Isolation and identification of dominant strains in the fermentation process of HuaFengDan YaoMu and investigation of their growth under different temperature and pH value / 中成药

AIM To isolate and identify the dominant strains in the fermentation process of the HuaFengDan YaoMu,and to investigate the effects of different temperatures and pH values upon their growth.METHODS During the 0-6 weeks fermentation process of HuaFengDan YaoMu in the selective medium,the cultured bacteria,the molds and the yeasts had the colonies counted,and the dominant strains isolated and purified,particularly the dominant bacteria and fungi determined by 16S rDNA and 26S rDNA sequences.The optimal growth temperature and pH value of Bacillus aryabhattai,Debaryomyces hansenii,Aspergillus oryzae were investigated respectively.RESULTS HuaFengDan YaoMu found its bacterial counts increased from 0 to 4 weeks and stabilized from 4 to 6 weeks;its yeast counts increased during 0-3 weeks,and decreased continuously during 4-6 weeks;and its mold counts remained unchanged at 0 week,but increased steadily from 1 to 4 weeks,and decreased at 5 to 6 weeks of the fermentation.In the fermentation process of HuaFengDan YaoMu,the dominant strains of Bacillus aryabhattai,Debaryomyces hansenii and Aspergillus oryzae,identified and screened out by the phylogenetic tree,adapted best to the growth temperature of 35℃,25-30℃and 35℃,at pH value of 8,6 and 7,respectively.CONCLUSION In the fermentation process of HuaFengDan YaoMu,the dominant microorganisms of Bacillus aryabhattai,Debaryomyces hansenii,Aspergillus oryzae are identifiable and separable.

Quality evaluation of Bolbostemmatis Rhizoma by UPLC fingerprint combined with QAMS / 中国中药杂志

The present study was performed to establish the UPLC fingerprints of Bolbostemmatis Rhizoma and determine the contents of three saponins by quantitative analysis of multi-components by single marker(QAMS), and provide basis for quality evaluation of Bolbostemmatis Rhizoma. The analysis was carried out on an analytical column of Waters Cortecs T3(2.1 mm×100 mm,1.6 μm)with gradient elution by acetonitrile-0.1% phosphoric acid solution, at a flow rate of 0.3 mL·min~(-1). The detection wavelength was 203 nm, the column temperature was 30 ℃ and the injection volume was 1 μL. The UPLC fingerprints of Bolbostemmatis Rhizoma were established and evaluated by similarity calculation, cluster analysis and principal component analysis. The relative calibration factors of toberoside B and toberoside C were determined with toberoside A as internal reference. The content was calculated by relative calibration factors to develop a method of QAMS. Comparing the results of QAMS with those of ESM, the accuracy and feasibility of one-eva-luation and multi-evaluation can be determined. RESULTS:: showed that the fingerprints of 19 batches of Bolbostemmatis Rhizoma have four common peaks with similarities ranging from 0.754 to 1.000. Cluster analysis and principal component analysis classified 19 batches of Bolbostemmatis Rhizoma into three categories, which was consistent with the similarity evaluation results. The relative deviation between the content of tubeicosides B and C in 19 batches of Bolbostemmatis Rhizoma determined by QAMS and ESM is less than 5.0%, indicating that there was no significant difference between the two methods. Therefore, the UPLC fingerprints combined with QAMS and similarity evaluation can be effectively used to evaluate the quality of Bolbostemmatis Rhizoma.

Expression profiles of IL-10, TNF-a, and SOCS3 in placenta of pregnant women with intrahepatic cholestasis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To detect the expression profiles of suppressor of cytokine signaling 3 (SOCS3), interleukin (IL)-10, and tumor necrosis factor-alpha (TNF-a) in the placenta of women with intrahepatic cholestasis of pregnancy (ICP) and determine the clinical significance of the differential expressions.</p><p><b>METHODS</b>Placentas were collected from 37 ICP gravidas who delivered through cesarean section at the First Teaching Hospital of Xingjiang Medical University from October 2010 to May 2011 and from 35 healthy pregnant women (controls). SOCS3, TNF-a, and IL-10 protein levels were detected by immunoblotting and the Envision immunohistochemical method.</p><p><b>RESULTS</b>TNF-a and IL-10 expression was detected in placentas of both groups, and was present mainly in the cytoplasm of trophoeytes. IL-10 expression was obviously lower in the ICP placentas than in the control placentas; meanwhile, TNF-a expression was obviously higher than in the control placentas (Z=-2.63, P less than 0.01). SOCS3 protein was significantly more abundant in the control placentas than in the ICP placentas. Furthermore, SOCS3 and IL-10 placental expressions were positively correlated (r=0.494, P less than 0.01), but there was a negative correlation between SOCS3 and TNF-a placental expressions (r=-0.472, P less than 0.01).</p><p><b>CONCLUSION</b>In ICP, an increase of the type 1 cytokine, TNF-a, is associated with decreases of the type 2 cytokine, IL-10, and of SOCS3, which may reduce the secretion of IL-10. Furthermore, SOCS3 may contribute to ICP pathogenesis by modulating the Th1/Th2 cytokine balance.</p>

Influential factors on psychosocial health of the migrant workers in Guangzhou / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the influential factors on psychosocial health of the migrant workers in Guangzhou.</p><p><b>METHODS</b>The Symptom Checklist 90 (SCL-90) and Eysenck Personality Questionnaire (EPQ) were used to investigate 518 migrant workers in Guangzhou.</p><p><b>RESULTS</b>The rate of migrant workers with psychosocial problems was 36.5%. The scores of SCL-90 and positive rates in migrant workers with the different personality types had significant difference (P < 0.01). The results of binary logistic regression analysis indicated that the working years, drinking, sex, P scores, E scores and N scores of EPQ were main predictors of the poor physical fitness status. The vocations, working years, P scores and N scores of EPQ were strong predictors of the somatization. he vocations, P scores and N scores of EPQ were strong predictors of the obsessive compulsive symptom. The smoking, P scores and N scores of EPQ were strong predictors of the interpersonal sensitivity. The working years, P scores of EPQ were strong predictors of the depression. P scores of EPQ was strong predictors of the anxiety. P scores, E scores and N scores of EPQ were strong predictors of the hostility. The working years, smoking, P scores, E scores and N scores of EPQ were strong predictors of the phobic anxiety. The working years, P scores of EPQ were strong predictors of the paranoid ideation. The working years, P scores and N scores of EPQ were strong predictors of the psychosis.</p><p><b>CONCLUSION</b>The level of mental health of the migrant workers was significantly associated with the personality. The results of present study indicated that different vocation, sex, working years, smoking and drinking might interfere with the psychological states. The migrant workers with the personality of psychoticism, neuroticism and introversion may have unhealthy mental reaction.</p>

Reversing effects of curcumin on multi-drug resistance of Bel7402/5-fu cell line / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the reversing effects of curcumin on hepatocellular carcinoma drug resistance Bel7402/5-Fu cell line.</p><p><b>METHODS</b>Through the exposure to gradual increased concentrations of 5-fluorouracil (5-Fu), the cell line Bel7402 was induced to establish a multi-drug resistant sub-cell line Bel7402/5-Fu. The sensitivity to 6 chemotherapeutics of Bel7402 and Bel7402/5-Fu were detected using methyl thiazolyl tetrazolium (MTT) assay. The 50% inhibitory concentration (IC50) and resistant index (RI) were calculated. The differences of the inhibition ratio of Bel7402/5-Fu by curcumin, 5-Fu, curcumin combined with 5-Fu were detected using MTT assay. The effects of curcumin, 5-Fu, curcumin combined with 5-Fu on the Bel7402/5-Fu apoptosis were detected using flow cytometry.</p><p><b>RESULTS</b>The Bel7402/5-Fu cell line showed multi-drug resistance (MDR) to various chemotherapeutics, with the highest RI shown of 5-Fu (being 109.55 +/- 14.30 times). The inhibition ratio of 5, 10, and 20 microg/mL curcumin combined with 5-Fu (50% IC50) was respectively 21.47% +/- 1.49%, 27.10% +/- 2.32%, and 59.37% +/- 2.45%. The Bel7402/5-Fu apoptosis ratio of 5, 10, and 20 microg/mL curcumin combined with 5-Fu (50% IC50) was 30.92% +/- 2.10%, 44.87% +/- 2.24%, and 50.36% +/- 2.58%, respectively, which was obviously higher than that of the curcumin group and the 5-Fu group. Besides, the apoptosis rate increased along with increased curcumin concentration in the range of 0 -20 microg/mL.</p><p><b>CONCLUSION</b>Curcumin could induce the apoptosis of Bel7402/5-Fu. Meanwhile, it showed favorable reversing effects on MDR.</p>

Effect of quercetin on neural stem cell proliferation in the subventricular zone of rats after focal cerebral ischemia-reperfusion injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of quercetin on the proliferation of neural stem cells in the subventricular zone (SVZ) of rats after focal cerebral ischemia.</p><p><b>METHODS</b>An adult rat model of middle cerebral artery occlusion (MCAO) model was established by placement of an intraluminal filament at the origin of the MCA. Quercetin was administered intraperitoneally in the rats at a dose of 50 mg/kg every 3 days starting at 6 h after MCAO, and BrdU (50 mg/kg daily) was also injected intraperitoneally starting at 4 h after MCAO. BrdU-positive cells in the SVZ were counted at 7, 14 and 21 days after MCAO.</p><p><b>RESULTS</b>Compared with the sham-operated group, the rats in the ischemic model group showed significantly increased BrdU-positive cells in the ipsilateral SVZ 7 days after MCAO, reaching the peak level on day 14 and beginning to decrease on day 21 (P<0.05). The number of ipsilateral BrdU-positive cells in quercetin group was significantly greater than that in the model group on days 7, 14 and 21 (P<0.05), and maintained the high level on day 21.</p><p><b>CONCLUSION</b>Quercetin can maintain a high level of neural stem cell proliferation in the SVZ after focal cerebral ischemia in adult rats.</p>

Protective effect of mild hypothermia on astrocytes with traumatic or ischemic injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effect of mild hypothermia on rat astrocytes with traumatic or ischemic injury.</p><p><b>METHODS</b>Rat astrocytes in primary culture were subjected to scratching or hypoxic injury and exposed to normothermia (37 degrees celsius;) or hypothermia (34 or 32 degrees celsius;) for 24 h. The morphology of the astrocytes was evaluated by live/dead staining, and the cell injury was measured by lactate dehydrogenase (LDH) release assay.</p><p><b>RESULTS</b>As the temperature reduced the LDH release rate from the cells in hypoxic group decreased significantly, to (11.48 - or + 1.53)% at 34 degrees celsius; and (3.79 - or + 0.45)% at 32 degrees celsius; as compared to that in normothermia [(33.02 - or + 3.58)%] in the absence of rat white blood cells (WBC) (P<0.001). LDH release rate of the hypoxic cells further decreased in the presence of rat WBC to (51.14 - or + 2.17 )% at 37 degrees celsius;, (19.53 - or + 4.37)% at 34 degrees celsius; and (16.68 - or + 1.47)% at 32 degrees celsius; (P<0.001). In the scratched cells, with or without WBC, LDH release rate showed no significant variation between the 3 temperatures (P>0.05).</p><p><b>CONCLUSION</b>Mild hypothermia offers obvious protective effects on rat astrocytes against ischemic damage but not against mechanical injury.</p>

Protective effects of quercetin on primary cultured rat astrocytes against oxidative stress / 中华神经医学杂志

Objective To study the protective effects of quercetin on primary cultured rat astrocytes against oxidative stress. Methods Oxidative stress in primary cultured astrocytes was induced by exposure to 2 mmol/L H2O2 for 6 h, and the protective effect of quercetin pretreatment at different concentrations (0, 50, 100, and 200 μmol/L) administered 24 h before H2O2exposure was assessed with lactic dehydrogenase (LDH) release assay and live-dead staining. Results Treatment with 2 mmol/L H2O2 caused obvious injuries to the astrocytes, resulting in significantly increased LDH release rate in comparison with that of the normal control cells [(90.27±2.68)% vs (3.89±+1.89)%, P<0.05] and significantly lowered cell survival rate [(59.73%±9.92)% vs (99.25±0.08)%, P<0.05]. Pretreatment with quercetin decreased the LDH release rate and increased the survival rate of the astrocytes exposed to H2O2; at the concentration of 50, 100, and 200 μmol/L, quercetin significantly decreased the LDH release rate of the exposed cells to (48.19±13.98)%, (27.81±9.33)% and (18.13±8.28)% (P<0.05), and increased the cell survival rate to (86.80±3.62)%, (88.32±5.77)% and (91.18±3.03)%, respectively (P<0.05). Conclusion Quereetin may produce protective effects on the primary cultured astrocytes against oxidative stress.

Establishment and application of an in vitro neruoinflammation model in astrocytes after ischemic injury / 中华神经医学杂志

Objective To establish a in vitro cell model of inflammation following ischemic injury in rat astrocytes, and study the effects of mild hypothermia on the response of astrocytes to ischemia and inflammation. Methods Primary cultured neonatal SD rat astrocytes identified by GFAP immunofluorescence staining were divided into normal control group (C), hypoxia group (H), and hypoxia and leukocyte treatment group (H+W). Hypoxic injury of the cells was induced by exposure to glucose-flee DMEM medium in 5% CO2 and 95% N2 for 4 h, and for leukocyte treatment, the astrocytes were co-cultured with leukocytes isolated from rat blood. The 3 groups of cells were subjected to hypothermia treatments at 37, 34, 32 and 30 ℃, and the cell viability was estimated by lactic dehydrogenase (LDH) release assay and live/dead assay. Results Ischemic injury occurred in the astrocytes after 4 h of hypoxic exposure. Hypoxic treatment of the cells at 37 ℃ resulted in increased LDH release in all the 3 groups, and the increment intensified in the order of groups C, H and H+W. Mild hypothermia at 34 ℃ and 32 ℃ significantly reduced the LDH release in groups H and H+W (P<0.05), but hypothermia at 30 ℃ significantly increased LDH release in comparison with that at 32 ℃ (P<0.05). Conclusion Mild hypothermia can significantly reduce the severity of ischemic and inflammatory injuries in rat astrocytes possibly mediated also by other mechanisms in addition to inhibited astrocyte metabolism due to low temperatures.

Construction of delta-pIRES2-EGFP plasmid and its expression in HEK293 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the delta-pIRES2-EGFP plasmid and investigate its expression in HEK293 cells.</p><p><b>METHODS</b>Full length cDNA of rat delta opioid receptor gene amplified from rat brain tissues using reverse transcription and nested PCR was cloned into pMD20 T vector. The delta cDNA was inserted into pIRES2-EGFP plasmid to construct the recombinant eukaryotic plasmid delta-pIRES2-EGFP, which was transfected into HEK293 cells via Lipofectamine2000. The expression of delta was examined under fluorescence microscope.</p><p><b>RESULTS</b>The recombinant delta-pIRES2-EGFP plasmid was successfully constructed, and high expression of delta was detected in HEK293 cells transfected by the plasmid.</p><p><b>CONCLUSION</b>delta-pIRES2-EGFP has been successfully cloned, which shows high expression of delta in HEK293 cells.</p>

Comparison between genotyping and serological phenotyping in RhCE blood group / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To genotype the RHCE gene of Hans, Xinjiang's Uigurs and Kazakstans in China, and to compare the results of RHCE genotyping with that of RhCc/Ee phenotyping.</p><p><b>METHODS</b>RHCE genes of 98 Hans with RhD positive and 230 Hans, 72 Uigurs and 18 Kazakstans with RhD/RHD negative were genotyped with PCR-sequence specific primer (SSP) technique.</p><p><b>RESULTS</b>The results of RHE/RHe genotyping from samples with RhD positive and negative were in accord with that of phenotyping. It would result in 4.44% error using C-->G polymorphism at nt48 of RHCE gene to genotype RHCE, and 4.05% failure of detection using the 109 bp insertion to detectRHCE gene in Chinese Hans. The results of RHE/RHe genotyping in unrelated 72 Uigurs and 18 Kazakstans with RhD phenotype were consistent with that of phenotyping, and false positive and false negative were not found in genotyping in Uigurs and Kazakstans tested.</p><p><b>CONCLUSION</b>The results of RHE/RHe and RHc genotyping were correct with PCR-SSP and accordant with that of phenotyping. Using the C48G polymorphism in exon 1 of RHCE to genotype RHC gene would result in false positive resulting from RHc mutation at this locus, and using the 109 bp insertion to genotype RHC gene would result in false negative because of the absence of the 109 bp. Therefore it is necessary to genotype RHC gene using more than two polymorphic loci.</p>

Construction and Identification of NBS1-targeting microRNA Expressing Eukaryotic Vector / 中国生物工程杂志

Objective:To construct NBS1 microRNA expressing eukaryotic recombinants,and identify biological activity of recombinants in Hela cell after transfection.Methods:According to sequence of NBS1mRNA,the NBS1 pre-microRNA was designed and synthesized,then cloned into the GFP reporter pcDNA6.2-GW/EmGFP-miR vector and transfected into Hela cell line.To detect integrity of inset fragment through colony PCR and sequencing analysis.The biological activity of recombinants through identify interference efficiency of NBS1 microRNA recombinants by way of Real-Time PCR and Western blot were determined.Results:Sequences of inset fragment in four microRNA expressing recombinants were correct.NBS1 mRNA and protein expression of four microRNA recombinants were decrease,which is the lowest in the NBS1mi-2 group.Conclusion:Four NBS1-targeting microRNA expressing recombinants all have biological activity in Hela cell line,and NBS1mi-2 recombinant has the most interference efficiency.The microRNA expressing plasmid which were successfully constructed and lay foundation for the studies on the tumor gene therapy of microrna targeting NBS1.

A comparative research on RHD gene structures of Chinese Han and Uigur population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To research comparatively on the RHD gene structures in unrelated RhD negative individuals of Chinese Uigur and Han population.</p><p><b>METHODS</b>The upstream, downstream, hybrid box and 10 exons of RHD gene were detected with sequence specific primer-PCR technique.</p><p><b>RESULTS</b>The results showed the genotypes of RhD negative individuals to have the significant difference between Chinese Uigur and Han population, that 94.44% Uigur individuals were with RHD(-)/RHD(-) genotype but just 61.40% Han population were with this genotype(94.44% versus 61.40%, P<0.01); 2.78% Uigur individuals were with RHD(+)/RHD(-) genotype but 34.21% Han population were with this genotype(2.78% versus 34.21%, P<0.01). However, there was significantly no RHD(+)/RHD(+) genotype difference between Chinese Uigur and Han population(2.78% versus 4.39%, P>0.05). In 78 cases of RhD negative Chinese Hans with single RHD gene, of which the RHD gene structure showed that 53(67.95%) cases were RHD(1-10) allele(of 53 RHD(1-10) alleles, 14 alleles were unexpressed); 15(19.23%) were RHD-CE(2-9)-D(2) allele; 5(6.41%) cases were RHD-CE(2-7)-D(2) allele; 2(2.56%) were similar to RHD-CE(3-6)D allele; 1(1.28%) case was RHD-CE(5-6)-D allele; and 2(2.56%) were RHD-CE(6)-D or point mutation respectively. Of 2 RhD negative Chinese Uigurs with RhD(-)/RHD(+) genotype, one carried RHD(1-10) allele, another carried RHD-CE(2-9)-D(2) allele.</p><p><b>CONCLUSION</b>The most frequently unexpressed RHD alleles were RHD-CE(2-9)-D(2), RHD(1-10) and RHD-CE(2-7)-D(2) respectively in Chinese Han population who carried single RHD allele with RHD(-) phenotype and RHD(+) genotype. It showed the confluent character of RH gene in Chinese Han and Uigur population that there existed unexpressed RHD-CE(2-9)-D(2) allele in Chinese Uigur nationality, which was infrequent in Chinese Uigur population but frequent in Chinese Han population.</p>

A method for Rhesus box test / 中国实验血液学杂志

To study the method for Rhesus box test and its significance, the sequence specific primers of upstream, downstream and hybrid Rhesus boxes were designed according to RhD gene sequence; the upstream, downstream and hybrid Rhesus boxes were determined by PCP-SSP and mismatched PCR. The results showed that this method was confirmed by DNA Standard test. It was shown that in unrelative RhD positive individuals RHD(+)/RHD(-), RHD(+)/RHD(+) genotype accounted for 9.00%, 91.00% respectively, and in RhD negative individuals RHD(+)/RHD(-), RHD(+)/RHD(+), RHD(-)/RHD(-) genotype were 26.14%, 3.92%, 69.94% respectively. It is concluded that the method of Rhesus box test was confirmed to be reliable and can be used for the identification of RhD haplotype gene structure, as well as for study on inheritance, clinical transfusion and neonatal hemolytic diseases.

Ex vivo expansion of T, NK and CD34+ cells from umbilical cord blood / 中国实验血液学杂志

Umbilical cord blood stem cell transplantation (CBSCT) has made significant progress in treatment of lethal congenital or malignant disorders. Both the incidence and severity of GVHD from CBSCT were lower than that from bone marrow and peripheral blood stem cell transplantation, particularly for adult patients, but these advantages were also associated with higher rates of relapse. The immune-mediated effect of natural killer and cytotoxic T cells against residual tumor cells were shown to prevent relapse and to induce remission after bone marrow transplantation. To explore possibility of ex vivo expansion of T, NK and CD34(+) cells from umbilical cord blood, cord blood was expanded ex vivo with different combinations of cytokines, T and NK cells proliferation and differentiation were observed. CB MNCs were separated in Ficoll-Isopaque column and cultured in IMDM for 14 days with different recombinant cytokines. Cultured cells were collected and analyzed for progenitor/stem cell immunophenotyping at day 0, 3, 7, and 14 by using flow cytometry. The results indicated that all test groups cultured with different combinations of SCF, IL-3, IL-6, IL-7, IL-2 showed significant expansion of UCB MNC, compared with the group without cytokines. All test groups showed expansion effects on CD34(+) cells, CD34(+) percentage went up from 1.6% in fresh CB to the highest 11.9% in group D (SCF + IL-3, IL-6, IL-2). The CD34(+) cells peak displayed at day 7 of culture in group A and D, while in other two groups B and C appeared at day 14 of culture. The expansion multiple of CD34(+) cells in all test groups at day 7 of culture were from 10 to 50. The average value of CD3(+) T cell in fresh UCB was 18.7 +/- 4.3%, the CD3(+) T cells decreased sharply in the medium without any interleukin, while obvious increase were observed in the other test groups containing different combinations of cytokines. The maximal expansion multiple of CD3(+) T cells reached 2 times of the fresh UCB level. CD56(+) cells amounted to 3.6 +/- 1.9% of fresh UCB, CD56(+) cell number increased significantly only in medium containing IL-2. It is concluded that T cells, NK cells as well as stem/progenitor cells can be expanded in the same time from CB-MNC with the combinations of cytokines.

Comparison of Rhesus boxes in Hans and Uighurs / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the difference and similarity between Hans and Uighurs in regard to Rhesus box and its significance.</p><p><b>METHODS</b>The sequence specific primers of upstream, downstream and hybrid Rhesus boxes were designed on the basis of RHD gene sequence. The upstream, downstream and hybrid Rhesus boxes were determined by polymerase chain reaction-sequence specific primer(PCP-SSP) and mismatched PCR.</p><p><b>RESULTS</b>The percentage of RHD-/RHD-, RHD+/RHD- and RHD+/RHD+ genotypes ascertained in the unrelated Hans with RhD(-) were 61.40%, 34.21% and 4.39% respectively, while those in the unrelated Chinese Uighurs with RhD(-) were 94.44%, 2.78% and 2.78% respectively. Furthermore, all 6 cases of some other minorities were RHD-/RHD- types. The percentage of RHD-/RHD- and RHD+/RHD- genotypes ascertained in the unrelated Chinese Uighurs were significantly higher than those in Chinese Hans (P < 0.01), whereas no statistically significant difference in the percentage of RHD+/RDH+ genotype between the two groups was observed (P > 0.05).</p><p><b>CONCLUSION</b>The Rh blood group of Uighurs in Xingjiang possesses both Oriental and Caucasian characteristics, which embodies a special ethnical aspect of the Chinese nation and is in accord with the anthropologic research results.</p>

Method of detection of soluble HLA-I and soluble HLA-I level alteration in storage blood / 中国实验血液学杂志

Aim of this study was to develop the detection method of soluble human leukocyte antigens I (sHLA-I) and to explore sHLA-I level alteration in storage blood and its significance. sHLA-I level in sera of 60 Guangdong normal individuals and sHLA-I concentration in blood components from 20 donors quantitatively were detected by sandwich ELISA. The results showed that sensitivity of this assay was 2.84 ng/ml. Coefficients of variation were 5.80% within assays and 9.00% between assays respectively. The recovery rate was >/= 98.57%. The sHLA-I level of normal individuals in Guangdong was (699.54 +/- 360.10) ng/ml. sHLA-I in red blood cells stored for 28 days and in random-donor platelets were significantly higher than that in other blood components and their amount was proportionate to the number of residual donor leukocytes and to the length of storage. In conclusion, sandwich ELISA assay for detection of sHLA-I is a sensitive, specific and stable technique. Blood components with different concentration of sHLA-I may be chosen for clinical transfusion.

Observation on gene polymorphism of Rh blood group in Chinese Han nationality / 中国实验血液学杂志

To observe the gene polymorphism of Rh blood group in unrelated random individuals and families for Chinese Han nationality, polymerase chain reaction-sequence specific primer (PCR-SSP) was used to amplify the Rh C/E gene, RhD gene, exons, intron 2 and 10, insert and Rh Box in 160 blood samples of RhD positive unrelated individuals and 71 samples of RhD negative unrelated individuals and 7 samples of families whose probands were RhD-negative. The results showed that RhD genes of RhD-negative individuals with C antigens were polymorphism, three forms were found for D exon including intact, partial deletion and complete deletion exons. Insert fragments and Rh Box were found in most cases of families whose probands were RhD-negative and its inheritance accorded with the Mendel's Law, and it did not affect the expression of RhD gene. "Normal" RhD exon 4 amplifying product was not found in all of the samples. It was concluded that gene structure of the RhD-negative in Chinese was polymorphism, intact, partial deletion and complete deletion exons were found in the individuals with C antigen and probably existed specific D (nf) Ce haplotype. The function of insert was uncertain. The Rh gene sequences of Chinese Han nationality are different from those of Caucasian and the Rh gene library based on Han nationality should be established.

Research of Typing for HLA-A, -B on Cord Blood Lymphocytes / 中国实验血液学杂志

Serological typing for HLA-A, -B has been used for a long time. Recently with the developing of molecular biology technologies, HLA-A, -B typing is now turning to genotyping methods. In our study, the capacity of PCR-SSP in solving problems in HLA-A, -B typing with serological methes was evaluated. With this aim the serological method was compared with PCR-SSP in 102 cord blood samples, and the results showed that 18.6% of 102 cord blood samples can't give a satisfactory detection, for 14 samples, give discrepant results with the 2 methods. It is mainly due to weak expression of HLA class I cord blood lymphocytes and the cross reaction of some antigens. About B 15 group, the further study was made, it was found that most of the B 15 splits is wrongly disassigned, especially among the B62-B75, B75/*1511(+)-B75/*1511(-), B46-*1511 antigens. It was concluded that DNA typing is more preferable than serological typing, about B 15 group, the subtyping or high resolution typing can be fulfilled at first in China.

Chemical camouflage of like human B antigens on Rhesus monkey red blood cells with methoxy polyethylene glycol / 中国输血杂志

Objective To observe the chemical camouflage effects of methoxy polyethylene glycol(mPEG) on like human B antigens on Rhesus monkey red blood cell(rRBC) and to study the safety of transfusion with mPEG modified monkey RBC(mPEG rRBC).Methods rRBC were modified with 3mg/ml mPEG. The rRBC blood group and the effects of mPEG modification on blood conversion were assayed by absorption and elution methods.The structural and functional changes of mPEG rRBC were also tested.The survival fraction of mPEG modified like B rRBC in like A recipient monkey were examined by flow cytometry.The transfusion effects of mPEG modified rRBC on recipient's blood and urine were also observed.Results mPEG have been covalently bound to the surface of Rhesus monkey RBC (like human B blood group),mPEG could cover rRBC like B antigen.Transfusion of mPEG modified like B rRBC to like A Rhesus monkey had no harmful effects on the recipient.Conclusion The present study suggests that transfusion of mPEG modified RBC could be safety in experimental animals