Protective effect and mechanism of serum containing Euonymus fortunei on rat pancreatic islet cells / 器官移植

Objective To investigate the protective effect and mechanism of serum containing Euonymus fortunei on the rat pancreatic islet cells. Methods Forty male SD rats were randomly divided into 5 groups (n=8 in each group), including the control group (normal rat islet cells were cultured with normal rat serum), ischemic preconditioning group (abdominal aorta was blocked first and then re-opened before the pancreas was obtained, and the pancreatic islet cells were cultured with normal rat serum), Euonymus fortunei treatment group (normal rat islet cells were cultured with rat serum containing Euonymus fortunei), Euonymus fortunei group and blank group (normal rats were administered orally with Euonymus fortunei extract or distilled water for the preparation of rat serum). Diphenylthiocarbazone (DTZ) staining was utilized to observe and calculate the quantity of islets. Acridine orange (AO)/propidium iodide (PI) staining was adopted to calculate the survival rate of islet cells. The insulin release experiment was performed to calculate the stimulation index (SI) and evaluate islet cell function. The concentration of glutathione (GSH) and nitric oxide (NO) in islet cells was detected using GSH and NO kits. The expression level of inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) was quantitatively measured by reverse transcription polymerase chain reaction (RT-PCR). Results Islet cells were observed in specifically scarlet color after DTZ staining. The quantity of islet cells did not significantly differ among different groups (all P>0.05). Along with the prolongation of culture time, the activity of islet cells in each group was gradually decreased. At 72 h after isolation and culture, compared with the control group, the survival rate of the cells was significantly higher in the Euonymus fortunei treatment group (P<0.05). The insulin release test results demonstrated that compared with the control group, the SI of the ischemic preconditioning and Euonymus fortunei treatment groups was significantly increased (both P<0.05). Compared with the control group, the GSH contents of pancreatic islet cells in the ischemic preconditioning and Euonymus fortunei treatment groups were considerably enhanced, the NO content was significantly decreased, and the expression level of iNOS mRNA was significantly down-regulated (all P<0.05). Conclusions Euonymus fortunei can increase the survival rate of islet cells and enhance the function of pancreatic islets by increasing the level of GSH, down-regulating the expression of iNOS and decreasing the NO production.

IPEX with congenital ichthyosiform skin lesions:a case report and literature review / 临床儿科杂志

Objective To explore the clinical phenotype, treatment and prognosis of immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome, and to improve pediatricians' knowledge of this disease. Methods Clinical data of a case of IPEX with congenital ichthyosiform skin lesions were retrospectively analyzed, and related literatures China were reviewed. Results The 2-month-11-day old boy came to our hospital due to ichthyosiform skin lesions accompanied by blood oozing in the head and feet exudatation, with severe sepsis and gastrointestinal perforation. He was died of multiple organ failure. DNA sequencing of whole-genome exon group showed a hemizygous mutation of c.1150G> A, p.A384> T in FOXP3 gene. His mother was a heterozygous mutation carrier, while his father was normal. Conclusions In addition to typical symptoms including early-onset refractory diarrhea, multiple endocrine disease and growth retardation, IPEX should be considered also in infants with ichthyosiform rash and severe infection. Gene sequencing will help diagnose the disease.

Ras-associated autoimmune leukoproliferative disorder: a report of 2 cases and literature review / 中华儿科杂志

Objective@#To investigate the clinical features and genetic characteristics of cases with Ras-associated autoimmune leukoproliferative disorder(RALD).@*Method@#Characteristics of clinical data and gene mutation of the first two cases in China with RALD were retrospectively analyzed. The related literature was searched by using search terms "NRAS" , "KRAS" or "RALD" .@*Result@#Case1, a seven-year-seven-month old girl, was admitted due to "thrombocytopenia and splenomegaly for three years" . Palpation showed enlargement of submandibular lymph nodes and hepatosplenomegaly.The platelet count fluctuated between 15×109/L and 60×109/L. Hemoglobin was as 57 g/L and Coomb's test was positive.Lung computed tomography revealed interstitial lung disease, bilateral pleural effusion, pericardial effusion, myocardial injury and ascites. Case2, a seven-year-five-month old girl, was admitted due to "recurrent thrombocytopenia for seven years, intermittent eyelid and abdominal swelling for three years" . Palpation showed enlargement of cervical and right inguinal lymph nodes, and hepatosplenomegaly.The number of platelet and monocyte were 9×109/L and 5.46×109/L, respectively. Bone marrow smear revealed an increase in the proportion of primitive immature cells (0.09 to 0.11). Lung computed tomography revealed interstitial lung disease, pericardial effusion, cardiac enlargement and pulmonary hypertension. The gene sequencing results showed KRAS gene c.38G> A somatic mutation in case1, and p.G12D and NRAS gene c.38G> A, p.G13D somatic mutation in case2. A total of 8 reports were retrieved including 23 cases caused by NRAS(10 cases) or KRAS(13 cases) gene somatic mutation. All the 23 cases showed hypergammaglobulinemia, splenomegaly, B cells hyperplasia or mononucleosis.@*Conclusion@#RALD often manifests as hepatosplenomegaly,lymphoproliferation, autoimmune hematocytopenia, B cells hyperplasia or mononucleosis, hypergammaglobulinemia. Gene sequencing analysis can help diagnose the disease.

Clinical pilot study on the rhinitis due to laryngopharyngeal reflux / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the relevance between nasal symptoms and laryngopharyngeal reflux disease in patients with allergic rhinitis.@*METHOD@#Thirty patients of laryngopharyngeal reflux disease were diagnosed in ENT outpatient department in our hospital. All patients have symptoms of sneeze, nasal discharge as chief complaint and they responded no effect for other normal treatment for nasal-sinusitis at least three months. Orally before meals, a dose of 5 mg Mosapride citrate each time, three times a day for 7 days. Orally before meals, a dose of 20 mg Esomeprazole each time, two times a. day for 2-3 months. Nasal spray, one spray of azelastine hydrochloride once, two times a day for 2 month.@*RESULT@#Laryngopharyngeal reflux symptom scores at four time points (the first visit, post treatment 15 days, 45 days, 75 days) were analyzed by repeated measures analysis of variance. There is a significant difference in four time points.@*CONCLUSION@#Laryngopharyngeal reflux disease has a strong association with allergic rhinitis. Patients who has allergic rhinitis nasal symptoms as chief complaint must be exclude, the laryngopharyngeal reflux disease first.

A case of chronic mucocutaneous candidiasis caused by STAT1 gain-of-function mutation and literature review / 中华实用儿科临床杂志

Objective To summarize the clinical characteristics,diagnosis and treatment of chronic mucocutaneous candidiasis(CMC).Methods The case diagnosed as CMC in the Department of Nephrology and Immunology of Shenzhen Children's Hospital in February 24,2014 was analyzed in terms of symptoms,signs,laboratory findings,gene tests and treatment process,and related literature was reviewed.Results The patient was a 14-year-old boy.The patient started to develop recurrent oral candida infection shortly after birth,then candida infection of skins and nails,which could be alleviated by antifungal agents,but easily relapsed.Since 4 years ago,autoimmune reactions such as autoimmune anemia,thrombocytopenia,leucopenia,proteinuria,and hypothyroidism had successively appeared,and cytopenia began to palliate after administering Glucocorticoid and Cyclosporin,but easily relapsed when the dosage was reduced.The genetic test showed the case was of signal transducer and activator of transcription 1 (STAT1) gain-of-function mutation.During the hospitalization,hemophagocytic syndrome (HPS) and stagnation of hematopoietic function successively occurred.The cytopenia did not improve and the patient suffered severe infection in spite of washing red blood cells 13 times and blood palate 3 times via infusion,together with high dosage of Dexamethasone and Cyclosporine.The peripheral blood cells and bone marrow gradually returned to normal after being treated by human granulocyte colony-stimulating factor combined with Dexamethasone and Cyclosporin.Retrieving the database in PubMed database,824 articles were found which were about CMC,and 39 of them were about the STAT1 gain-of-function mutation,including 120 cases.But there was only 1 domestic case in 2012,who was a three-year-old child,manifesting recurrent fungal infection of the skin.Conclusions STAT1 gain-of-function acquired mutation is one of the reasons that can lead to CMC.Autoimmune reactions prominently represented by cytopenia occur in a few patients with CMC.It should be alert on those who are with CMC and simultaneously with autoimmtne reaction of blood system.And gene tests facilitate the early diagnosis.

The association between concentration of particulate matt(PM2.5) and asthma attack in children: a Meta-analysis / 中华实用儿科临床杂志

Objective To evaluate the association between the concentration of particulate matter (PM2.5)and asthma attack in children.Methods The computer and databases were used to search the key wordsAsthma andChild were searched by means of the subject headings ( Asthma andChild) and free words (Asthma * , Child *andPediatric * etc.),moreover PM2.5 was searched on the basis of free words such asPM2.5, Particulate Matter *which were linked withAND in PubMed and Medline database.Furthermore,the key words asthma AND (Child OR Pediatric OR Infant) AND (PM2.5 OR Particulate Matter OR Inhalable Particles) for keywords were searched in CBM,CNKI,VIP and Wanfang databases.The literatures included those from the initial computer database up to those of December 2014 from computer database.Meanwhile manual research was added to screen literatures according to the standards of inclusion and exclusion.Newcastle-Ottawa Scale was employed to assess the quality of the included literatures including selection,comparability and outcomc.Data wcre synthesized and analyzed by Rcvman5.3 and Stata12.0 version for statistical analysis,heterogeneity test,sensitivity analysis,subgroup analysis,and so on.Results A total of 17 literatures involving 1326573 children were included.The findings from quality assessment involved 2 studies for 8 scores,2 for 7 scores,7 for 6 scores,5 for 5 scores,and 1 for 4 scores,respectively.The result of Meta-analysis showed that the combined the odds ratio (OR) of PM2.5 with childhood asthma attack was 1.06,and 95％ confidence interval(CI) was 1.03-1.08.Subgroup analysis showed that the OR on the asthma hospital visit equaled to 1.03 (95％ CI 1.01-1.06),and the OR on the asthma symptom equaled to 1.11 (95％ CI 1.06-1.16),moreover,subgroup analysis between developed and developing countries showed that the OR of the asthma on developing countries corresponded to 1.04 (95％ CI 1.02-1.06),and the OR of the asthma on developed countries corresponded to 1.06 (95％ CI 1.03-1.09).Conclusion There is a positive correlation between concentration of PM2.5 and asthma attack in children.

Clinical effect evaluation of the recombinant Ⅱ type of tumor necrosis factor receptor antagonists treatment of juvenile idiopathic arthritis / 中华实用儿科临床杂志

Objective To observe the short-term,long-term clinical results and complications in refractory juvenile idiopathic arthritis (JIA) treating with TNF-α inhibitors,and to compare 2 evaluation systems.Methods A retrospective review of 52 cases of patients with refractory JIA in Shenzhen Children's Hospital was performed.With reference to International Leagne of Associations for Rheumatology(ILAR)2001 diagnostic criteria,the patients were divided into 4 groups:26 polyarticular JIA patients,14 systemic JIA patients,9 oligoarticular JIA patients and 3 other types of patients.The children with JIA were based on the conventional treatment such as Methotrexate,combination of TNF-α inhibitors treatment.The short-term and long-term clinical outcomes were evaluated and compared with American College of Rheumatology (ACR) and Juvenile Arthritis Disease Activity Score (JADAS).Complications in each group were recorded.Results (1) Short-term outcome assessment:ACR 50 were achieved in 69.2％ of the polyarticular JIA,66.7％ in oligoarticular and 35.7％ in systemic JIA patients on the third month;and by the time of the sixth month it reached to 73.0％ in polyarticular JIA,77.7％ in oligoarticular JIA and 14.3％ in systemic JIA patients on the sixth month.Significant improvement of JADAS after the treatment was observed in each type of JIA,and the differences were statistically significant(all P ＜ 0.05).(2) Long-term outcome assessment:except for the cases missing follow-ups and withdrawal cases,46 patients were able to complete 2 years assessments:according to ACR,effective rate was 84.0％ in polyarticular JIA (21/25 cases),50.0％ in oligoarticular JIA (4/8 cases) and 7.7％ in systemic (1/13 cases) JIA patients;JADAS was significantly decreased in polyarticular JIA patients (76.0％,19/25cases) (P ＜ 0.05),while significant improvement was not observed in oligoarticular JIA and systemic JIA patients(P ＞ 0.05).(3) Complications of upper respiratory tract infection (23.0％,12 cases) and local reaction in injection site (7.6％,4 cases) were noticed.Higher risks of tuberculosis infection and malignancy were not observed.Conclusions (1) TNF-α inhibitors treatment showed a better short-term and long-term outcome in polyarticular and oligoarticular JIA patients,and it may also improve short-term outcome in systemic JIA but with poorer long-term outcome.(2)Two evaluated systems (ACR and JADAS) were well relative,but ACR was capable to compare clinical course between different types of JIA.(3) TNF-α inhibitors treatment was relatively safe with unremarkable adverse reactions.

Tyk2 gene mutations in primary immunodeficiency disease：a case report and literature review / 临床儿科杂志

Objective To explore the clinical features and diagnosis of primary immunodeifciency disease caused byTyk2 gene mutations.Methods Clinical data from the ifrst case in China diagnosed of primary immunodeifciency disease caused by Tyk2 gene mutation were retrospectively analyzed, and related literature was reviewed.Results One year and 3 month old boy suffered with repeated pulmonary infection, chronic otitis media, intractable eczema like rash, repeated skin abscess, HSV infection, intracellular bacterial infection, and remarkedly increased total IgE. It was detected that compound heterozygous mutations of c.2269C>G in No. 16 exon and c.149delC in No. 3 exon inTyk2 gene. Literature searching found other 8 cases (5 males and 3 females) of immune deifciency patients caused byTyk2 gene defects, all of which hadTyk2 gene homozygous mutations and presented with repeated infection of paranasal sinus and lung. In the 8 cases 6 cases were combined with mycobacterium tuverculosis infection, 4 cases had repeated virus infection, 4 cases had meningonecephalitis, 3 cases had intractable eczema like rash, 2 cases had salmonella enteritis, 1 case had remarkedly increased total IgE, one case had elevated eosinophils, 5 cases were born in intermarriage family and 1 case died of meningitis caused by unknown etiology.Conclusions When patients have repeated paranasal sinus infection and lung infection, combined with intracellular bacterial infection (including mycobacterium tuberculosis infection), and repeated virus infection or intractable eczema like rash, with or without increased total IgE, immunodeifciency disease caused by Tyk2 gene defection should be considered. Gene sequence analysis can assist in early diagnosis.

Expression and kinetic analysis of catalytic domain of protein tyrosine phosphatases SHP-1/SHP-2 / 生物工程学报

In order to express and purify the catalytic domain of SHP-1/SHP-2 (named as D1C and D2C respectively) and determine their kinetics, the constructed D1C and D2C plasmids were transformed into Escherichia coli BL21 and the expression was induced with IPTG. The harvested cells were suspended in extraction buffer. After sonication, the solution was applied to HPLC and the results were confirmed by SDS-PAGE. The purified peptides were further subjected to kinetic specificity study using synthetic phosphotyrosine (pY) as substrate by malachite green method and analyzed by Lineweaver-Burk plot calculation. From this study, we found D1C and D2C were expressed successfully in soluble state in Escherichia coli BL21 and purified efficiently with HPLC system. The molecular weight of D1C was 34.6 kD, and its Michaelis constant (K(m)) was 2.04 mmol catalytic constant (K(cat)) was 44.98 s(-1), specific constant (K(cat)/K(m)) was 22.05 L/(mmol x s); the molecular weigh of D2C was 35.3 kD, and its Michaelis constant (K(m)) was 2.47 mmol, catalytic constant (K(cat)) was 27.45 s(-1), specific constant (K(cat)/K(m)) was L/(mmol x s). The enzyme activity of D1C is stronger than that of D2C.

Advantage of modified abdomen pathway for excision of huge pheochromocytoma / 中华泌尿外科杂志

Objective To analyze the application of modified abdomen pathway for excising the adrenal huge pheoehromocytoma.Methods One patient(male,42-year-old)had adrenal huge pheochromocytoma.The pheochromocytoma was about 15.0 cm×8.0 cm×7.0 cm.After 3 weeks' preparations,the patient was operated.The operation was made through the modified abdomen pathway for excision,without excising the transverse on peritoneal,and the interference to organs of peritoneal was reduced.The tumor on adrenal gland adhered kidney very tightly.Expanded radical excision including tumor,kidney,adrenal gland was applied.Results The operative time was 300 rain and the volume of bleeding was about 1000 ml.In the operation process,blood pressure of the patient was stable,the visual field of operation was satisfactory.Blood pressure of the patient returned normal 6 months postoperatively.And there was no indication of tumour relapse or matastasis.Conclusion The modified abdomen pathway can expose the satisfactory visual field,and is safe and effective for adrenal gland tumor operation.

Cold ischemia time and blood compatibility associated with activity of transplanted islet cells / 中国组织工程研究

BACKGROUND: The quantity and bioactivity of isolated islet are vital to islet transplantation; while, the cold ischemia time and human leucocyte antigen (HLA) typing are key factors to islet quantity and bioactivity which inflect islet transplantation.OBJECTIVE: To observe the effects of cold ischemia time and blood compatibility on quantity and bioactivity of islet cells.DESIGN: Observational study.SETTING: Ruikang Affiliated Hospital of Guangxi College of Traditional Chinese Medicine.MATERIALS: Organs from voluntary donors died of irreversible coma were adopted whose blood-type and HLA typing had been known, pancreases acquisition was carried on after other organs ablation or in the meantime. The blood of identical ABO and HLA matching conformity, or HLA cross-matching hypersensitization (missmatching over 3 Iocuses),or panel reaction antibody (PRA) ＞ 50%, or lymphocyte cytotoxin crossmatching test positive. The isolated and purified islet suspension was filtered by 70 μm filter, which result in preparing 1.2×105/L islet suspention.METHODS: Hypertonic citrate adenine solution was perfused into aorta, and kidney-pancreases and kidney-pancreases-liver were cut together or kidney-pancreases-liver was cut separately. Islet activity was judged by diphenylthiocarbazone (DTZ) dyeing and acridine orange (AO) dyeing; meanwhile, twelve pancreases far from contamination were aquired, mean ablation time was 15 minutes; cold ischemia time ranged from 2.5 to 8 hours. Cold ischemia time of nine pancreases was controlled in 5 hours, warm ischemia time was 0-3 minutes, and peptic time was (15±2.4) minutes, correlation of islet cells and histocompatibility with survival of islet cells was analyzed.MAIN OUTCOME MEASURES: Survival rate of islet cells; counts of platelet, heterophil granulocyte and monocyte.RESULTS: The cutting of kidney, pancreases and liver were successful. If the cold ischemia time was controlled within 5 hours, activity of islets was above 80%. Pancreatic gland used for islet transplantation and cutting of other organs could not affect activity of islet cells. When human islets were exposed to human blood, it would induce a rapid consumption of platelets, neutrophils, monocytes and lymphocytes in the blood. Consumption of blood cells was more in the HLA typing groups than that in the control group. After adding heparin, there was significant difference in cell account among the three groups (P ＜ 0.05) and the reaction was relieved obviously. After 24-hour cultivation, there were significant difference in active islets quantity between HLA typing compatibility group and HLA typing incompatibility group (P ＜ 0.05).CONCLUSION: Pancreatic gland obtained under the cold ischemia time ＜ 5 hours can be used in clinical transplantation of islet cells; a good histocompatibility can raise successful rate of transplantation of islet cells.

Analysis of human leucocyte antigen allele and haplotype in 36 cases receiving living-related donor kidney transplantation / 中国组织工程研究

BACKGROUND:It has great significance to understand individual human leucocyte antigen allele(HLA) haplotype and genotype in organ allotransplantation.OBJECTIVE:To analyse HLA allele frequency,haplotype frequency and characteristices in 36 cases of living-related donor kidney transplantation.DESIGN,TIME AND SETTING:The gene polymorphism analysis was performed in the Tissue Typing Laboratory of Organ Transplantation of Ruikang Hospital of Guangxi Traditional Chinese Medical University from January 2007 to June 2008.PARTICIPANTS:Thirty-six cases of living-related donor kidney transplantation,who were relatives of Guangxi Han population.METHODS:HLA-Ⅰantigen was detected by dry plates of monoclonal antibodies,and HLA-Ⅱ antigen was subjected to genotyping using PCR-SSP technique.And then the allele and haplotype frequencies of HLA-A,B,DRB1 were calculated.MAIN OUTCOME MEASURES:HLA haplotype and genotype of Guangxi Han population.RESULTS:A total of 11 HLA-A,24 HLA-B and 13 HLA-DR alleles were detected among 36 cases,showing a rich polymorphism.A2-B46,A11-B60,B46-DRB1*09,B60-DRB1*15 and A2-B46-DRB1*09 among 144 haplotypes were informative with frequency higher than 0.05.A total of 10 HLA-A-B haplotypes and 9 HLA-B-DRB1 haplotypes were in strongest linkage disequilibrium.Distribution of HLA-A,B,DRB1 alleles and haplotypes in Guanxi population is similar to Han population in South China,such as Hunan and Taiwan,but had its own characteristics.CONCLUSION:HLA alleles have richer polymorphisms with exhibit geographic genetic characteristics.