To study the correct method for determining ABO blood types in infants and its influencing factors, blood types of 33 infants under 6 months old were determined by routine serological method, micro-column gel typing system and PCR-SSP genotyping method. Of the 33 cases with discrepant results of ABO blood type by different methods, the blood types of 32 cases were discrepant between red cell and serological typings in the routine serological method, and a false coincidence in 1 case was caused by bacterial infection resulting in B-like antigen. Correct blood typing was obtained in 27 cases with a correct rate of 84.4% (27/32) by using micro-column gel typing system. PCR-SSP method gave correct results in all of 33 cases. There was a significant difference between the results of micro-column gel typing system and PCR-SSP. It is concluded that to determine ABO blood type for infants < 6 months old, it is recommended to adopt micro-column gel typing system method, and what must be taken into account is the possible false coincidence caused by bacterial infection resulting in B-like antigen. In micro-column gel typing system, if the results of red cell and serological typing are identical, the principle is that blood transfusion must be performed with same ABO blood type between recipient and donor. If not, washed O red blood cells should be used for infants, and then change to transfusion with identical blood group according to PCR-SSP typing results.
ABO Blood-Group System ; genetics ; Blood Grouping and Crossmatching ; methods ; Blood Transfusion ; DNA ; genetics ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Polymerase Chain Reaction ; methods ; Reproducibility of Results

OBJECTIVETo evaluate the predictive value of serum inhibin B (INH B) levels as an indicator of the presence of testicular spermatozoa in nonobstructive azoospermia.

METHODSForty patients with nonobstructive azoospermia (NOA), 20 patients with obstructive azoospermia (OA), and 10 fertile volunteers were involved in this study. A chemoluminescence method was used to measure the levels of FSH; Inhibin B was analysed by using sandwich enzyme-linked immuno-sorbent assay.

RESULTSPatients with nonobstructive azoospermia has significantly higher levels of serum FSH [(21.34 +/- 12.15) IU/L] and significantly lower levels of inhibin B [(53.15 +/- 58.74) ng/L] than patients with obstructive azoospermia [FSH: (3.94 +/- 1.52) IU/L, INH B: (162.49 +/- 78.38) ng/L, P < 0.01] and fertile volunteers [FSH: (4.27 +/- 2.84) IU/L, INH B: (228.49 +/- 110.68) ng/L, P < 0.01]. Mean serum inhibin B were significantly higher in patients with nonobstructive azoospermia who had spermatozoa on TESE than in those in whom no spermatozoa was found on TESE [INHB: (90.31 +/- 72.18) ng/L vs (19.54 +/- 20.38) ng/L, r = 0.528, P < 0.01], but mean FSH levels did not have similar predictive power (P > 0.05).

CONCLUSIONSerum INH B level seems to be more accurate than serum FSH in the prediction of presence of testicular spermatozoa in patients with nonobstructive azoospermia. Serum inhibin B determination may be substitute of TESE as a diagnostic index.

Adult ; Azoospermia ; blood ; diagnosis ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Follicle Stimulating Hormone ; blood ; Humans ; Inhibins ; blood ; Luminescent Measurements ; Male ; Predictive Value of Tests ; Spermatozoa ; physiology ; Testis ; physiology ; surgery

OBJECTIVETo study the difference and similarity between Hans and Uighurs in regard to Rhesus box and its significance.

METHODSThe 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.

RESULTSThe 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).

CONCLUSIONThe 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.

China ; Genetics, Population ; Genotype ; Humans ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System ; genetics

OBJECTIVETo 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.

METHODSRHCE 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.

RESULTSThe 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.

CONCLUSIONThe 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.

Ethnic Groups ; genetics ; Genotype ; Humans ; Phenotype ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; blood ; genetics ; Serologic Tests ; methods

OBJECTIVETo evaluate the underlying mechanism of Jianpi Jiedu Recipe (, JJR) in the reversion of multidrug resistance concerning colorectal cancer in vitro and in vivo.

METHODSMice were treated orally with JJR at a daily 4.25 g/(kg·day) or injected with vinblastine (VCR) 2.5 mg/(kg·day) for 3 weeks after having been inoculated with HCT8/V cells; tumor tissues were assayed by hematoxylin and eosin staining. Firstly, the effects of JJR on the expression of cyclooxygenase-2 (COX-2) were tested by real-time polymerase chain reaction (PCR) technique and COX-2 gene silenced by siRNA. Secondly, the variation of intracellular concentration of oxaliplatin (L-OHP) was evaluated by the inductively coupled plasma mass spectroscopy (ICPMS) in HCT8/V and its COX-2 siRNA cells; the concentration of JJR combined with chemotherapeutic drugs and the reverse effect of multidrug resistance (MDR) in HCT8/V cells was evaluated by the MTT assay. Thirdly, real-time quantitative PCR and Western blot analysis were used to detect the multidrug resistance gene 1 (MDR1) mRNA and P-gp expression.

RESULTSJJR had an inhibitory effect on the growth of tumors in vivo, and it, in combination with chemotherapeutic drugs, could reverse the drug-resistance of HCT8/V cells and increase the sensitivity of HCT8/V cells to VCR, DDP, 5-Fu, and THP. ICP-MS results showed that JJR could increase the concentration of drugs in HCT8/V cells (P<0.01). Furthermore, it was shown that JJR could reverse drug resistance of colorectal cancer cells by decreasing MDR1 expression and P-gp level via downregulation of COX-2, which has been represented as one of the major mechanisms that contributes to the MDR phenotype (P<0.01).

CONCLUSIONJJR reversed multidrug resistance and enhanced the sensitivity to chemotherapy, which could be attributed to the down-regulation of COX-2 in MDR1/P-gp-mediated MDR colorectal cancer after chemotherapy.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Animals ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; drug therapy ; enzymology ; pathology ; Cyclooxygenase 2 ; genetics ; metabolism ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Green Fluorescent Proteins ; metabolism ; Humans ; Intracellular Space ; metabolism ; Mice, Inbred BALB C ; Organoplatinum Compounds ; metabolism ; RNA, Small Interfering ; metabolism ; Signal Transduction ; drug effects ; Vinblastine ; pharmacology ; therapeutic use ; Xenograft Model Antitumor Assays

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.
Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Humans ; Introns ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; genetics

The objective of this study was to investigate the method and effect of blocking the specific reaction between lymphocyte HLA-I antigen and its antibody. The lymphocytes were disposed with 12 mmol/L methoxypolyethelene glycol benzotriazol carbonate (mPEG-BTC) in concentration gradient in PBS (pH 7.4) at 22 degrees C. The effect of the modified lymphocytes was detected by microlymphocytotoxicity assay. The results showed that lymphocytes modified by mPEG-BTC did not react with related HLA-I antibodies in microcytotoxicity test. It is suggested that the specific reaction between HLA-I antigen of lymphocyte and HLA-I antibodies can be completely camouflaged by mPEG-BTC in PBS (pH 7.4) under 22 degrees C room temperature.
Antigen-Antibody Reactions ; Cytotoxicity, Immunologic ; Histocompatibility Antigens Class I ; immunology ; Humans ; Lymphocytes ; immunology ; Polyethylene Glycols ; pharmacology ; Triazoles ; pharmacology

OBJECTIVETo detect micrometastasis in regional lymph nodes using RT- PCR assay and evaluate the significance of the new assessment of nodal status in determining pN staging in gastric carcinoma.

METHODSIn addition to HE,RT- PCR assay for cytokeratin-20 gene marker was used to detect micrometastasis in 850 lymph nodes from 30 patients with gastric carcinoma who underwent gastrectomy with lymphadenectomy from Dec. 2003 to Apr. 2004.

RESULTSThe detection rate of HE staining was 27.1% (233/850), the detection rate of RT-PCR assay was 36.5% (310/850) (Pa< 0.01). Lymph node micrometastasis was further detected in 77 nodes from 14 patients. The detection rate of RT- PCR assay from the routine negative lymph nodes was 12.5% (77/617). Seven of those patients were up- staged (from IB stage to II stage, from IB stage to III A stage, from I stage to III A stage, from III A stage to III B stage, from III A stage to IV stage in one patient respectively, and from III B stage to IV stage in 2 patients).

CONCLUSIONRT- PCR assay can increase the detection rate of lymph node metastasis and have a significant impact on the staging system of gastric carcinoma.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; pathology

BACKGROUNDMutations in the cationic trypsinogen gene (PRSS1) have been detected in patients with hereditary pancreatitis (HP). This study investigated the prevalence of the R122H (c.365 G > A), A121T (c.361 G > A) and D162D (c.488 C > T) mutations or polymorphisms in the common, non-hereditary forms of chronic pancreatitis and in an HP family.

METHODSDNA was prepared from blood samples of 54 patients with chronic pancreatitis (35 alcoholic, 17 idiopathic and 2 hereditary) and 120 normal controls. The PRSS1 genes were amplified by polymerase chain reaction (PCR) and their products were analyzed by sequencing and related clinical data were also collected.

RESULTSA new polymorphism (c.488 C > T) of PRSS1 was found in 25 patients with chronic pancreatitis (including one affected member of the HP family) and six members of the normal controls. The C/T genotype was significantly increased in chronic pancreatitis (OR: 16.379, 95% CI: 5.7522 - 52.3663), the frequency of c.488 C > T change was in according with the Hardy-Weinberg equilibrium, but it doesn't affect the clinical phenotype. The commonly reported change of R122H (c.365 G > A) was not detected in any of the study subjects. c.361 G > A was found in 2 affected members and one unaffected carrier in an HP family. One of the affected members of an HP family had c.361 G > A mutation and polymorphism (c.488 C > T) in the PRSS1 gene at the same time. The patient's clinical values (C3, C4, CA19-9 and HbA1c) were higher than those of the other patients with chronic pancreatitis. The two patients with HP developed diabetes mellitus and their father died with pancreatic cancer.

CONCLUSIONA new polymorphism (c.488 C > T) in the PRSS1 gene is associated with chronic pancreatitis, but it did not affect the clinical phenotype while the A121T (c.361 G > A) mutation in the gene shows a significant correlation in the patients with HP.

Female ; Humans ; Male ; Mutation ; Pancreatitis ; genetics ; Pancreatitis, Chronic ; genetics ; Polymorphism, Genetic ; Trypsin ; Trypsinogen ; genetics

OBJECTIVETo research comparatively on the RHD gene structures in unrelated RhD negative individuals of Chinese Uigur and Han population.

METHODSThe upstream, downstream, hybrid box and 10 exons of RHD gene were detected with sequence specific primer-PCR technique.

RESULTSThe 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.

CONCLUSIONThe 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.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Ethnic Groups ; ethnology ; genetics ; Exons ; genetics ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Population Groups ; Rh-Hr Blood-Group System ; genetics