The impact of HLA high resolution typing mismatching of donor-recipient pairs on outcome of unrelated donor hematopoietic stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the impact of various human leukocyte antigen (HLA) high resolution typing mismatching of donor-recipient pairs on prognosis of unrelated donor hematopoietic stem cell transplantation.</p><p><b>METHODS</b>835 donor-recipient pairs of CMDP data from 2005 to 2010 were analyzed retrospectively. HLA-A, B, C, DRB1 and DQB1 typing were performed using SBT, SSOP and SSP methods. The diseases involved in acute myeloid leukemia (AML) (n = 288), acute lymphoid leukemia (ALL) (n = 227), chronic myeloid leukemia (CML) (n = 187), myelodysplastic syndrome (MDS) (n = 52), non-hodgkin's lymphoma(NHL) (n = 25), aplastic anemia(AA) (n = 42) and thalassemia (n = 14). Of 835 donor-recipient pairs, 362 were completely matched, 159 had a mismatch for a single allele, 125 had a mismatch for a single antigen, 95 had mismatched for both single allele and single antigen, 29 were mismatched at double allele, 20 at double antigen, 45 at multiple allele and antigen. The follow-up assessment was completed before March 2011.</p><p><b>RESULTS</b>HLA-matched pairs had higher overall survival (OS) than HLA-mismatched pairs (79.83% vs 73.15%), but there was no statistically significant differences (P > 0.05). HLA mismatch for a single allele plus a single antigen was a significantly risk factor for OS, disease free survival (DFS) and transplant-related mortality (TRM). The OS from high to low in different diseases were thalassemia, AA, CML, MDS, AML, NHL, and ALL. OS of HLA locus mismatch were DRB1 (94.4%), DQB1 (83.3%), B (75%), A (74.4%) and C (71.4%), respectively. OS of single allele mismatch at HLA locus from high to low were DRB1, C, A, B and DQB1.HLA-A, B, C locus mismatch were statistically significantly associated with lower OS and grade II-IV acute GVHD compared with HLA-matched pairs (P < 0.05). The donor-recipient pairs with HLA-B*15:01/B*15:05, DRB1*12:01/DRB1*12:02, C*04:01/C*03:04, DQB1*03:02/DQB1*03:03 alleles mismatch were given priority. But the donor-recipient pairs with HLA-B*39:01/B*39:05, C*15:02/C*14:02, C*08:01/C*03:04, C*07:02/C*15:02 alleles mismatch were risk factors for influence of OS and aGVHD.</p><p><b>CONCLUSION</b>The high resolution typing for HLA-A, B, C, DRB1, DQB1 can be identified nonpermissive mismatch, which is beneficial for the selection of a suitable donor improves survival on unrelated donor HSCT.</p>

A study on apoptosis and apoptotic mechanisms of HL-7702 cell line induced by methylmercury / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the apoptotic effect and mechanisms of methylmercury (MeHg) on HL-7702 cell line in vitro.</p><p><b>METHODS</b>In this study, the cell apoptosis was observed by AO/EB method and FCM method; the mitochondrial membrane potential was detected by FCM; and the expression of proteins related to apoptosis was measured by immunocytochemical method.</p><p><b>RESULTS</b>After exposure to MeHg for 24 h in different doses, apoptotic rate ascended with the increasing of MeHg concentration. By AO/EB method, cell apoptotic ratio of negative control group was (2.62 +/- 0.19)%, cell apoptotic ratio of 10-50 micromol/L exposure groups were (7.97 +/- 0.64)%, (12.66 +/- 0.76)%, (19.16 +/- 0.87)%, (18.42 +/- 0.88)%, and (11.52 +/- 0.63)%, there were significant differences between the exposure and negative control groups (q values were 17.057, 32.009, 52.732, 50.373, 28.375; P<0.05). Mitochondrial membrane potential descended with the increase of MeHg, mitochondrial membrane potential of negative control group was (10.23 +/- 3.43) mV, mitochondrial membrane potential of 10-50 micromol/L exposure groups were (3.25 +/- 0.66), (3.03 +/- 0.35), (1.68 +/- 1.26), (1.69 +/- 1.13) and (1.77 +/- 0.88) mV, and there was significant differences between exposure and negative control groups (q values were 9.569, 9.871, 11.722, 11.708, 11.598; P<0.05). The expression of Bax, Bcl-2, CytC, Caspase-3 and AIF enhanced with the increase of MeHg, Bax/Bcl-2 ratio also appeared a trend of increase. Bax expression integral optical density (IOD) of negative control group was (21295.86 +/- 1969.81), Bax expression IOD of 10, 20, 30 micromol/L groups were 42807.87 +/- 4416.64, 55651.65 +/- 4662.72, and 72708.56 +/- 910.10, there were significant differences in Bax expression between 10, 20, 30 micromol/L groups and negative control group (q values were 14.191, 14.320, 33.917; P<0.05); Bcl-2 expression IOD of negative control group was (12588.33 +/- 4091.02), Bcl-2 expression IOD of 10, 20, 30 micromol/L groups were 20539.16 +/- 4906.09, 23689.97 +/- 2281.42, and 28692.80 +/- 4655.86, there were significant differences in Bcl-2 expression between 10, 20, 30 micromol/L groups and negative control group (q values were 4.322, 6.035, 8.754; P<0.05); and AIF expression IOD of negative control group was (12942.72 +/- 457.94), AIF expression IOD of 10, 20, 30, 40 micromol/L groups were 16973.57 +/- 1922.87, 29998.91 +/- 6803.58, 52467.16 +/- 1916.25 and 106342.53 +/- 1273.19, there were significant differences in AIF expression between 20, 30 and 40 micromol/L groups and negative control group (q values were 11.449, 26.530, 62.692; P<0.05).</p><p><b>CONCLUSION</b>MeHg could induce apoptosis on HL-7702 cell line in vitro. The mechanisms could be related to mitochondrial pathway in apoptosis.</p>

Toxicity of nano silica particles on rat type I-like alveolar epithelial cells / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To access rat lung toxicity of nano silica particles.</p><p><b>METHODS</b>Transmission electron microscope was used to observe size, shape and dispersibility of two silica particles; Size of two particles in water and RPMI 1640 containing 1% FBS were measured using Zeta Potential Analyzer. LDH activities of rat type I-like alveolar epithelial cell line R3/1 cells after 6, 24 and 48 h incubation with 2.5, 5.0, 10.0, 20.0 microg/ml of two silica nano particles were detected by spectrophotometric method; protein carbonylation and MIP-2 release of R3/1 cells after 24 h incubation with 2.5, 5.0, 10.0, 20.0 microg/ml of two silica nano particles were measured using ELISA kits.</p><p><b>RESULTS</b>TEM image showed nano silica particles were round and dispersed evenly; the average sizes of the two silica particles were (43+/-4.2) and (68+/-5.7) nm. Two silica particles had similar size in water and RPMI 1640 containing 1% FBS, respectively. Both nano silica particles in 2.5 approximately 20.0 microg/ml dose range did not cause significant increase of LDH activities (P>0.05), did not elevate protein carbonylation and MIP-2 levels in R3/1 cells (P>0.05).</p><p><b>CONCLUSION</b>Two nano silica particles do not have lung toxicity in 2.5 approximately 20.0 microg/ml dose range.</p>

Effects of over-expressed Smac gene coupling with cisplatin on proliferation and apoptosis of hepatocarcinoma cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects of over-expressed Smac gene combined with cisplatin (CDDP) on proliferation and apoptosis of hepatic carcinoma cells.</p><p><b>METHODS</b>The recombinant plasmid pcDNA3.1+-hSmac was introduced into the human hepatic carcinoma SMMC-7721 cells using a liposome-mediated method. The expression of Smac protein was detected by Western blot and flow cytometry. The cells were treated with three different doses of CDDP, 5, 15 and 25microg/ml, for 24 hours after the transfection. MTT colorimetry was used to detect the cellular growth-inhibitory effects; acridine orange-ethidium bromide fluorescent staining (AO/EB) and flow cytometry with annexin V-PI double staining</p><p><b>METHODS</b>were used to detect the changes of cell apoptosis.</p><p><b>RESULTS</b>Western blot and flow cytometry results demonstrated that the Smac protein level in SMMC-7721 cells was significantly increased after the transfection (P less than 0.01). Compared with that of the control group, the over-expressed Smac gene inhibited the cell growth and induced cell apoptosis (P less than 0.01). After being treated with CDDP, the inhibitory rates were increased significantly with increasing concentrations of CDDP compared with that of the control group, and the inhibitory rate of the CDDP-treated plus Smac group was significantly higher than that of the CDDP-treated group (P less than 0.01). The results detected by AO/EB and flow cytometry demonstrated that the apoptotic rates of CDDP-treated plus Smac group were higher than those of the CDDP-treated group (P less than 0.01). The results demonstrated that the Smac over-expression enhanced the effects of cell growth inhibition and apoptotic promotion induced by CDDP.</p><p><b>CONCLUSION</b>The pro-apoptotic Smac gene could be over-expressed in hepatocarcinoma SMMC-7721 cells and inhibit cell growth and induce apoptosis. Moreover the over-expressed Smac could enhance the chemotherapeutic sensitivity of SMMC-7721 to cisplatin. This experimental work may help in further study on the regulatory mechanism of Smac in apoptosis and improve the chemotherapeutic effect on hepatoma.</p>

Breeding of High-yielding ?-galactosidase Strains from Protoplast of Aspergillus niger / 微生物学通报

The protoplasts of original Aspergillus niger strain Uco-3 were treated with the cooperation of UV and ?-ray to obtain the high-yielding strain producing the thermostable ?-galactosidase. Under the optimum conditions of formation and regeneration protoplasts were prepared. According to the interaction of positive mutation rate and radiation dose,the optimum condition was determined. The optimum dose of UV was 4 minutes and the optimum dose of ?-ray was 500 Gy. After mutagenetic treatment of protoplasts and selection from a lot of mutants,a mutant DL116 producing the thermostable ?-galactosidase was obtained. The ?-galactosidase activity of DL116 was increased from 16.27 U/mL to 44.37 U/mL,which was higher than that of strain Uco-3.