L11P, I590F and Y591S), two nonsense (W281X, and Q234X) mutations resulting in premature stop codons. A 10-base pair nucleotides duplicated insertion located between the nucleotide 596 and 597 resulting in frameshift, and a 8 base pair deletion at the nucleotide position 472 resulting in frameshift. Four of the 7 mutations are novel mutation types and have not been reported. Four of 7 mothers were analyzed, 3 of them were carrier and 1 was normal.

CONCLUSIONThe patients enrolled in this study had classical clinical features of XLA. All the 7 identified mutations located at BTK coding region and 4 of them were novel mutations. Genetic analysis can be used for diagnosis of XLA and distinguish it from other hypogammaglobulinemia.

Adolescent ; Agammaglobulinemia ; diagnosis ; genetics ; Base Sequence ; Child ; China ; Codon, Nonsense ; DNA, Complementary ; Genetic Diseases, X-Linked ; diagnosis ; genetics ; Genotype ; Humans ; Male ; Mutation ; Mutation, Missense ; Protein-Tyrosine Kinases ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult

OBJECTIVETo evaluate the influence of VPA treatment on neutrophils' oxidative metabolism and oxidant status in epileptic children.

METHODTwenty-six newly diagnosed epileptic children with idiopathic epilepsy and 30 healthy children were included in the study. The activation rates of neutrophils and stimulation indexes were detected in patients before and 6 months and 12 months after VPA treatment respectively and in all the healthy children by flow cytometry with dihydrorhodamine as fluorochrome. The activities of myeloperoxidase from neutrophils were also detected. Malondialdehyde as an indicator of lipid peroxidation and antioxidant enzymes including superoxide dismutase, catalase, and glutathione peroxidase were measured in plasma respectively.

RESULTThe activation rates of neutrophils in patients treated with VPA after 6 and 12 months were (11.50 ± 6.52)% and (14.31 ± 5.76)% respectively, which were significantly higher than the data of control group (5.90 ± 3.77)% and pretreatment level (7.42 ± 3.15)%. The stimulation indexes 6 and 12 months after VPA therapy were (474.88 ± 118.98) and (416.31 ± 110.00) respectively, which were lower than the data of control group (544.83 ± 140.83) and pretreatment level (535.23 ± 111.55). The plasma MPO activities and levels of malondialdehyde in VPA treated patients were also higher while the activities of SOD and CAT were significantly lower than the control and untreated groups. GSH-Px levels did not differ between the groups. Multiple linear regression analysis showed that the time of treatment and the activation rates of neutrophils were indicators which had positive correlation with the levels of plasma MDA and that SOD activities were inversely correlated with MDA levels.

CONCLUSIONVPA which is frequently used in childhood epilepsy may activate the neutrophils of patients and cause oxidative stress and prolonged treatment may aggravate it.

Anticonvulsants ; pharmacology ; therapeutic use ; Antioxidants ; pharmacology ; Case-Control Studies ; Catalase ; blood ; Child ; Child, Preschool ; Epilepsy ; blood ; drug therapy ; Female ; Glutathione Peroxidase ; blood ; Humans ; Lipid Peroxidation ; drug effects ; Male ; Malondialdehyde ; blood ; Neutrophils ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Superoxide Dismutase ; blood ; Valproic Acid ; pharmacology ; therapeutic use

OBJECTIVETo explore safety, indications and advantages of mapping and ablation of arrhythmia in children guided by Carto and Ensite system.

METHODSGuided by Carto system, radiofrequency catheter ablation (RFCA) was performed on 8 pediatric patients with tachycardia whose mean age was (6.2 + or - 1.7) years, mean weight was (18.0 + or - 2.0) kg. Guided by Ensite system, RFCA was performed on 10 pediatric patients with arrhythmia, 8 of them were ablated guided by Ensite Array system: 6 cases with premature ventricular contractions (PVCs), 2 cases with right atrial tachycardia, their mean age was (11.3 + or - 1.2) years, and mean weight (40.0 + or - 5.0) kg. The other two cases with W-P-W syndrome were ablated guided by Ensite Navx system.

RESULTGuided by Carto system, 8 cases were successfully mapped and ablated: 6 cases had incision atrial tachycardia, 1 case had left atrial tachycardia and 1 case had right atrial tachycardia. In 1 case with incision atrial tachycardia the condition recurred after 3 months, and was ablated again successfully. Guided by Ensite Array system, 6 cases with PVCs (in 2 originating from the right ventricular inflow tract and in 4 originating from the right ventricular outflow tract) and 2 cases with right atrial tachycardia were successfully mapped and ablated, PVCs of the first 6 cases were reduced from (32 333 + or - 4509) 24 h to (0-4)/24 h after ablation. In 1 case with automatic atrial tachycardia, mapping could not be done by Ensite Array system, because P wave could not be identified from T wave. Single bolus of adenosine 20 mg was given within 30 s to let ventricles stop for 2 s (cardio-ventricular pacing standby) until T wave vanished, mapping and ablation were operated again successfully, but another atrial tachycardia occurred 1 day later. Guided by Ensite Navx system, 2 cases with W-P-W syndrome were successfully ablated, operation under X-rays lasted for 8 and 10 min. In none of the 9 patients the disease recurred after follow-up for 6 months.

CONCLUSIONCarto system is suitable for mapping and ablation in pediatric patients with continuous tachycardia, especially with incision atrial tachycardia; Ensite Array system fits children older than 10 years with right heart discontinuous arrhythmia; and Ensite NavX system can set up model and display endocardial anatomic structure quickly. Compared with two-dimensional mapping system, the three-dimensional mapping system (Carto and Ensite) can display the origin of arrhythmia and activation sequence clearly, decrease difficulty of operation efficiently and diminish operation time under X-ray.

Arrhythmias, Cardiac ; physiopathology ; surgery ; Catheter Ablation ; methods ; Child ; Child, Preschool ; Electrophysiologic Techniques, Cardiac ; methods ; Humans ; Imaging, Three-Dimensional ; Treatment Outcome

Objective To study the relativity between imageology and pathology during lung cancer,and estimate whether the lung cancer is preinvasive lesions,which can support evidences for the operation methods.Methods Clinical data of 624 patients who were diagnosed as lung adenocarcinoma and had solitary pulmonary nodule(diameter≤3 cm) were collected,all of them were scanned by thin layer CT scan(1 mm).The correlation between imageology and pathology data were analyzed.Results In 125 cases of GGO,the ratio of invasive lesions were 0 (0/72),6.1％ (3/49) and 100％ (4/4) in stage T1a,T1b and T1c respectively.In 285 cases of mGGO,if solid component was less than 0.5 cm,the ratio of invasive lesions were 1.7％ (1/58),6.9％ (2/29) and 50.0％ (2/4) in stage T~,T1b and Tic;but the ratio of invasive lesions were 81.3％ (13/16),94.1％ (96/102) and 97.4％ (74/76) respectively when the solid component was more than 0.5 cm.In 214 cases with solid nodules,the ratio of invasive lesions were 87.1％ (27/31),98.8％ (84/85) and 99.0％ (97/98) in stage T1 a,T1b and T1c.Conclusion The ratio of invasive lesions and solid component increased gradually along with the growing of tumor diameter in stage T1 lung cancer.CT imaging was highly correlated with the pathology diagnosis of preinvasive lesions and invasive lesions,which can be used as the guidance for operation methods.