Two Series of Main-Group Heterometallic Selenides Synthesized in Two Different Types of Ionic Liquids.

Imidazolium-based ionic liquids have been widely applied in the synthesis of organic hybrid chalcogenidometalates, while the other types of ionic liquids are rarely tried. Reported here is the first application of a pyridinium-based ionic liquid in the preparation of two main-group heterometallic selenides featuring isomorphic three-dimensional frameworks. Of particular interest is that three gallium-tin selenides possessing another type of three-dimensional framework have been prepared by replacing the pyridinium-based ionic liquid with imidalolium-based ionic liquids under the same reaction conditions.

[Correlation study between Chinese myasthenia gravis patients from Guangdong province and the polymorphism of HLA immunogene].

OBJECTIVE: To explore the correlation between Chinese myasthenia gravis (MG) patients from Guangdong province and the polymorphism of HLA immunogene. METHODS: The genotypes of HLA-A, B and DRB1 alleles in 104 MG patients and 121 healthy blood donors were detected by PCR-SBT (polymerase chain reaction-sequencing-based typing). RESULTS: (1) There were 15 alleles at A locus, 32 at B locus and 23 at DRB1 locus in MG group. (2) The frequency of HLA-A*02:07(P = 0.000, RR = 3.715), -B*46:01(P = 0.000, RR = 5.698), -DRB1*04:03(P = 0.033, RR = 6.312), -DRB1*09:01(P = 0.000, RR = 5.884) in MG patients was higher than that in healthy controls. (3) There were positive associations of HLA-DRB1*09:01(P = 0.000, RR = 1.349) with juvenile-onset ocular MG. CONCLUSION: There is susceptibility association of HLA-A*02:07, -B*46:01, -DRB1*04:03, -DRB1*09:01 with Chinese MG patients from Guangdong province. There is a close genetic and immunological correlation between HLA alleles and the pathogenesis of MG. It has directional significance in the race and region incidence study, clinical classification, differential diagnosis, treatment and prognosis of MG.

[Genotyping of RhD-negative blood samples diagnosed by serological tests from patients waiting for kidney transplantation].

OBJECTIVE: To compare the accuracy of serological and molecular approaches to identification of RhD-negative patients waiting for kidney transplantation. METHODS: A total of 103 RhD-negative blood samples by serological test were collected from patients waiting for kidney transplantation between January, 2006 and January, 2016. Quantitative PCR and sequencing were used to verify the results of RHD genotyping, and the false negative rates of the serological and molecular methods for RhD genotyping were compared. RESULTS: Among the 103 blood samples, true RhD negativity (with all the 10 exons missing) was found in 56 samples (54.5%), and false RhD negativity (RhD positivity with loss, repetition, or missense mutation in the 10 exons) in 47 samples (45.6%). In the 47 false RhD-negative cases, weak D was detected in 1 case (2.1%), partial D in 13 cases (27.7%), and D-elution in 33 cases (70.2%). The detection rates of RhD negativity differed significantly between the serological and molecular methods (P<0.05). CONCLUSION: Serological test is associated with a high false negative rate in detecting RhD blood group, and the use of the molecular approach has important clinical significance in accurate RhD genotyping for patients waiting for renal transplantation.

[The outcomes of the thirty patients with refractory leukemia treated with related HLA haploidentical stem cells transplantation].

OBJECTIVE: To assess the outcomes of the therapy for patients with refractory leukemia with HLA haploidentical stem cells transplantation. METHODS: To analyze the outcomes of 30 patients with refractory leukemia who underwent HLA haploidentical peripheral blood stem cells transplantation from August 1998 to August 2004. RESULTS: Thirty refractory leukemia patients including 13 cases of acute non-lymphocytic leukemia, 10 cases of acute lymphocytic leukemia (ALL), 6 cases of chronic myeloid leukemia and 1 case of phase IV non-Hodgkin's lymphoma underwent HLA haploidentical peripheral blood stem cells transplantation. The median age was 25 years old (3-52 years old). Twelve patients received stem cells from parent donors, four from daughter or son donors and the remaining from sibling donors. Three HLA loci mismatched in twelve cases, two HLA loci mismatched in thirteen cases and one HLA locus mismatched in five cases. The conditioning regime consisted of fludara (25 mg/m(2) x 5 d), busulfan (4 mg/kg x 4 d) and cyclophosphamide (60 mg/kg x 2 d). Rabbit anti-human lymphocyte globulin (5 mg/kg x 5 d) was added in some patients in the conditioning regime. A mean of 5.0 (2.9-8.0) x 10(8)/kg mononucleated cells was grafted. The number of mean CD(34)(+) cells was 5.5 (3.0-6.5) x 10(6)/kg. Twenty-seven patients were successfully grafted, one failed to graft, one died from severe fungal infection at day 2 and one died from severe veno-occlusive disease at day 28. The mean time of white cell count more than 1.0 x 10(9)/L was 14 (11-18) days and platelet count more than 20 x 10(9)/L was 15 (11-18) days. ALL the 27 successfully grafted patients got complete remission. Severe acute graft versus host disease occurred in six patients and four of them died. Seven patients suffered from chronic graft versus host disease. Seven patients relapsed and died. The median relapse time was 10 (3-24) months. Fourteen patients are still surviving, and ten have disease free survival. CONCLUSION: It is concluded from our observation that HLA haploidentical peripheral blood stem cells transplantation may be an effective therapy for refractory and relapse leukemia. Some patients with refractory and relapse leukemia treated with HLA haploidentical stem cells transplantation may have disease free survival. Graft versus leukemia effect may be strong in patients receiving HLA haploidentical blood stem cells transplantation and leukemia will probably be relapsed when the patient without complete remission was treated with this therapy.

[ROC curve-based evaluation of platelet parameters for predicting delayed renal graft function].

OBJECTIVE: To investigate the value of evaluating 5 platelet parameters in predicting delayed graft function (DGF) in patients following kidney transplantation. METHODS: We retrospectively analyzed the pre- and postoperative (within 2 months) data of 330 renal transplant recipients. The cases with DGF and those without were analyzed to assess the association between relationship between DGF following transplantation and the variations of blood platelet parameters including platelet count (PLT), large platelet ratio (P-LCR), mean platelet volume (MPV), platelet volume distribution width (PDW) and platelet hematocrit (PCT). RESULTS: The DGF and non-DGF cases were comparable for the platelet parameters before the operation. On postoperative day 7 when the diagnosis of DGF was made, PLT (P<0.05) and PCT (P<0.02) were significantly lower while MPV (P<0.01), PDW (P=0.036) and P-LCR (P=0.01) significantly higher in DGF group than in non-DGF group. The AUCs of P-LCR (0.611±0.047), PDW (0.603±0.048) and MPV (0.762±0.037) were significantly higher than the reference area (P<0.05) with cut-off values of 34.80%, 12.95fl and 11.55fl, respectively. MPV showed a high sensitivity, specificity and Youden index for predicting DFG; PDW and P-LCR had a high sensitivity but a low specificity for predicting DFG with a modest diagnostic value. PLT and PCT, with AUCs of were 0.37 and 0.38, respectively, did not have a predictive value for DGF. CONCLUSIONS: Significant variations in platelet parameters occur in the event of DGF in renal transplant recipients, and monitoring the postoperative changes in MPV, PDW, and P-LCR can help in early diagnosis and treatment of DGF. MPV has a moderate value (0.7-0.9) in predicting DGF, and a MPV>11.55 fl suggests the risk of DGF.

[HLA class I and II polymorphism and haplotypes in Guangdong Han population].

OBJECTIVE: To analyze the polymorphism and haplotypes of HLA class I and II in Guangdong Han population and detect the HLA-A, B, Cw and DRB1 allele frequencies. METHODS: An auto semi-quantitative PCR-sequence speacific oligonucleotide probe(PCR-SSOP) method was adopted in exploring the HLA-A, B, Cw and DRB1 genotypes of the samples from 160 bone marrow donors. RESULTS: Twelve HLA-A, 23 B, 11 Cw and 13 DRB1 alleles were obtained. A total of 9 HLA-A-B, 20 Cw-B, 7 A-Cw, and 8 A-DRB1, 9 B-DRB1, 10 Cw-DRB1 haplotypes were found. CONCLUSION: HLA class I and II alleles in Guangdong Han population have plenty of polymorphisms. The haplotype distribution possesses territory characteristic.

[Therapy of Duchenne muscular dystrophy with umbilical cord blood stem cell transplantation].

OBJECTIVE: To analyze a Duchenne muscular dystrophy(DMD) patient's muscular regeneration, dystrophin expression and locomotive variation before and after he underwent umbilical cord blood stem cell transplantation in order to assess the therapeutic effect. METHODS: A 12-year-old DMD boy who could not walk for 3 years was confirmed by gene analysis and dystrophin protein immune test on his muscle. He had no other chronic disease. By HLA matching, a piece of umbilical cord blood stem cell with 6 HLA sites matching to the boy was found in Guangdong Umbilical Cord Blood Bank. The number of the nucleated cells of the umbilical cord blood stem cell was 24.08x 10(8). After pretreatment for the DMD boy with busulfan, cyclophosphamide and rabbit anti-human thymocyte globulin, the allergenic cord blood stem cells were transplanted into him by intravenous injection. Cyclosporin A, methylprednisolone, MMF, prostaglandin E1 and ganciclovir were given after the transplantation. At the same time, Gran, the granulocytic cell stimulating factor, and gamma globulin were administered. The biochemistry profile including serum creatine kinase (CK), the reconstruction of blood making, the deletion exon of DMD gene, the regenerating muscles, the dystrophin protein expression, and the locomotive function of the DMD boy were tested regularly. RESULTS: (1) The white blood cells (WBC) of peripheral blood decreased gradually to zero after pretreatment. In a period of 15 days after transplantation, the neutrophil increased to 0.5x 10(9)/L; at 25 days, WBC increased to normal level. Blood platelet was more than 20x 10(9)/L at 22 days. The hemoglobin rose to 85-100 g/L. At 140 days, sternal puncture revealed the rapid growth of neutrophil, blood platelet and hemoglobin. (2)At 140 days, the blood type of the DMD boy transformed from type O to type AB (the donor's blood type being AB). There was no grafe versus host reaction. (3) At 18, 30, 43, 55, 74 and 233 days after transplantation, the PCR-short tandem repeat test of the boy's peripheral blood DNA showed that his genotype was completely the same as the donor's. The results of PCR-short tandem repeat tests of the bone marrow cells DNA by sternal puncture at 140, 183 and 235 days were the same as those of the blood DNA. (4) At 60 days, DMD gene analysis by PCR showed that the defected DMD gene (exon 19 deletion) had been corrected by the umbilical cord stem cells transplantation. (5) At 75 days, the biopsy of calf muscle showed there were myoblast cells and muscular tubes growing. The dystrophin expressions were weak, but a few of them were strong. DNA analysis showed that the donor's gene DNA accounted for 1%-13%. At 126 days, obviously increased dystrophin positive muscular fibers of the boy were found. The donor's fibers rose to 2.5%-25%. (6) The serum CK of the boy declined from 5735 U/L to 274 U/L. (7) At 100 days, physical examination revealed improvement in his arms and legs. CONCLUSION: The therapy of Duchenne muscular dystrophy with allogeneic umbilical cord blood hematopoietic stem cell transplantation may reset up the blood-making function, decrease the serum CK level, restore the dystrophin in muscles, and improve the locomotive function of the DMD boy. These data suggest that the allogeneic umbilical cord blood hematopoietic stem cell transplantation may benefit the DMD boys.

[Allogeneic umbilical cord blood stem cell transplantation in Duchenne muscular dystrophy].

OBJECTIVE: To study the feasibility of treatment of Duchenne muscular dystrophy (DMD) with umbilical cord stem cell transplantation. METHODS: HLA matching was conducted for a 11-year-old DMD boy with family history was underwent umbilical cord blood stem cell transplantation and a sample of umbilical cord stem cells with 5 matched HLA sites was found in the cord blood bank with 27.32 x 10(8) nucleated cells, about 2.6 times that of the treatment dosage for him. After pretreatment with busulfan 14 mg/kg.d, cyclophosphamide 50 mg/kg.d, and rabbit anti-human thymocyte globulin 10 mg/kg.d, the allogeneic cord blood stem cells were transplanted intravenously. Cyclosporin A, methylprednisolone and MMF were used after the transplantation so as to prevent graft versus host reaction. Prostaglandin E1 was used to prevent Budd-Chiari syndrome, and ganciclovir was used to prevent cytomegalovirus infection. At the same time, Gran, granulocytic cell stimulating factor, and gammaglobulin were also used. Biochemistry test, including serum creatine kinase (CK), was conducted. Evidence of reconstruction of blood making, including conversion of blood type, was observed. PCR-STR analysis was used to observe the status of implantation of the donor umbilical cord blood stem cells. RESULTS: (1) 12 days after transplantation, the white blood cells (WBC) of peripheral blood were 0.5 x 10(9)/L, 14 days after, the numbers of WBC and neutrophils were 1.0 x 10(9)/L and 0.6 x 10(9)/L respectively. In 37 days, granulocytic cell stimulating factor was no more used, the peripheral blood WBC fluctuated around 3.34 approximately 12.2 x 10(9)/L. In the 27th day, the number of blood platelets was more than 20 x 10(9)/L and hemoglobin rose to 88 g/L. On the 24th day red blood cells transfusion was stopped. (2) In the 42nd day, the blood type of the patient transformed from type A before transplantation to type AB (the blood type of transplanted stem cells is type B). (3) PCR-STR test of the peripheral blood made 17, 26, and 42 days after transplantation showed that the gene type of the patient was mixed mosaic: The ratio of donor gradually increased from 40% approximately 45% to 55% approximately 65%. (4) In the 38th day I degrees GVHD appeared. (5) serum CK level declined from 6000 U/L to 600 approximately 2200 U/L. (6) In the 42nd day, physical examination revealed obviously improvement in walking, turning the body over, and standing up. CONCLUSION: This is first case of prospective clinical transplantation on DMD by allogeneic cord blood stem cell. Umbilical cord stem cell transplantation helps re-build blood-making function, and improve locomotive function with a mild GVHD reaction. The genotype of rebuilt blood is mosaic but the ratio of gene mosaic gradually turn from recipient gene type > donor gene type to recipient gene type < donor gene type. The serum CK level decreases significantly after transplantation, which may slow down the necrosis of muscle cell. DMD patient will be benefited by stem cell transplantation.

[Induced differentiation of human cord blood monocytes into neuron-like cells in vitro].

OBJECTIVE: To probe the conditions for inducing human cord blood monocytes to differentiate into neuron-like cells. METHODS: The mononuclear cells were isolated from human umbilical cord blood samples and plated in 25-mm culture flasks containing DMEM/F12 medium. The fifth passage of mesenchymal stem cells (MSCs) were induced by beta-mercaptoe- thanol (beta-ME), dimethyl sulfoxide (DMSO) and conditioned medium for neuron induction, respectively, to differentiate into neuron-like cells. The expression of nestin, neuron-specific enolase (NSE) and neurofilament (NF) on the treated cells were detected by immunocytochemical method. RESULTS: Nestin expression were found in 6.7% of the primary, 12.4% of the second passage, 20.8% of the fifth passage of MSCs, respectively. Thirty-six percent of cell cultures treated with the conditioned medium for neuron induction were immunoreactive for NSE and NF, and 33% of the cells induced by beta-ME and 25% of the cells by DMSO also expressed NSE and NF. CONCLUSION: Cord blood MSCs possess some features of neural stem cells, and have the capacity to differentiate into neuron-like cells under proper conditions.

[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.

[HLA-Cw alleles polymorphism and haplotypes in Guangdong Han population].

OBJECTIVE: To analyze the polymorphism and haplotypes of HLA-Cw and detect HLA-A, B, Cw and DRB1 allele frequencies in Guangdong Han population. METHOD: An auto semi-quantitative PCR with reverse sequence-specific oligonucleo- tide was adopted to explore the HLA-A, B, Cw and DRB1 genotypes of 185 bone marrow donors. RESULT: Eleven HLA-Cw alleles were obtained in which Cw*03 (0.2580), 07 (0.1887), 01 (0.1732), and 08 (0.1071) had much higher allele frequencies. A total of 7 HLA-Cw-A, 20 HLA-Cw-B and 10 HLA-Cw-DRB1 haplotypes were found. CONCLUSION: HLA-Cw alleles have richer polymorphisms and their linkage disequilibrium with HLA-A, B, DRB1 exhibits geographic genetic characteristics.

[Killer immunoglobulin-like receptor gene distribution in Guangdong Han population].

OBJECTIVE: To investigate the distribution of killer immunoglobulin-like receptor (KIR) gene in Guangdong Han population. METHODS: KIR phenotype was examined by PCR with sequence-specific primers in 96 subjects of Han nationality in Guangdong Province of China, and KIR frequency was calculated and compared with those in Caucasian, north Indian and Japanese populations. RESULT: The gene expression frequency of KIR in Guangdong Han people was 2DL1(0.85), 2DL2(0.12), 2DL3(0.58), 2DL4(1), 2DL5(0.24), 3DL1(0.96), 3DL2(1), 3DL3(1), 2DP1(0.97), 2DP2(0.98), 2DS1(0.10), 2DS2(0.30), 2DS3(0.02), 2DS4(0.28), 1D(0.65), 2DS5(0.19), and 3DS1(0.23) respectively. Comparison of the KIR recognizing the same HLA ligand suggested significantly higher expression frequency of inhibitory KIR than that of activating KIR. Compared with Caucasian and north Indian populations, Guangdong Han population had significantly lower expression frequency of activating KIR gene with the exception of KIR2DS4. CONCLUSION: Different KIR genes have different expression frequencies in Guangdong Han population, and KIR gene distribution varies between populations of different races.

[Panel reactive antibody screening and human leukocyte antigen typing in 1 700 cases of renal transplantation].

OBJECTIVE: To evaluate the role of panel reactive antibody (PRA) screening and human leukocyte antigen (HLA) typing in renal transplantation. METHODS: PRA screening and HLA typing were performed in 1 700 patients eligible for the first group of renal transplantation who had 3 to 6 HLA matches in HLA-A, B and DR with the donor, and in cases positive for PRA, plasma exchange was conducted. Another 423 patients who did not receive PRA screening or HLA typing constituted the second group. The changes of immune variables, incidences of acute rejection and the effect of HLA-A, B, DR matching on long-term graft survival were observed. RESULTS: In 1 700 cases of group 1, post-transplantation CsA dose was reduced to 5 to 7 mg*kg(-1)*d(-1) and the graft function recovery time ranged from 2 to 16 d, averaging 5 d. Acute graft rejection occurred in 252 (14.8%) cases, but no hyper-acute rejection was observed. The 1-, 3- and 5-year patient/graft survival rates were 98.6%/96.7%, 93.1%/87.3% and 88.1%/83.6% respectively. In group 2, CsA dose ranged from 8 to 12 mg*kg(-1)*d(-1) and the graft function recovery time was 4 to 30 d, averaging 13 d. Hyper-acute rejection occurred in 9 (2.1%) and acute rejection in 198 (46.8%) cases, and the 1-, 3- and 5-year patient/graft survival rates were 86.7%/76.3%, 72.5%/67.9% and 69.5%/59.3% respectively. CONCLUSIONS: Negative PRA and good HLA matching can eliminate the incidences of hyper-acute rejection, decrease the rate of acute rejection and improve both patient and graft survival rates.

[Relationship between polymorphism of HLA-A, -B, -DRB1 alleles and susceptibility of children to acquired aplastic anemia].

The aim of this study was to identify the relationship between susceptibility of children to acquired aplastic anemia (AA) and HLA-A, -B, -DRB1 alleles. 80 children with AA were enrolled in this study. Among of them, 34 patients collected from tissue typing test centers of Nanfang Hospital; 46 patients were diagnosed at Department of Pediatrics of Sun Yat-Sen Memorial Hospital. In these patients, 48 were males, 32 were females, and with average age 8.1 years old, 6 cases were non-severe AA (nSAA), 74 case were severe AA (SAA). The healthy control group consisted of 109 donors who were from the same area. All the patients and healthy controls were of Han Chinese, and all were unrelated individuals. The polymerase chain reaction sequence specific primers (PCR-SSP) was used to analyze the polymorphism of HLA-A, -B and -DRB1 alleles. Pearson Chi-square or continuity correction or two-sided Fisher's exact test were used. The results showed that the genotype frequency of HLA-B*48:01 and DRB1*09:01 were significantly higher in children with AA as compared with healthy controls (P < 0.05). The genotype frequency of HLA-B*51:01, DRB1*03:01 and DRB1*11:01 were significantly lower in children with AA as compared with healthy controls (P < 0.05). Besides, the results also demonstrated that the genotype frequencies of HLA-B*48:01 and DRB1*09:01 were significantly higher in SAA as compared with controls, the genotype frequencies of B*51:01, DRB1*03:01 and DRB1*11:01 were significantly lower in SAA, as compared with controls. In conclusion, HLA-B*48:01 and DRB1*09:01 are related with children AA, and may be susceptible alleles to the development of children AA. Besides, the expression of HLA-B*51:01, DRB1*03:01 and DRB1*11:01 are low in children with AA, whether they are relative protection alleles of children needs to be further studied.

[Anti-MICA antibodies: risk factors for sensitization and the impact on renal transplantation outcomes].

OBJECTIVE: To investigate the risk factors for sensitization of anti-MICA antibodies and their impact on the outcomes of renal transplantation. METHODS: Luminex flow cytometry were used to identify 10 MICA antibodies and evaluate the antibody specificity in 98 uremic patients positive or negative for anti-MICA antibodies undergoing kidney transplantation. The factors contributing to MICA sensitization were analyzed, and the incidence of acute rejection and graft function recovery time were compared between the positive and negative cases for anti-MICA antibodies. RESULTS: Of the 98 uremic patients, 16 (16.3%) were positive for anti-MICA antibodies. The positive and negative cases showed significant differences in the history of blood transfusion, pregnancy, transplantation, and PRA status (P<0.05). In the 38 renal transplant recipients, 6 experienced acute graft rejection, which was reversed by methylprednisolone pulse therapy; of the 10 recipients positive for anti-MICA antibodies, 4 showed acute graft rejection as compared to 2 out of the 28 recipients negative for anti-MICA antibodies (P=0.031). The cases positive for anti-MICA antibodies showed a significantly longer graft function recovery time than the negative cases (14.6∓4.7 vs 8.2∓4.5 days, P=0.001). CONCLUSIONS: Blood transfusion, pregnancy, and transplantation all contribute to the production of anti-MICA antibodies. Patients positive for anti-MICA antibodies may require strict HLA matching and more potent immunosuppressive drugs to prevent renal graft rejection and improve graft survival.

[KIR/HLA ligand matching and acute rejection after kidney transplantation].

OBJECTIVE: To explore the effect of KIR/HLA ligand matching which mediates activated or inhibitory signal pathways on acute rejection (AR) after kidney transplantation. METHODS: HLA and KIR genotype assortments were analyzed in 53 donor/recipient pairs of kidney transplantation. The recipients were divided into AR group (GI, n=19) and stable renal function group (GII, n=34) based on the early graft function. The impact of donor HLA, recipient KIR and distinct KIR/HLA class I ligand combinations on acute rejection after kidney transplantation was studied. RESULTS: No significant differences were found in donor HLA-C1/2, HLA-A3, HLA-A11, or HLA-Bw4 between GI and GII groups. The frequency for KIR2DL2/2DS2 and KIR genotype assortment (AA) of the recipients in GI group were significantly lower than that in GII group (26.3% vs 55.9%, P=0.038; 31.6% vs 67.6%, P=0.011). The incidence of AR was significantly lower in donor HLA-C1/1 than in non-C1/1 (31.6% vs 46.7%, P>0.05), and lower in recipient KIR genotype assortment (AA) than in non-AA (20.7% vs 52.2%, P=0.011). A significant higher number of matches for the KIR2DL2/ HLA-C1 and KIR2DL3/HLA-C1 were observed in GII group (P=0.030, P=0.028). CONCLUSION: Distinct KIR/HLA class I ligand combinations between the donor and recipient (such as KIR2DL2/ HLA-C1 and KIR2DL3/HLA-C1) may reduce the incidence of AR. A good KIR/HLA class I ligand matching will benefit the survival of the renal allograft.

[Analysis of natural killer cell immunoglobulin-like receptor genes in Chinese].

OBJECTIVE: To investigate the genotypes of natural killer cell immunoglobulin-like receptor (KIR) genes and their frequencies in Chinese subjects and explore the mechanism of the actions of nature killer cells. METHODS: The DNA samples were obtained from 67 randomly selected unrelated Chinese Han individuals for genotyping of the KIR genes using PCR with sequence-specific primers (PCR-SSP), and the frequencies of the KIR genes in these Chinese subjects were compared with the reported frequencies in populations of other nationalities. RESULTS: Sixteen KIR genes were identified in these Chinese subjects, and 87.5% of these genes were expressed at frequencies above 0.35. Fourteen functional KIR genes combined into 25 KIR genotypes, among which the most frequent genotype KIR-2DL1-2DL3-2DL4-3DL1-3DL2-3DL3-2DS4 showed a frequency of 0.373, while the frequencies of all the other genotypes were no greater than 0.09. Comparison of the KIR combinations in Chinese Han population with those of Japanese, Korean, and Caucasians populations identified 8.93% of the KIR combinations shared by all these populations; the Chinese, Koreans and Caucasians shared 5.36% common KIR combinations, whereas only 1.79% common combinations were found in Chinese and Caucasians. In this study, 16 new gene combinations were identified (25.28%). CONCLUSION: This study shows the high-frequency distribution of a single KIR gene polymorphism. The KIR combination KIR-2DL1-2DL3-2DL4-3DL1-3DL2-3DL3-2DS4 has the highest frequency in Chinese, Japanese, Korean and Caucasian populations, indicating that inhibitory signal transduction pathway plays an important role in the function of the natural killer cells. This study provide clues for new approaches for improving the prognosis of kidney transplantation by enhancing or inhibiting the function of the natural killer cells instead of life-time usage of immunosuppressive agents.

[Expression of major histocompatibility complex class I-related chain A antibody in patients with end-stage renal disease and its clinical implications].

OBJECTIVE: To study the frequency of major histocompatibility complex class I-related chain A (MICA) antibody in patients with end-stage renal disease (ESRD). METHODS: Luminex flow cytometry and beads loaded with 11 MICA antigens were used to identify the MICA antibody and evaluate the antibody specificity in 110 patients with ESRD. RESULTS: The positivity rate of MICA antibody was 40% (12/30) in PRA-positive patients, significantly higher than the rate of 17.5% (14/80) in PRA-negative patients (chi(2)=6.120, P=0.013). MICA-specific antibodies against 10 of the 11 MICA antigens were detected in 26 MICA antibody-positive patients, and 26.92% of the MICA antibody-positive patients had antibodies with single-specificity and 73.08% had polyspecific antibodies. Three MICA antibody-positive patients with cadaveric kidney transplantation showed good function of the graft without acute rejection 2 months after the operation. CONCLUSION: The positivity rate of MICA antibody is significantly higher in PRA-positive patients, suggesting a strong correlation between MICA and PRA positivity. The MICA antibodies are polyspecific and probably consist of IgM and IgG. These data can be used as prospective data for these ESRD patients considering potential renal transplantation, and may facilitate further investigation of the association of MICA with renal transplantation.

[Factors responsible for inter-individual variations in dosage/concentration of tacrolimus in renal transplant recipients].

OBJECTIVE: To identify the factors responsible for the inter-individual variations in the dosage/concentration of tacrolimus in renal transplant recipients. METHODS: This study involved renal transplant recipients receiving immunosuppressive therapy with the tacrolimus, mycophenolate and prednisone regimen after the operation. The gender, age, height, body weight, tacrolimus dosage, hormone dosage, diarrhea, blood lipids, liver function, renal function, albumin, and hematocrit of the patients were recorded at different time points, namely in early stage (3, 7, 14, and 30 days postoperatively, 118 cases), at 3 months (103 cases), 6 months (75 cases) and over one year (119 cases) after the operation. The concentrations of tacrolimus and gene polymorphisms at CYP3A5, MDR1 3435, MDR1 2677 and MDR1 1236 were also determined in these patients. Multiple linear regression was used for analysis of these factors with tacrolimus concentration/dosage*body surface area as the independent variable. RESULTS: Patients in early stage following renal transplantation showed rather poor fitting of the stepwise regression model, which increased obviously 3 months after the operation and further increased till reaching a stable level at 6 months. Multiple factors were found to affect tacrolimus concentration/dosage in the early postoperative stage, during which period these factors underwent drastic variations and became stable 3 months later. In terms of pharmacogenomics, the major factors affecting tacrolimus concentration/dosage included MDR1 3435, MDR1 2677 and MDR1 1236 polymorphisms, which vastly varied between the patients early after the operation. Of these polymorphic sites, CYP3A5 produced only minor effects on tacrolimus concentration/dosage, and was not included as an active factor until the stable phase (over 1 year) following the transplantation; MDR1 3435 was found to be the predominant factor affecting tacrolimus metabolism in the stable phase. Age, liver function, albumin and hematocrit were found to be positively correlated to the independent variable tacrolimus concentration/dosage*body surface area, and identified as important factors responsible for the intra-individual variation of tacrolimus dosage/concentration. CONCLUSION: The variations in the factors affecting tacrolimus dosage/concentration after renal transplantation are consistent with the clinical features of the patients, and these factors vary with the postoperative stages. Pharmacogenomic factors produce the most conspicuous effect on tacrolimus dosage/concentration, and agents that may interfere with tacrolimus metabolism should be avoided after the operation. Age, liver function, albumin and hematocrit are also important factors responsible for the variation of tacrolimus dosage/concentration.