Biomedical subjects

Sheng Zhang

Publications and source records attributed to Sheng Zhang.

At least 55 records · Page 3Linked to original sources

PubMed2004

Characterization of protein glycosylation using chip-based infusion nanoelectrospray linear ion trap tandem mass spectrometry.

Mass spectrometry (MS) has the potential to revolutionize structural glycobiology and help in the understanding of how post-translation events such as glycosylation affect protein activities. Several approaches to determine the structure of glycopeptides have been used successfully including fast atom bombardment, matrix-assisted laser desorption ionization, and electrospray ionization with a wide variety of mass analyzers. However, the identification of glycopeptides in a complex mixture still remains a challenge. The source of this challenge is primarily due to the poor ionization efficiency and rapid degradation of glycopeptides. In this report we describe the use of a chip-based infusion nanoelectrospray ionization technique in combination with a recently developed linear ion trap for identification and characterization of glycosylation in complex mixtures. Two standard synthetic glycans were analyzed using multiple-stage fragmentation analysis in both positive and negative ionization modes. In addition, the high mannose type N-glycosylation in ribonuclease B (RNase B) was used to map the glycosylation site and obtain the glycan structures. We were able to map the glycosylation site and obtain the glycan structures in RNase B in a single analysis. The results reported here demonstrate that the fully automated chip-based nanoelectrospray linear ion trap platform is a valuable system for oligosaccharide analyses due to the unique MS/MS and MS(n) capability of the linear ion trap and the extended analysis time provided by the ionization technique.

Amino Acid Sequence↗

PubMed2004

[Dual effect of angiopoietin-2 on angiogenesis in gastric cancer].

OBJECTIVE: To explore the effect of Ang-2 on angiogenesis in gastric cancer. METHODS: The expression of Ang-2 mRNA was analyzed by RT-PCR and the expression of VEGF and CD34 was detected by immunohistochemistry in 36 cases of gastric cancer tissues and their paired adjacent gastric mucosa. RESULTS: The expression of Ang-2 mRNA was found both in gastric cancer and their paired adjacent gastric mucosa; the correlationship between the general expression levels of Ang-2 mRNA and microvessel density (MVD) in gastric cancer tissues was not found. However, in 27 cases whose Ang-2 mRNA expression levels in cancer tissues were lower than those in adjacent gastric mucosa. A significant positive correlation between the expression level of Ang-2 mRNA and MVD in the tumor tissues was found (r = 0.411, P < 0.05). In these 27 cases, the MVD in the gastric cancer tissues with positive VEGF expression (45.45 +/- 10.30) was higher than that with negative VEGF expression (30.15 +/- 8.69, P < 0.05), whereas in the other 9 cases whose expression levels of Ang-2 mRNA in cancer tissues were higher than those in adjacent gastric mucosa, a significant negative correlation between expression level of Ang-2 mRNA and MVD in the tumor tissues (r = -0.758, P < 0.05), but without correlation between the MVD and VEGF. CONCLUSION: Under the conditions that the expression of Ang-2 mRNA in cancer tissues was lower than that in adjacent gastric mucosa, VEGF could promote the sprouting of new vessels along with Ang-2 upregulation. But under the conditions that the expression of Ang-2 mRNA in cancer tissues was higher than that in adjacent gastric mucosa, Ang-2 inhibited angiogenesis. Angiopoietin-2 may play a dual effect on angiogenesis in gastric cancer.

Angiopoietin-2↗

PubMed2004

CD4+ T cells tumor specific response exists in L615 leukemia mice: adoptive transfer in combination with cyclophosphamide.

AIM: L615 leukemia cell line is a transplantable acute lymphocytic leukemia model with the CD4 positive phenotype. In this study, we explored whether tumor response specific T cells can be separated from the live leukemia mice or not. METHODS: The mutant HGPRT- L615 cell line was first established. The splenocytes from HGPRT- L615 leukemia mice were cultured and expanded in mixed tumor-lymphocytes culture manner. The expanded T cells were sorted with FACScan. Then their killing capacity, IFN-gamma release as well as antitumor capacity in adoptive transfer experiments were analyzed. RESULTS: The expanded response T cells are mostly CD4 positive. The CD4 positive T cells showed high release of IFN-gamma upon stimulation but lacked significant cytotoxicity. In immunochemotherapy model, these CD4 positive T cells can cure most leukemia mice. CONCLUSIONS: We demonstrated the feasibility of separation of tumor response specific CD4+ T cells from CD4+ L615 leukemia mice. These CD4+ T cells can cure leukemia mice upon adoptive transfer in combination with cyclophosphamide pretreatment.

Adoptive Transfer↗

PubMed2004

[Comparison on the performance of biostyr and bio-ceramic filter used for biological pretreatment of raw water].

A new biological filter called biostyr using biostyrene as media to purify raw water was compared with the widely studied bio-ceramic filter. The raw water was taken from a reservoir located in Northern China. It was shown this new biological filter could obviously improve the raw water quality. As far as the raw water investigated in this study was concerned, the removal percentage of COD(Mn), NH4+-N in the filter was 5%-20%, 80%-95%, respectively, and turbidity of the raw water decreased correspondingly, which indicated the feasibility that this filter could be applied in the field of biological pretreatment of raw water. It was also shown that under the uniform operating conditions, the performance of this new filter on the removal of contaminants, especially organics and turbidity, was slightly inferior to the bio-ceramic filter and would be more remarkably affected adversely by backwashing than bioceramic filter.

Bioreactors↗

PubMed2004

[Studies on clogging of the constructed wetland].

The parameters of substrate related to clogging, such as the grain size, the filtration rate and organic substance in the substrate, were investigated in the integrated vertical-flow constructed wetland (IVCW). The substrate of IVCW was collected by a specific sampling tube in situ and the infiltration experiment showed that the filtration rate of substrate increased with the depth of substrate and the minimum filtration rate existed 15-30 cm below the surface of substrate. The amount of organic substances in upper 15 cm layer of the substrate was limited and even less in deeper layer after fifty two-month running, and this revealed that IVCW was operating steadily and no clogging was observed. Considering the factors related to clogging, the strategy to prevent and recover the clogging phenomenon was put forward.

Ecosystem↗

PubMed2004

Antitumor reactivity of splenocytes primed in vivo with dendritic-cell-based vaccine and secondarily activated with a cocktail of cytokines in vitro.

AIM: To evaluate the antitumor reactivity of splenocytes primed in vivo with dendritic cell (DC) based vaccine and secondarily activated with cocktail of cytokines in vitro. METHODS: Tumor lysate pulsed DC were used to generate effector T cells in the murine L615 leukemia model. DC were induced and pulsed with tumor lysate. The splenocytes from the DC based vaccine primed mice were termed VPS. Some VPS were activated with anti-CD3 alone (VPS-CD3), the other cells were activated with a cocktail of cytokines including IFN-gamma, IL-1beta, anti-CD3 and IL-2 (VPSC). RESULTS: Both VPS and VPS-CD3 showed moderate IFN-gamma release to stimulatory tomor cells in vitro and antitumor capacity in vivo. However, VPSC showed much higher IFN-gamma release and far superior antitumor capacity than the other cells in this leukemia model. CONCLUSION: The result here indicates that tumor lysate pulsed DC can be used as a strategy to generate effector T cells which can be further activated with a cocktail of cytokines for adoptive immunotherapy.

Animals↗

PubMed2004

[Treating of algae-laden raw water with GAC-sand dual media deep bed dissolved air flotation/filtration].

GAC-sand dual media deep bed dissolved air flotation/filtration (GSDDB-DAFF) is a new integrated process, its characteristics include: integration of dissolved air flotation and filtration, GAC-sand dual media deep bed filtration, integration of general treatment and advanced treatment. When algae-laden raw water was treated with the new process. The result showed that removal percentage of algae was 95.1%, outlet algae-count was 4.30 x 10(5) cell/L. Removal percentage of chlorophyll-a was 92.2%, outlet chlorophyll-a was 0.88 microg/L. Outlet turbidity was 0.18 NTU, removal percentage of UV254 was 54.3%, outlet UV254 was 0.016 cm(-1), Removal percentage of OC was 63.6%, outlet OC was 0.78 mg/L; outlet had no odor; removal percentage of color was 86.4%, outlet color was 3; outlet Al-residual was 0.011 mg/L which accorded with the quality standard of drinking water. Run time of filter was 36 h, UFRV (Unit Filter Run Volume) was 504 m3/m2.

Charcoal↗

PubMed2003

Amphiphilic diblock dendrimers with a fullerene core.

Amphiphilic dendrimers with a C(60) core have been obtained by cyclization of dendritic 1,3-phenylenebis(methylene)-tethered bis-malonate derivatives at the carbon sphere. The relative position of the two cyclopropane rings in the resulting bis-methanofullerene derivatives has been determined based on the molecular symmetry (C(s)()) deduced from the (1)H and (13)C NMR spectra. The hydrophobic-hydrophilic balance of these dendrimers has been systematically modified by changing the size of the polar headgroup in order to investigate the role of the amphiphilicity both at the air-water interface and during deposition onto solid substrates. Langmuir studies have revealed a conformational change in the dendritic structure with the size of the polar headgroup. Because of a better anchoring onto the water surface, the compounds with the largest polar headgroup adopt a more compact structure and the dendritic branches are forced to wrap the fullerene core. This model is nicely confirmed by the amount of fullerene-fullerene interactions within the Langmuir-Blodgett films as deduced from their absorption spectra.

Journal Article↗

PubMed2003

Pancreatic islets and insulinoma cells express a novel isoform of group VIA phospholipase A2 (iPLA2 beta) that participates in glucose-stimulated insulin secretion and is not produced by alternate splicing of the iPLA2 beta transcript.

Many cells express a group VIA 84 kDa phospholipase A(2) (iPLA(2)beta) that is sensitive to inhibition by a bromoenol lactone (BEL) suicide substrate. Inhibition of iPLA(2)beta in pancreatic islets and insulinoma cells suppresses, and overexpression of iPLA(2)beta in INS-1 insulinoma cells amplifies, glucose-stimulated insulin secretion, suggesting that iPLA(2)beta participates in secretion. Western blotting analyses reveal that glucose-responsive 832/13 INS-1 cells express essentially no 84 kDa iPLA(2)beta-immunoreactive protein but predominantly express a previously unrecognized immunoreactive iPLA(2)beta protein in the 70 kDa region that is not generated by a mechanism of alternate splicing of the iPLA(2)beta transcript. To determine if the 70 kDa-immunoreactive protein is a short isoform of iPLA(2)beta, protein from the 70 kDa region was digested with trypsin and analyzed by mass spectrometry. Such analyses reveal several peptides with masses and amino acid sequences that exactly match iPLA(2)beta tryptic peptides. Peptide sequences identified in the 70 kDa tryptic digest include iPLA(2)beta residues 7-53, suggesting that the N-terminus is preserved. We also report here that the 832/13 INS-1 cells express iPLA(2)beta catalytic activity and that BEL inhibits secretagogue-stimulated insulin secretion from these cells but not the incorporation of arachidonic acid into membrane PC pools of these cells. These observations suggest that the catalytic iPLA(2)beta activity expressed in 832/13 INS-1 cells is attributable to a short isoform of iPLA(2)beta and that this isoform participates in insulin secretory but not in membrane phospholipid remodeling pathways. Further, the finding that pancreatic islets also express predominantly a 70 kDa iPLA(2)beta-immunoreactive protein suggests that a signal transduction role of iPLA(2)beta in the native beta-cell might be attributable to a 70 kDa isoform of iPLA(2)beta.

Alternative Splicing↗

PubMed2003

Dual catalytic apparatus of the thiamin diphosphate coenzyme: acid-base via the 1',4'-iminopyrimidine tautomer along with its electrophilic role.

It was recently reported (Jordan, F.; Zhang, Z.; Sergienko, E. A. Bioorg. Chem. 2002, 30, 188-198) that addition to the E477Q active-center variant of yeast pyruvate decarboxylase of (a) pyruvate on a rapid-scan UV stopped-flow, or (b) acetaldehyde or benzoylformate on a circular dichroism (CD) instrument, generates a new band with lambda(max) near 300-310 nm. A chemical model demonstrated that the wavelength is appropriate to the 1',4'-iminopyrimidine tautomer of the 4'-aminopyrimidine ring in thiamin diphosphate. Herein, we report the formation of a new positive CD band centered at 305 nm when the Escherichia colipyruvate dehydrogenase complex first E1 subunit and its variants are exposed to phosphonolactylthiamin diphosphate, a stable analogue of the covalent adduct formed between the substrate pyruvate and the C2 atom of thiamin diphosphate. The behavior of this CD band, whether it suggests saturation of the enzyme by phosphonolactylthiamin diphosphate, or its very existence (the band is not seen with the E571A E1 variant, where E571 is hydrogen bonded to the N1' atom of the 4'-aminopyrimidine ring), as well as its position are consistent with its assignment to the 1',4'-imino thiamin diphosphate tautomer on the enzyme, chiral by virtue of its fixed V conformation. The mechanism of binding of phosphonolactylthiamin diphosphate closely resembles that of thiamin diphosphate itself.

Binding Sites↗

PubMed2003

An engineered chorismate mutase with allosteric regulation.

Besides playing a central role in phenylalanine biosynthesis, the bifunctional P-protein in Eschericia coli provides a unique model system for investigating whether allosteric effects can be engineered into protein catalysts using modular regulatory elements. Previous studies have established that the P-protein contains three distinct domains whose functions are preserved, even when separated: chorismate mutase (residues 1-109), prephenate dehydratase (residues 101-285), and an allosteric domain (residues 286-386) for feedback inhibition by phenylalanine. By deleting the prephenate dehydrase domain, a functional chorismate mutase linked directly to the phenylalanine binding domain has been engineered and overexpressed. This manuscript reports the catalytic properties of the mutase in the absence and presence of phenylalanine.

Allosteric Regulation↗

PubMed2003

Quantitative determination of noncovalent binding interactions using automated nanoelectrospray mass spectrometry.

Electrospray ionization mass spectrometry (ESI-MS) has proven to be an extremely powerful tool for studying biomolecular structures and noncovalent interactions. Here we report a method using a fully automated, chip-based nanoESI-MS system to determine the dissociation constants (Kd) for the complexes of two different proteins with their ligands. The automated nanoelectrospray system, consisting of the NanoMate and ESI chip, serves functionally as a combination of autosampler and nanoelectrospray ionization source. This system provides all the advantages of conventional nanoelectrospray plus automated, high-throughput analyses without carryover. The automated nanoESI system was used to investigate quantitative noncovalent interactions between ribonuclease A (RNase A) and cytidylic acid ligands (2'-CMP, CTP), a well-characterized model protein-ligand complex, and between an inactive endocellulase mutant (Thermobifida fusca Cel6A D117Acd) and four oligosaccharide ligands (cellotriose, cellotetraose, cellopentaose, cellohexaose). Both titration and competitive binding approaches were performed prior to automated nanoESI-MS analysis with a Q-TOF mass spectrometer. Dissociation constants for each complex were calculated from the sum of ion peak areas of free and complexed proteins during the titration and competition experiments. The measured Kd values for the RNase A-CMP and Cel6A D117Acd-G3 complexes were found to be in excellent agreement with the available published values obtained by standard spectroscopic titration techniques. To our knowledge, this is the first report of using an ESI-MS approach to study the interactions between a cellulase and oligosaccharides. The results provide new insights for understanding the nature of cellulase-cellulose interactions.

Animals↗

PubMed2003

[Experimental study of magnetic pingyangmycine-bearing microspheres in the treatment of cavernous hemangioma].

OBJECTIVE: To investigate the probability and mechanism magnetic pingyangmycine-bearing microspheres(MPMs) in the treatment of cavernous hemangioma. METHODS: Twenty rabbits were divided into four groups. MPMs, pingyangmycine and magnetic nonpingyangmycine-bearing microspheres were injected into marginal ear veins respectively. All animals were killed after 21 days and histological varations of the veins were observed under the light microscope and transmission electron microscope. RESULTS: The occlusion of the veins in the group of MPMs with the external magnetic field was more prominent than that in the other groups. CONCLUSION: MPMs can occlude veins by means of its embolized effect and PYM drug effect. MPMs may be better used in the therapy of cavernous hemangioma.

Animals↗

PubMed2003

Automated chip-based nanoelectrospray-mass spectrometry for rapid identification of proteins separated by two-dimensional gel electrophoresis.

We report a method using a fully automated chip-based nanoelectrospray system for two-dimensional (2-D) gel sample analyses with mass spectrometric detection. The automated nanoelectrospray system, consisting of the NanoMate and electrospray ionization (ESI) chip, serves as both an autosampler and nanoESI source. This infusion system aspirates samples from a 96-well plate using disposable pipette tips and then delivers these samples sequentially to an ESI chip. This chip is a fully integrated monolithic device consisting of a 10x10 array of nozzles. The automated nanoelectrospray system is easily controlled through software, permitting the user to select the number of samples to be analyzed, the volume of sample to aspirate, the spray voltage, and analysis time. The system offers all the advantages of conventional nanoelectrospray plus automated, high-throughput analyses without analyte carryover. The system was used for a protein identification study of 2-D gel spots of both Escherichia coli and yeast crude cell extracts. The identification of 50 spots from E. coli crude cell extract and 27 spots from yeast extract is presented, demonstrating the powerful combination of the automated nanoESI system, the Thermo Finnigan LCQ Deca ion-trap mass spectrometer, and SEQUEST search software. In addition, the effects of silver staining and colloidal Coomassie blue staining of 2-D gel spots on the detection sensitivity and protein sequence coverage are compared and discussed. Furthermore, the comparison results using the multiwell microscale preparation kit versus manual extraction for in-gel samples are presented.

Animals↗

PubMed2003

A fully automated nanoelectrospray tandem mass spectrometric method for analysis of Caco-2 samples.

Caco-2 cells offer a means to rapidly screen permeability of drug candidates, allowing pharmaceutical companies to eliminate candidates unable to cross the intestinal barrier early in the discovery process. This screening process is typically performed by conventional liquid chromatography/tandem mass spectrometry (LC/MS/MS), which can require time-consuming method development. An alternative to LC/MS/MS, automated nanoelectrospray tandem mass spectrometry (nanoESI-MS/MS), is introduced. This novel approach requires an off-line ZipTip desalting step followed by automated nanoESI-MS/MS, using the NanoMate 100 and ESI Chip. In addition to reduced method development time, automated nanoESI-MS/MS also offers no carry-over between samples, low sample consumption, and ease-of-use as compared with conventional pulled-capillary nanoelectrospray. Furthermore, the infusion system described has the potential to be high-throughput. A comparison of Caco-2 samples analyzed both by LC/MS/MS and by automated nanoESI-MS/MS is presented. The permeability and recovery data of the two compounds analyzed in this study obtained from conventional LC/MS/MS and by automated nanoESI-MS/MS were in excellent agreement.

Algorithms↗

PubMed2003

A novel mutation in the RDS/Peripherin gene causes adult-onset foveomacular dystrophy.

PURPOSE: To describe a novel mutation in the RDS/Peripherin gene that results in a moderately severe form of adult-onset foveomacular dystrophy. DESIGN: Observational case series. METHODS: Selected members of a family with adult-onset foveomacular dystrophy underwent complete ophthalmic evaluation, including fundus photography and fluorescein angiography, in a tertiary care referral center. The study population consisted of 12 members of a Caucasian kindred. After providing informed consent, patients donated blood for genomic DNA extraction and mutational screening using standard techniques. The main outcome measure were the presence of a RDS/Peripherin gene mutation in a patient with the disease and its absence in unaffected family members and controls. RESULTS: Eight affected family members and no unaffected family members demonstrated a single guanine base deletion at nucleotide 112 that led to premature termination at amino acid 38 of RDS/Peripherin polypeptide. This frameshift mutation results in truncation of nearly 90% of the gene product, thus probably representing a null allele. That results in a relatively severe phenotype, with choroidal neovascularization developing in two patients and geographic atrophy involving the macula in three patients. CONCLUSIONS: We describe a frameshift null mutation in the RDS/Peripherin gene associated with a relatively severe manifestation of adult-onset foveomacular dystrophy in affected family members.

Adult↗

PubMed2003

Imbalance between expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in invasiveness and metastasis of human gastric carcinoma.

AIM: The expressive balance between matrix metalloproteinase-9 (MMP-9) and its tissue inhibitor of metalloproteinase-1 (TIMP-1) plays a critical role in maintaining the degradation and synthesis of extracellular matrix. Loss of such balance is associated with invasion and metastasis of tumors. This study aimed to determine the expression of MMP-9 and TIMP-1 in gastric carcinoma, and the association of the expressive imbalance between MMP-9 and TIMP-1 with the invasion and metastasis and prognosis of gastric carcinoma. METHODS: We used immunohistochemistry to determine the expressions of MMP-9, TIMP-1 and proliferating cell nuclear antigen Ki-67 in the gastric specimens taken from 256 patients with primary gastric carcinoma. The patients were followed-up for up to 96 months. RESULTS: No association between the expression of MMP-9 and TIMP-1 and patients' sex and age, tumor size and location of gastric carcinoma was observed. The incidence of the positive expression of MMP-9 in cases with tumors invasion to muscularis propria and visceral peritoneum (70.13 % and 69.09 %, respectively) was significantly higher than that in cases with tumor invasion only to lamina propria or submucosa (42.50 %, P=0.0162). The positive correlation between MMP-9 expression and the depth of tumor invasion was observed (Pearson correlation coefficient=0.2129, P=0.016). Along with the increase of the metastatic station of lymph nodes, the incidence of the MMP-9 expression was increased by degrees; a positive correlation between them was observed (Pearson correlation coefficient=0.2910, P=0.0001). There was also a significant correlation between MMP-9 expression and the TNM stage in gastric carcinoma (Pearson correlation coefficient=0.3027, P<0.0001). The incidence of MMP-9 expression in stage II and III/IV (75.00 % and 76.15 %, respectively) was significantly higher than those in stage I (46.15 %, P<0.0001). A negative correlation between TIMP-1 immunoreactivity and the depth of invasion, status of lymph node metastasis and TNM stage was observed (Pearson correlation coefficient =-0.1688, -0.3556 and -0.3004, P=0.023, <0.0001 and <0.0001, respectively). Four types of co-expression of MMP-9 and TIMP-1 were observed; i.e. MMP-9 positive but TIMP-1 negative (n=115), both positive (n=52), both negative (n=62) and MMP-9 negative but TIMP-1 positive (n=27). The frequency of serosal invasiveness was significant higher in patients with MMP-9 but without TIMP-1 expression than those with other types of the co-expression (P=0.0303). The incidence of lymph node metastasis was highest in patients with MMP-9 but without TIMP-1 expression, and lowest in those with TIMP-1 but without MMP-9 expression (P<0.0001). The survival rate in patients with MMP-9 but without TIMP-1 expression was lower than that in those with TIMP-1 but without MMP-9 expression (P=0.0014). CONCLUSION: Our results in gastric carcinoma demonstrated a significant positive association of MMP-9 over-expression with proliferation of tumor cells, the depth of invasiveness, lymph node metastasis and TNM stage, suggesting MMP-9 can serve as a molecular marker of tumor invasion and metastasis. We also demonstrate a significant negative relationship of TIMP-1 expression with the depth of invasiveness and lymph node metastasis, which provide a new idea in the tumor biological and genetic treatment. The interaction between MMP-9 and TIMP-1 in the processes of tumor invasion and metastasis is that MMP-9 mainly promotes tumor invasion and metastasis and TIMP-1 inhibits functions of MMP-9. The imbalance between MMP-9 and TIMP-1 expression may suggest the occurrence of tumor invasion and metastasis, predict poor prognosis. For patients with imbalanced MMP-9 and TIMP-1 expression, the optimal treatment scheme needs to be selected.

Adult↗

PubMed2003

[HLA genotyping by automatic semi-quantitative polymerase chain reaction-reverse sequence specific oligonucleotide].

OBJECTIVE: To explore a HLA genotyping method that can be used for organ transplantation tissue typing, and especially for establishing hemopoietic stem cell bank and the cord blood stem cell bank by analyzing the PCR-DNA. METHODS: Modified automatic method based on semi-quantitative amplification system of HLA-I, I allele genotyping was established using reverse sequence-specific oligonucleotide with polymerase chain reaction (PCR-RSSO), and was compared with PCR (using sequence specific primer, PCR-SSR) and manual PCR-RSSO in terms of the accuracy, resolution, and the quantity of DNA consumption. A total of 635 blood samples were genotyped with HLA-A, B, C, DR and DQ alleles using auto semi-quantitative PCR-RSSO, 166 of which were also examined using PCR-SSP and manual PCR-RSSO simultaneously. RDSULTS: The success rate of automatic semi-quantitative PCR-RSSO, PCR-SSP and manual PCR-RSSO was 98.4% (3 124/3 175), 98.8% (656/664) and 88.3% (732/830) respectively, with no significant difference between the former 2 as indicated by X2 test (P>0.05). Auto semi-quantitative PCR-RSSO, however, yielded significantly higher success rate than manual PCR-RSSO (P<0.05). CONCLUSION: PCR-RSSO is capable of identifying 706 alleles of HLA-I, II antigens which amounts to 75.43% of the 936 alleles published by WHO in 2000, with intermediate to high resolution, even in the case of the homozygote. The hybridization results documenting the original data can be conveniently preserved. This method therefore possesses the merits of low expenses, low labor intensity, and rapid processing of large number of samples.

Alleles↗