[A clinical study of haploidentical hematopoietic stem cell transplantation in the treatment of pediatric patients with acquired severe aplastic anemia: single center experience].

Objective: To investigate the efficacy of haplotype hematopoietic stem cell transplantation in the treatment of acquired severe aplastic anemia (SAA) in children. Methods: The clinical characteristics of 59 pediatric patients with SAA, including 26 cases VSAA, 37males and 22 females, 47 cases typeⅠ and 12 cases typeⅡ, undrerwent haplo-HSCT in our hospital between December 1st, 2011 and December 1st, 2017 were retrospectively analyzed. Among 59 patients, 56 patients with a median age of 4.5 (1.2-14.8) years and median weight of 43 (12-80) kg underwent their first HSCT and 3 patients underwent their second HSCT. All patients received the following conditioning regimen: busulfan, cyclophosphamide, and rabbit ATG or Bu (-, CTX) , fludarabineand rabbit ATG. The prophylaxis of acute graft versus host disease (aGVHD) was cyclosporine (CsA) , MMF and methotrexate. All patients received bone marrow transfusion on day 01 and peripheral stem cell transfusion on day 02 from haploid donor. The median dose of donor mononuclear cell counts was 15.60 (7.74-21.04) ×108/kg of recipient weight and CD34+ cell counts was 4.86 (3.74-7.14) ×106/kg of recipient weight. Results: Neutrophils and platelets of all 59 children were implanted. The median implantation time of granulocytes and platelets were 13 (10-19) d, 19 (9-62) d, respectively. The incidence of grade Ⅰ-Ⅱ aGVHD was 45.76% (27 cases) and grade Ⅲ/Ⅳ 13.56% (8 cases) , The incidence of chronic GVHD was 8.47% (5 cases) , The incidences of CMV and EBV viremia were 59.32% (35 cases) and 28.81% (17 cases) , respectively. The median follow-up was 30 (8-80) months, 57 patients survived with disease free, 2 patients died of GVHD. Both of the estimated 5-year OS and DFS rates were (96.4±2.5) %. Conclusion: Haplo-HSCT could improve the outcomes of SAA children.

Dynamic Virus-Bacterium Interactions in a Porcine Precision-Cut Lung Slice Coinfection Model: Swine Influenza Virus Paves the Way for Streptococcus suis Infection in a Two-Step Process.

Swine influenza virus (SIV) and Streptococcus suis are common pathogens of the respiratory tract in pigs, with both being associated with pneumonia. The interactions of both pathogens and their contribution to copathogenesis are only poorly understood. In the present study, we established a porcine precision-cut lung slice (PCLS) coinfection model and analyzed the effects of a primary SIV infection on secondary infection by S. suis at different time points. We found that SIV promoted adherence, colonization, and invasion of S. suis in a two-step process. First, in the initial stages, these effects were dependent on bacterial encapsulation, as shown by selective adherence of encapsulated, but not unencapsulated, S. suis to SIV-infected cells. Second, at a later stage of infection, SIV promoted S. suis adherence and invasion of deeper tissues by damaging ciliated epithelial cells. This effect was seen with a highly virulent SIV subtype H3N2 strain but not with a low-virulence subtype H1N1 strain, and it was independent of the bacterial capsule, since an unencapsulated S. suis mutant behaved in a way similar to that of the encapsulated wild-type strain. In conclusion, the PCLS coinfection model established here revealed novel insights into the dynamic interactions between SIV and S. suis during infection of the respiratory tract. It showed that at least two different mechanisms contribute to the beneficial effects of SIV for S. suis, including capsule-mediated bacterial attachment to SIV-infected cells and capsule-independent effects involving virus-mediated damage of ciliated epithelial cells.

[Role of STAT1 on the regulation the human hsp90 alpha gene expression].

OBJECTIVE: To investigate the role of STAT1 on the regulation of human hsp90 alpha gene expression. METHODS: We first transfected Jurkat cells with the STAT1 expression construct and analyzed the expression of hsp90 alpha gene expression via quantitative RT-PCR system. Then we co-transfected the STAT1 expression construct and the CAT reporter gene driven by different length of 5' flanking sequence of hsp90 alpha gene. Western blot was carried out to detect the level of tyrosine phosphorylation in Jurkat cells with and without heat shock treatment (42 degrees C 1 h). By electrophoretic mobility shift assays (EMSA), we evaluated the DNA binding activity of a STAT1 responsible element located in the regulatory region of hsp90 alpha gene in Jurkat cell nuclear extracts. RESULTS: The mRNA level of hsp90 alpha gene in Jurkat cells was decreased when transfected by STAT1 expression construct, over-expression of STAT1 down-regulates the expression of CAT reporter gene with the present of a distal fragment from -1756 to -1463 within the 5' flanking regulatory sequences of hsp90 alpha gene. The tyrosine phosphorylation of STAT1 was detectable in Jurkat cells and increased when subjected to heat shock. Electrophoretic mobility shift assays (EMSA) results showed that STAT1 could bind to its responsible element in the regulatory region of hsp90 alpha gene. CONCLUSION: STAT1 could negatively regulate the human hsp90 alpha gene expression.

[Gene expression changes after heat shock by cDNA array analysis].

OBJECTIVE: To study gene expression changes of cells in response to heat stress, we isolated total RNA from Jurkat cells before and after heat shock treatment. METHODS: cDNA was labeled with alpha-32P-dATP during reverse transcription of RNA and then used as probe to cDNA expression array. Autoradiogram images were analyzed by ESTblot software. RESULTS: After heat shock, the expression level of some forty genes increased while that of sixteen genes decreased. In addition to the elevated expression of heat shock genes, expression of c-Jun and CLK-1 increased most remarkably. The genes with notably decreased expression were integrin alpha-4 and transforming growth factor beta. The elevated expression of c-Jun and hsp90 alpha was further confirmed by Northern blot analysis. CONCLUSIONS: The expression of some genes in Jurkat cells changes after heat shock treatment. Heat shock induces elevated expression of hsp, c-Jun, CLK-1 gene while decreases expression of integrin alpha-4 and transforming growth factor beta.

Hepatitis B genotypes and the response to interferon therapy.

BACKGROUND/AIMS: Possible pathogenic differences among hepatitis B virus (HBV) genotypes have been observed; however, the response to interferon therapy among HBV genotypes remains unknown. We therefore analyzed the efficacy of interferon alfa in the treatment of chronic hepatitis B patients with different HBV genotypes. METHODS: Fifty-eight genotype B or C infected chronic hepatitis B patients who had been treated with interferon alfa-2b were retrospectively studied. The response to interferon was defined as normalization of serum aminotransferase level, loss of hepatitis B e antigen and HBV DNA 48 weeks post-treatment. RESULTS: Baseline data of both groups of patients were comparable; however, genotype C patients had a higher serum aminotransferase level and a higher frequency of core promoter mutation. The response rate was 41% and 15% in genotype B and C patients, respectively (p=0.045). In those with higher serum aminotransferase levels, the response rate was 50% and 17%, respectively (p=0.025). Additionally, younger age and genotype B infection may predict a better response to interferon alfa. CONCLUSIONS: HBV genotype C, compared to genotype B, is associated with a higher frequency of core promoter mutation, and a lower response rate to interferon alfa therapy.

Regulation of human hsp90alpha gene expression.

Mammalian HSP90alpha and HSP90beta are encoded by two individual genes. On the basis of the upstream sequences of the human hsp90alpha gene, GenBank accession number U25822, we have constructed CAT reporter plasmids driven by individual fragments of the hsp90alpha gene. We found that (1) the proximal heat shock element complex located at -96/-60 enhances hsp90alpha promoter expression; (2) heat shock induction depends upon the coexistence of distal heat shock element at -1031/-1022 and the proximal heat shock element complex of the hsp90alpha gene; (3) unlike hsp90beta, downstream sequences of the transcription start site inhibit hsp90alpha expression. We conclude that the regulatory mechanisms for the expression of hsp90alpha and hsp90beta genes are different.

A different experience of hepatic adenoma in Taiwan with emphasis on clinical manifestations and ultrasonographic features.

BACKGROUND/AIMS: Hepatic adenoma is a rare benign hepatic tumor and it is difficult to differentiate it from other focal hepatic tumors. Ultrasonography has become the choice of methods to detect focal hepatic lesions. The study aims at analyzing ultrasonographic features of hepatic adenoma. METHODOLOGY: A total of 8 patients with pathologically proven hepatic adenoma were studied retrospectively during an 8-year period. The ultrasound scanners used were Toshiba SSA-100A, Toshiba SSA-240 and Aloka 630. The ultrasonographic features and clinical data were analyzed. RESULTS: There were 7 males and 1 female. The mean age was 50 years. Of the 8 cases, 2 symptomatic cases had a tumor larger than 10 cm. The remaining 6 cases were asymptomatic and had tumors smaller than 5 cm. The echogenicity was variable in these tumors. An irregular sonolucent was only noted in a 15 cm tumor and was histologically proven to be internal bleeding. All the tumors were well-defined, however, a hypoechoic rim was obvious only in the isoechoic and hyperechoic tumors. CONCLUSIONS: Ultrasonographic features of some hepatic adenomas are different from those of hepatocellular carcinomas and hemangiomas, although the differential diagnosis cannot be made in small hypoechoic tumors. When ultrasonography is used more widely, more asymptomatic patients with small-sized hepatic adenoma will be detected, even in male subjects. The concept about the pathogenesis of hepatic adenomas may be changed.

RAPD analysis of natural populations of Acanthopanax brachypus.

Random Amplified Polymorphic DNA (RAPD) analysis is a new technology of molecular marking which has proved very powerful in detecting genetic diversity at the level of population. The genomic DNAs used in our experiment were extracted from fresh leaves taken from 59 individuals sampled from three natural populations in Yan An, Shanxi Province. Through more than 2,000 PCRs, deep-going RAPD analysis was carried out on DNA samples from 49 individuals. The percentage of polymorphic RAPD loci found in these three populations were respectively 27.2%, 18.6% and 5.4%; the average genetic distances within population, 0.055, 0.036 and 0.008; the average genetic distances between populations (I-II), (I-III) and (II-III), 0.105, 0.096 and 0.060. The genetic diversity of A. brachypus within and between populations was found, for the first time, to be rather poor, thus revealing innate factors as the cause contributing to its endangered status. In addition, our work also provides basic materials for elucidating the underlying cause of its endangerment and for its protection biology.

Factors associated with orthostatic hypotension in the Chinese population in Taiwan.

Factors associated with orthostatic hypotension are heterogeneous, and some of the factors are interrelated and interdependent, which may confound their relationships to orthostatic hypotension. To investigate the factors that were most likely related to orthostatic hypotension, a study of community-dwelling persons (419 men and 309 women) was conducted. Blood pressures and heart rates were measured after the subjects had been recumbent for 5 min and upright for 1 min. A total of 119 persons (16.3%) experienced orthostatic hypotension. Univariate analysis showed that orthostatic hypotension was associated with the following variables: hypertension, diabetes mellitus, cerebrovascular disease, proteinuria, abnormal renal function, or medications use. Those patients with orthostatic hypotension were older in age and had a higher body mass index, seated blood pressure, plasma creatinine, hemoglobin A1c, fasting and 2-h postload glucose levels than those without orthostatic hypotension. Multivariate analysis revealed that diabetes mellitus, hypertension, and age were independently associated factors for orthostatic hypotension. The higher the level of plasma hemoglobin A1c (%) elevation, the higher the likelihood of orthostatic hypotension manifestation. Clinically, elderly persons or patients with hypertension or diabetes mellitus should receive regular monitoring of supine and upright blood pressure in order to detect orthostatic hypotension and prevent its complications.