China ; Hearing Tests ; Humans ; Infant, Newborn ; Neonatal Screening

Adenovirus vectors are promising delivery systems for gene therapy. We established a new process for clinic trial of recombinant adenovirus vectors using a novel disposable bioreactor. The suspension HEK293 cells were inoculated into a 5 L disposable bioreactor with parameters control of pH, DO, agitation and temperature. After 6 days of a fed-batch culture, the final cell density reached 2.0 x 10(6) cells/mL. The culture was infected with Ad-IFNγ at an MOI of 30. The harvest was performed at approximately 48 h post-infection and crude viral lysate was obtained after 3 freeze/thaw cycles and centrifugation. The maximum titers of crude viral lysate was 1.49 x 10(13) Infectious units (IFU) and the bulk product specific was 3,800 IFU/cell. Purified Ad-IFNγ by anion-exchange chromatography and the final recovery of infectious unit reached 35.9%. The result demonstrates that an efficient and stable process of producing Ad-IFNγ using a disposable fed-batch bioreactor is established.
Adenoviridae ; growth & development ; Batch Cell Culture Techniques ; Bioreactors ; Cell Count ; Genetic Therapy ; Genetic Vectors ; HEK293 Cells ; Humans ; Virus Cultivation ; methods

Objective To observe the changes in the expression of renal tissue TNF-α , NF-κB and the interrelation to renal cell apoptosis, and their influences of Inula Britannica(an inhibitor of inflammatory signal pathway) in exhausted swimming rats, and to investigate the role of inflammatory signal pathway. Methods Forty-eight male Wistar rats were randomly divided into three groups: control group (CN, n=8), exhaustive swimming group (ES, n=24) and Inula Britannica group (IB, n=16). The rats of CN were quiet without swimming. The rats of ES swam to exhaustion and were sacrificed at immediately (ESI, n=8), 6 hour (ES 6 h, n=8) and 24 hour (ES 24 h, n=8) after exhanstiing swimming. The rats of IB group took orally Inula Britannica at the dose of 25 ml/kg body weight at 24 h before swimming and then swam to exhaustive state. The rats of IB group were sacrificed at 6 hour (IB 6 h, n=8) and 24 hour (IB 24 h, n=8) after exhaustiing swimming. The renal cell apoptosis was measured by the method of terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL). The expression of TNF-α in renal tissue was examined by immunohistochemistry. The changes of NF-κB in renal tissue were measured by flow cytometry and immunnhistochemistry. The interrelation between TNF-α and NF-κB was analyzed by Pearson method, and the interrelation between TNF-α, NF-κB and renal tissue cell apoptosis was analyzed by Spearman method. Resulls The number of renal tissue apoptotic cells was increased progressively from ESI to ES 24 h rats (P <0.05). Immunohistochemistry staining showed that the positive expressions of renal tissue TNF-α and NF-κB were increased progressively at 0 h (0.136±0.009, 0.129±0.011), 6 h (0.171±0.011, 0.166± 0.009) and 24 h (0.229±0.008, 0.218±0.019) after exhaustiing swimming in ES compared with control group (0.109±0.010, 0.095±0.010) ( all P<0.05). The similar changes of renal tissue NF-κB was also revcalved by flow cytometry. The expression of TNF-α was positively correldted with NF-κB (r=0.955, P<0.01 ), and renal cell apoptosis was also positively correlated with TNF-α and NF-κB (r =0.953, r=0.939, P<0.01) in ES rats. Pretreatment with Inula Britannica, inhibited the up-regulation of expressions of renal tissue TNF-α (6 h:0.142±0.012, 24 h:0.130±0.010) and NF-κB (6 h:0.138±0.010, 24 h:0.136±0.011 ) induced by exhausting swimming. Conclusion Overtraining can induce the up-regulating expressions of renal tissue TNF-α and NF-κB, and Inula Britannica can partly counter the above changes in exhaustied swimming rats, which may be one important mechanisms of overtraining-induced renal tissue cell apoptosis and the anti- apoptosis effect of Inula Britannica.

ObjectiveTo evaluate the effect of losartan on ventricular arrhythmia in elderly patients with hypertension.Methods The 78 hypertension subjects with ventricular arrhythmia aged 65 to 89 years were randomly assigned to treatment with losartan or placebo (enalapril) for 12 months. The blood pressure, left ventricular mass index and ventricular arrhythmia were observed and analyzed.ResultsAfter 12 months treatment, the pressure lowering effect was similar in losartan versus enalapril groups, there were no differences in systolic pressure decrement and diastolic pressure decrement between the two groups (P＞0. 05). Left ventricular mass index was lower in enalapril groupthaninlosartangroup [(109.2±15. 4) g · m-2 vs. (128.5±16. 7) g · m-2, t=2.015, P＜0. 05].However, the prevalence of ventricular arrhythmia was lower in losartan group than in enalapril group [(628. 5±176.8)/24 h vs. (852.9±215.7)/24 h, t=2.417,P＜0.05]. No Pearson's correlations of reduction of ventricular arrhythmia with reduction of blood pressure (systolic: r=0. 094, P＞0. 05; diastolic: r= 0.08, P＞0. 05) and reduction of left ventricular mass index were found. in losartan group.Conclusions Initiation of antihypertensive treatment with losartan in elderly subjects appears to cause more reduction of ventricular arrhythmia than with enalapril, despite similar reduction of blood pressure.

Objective To investigate the molecular basis for anti-ventricular arrhythmic effects by losartan through measuring alteration in mRNA and protein levels of key K+ α channel-and-β subunits (Kv4.2,Kv4.3 and KChIP2) in ventricular myocytes in spontaneously hypertensive rats (SHRs).Methods SHRs were randomly assigned to losartan[10mg·kg-1·d-1,n=12] or placebo (n= 12) with age-and weight-matched WKY rats (n = 12) as control.After 8 weeks of treatment,cardiomyocytes were isolated by enzymolysis.Action potential of cardiomyocytes Ito was recorded,mRNA and protein levels of Kv4.2,Kv4.3 and KChIP2 were assessed by reverse transcriptase polymerase chain reaction and Western blot.Results The action potential duration (APD) measured at 50% and 90% repolarization was shorter in losartan group [(16.82 ± 3.79) ms and (68.49±13.25) ms] than in SHR control group [(24.56±4.59) ms and (73.26±15.47) ms,all P<0.01].Losartan increased Ito current density associated with significant increases in the mean levels of mRNA and protein of Kv4.2 and Ky4.3,and with significant decreases in the mean levels of mRNA and protein of KChIP2 compared with those in placebo SHR (all P<0.01).Conclusions Chronic blockade of AT1 receptors with losartan reverses cardiomyocytes electrical remodeling in SHR,resulting in the shortening of APD,which is associated with increasing Ito density by increasing mRNA and protein expression of Kv4.2,Kv4.3 and by decreasing mRNA and protein expression of KChIP2.

Objective To evaluate the diagnostic value of myocardial enzymes on the cardiac muscle injury for the children with acute mycoplasma pneumonia(MP).Method The levels of phosphocreatine kinase-MB(CK-MB),lactate dehydrogenase(LDH),aspartate aminotransferase(AST)in 48children with MP(MP group)were measured,48 normal children served as control group.Results The levels of AST,LDH,CK-MB were(37.18±6.15),(294.10±46.20),(84.50±13.70)U/L in acute stage in MP group,they were higher than those in control group[(25.14±10.50),(140.50±76.21),(19.50±4.10)U/L](P＜0.01).All the values above was descended in recovery phase in MP group,but the values of CK-MB[(27.80±8.40)U/L]and LDH[(189.50±69.73)U/L]were still higher than those in control group(P＜0.01 or＜0.05).Conclusions Children with MP may combine myocardial injuries and CK-MB isa parameter to diagnose the damage.Protection therapy for cardiac muscle should be performed on the children with MP.

B7H1 is a recently described B7 family member.Ligation of PD-1 receptors with B7hl on the surface of activated T cell inhibits T cell proliferation and cytokine production. B7H1 plays a negative regulatory role in peripheral tolerance, autoimmune diseases and chronic infections.B7H1 also has been found to be largely expressed on a broad range of cancers and is thought to contribute to immune evasion by cancers. Blockade of B7H1/PD-1 pathway may contribute to the treatment of autoimmune diseases and maligant tumors.

Calreticulin (CRT) participates in the progression and apoptosis of many kinds of tumor cells. Recent studies indicate expression of CRT on tumor cells acts as an "eat me" signal for dendritic cells,allowing them to phagocytose tumor cells. It can induce immunogenic tumor cell death by adsorbing recombinant calreticulin to tumor cells or by enforcing the translocation of endogenous calreticulin to the cell surface. In animal experiments,CRT related tumor vaccine has good therapeutic effects in some tumor bearing mice,which can be used in tumor immunotherapy and targeted therapy.

Objective To clarify the morphological features, types, regional distributions and cell densities of gastrointestinal (GI) endocrine cells in various parts of the digestive track (DT) of Eumeces elegans. Methods Using immunohistochemical techniques of streptavidin-peroxidase(S-P) method. Results The endocrine cells found in the DT were round or oval, spindle, shuttle, pyramid, flask and bacilliform in shape. Three types of endocrine cells, namely 5-hydroxytryptamine (5-HT) positive cells, somatostatin (SS) positive cells and gastrin (GAS) positive cells were found. No substance P(SP)-, glucagon (GLU)-, pancreatic polypeptide ( PP)- and insulin(INS) positive cells were detected. 5-HT positive cells distributed throughout the whole DT at various densities and they were most predominant cell types in the DT. SS positive cells were detected only in stomach. GAS positive cells showed a restricted distribution and were only demonstrated in the pylorus and duodenum. In the DT of E. elegans the region with the highest degree of cell type heterogeneity was pylorus and all types of endocrine cell along the DT showed peak density in the pylorus as well. Conclusion Some common features of the distribution of different types of GI endocrine cells are found between E. elegans and other reptiles. This common trait may reflect the similarity in digestive physiology of various vertebrates. On the other hand, some species-dependent unique distribution features of endocrine cells in DT were also detected in E. elegans.

A patient with sparganosis mansoni was treated with praziquantel and cured with little side effect.