Objective To investigate the correlation of plasma N‐terminal pro‐B‐type natriuretic peptide (NT‐proBNP) level with echocardiographic indicators and P wave terminal force of lead V 1 (PtfV1) in the patients with hypertension and paroxysmal atrial fibrillation(PAF) .Methods Fifty‐six outpatients and inpatients with hypertension were divided into the PAF group (n=26) and the sinusrhythm group (n=30) .Thirty age‐matched and gender‐matched healthy volunteers were taken as the control group . The plasma NT‐proBNP level was determined .Left ventricular enddiastolic diameter (LVEDD) ,left atrial diameter(LAD) and left ventricular ejection fraction(LVEF) were examined by echocardiography .the 12‐lead electrocardiogram was routinely performed Pt‐fV1 was calculated .Results The plasma NT‐proBNP level in the PAF group was higher than that in the sinusrhythm group and the control group ,the difference was statistically significant (P<0 .05) .The plasma NT‐proBNP level in the PAF group was de‐creased significantly after successful cardioversion .The plasma NT‐proBNP level in the PAF group was positively correlated with LVEDD(r=0 .543 ,P<0 .05) and LAD (r=0 .606 ,P<0 .01) .The plasma NT‐proBNP level was negatively correlated with LVEF (r= -0 .750 ,P<0 .01) and positively correlated with the PtfV 1 absolute value (r= 0 .513 ,P< 0 .01) .Conclusion The plasma NT‐proBNP level can better reflect the heart structure and function in the patients with atrial fibrillation .Detecting the plasma NT‐proBNP level combined with echocardiographic indicators and PtfV 1 is conducive to comprehensively assess the heart function in the patients with hypertension and PAF .

Stirred culture offers a number of advantages over static systems as it maintains a stable, homogeneous culture environment and is easy to scale-up. This paper focused on the development and application of stirred tank bioreactor to culture hematopoietic cells. Preliminary study of stirred culture of hematopoietic cells was carried out in cord blood mononuclear cells culture in spinner flask. The results showed that the amplification rates of total cell, CFU-GM and BFU-E, with the exception of CFU-Mk, were greater in spinner flask than T-flask. The number of total cells increased 20 fold after 14 days incubation in spinner flask. The amplification rates of CFU-GM, CFU-Mk and BFU-E reached maximum at 10th day, 10th day and 7th day respectively, and the maximal amplification rates were 9.2-fold, 5.5-fold and 2.4-fold respectively, whereas the rate of CD34+ cells in spinner flask was (6.7 +/- 4.0)-fold at day 10. These results indicated that the stirred culture system is better than the static culture systems for hematopoietic cell proliferation. The biocompatibility of cord blood MNC to different types of materials used in bioreactors was also tested. The results showed that glass, stainless steel 316L and polytetraflouroethylene (PTFE) supported the growth of hematopoietic cells well. A higher cell density was reached in stirred bioreactors with controlled pH and DO than static culture. These findings suggested that the controlled large-scale culture could be used to overcome the clinical shortage of hematopoietic cells.
Antigens, CD34 ; metabolism ; Bioreactors ; Cell Culture Techniques ; instrumentation ; methods ; Erythroid Precursor Cells ; cytology ; Fetal Blood ; cytology ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Humans ; Polytetrafluoroethylene ; Stainless Steel

To optimize the culture environment and protocol of hematopoietic cells' expansion, avoiding the fluctuation caused by medium changing in stirred culture and concentration gradient in static culture, the hematopoietic cells from cord blood (CB) were cultured in a stirred bioreactor connected with a cell retention system, which is a gravity sedimentation settler designed for hematopoietic cell. Total cells expanded 11.5 and 18.6 fold respectively in the twice perfusion stirred cultures, in which CFU-Mix was expanded 23.2 and 20.4 fold, CFU-GM 13.9 fold and 21.5 fold, BFU-E 8.0 fold and 6.9 fold, CD34+ cells 17.1 fold and 15.4 fold. After 12-day culture, it was obtained that 1082 x 10(6) total cells, 6.31 x 10(6) CFU-GM, 6.2 x 10(6) CFU-Mix and 23 x 10(6) CD34+ cells from 267 x 10(6) CB mononuclear cells (MNC) in the first culture, and 1080 x 10(6) total cells, 4.65 x 10(6) CFU-GM, 11.0 x 10(6) CFU-Mix, and 25.0 x 10(6) CD34+ cells from 180 x 10(6) CB MNC. These two cultures met to the clinical scale. Due to the optimized dissolved oxygen (DO) and stable culture environment, the rate of stem/progenitor cells to total cells in the perfusion culture was higher than that in T-flask cell-retention feeding culture. But the cell growth was inhibited in the later phase of perfusion culture, when the cell density is high. The inhibition should be attribute to the high cell density itself. The perfusion culture environment in bioreactor with optimal DO and pH controlling is more favorable for stem/progenitor cells' maintenance and expansion, and the expanded cells' number has reached a clinical scale. But the high cell density in the later phase of perfusion culture caused inhibition to mature hematopoietic cell's growth.
Bioreactors ; Cell Culture Techniques ; methods ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans

OBJECTIVETo establish a Real-time Taqman probe technique system to detect the mtDNA 1555A > G mutation in deaf population.

METHODSPrimers and Taqman probes for mtDNA 1555A > G mutation were designed and synthesized. The technique system for detecting mtDNA 1555A > G mutation using Real-time Taqman probes was established. Then the reliability of the technique was tested in 132 patients with severe to profound hearing loss who were detected for the mtDNA 1555A > G mutation by sequencing, Kit method and Real-time Taqman probe technique at the same time. Finally, the results by the above three ways were compared.

RESULTSThirty-two cases with mtDNA 1555A > G mutation were found by the technique of Real-time Taqman probe. These findings coincided with the results from sequencing and Kit method completely. Both the false positive rate and the false negative rate were zero.

CONCLUSIONSThe technique possesses the merits of accuracy, convenience, high sensitivity, high specificity and intuitionistic results, etc. Importantly, the Real-time Taqman probe technique only needs 1.5 hours to detect the 1555A > G mutation and it saves 4.5 hours for one reaction compared with the Kit method popularly used nowadays. The technique system of detecting mtDNA 1555A > G mutation is reliable. It's suitable for large-scale detecting and preventive diagnosis of mtDNA 1555A > G mutation.

Adolescent ; Adult ; Child ; Child, Preschool ; DNA Primers ; genetics ; DNA, Mitochondrial ; genetics ; Female ; Hearing Loss ; diagnosis ; genetics ; Humans ; Male ; Point Mutation ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Young Adult

Objective To analyze the imaging and pathological characteristics, as well as treatment strategies of intracranial hemangioblastomas (HBs),and explore the advancement of diagnosis,etiopathogenisis and treatment of HBs. Methods Thirty-five patients with intracranial HBs,admitted to our hospital and performed tumor resection from January 2005 to January 2010,were chosen in our study; all patients were divided into type of big cystic HBs with a small mural nodule (n=19),type of small cystic HBs with a big nodule (n=9) and type of solid HBs (n=7) by imaging features.The clinical manifestations,imaging findings and surgical methods were retrospectively analyzed; the expressions of NSE and CD34 in these tumor samples were detected by HE staining and immunohistochemical staining.Results All patients were treated by surgery; total resection was achieved in 34 and subtotal resection in 1; no death occurred after the surgery.Twenty-eight patients were followed up for 3 months to 3 years after discharge; recurrence appeared in 1 patient with big cystic HBs with a small mural nodule and Gamma knife treatment was performed.No significant difference was observed in the numbers ofCD34+cells between each 2 types of patients (P＞0.05).The numbers of NSE positive cells between each 2 types were statistically significant (P＜0.05). Conclusion There were no specific clinical manifestations of HBs.Diagnosis was mainly according to imaging features.Treatment of HBs with total resection is just the first selection and the key to reduce palindromia; the formation of HBs cysts is closely related to tumor stromal cells.

OBJECTIVETo analyze the molecular pathogenesis of deaf couples by means of genetic testing. To provide accurate genetic counseling and instruction for deaf couples with different etiology based upon results of genetic testing.

METHODSFour deaf families from July 2005 to May 2006. Each subject was with moderate to profound hearing loss. Genomic and mitochondrial DNA (mtDNA) of each subject were extracted from whole blood. Genetic testing of GJB2, SLC26A4 (PDS) and mtDNA A1555G mutation were offered to each individuals.

RESULTSThe husband from family 1 didn't carry GJB2, SLC26A4 and mtDNA A1555G mutation while his wife was confirmed to carry compound SLC26A4 mutations. The possibility of their offspring's to be SLC26A4 single mutation carrier was 100%. The couple from family 2 both didn't carry GJB2, SLC26A4 and mtDNA A1555G mutation. The possibility of their offspring's having hereditary deafness caused by GJB2, SLC26A4 and mtDNA A1555G mutation was excluded. The husband from family 3 was confirmed to carry homozygous GJB2 mutations and a single SLC26A4 mutation while his wife who was diagnosed with enlarged vestibular aqueduct syndrome (EVAS) by CT scan was proven to carry a single SLC26A4 mutation. The risk of their offspring's suffering EVAS was 50%. The husband from family 4 was mtDNA A1555G positive while his wife who was diagnosed with cochlear malformation by CT scan didn't carry GJB2, SLC26A4 and mtDNA A1555G mutation. The risk of their offspring's having hereditary deafness caused by GJB2, SLC26A4 and mtDNA A1555G mutation was excluded.

CONCLUSIONSGenetic testing could be applied to offer the more accurate genetic counseling and instruction to deaf couples.

Connexin 26 ; Connexins ; genetics ; DNA, Mitochondrial ; analysis ; Deafness ; diagnosis ; genetics ; prevention & control ; Female ; Genetic Counseling ; Genetic Diseases, Inborn ; diagnosis ; genetics ; Genotype ; Humans ; Male ; Membrane Transport Proteins ; genetics ; Mutation

OBJECTIVETo evaluate the diagnostic utility of Warthin-Starry silver stain, immunohistochemistry and transmission electron microscopy in the detection of human Bartonella henselae infection and pathologic diagnosis of cat scratch disease (CSD).

METHODSThe paraffin-embedded lymph node tissues of 77 histologically-defined cases of cat scratch disease collected during the period from January, 1998 to December, 2008 were retrieved and studied using Warthin-Starry silver stain (WS stain) and mouse monoclonal antibody against Bartonella henselae (BhmAB stain). Five cases rich in bacteria were selected for transmission electron microscopy.

RESULTSUnder electron microscope, the organisms Bartonella henselae appeared polymorphic, round, elliptical, short rod or bacilliform shapes, ranged from 0.489 to 1.110 microm by 0.333 to 0.534 microm and often clustered together. Black short rod-shaped bacilli arranged in chains or clumps were demonstrated in 61.0% (47/77) of CSD by WS stain. The organisms were located outside the cells and lie mainly in the necrotic debris, especially near the nodal capsule. In 72.7% (56/77) of the cases, dot-like, granular as well as few linear positive signals were observed using BhmAB immunostain and showed similar localization. Positive results for both stains were identified in 59.7% (46/77) of the cases. When applying both stains together, Bartonella henselae was observed in 74.0% (57/77) of the case. The difference between the results obtained by WS stain and BhmAB immunostain was of statistical significance (P < 0.05).

CONCLUSIONSBartonella henselae is the causative pathogen of cat scratch disease. WS stain, BhmAB immunostain and transmission electron microscopy are helpful in confirming the histologic diagnosis. Immunostaining using BhmAB can be a better alternative than WS stain in demonstrating the organisms.

Adolescent ; Adult ; Aged ; Antibodies, Bacterial ; blood ; Bartonella henselae ; immunology ; isolation & purification ; ultrastructure ; Cat-Scratch Disease ; diagnosis ; microbiology ; pathology ; Child ; Child, Preschool ; Humans ; Immunohistochemistry ; methods ; Infant ; Lymph Nodes ; pathology ; ultrastructure ; Microscopy, Electron, Transmission ; Middle Aged ; Paraffin Embedding ; Staining and Labeling ; methods ; Young Adult

OBJECTIVETo carry out molecular epidemiology study of SLC26A4 IVS7-2 A > G mutation in large Chinese deaf population and to provide evidence for fast screening and gene diagnosis of enlarged vestibular aqueduct syndrome (EVAS).

METHODSA total of 1979 patients with non-syndromic hearing loss(NSHL) underwent questionnaire and PCR for IVSA > G mutation detection of SLC26A4 gene.

RESULTSAll 245 patients (12.38%) with homozygotes and heterozygotes IVS7-2 A > G mutation were found among the 1979 NSHL It showed statistically significant difference among north and northeast, northwest, east and southeast, southwest and central area in China. (chi2 = 34.4899, P < 0.05). Carrier frequency of the central area (27.52%) was notably higher than southwest area (6.69%). The IVS7-2 A > G mutation was most frequently found in Han deaf groups (13.88%). Tibetan, Hui, and other western minorities were lower than Han deaf population (chi2 = 35.4456, P < 0.05).

CONCLUSIONSA high SLC26A4 IVS7-2 A > G mutation frequency for deafness in Chinese patients was found. Detection of the pathogenic mutations was bringing the possibility to detect EVAS at an early stage. Moreover, it might help to establish diverse diagnostic strategies toward differently ethical deaf population in different region of China.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; China ; epidemiology ; Ethnic Groups ; Gene Frequency ; Genotype ; Hearing Loss, Sensorineural ; epidemiology ; genetics ; Humans ; Membrane Transport Proteins ; genetics ; Point Mutation ; Young Adult

During the pathogensis of rheumatoid arthritis (RA), activated RA fibroblast-like synoviocytes (RA-FLSs) combines similar proliferative features as tumor and inflammatory features as osteoarthritis, which eventually leads to joint erosion. Therefore, it is imperative to research and develop new compounds, which can effectively inhibit abnormal activation of RA-FLSs and retard RA progression. Neohesperidin (Neo) is a major active component of flavonoid compounds with anti-inflammation and anti-oxidant properties. In this study, the anti-inflammation, anti-migration, anti-invasion, anti-oxidant and apoptosis-induced effects of Neo on RA-FLSs were explored to investigate the underlying mechanism. The results suggested that Neo decreased the levels of interleukin IL-1β, IL-6, IL-8, TNF-α, MMP-3, MMP-9 and MMP-13 in FLSs. Moreover, Neo blocked the activation of the MAPK signaling pathway. Furthermore, treatment with Neo induced the apoptosis of FLSs, and inhibited the migration of FLSs. It was also found that Neo reduced the accumulation of reactive oxygen species (ROS) induced by TNF-α. Taken together, our results highlighted that Neo may act as a potential and promising therapeutic drug for the management of RA.
Arthritis, Rheumatoid/drug therapy* ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; Hesperidin/analogs & derivatives* ; Humans ; Synoviocytes ; Tumor Necrosis Factor-alpha/genetics*