AIM:To explore the effect of glucose-regulated protein 78 (GRP78) on the gastric carcinogenesis. METHODS:GPR78 expression patterns were examined in 34 specimens from gastric carcinoma patients using the immu-nohistochemistry (IHC) assay, and in 10 specimens using Western blotting analysis .In addition, the expression of GPR78 and cyclin D1 was detected in human gastric cancer cell lines SGC 7901 and SGC7901-H78 (overexpressing GRP78) by Western blotting.RESULTS:By IHC assay, GRP78 was found to be highly expressed in the cytoplasm of gastric carcino-mas as compared with the adjacent non-malignant tissues and corresponding normal tissues .GRP78 expression was positive-ly correlated with gender and histological differentiation (P<0.05), but not with age, tumor stage and lymph node metas-tasis (P>0.05).Furthermore, we found that with the increased expression of GRP 78 in SGC7901-H78 cells, the expres-sion of cyclin D1 was also elevated .CONCLUSION:GRP78 might be a key player to be involved in the growth of gastric cancer.

Objective To study the inter-allelic recombination event occurring in the HLA-C locus in a family of Chinese Han nationality, and to evaluate the molecular genetic background of the new HLA allele.Methods Peripheral blood samples were collected from a Chinese leukemia woman patient, as well as her healthy parents and two brothers.HLA-A, C, B, DRB1 and DQB1 alleles were typed by high-resolution PCR-sequence-based typing (SBT) method using Atria Genetic AlleleSEQR HLA SBT kits.The Protrans S4 HLA-C single allele-specific sequencing strategy was used to separate the two HLA-C alleles and to determine novelty of the allele.The full length sequences of HLA-C alleles of the patient and her parents were further analyzed using cloning and haplotype sequencing method. The HLA five loci linked haplotypes and the recombination site were analyzed by family study, meanwhile the full length sequences of the five HLA-C alleles were compared with the IMGT/HLA database by the program BLAST.Results The two haplotypes of the father and mother were a:A*0207-C*010201-B*550201-DRB1*090102-DRQ1*030302 and b:A*240201-C*120202-B*5201-DRB1*1502-DRQ1*0601, c:A*300101-C*060201-B*130201-DRB1*0405-DRQ1*0401 and d:A*110101-C*070201-B*4001-DRB1*080302-DRQ1*0601,respectively.The two brothers inherited their parent′s haplotypes a, d and b, c respectively.The two haplotypes of the patient were the maternal c and paternal recombinant a/b haplotype.The recombinant a/b haplotype A*240201-C*new-B*550201-DRB1*090102-DRQ1*030302, A*240201 came from the paternal haplotype b,while B*550201-DRB1*090102-DRQ1*030302 came from the other paternal haplotype a.When comparing the full length sequences of the HLA-C new allele with the father′s allele C*010201 and C*120202, it could deduce that the recombinant a/b haplotype derived from a recombination event occurring between the paternal chromosome 6 during meiosis.The crossover site was between genomic nt273 and nt330 of HLA-C alleles, which created a HLA-C new allele and the fifth haplotype of the family, and inherited it to the patient.The full length sequences of the new allele had been submitted to Genbank, and officially named C*0121 by WHO nomenclature committee.Conclusion This study demonstrates a rare inter-allelic recombination event occurring in the HLA-C locus within a Chinese Han family and illustrates the process of novel allele and haplotype, and provides direct theory for further studying the mechanisms of gene recombination and HLA polymorphism.

Objective To investigate the features and main reasons of blindness induced by uveitis in China. Methods A retrospective analysis was performed on the data from 1 214 patients with uveitis, referring to Zhongshan Ophthalmic Center, with special respect to the incidence of blindness in different uveitis entities, the characteristics of blindness, and possible causes for the blindness. Results In the affected 1 892 eyes of 1 214 patients with uveitis, 355 eyes (18.83%) were blind. The mean age at the onset of blindness was 34.38 years and the gender ratio of male to female was 1.52:1. The blindness led by panuveitis was found in 248 eyes (26.27%), including 128 (51.61%) and 73 (29.44%) blind eyes caused by Behcet's disease and Vogt-Koyanagi-Harada syndrome. Complicated cataract, vitreous opacity and secondary glaucoma were responsible for the blindness of the patients with panuveitis [89(35.89%), 53 (21.37%), and 30 eyes (12.10%), respectively]. Blindness caused by anterior uveitis was noted in 79 eyes (10.73%) with the main reasons of complicated cataract [56 eyes (70.89%)] and secondary glaucoma [16 eyes (20.25%)], posterior uveitis in 15 eyes (15.63%) with the main reason of vitreous opacity [9 eyes (60.00%)], macular diseases in 3 eyes (20.00%), intermediate uveitis in 13 eyes (11.21%) with the main reasons of vitreous opacity [8 eyes (61.54%)], and complicated cataract in 5 eyes (38.46%). Conclusions Uveitis is one of the important causes leading to blindness, especially in the young adults. Panuveitis, especially Behcet's disease and Vogt-Koyanagi-Harada syndrome, are the most common entities responsible for blindness in patients with uveitis. Complicated cataract and secondary glaucoma are the main causes of blindness in uveitis.

Objective To prepare B-lymphoblastoid cell lines of HLA novel allele B*5610 in a family for further study and identification . Method Isolate mononuclear cells under aseptic conditions from the peripheral blood. After infection with Epstein-Barr virus, the cells were cultured in 20% FBS, 2?g/ml CsA RPMI 1640. Results Immortalized B-lymphoblastoid cell lines of five B *5610 carriers in a family were achieved, and the new genes were inherited stably. Conclusion Our work is important for storing and breeding the precious material of biomedicine because the B *5610 genes in the immortalized B-lymphoblastoid cell lines were inherited stably.

BACKGROUND: Umbilical cord blood (UCB) with limited karyocytes is mainly used in child patients. Recently, physicians have tried to mix two units of cord blood in the treatment of adults with hematological system diseases.OBJECTIVE: To monitor quantitatively the dynamic changes and the development rules of engraftment, chimera types and relative amount after allogeneic transplantation of mixed UCB from two units in adults with leukemia.DESIGN: Donors and the recipient were regarded as observational subjects in umbilical cord blood transplantation (UCBT). DNA extracted from blood samples of donors and the recipient before and after transplantation was considered as detecting samples. Short tandem repeat (STR) loci were as observational measures.SETTING: Key Laboratory of Immunology and Genetics of Institute of Transfusion Medicine of Shenzhen Blood Center.PARTICIPANT: A 43-year male patient with acute myeloid leukemia (AML), 75 kg, who was hospitalized at Shenzhen Hospital of Peking University, was enrolled in June 2005. The patient received two units of human leucocyte antigen (HLA), one locus mismatched unrelated UCBT (2.5×107 kg-1 karyocytes in UCB 1, and 1.53×107 kg-1 karyocytes in UCB 2) at month 6 after complete remission from first chemotherapy. UCB was collected from Guangzhou umbilical cord blood bank. The patient signed the informed consent.METHODS: The adult with AML received two units of HLA, one locus mismatched unrelated UCBT (2.5×107 kg-1 karyocytes in UCB 1, and 1.53×107 kg-1 karyocytes in UCB 2). Nine STR loci of the blood sample were determined before and after transplantation by quantitative technique of fluorescence labeling with multiplex polymerase chain reaction (MPCR), while the engraftment and chimera types were qualitatively evaluated by comparing differential loci between the recipient and the donors. The relative amount of two units of UCB was calculated in the patient after transplantation according to the differential gene peak areas of two donors with 377XL DNA sequencer after fluorescence scanning. The engraftment level and the development rules of donors' cells were analyzed quantitatively. In addition, the results were also compared with that of HLA loci distinct analysis for engraftment.MAIN OUTCOME MEASURES: After UCBT, transition process of nine STR loci of the recipient and two donors was observed, and engraftment was quantitatively and qualitatively described.RESULTS: Two units of UCB at day 15 after transplantation were engrafted simultaneously and revealed a complete chimera of the two. The relative amounts of UCB 1 and UCB 2 were 51.3% and 48.7%, respectively. Subsequently, UCB 1 went up to 70.0% and UCB 2 declined to 30.0% at day 30. However, only the genotype of UCB 1 was detected at day 52, and engraftment turned to a complete chimera of a single donor. The one with fewer karyocytes was rejected and the one with more karyocytes was engrafted for a long term.CONCLUSION: To detect quantitatively STR chimera with fluorescence labeling and MPCR can show precisely the engraftment level and the change of two units of UCB. It provides an accurate and reliable experimental basis for clinical UCB application and donor selection. It is proved that adult transplantation at the same time with mixed UCB from two units HLA one locus mismatched unrelated donors is feasible.

BACKGROUND: The special genetic law of short tandem repeat on chromosome X (X-STR) makes it incomparable with autosome markers in forensic identification. However, the population genetics data is far less than that of the autosome STR, and especially the haplotype data are rarely reported.OBJECTIVE: To study the genetic polymorphism of 12 X-STR loci in Shenzhen area by pedigree analysis, aiming to provide scientific and effective data for the application of X-STR in forensic medicine and genetics. METHODS: The blood samples of 118 families were taken to extract DNA by Chelex-100, followed by PCR amplification using Investigator Argus X-12 kit. The frequency of alleles of 231 unrelated individuals was counted by direct counting method and Excel software. Hardy-Weinberg equilibrium test was performed on 12 X-STR loci of female samples by chi-square test. Discrimination power and mean exclusion chance were calculated according to the formula. Pedigree analysis was done to identify haplotypes of female samples and the haplotype frequencies of 4 linkage groups in 111 fathers and 119 mothers were calculated using direct counting method and Excel software.RESULTS AND CONCLUSION: In this study, 349 haplotypes were obtained. There were 238, 139, 153 and 157 haplotypes in linkage groups X1-X4, respectively. The polymorphism of DXS10135 locus was the highest with 21 alleles,while the polymorphism of DXS7423 locus was the worst with only 4 alleles. The combined discrimination power was 0.99999999 in males and 0.99999999 in females. The combined mean exclusion chance was 0.99999999 in trio cases,and 0.99999811 in duo cases. These findings indicate that the X-12 detection system has high polymorphism in Shenzhen Han population, and has important application value in forensic individual identification and paternity testing.

Objective To evaluate the heterozygous ambiguity resolution primers (HARPs) method in resolving ambiguous genotyping results of human leukocyte antigen (HLA) genes in Chinese Hart population, and choose some appropriate HARPs primers. Methods HLA-A, HLA-B and HLA-DRB1 genes of 416 southern Chinese Han individuals were genotyped by sequence-based-typing(SBT) method and then the ambiguous genotyping samples were sequenced again by HARPs primers provided by American Atria company. Results The percentage of ambiguous genotyping samples resolved by HARPs for HLA-A, HLA-B and HLA-DRBI locus was 86.3% (132/153), 73.9% (130/176) and 38.1% (85/223) respectively. Among them, 48.5% (64/132)HLA-A, 80.0% (104/130)HLA-B and all HLA-DRB1(85/85)samples only need one primer, 47.7 % (63/132)HLA-A and 20.0% (26/130)HLA-B samples need two primers. Three to six different HARPs primers can resolve more than 90% ambiguities. Conclusion HARPs is a convenient method and could be a routine method to resolve ambiguities for HLA-A, HLA-B and HLA-DRB1 genes genotyped by SBT in Chinese Han population.

BACKGROUND: The judgment of the engraftment of hematopoietic stem cells after transplantation mainly depends on various genetic labeling in vivo, which are different in sensitivity and effectiveness, thus a method with powerful differential ability, high sensitivity and not restricted by sex is to be established.OBJECTIVE: To observe the DNA genetic loci of short tandem repeat in the blood samples of both donors and recipients before allo-hematopoietic stem cell transplantation and those of recipients at different time points after transplantation.DESIGN: An observation measurement.SETTING: Laboratory of Immunogenetics, Shenzhen Institute of Transfusion Medicine, Shenzhen Blood Center.PARTICIPANTS: Blood samples of 18 pairs of donors and recipients, who were successfully matched and accepted hematopoietic stem cell transplantation, were selected from the Laboratory of Immunogenetics, Shenzhen Institute of Transfusion Medicine, Shenzhen Blood Center from February 2004 to December 2005. Among the 18 patients, there were 10 males and 8 females, with a mean age of 35 years old, including 6 cases of them were donated by relatives with blood relationship, and 12 cases by volunteers without blood relationship. Informed consents were obtained from all the participants.METHODS: The blood samples of both donors and recipients before transplantation and the blood samples of recipients after transplantation were collected, and the fluorescence labeling short tandem repeat technique was used to detect the 15 loci for short tandem repeat and Amelogenin sex locus, so that the differential loci between the donor and recipient could be screened. The engraftment and dynamic changes of the short tandem repeat genes of the donors in the recipients after transplantation were observed, the times for the earliest occurrences of short tandem repeat genes of the donors and the complete chimerism were recorded.MAIN OUTCOME MEASURES: ① Differential genes between the donors and recipients before transplantation;②Times for the earliest occurrences of short tandem repeat genes of the donors and the complete chimerism.RESULTS: All the 18 pairs of donors and recipients were involved in the final analysis of results. Satisfactory results of the typing at the 15 loci for short tandem repeat and 1 sex locus in the 18 pairs of samples of both donors and recipients before transplantation and the sample of the recipients after transplantation respectively. Averagely 12.4 (8-15) differential loci for short tandem repeat could be distinguished between the donors and recipients. ②After transplantation, short tandem repeat genes could be detected the earliest at 8 (5-14) days averagely, It took 14 (9-23) days averagely for short tandem repeat loci to convert from recipient type completely into donor type, and the engraftment converted from the recipient chimerism types completely into the donor types.CONCLUSION: The fluorescence labeling compound amplification of short tandem repeat technique can precisely measure the number of PCR products, describe the engraftment of hematopoietic stem cells and the whole process of development. It can also provide accurate and timely information for the early judgement of engraftment, predicting failure of transplantation and controlling recurrence.

ObjectiveTo explore the clinical effects of cold therapy in treatment of inflammatory hemorrhoids.MethodsSixty patients were separated into two groups according to the visiting sequence with 30 cases each.The treatment group accepted cold therapy,meanwhile the control group took the tablets diosmin and accepted hot compress with 50％ magnesium sulfate.ResultsThe symptoms evaluation score was ( 10.5 ± 1.3 ) scores and (6.4 ± 1.2) scores before and after treatment in treatment group,while ( 10.3 ±1.4) scores and(9.4 ± 1.3) scores in control group,there was no significant difference before treatment between two groups (P＞ 0.05 ),but there was significant difference after treatment between two groups (P＜0.05).After 3 days treatment,cure rate was 60.0％(18/30),eifficient rate was 90.0％(27/30) in treatment group,while 10.0％ (3/30),53.3％ ( 16/30 ) in control group,there were significant differences between two groups(P＜ 0.05).ConclusionCold therapy is good for inflammatory hemorrhoids.

ObjectiveTo observe the clinical characteristics and influential factors of pulmonary hypertension (PH) in maintenance hemodialysis (MHD) patients.MethodsOne hundred and two patients with MHD patients whose therapy time were all more than one year were enrolled from June 2008 to May 2009.Pulmonary arterial systolic pressure (PASP) and parameters related with cardiac structure and function were checked out by echocardiography.These patients were divided into two groups according to PASP:PH group[PASP≥35 mm Hg( 1 mm Hg =0.133 kPa)] and non-PH group(PASP ＜ 35 mm Hg).Gender,age,dialysis time,dialysis access and urine volume during dialysis was recorded.The blood biochemistry,parathyroid hormone(PTH),hemoglobin,postdialysis urea nitrogen was recorded too and Kt/V was calculated.ResultsPH was detocted in 26 MHD patients (25.5％,26/102).Right atrium transverse diameter,right atrium long diameter and pulmonary artery internal diameter in PH group [(32.00 ± 8.13),(41.00 ±9.15),(24.38 ± 3.88) mm] were higher than those in non-PH group [(27.44 ± 4.43),(36.45 ±8.88 ),(21.78 ± 2.46) mm] and LVEF in PH group [(55.59 ± 7.88 )％] was lower than that in non-PH group [(59.09 ± 5.69)％] (P ＜ 0.01 or ＜ 0.05).There were significant differences in calcium-phosphorus product,PTH,serum phosphorus and hemoglobin between PH group and non-PH group [(61.36 ± 17.83)mg2/dl2 vs.(53.24 ± 18.28) mg2/dl2,(365.95 ± 178.65) ng/L vs.(287.02 ±209.91 ) ng/L,(2.24 ±0.60)mmol/L vs.( 1.95 ± 0.68) mmol/L,(98.65 ± 25.69) g/L vs.( 114.29 ± 22.31 ) g/L] (P ＜ 0.05 ).Logistic regression found that PTH levels,hemoglobin and calcium-phosphorus product were the major risk factors for PH.ConclusionsMorbidity of PH in MHD patients is obviously high.Secondary hyperparathyroidism,anemia and high calcium-phosphorus product level may take part in the pathogenesis of PH in MHD patients.