Experimental observation of effects of acupoints, cone numbers and durations of moxibustion with different moxibustion methods on skin surface and inside temperature / 针灸推拿医学（英文版）

Objective: To observe the effects of acupoints, cone numbers and durations of moxibustion with different moxibustion methods on skin surface and inside temperature, and to provide references for the clinical standardization of moxibustion amount. Methods: The 42 big-ear white rabbits were divided into 6 groups according to the random number table method, a 1-cone direct moxibustion group, a 2-cone direct moxibustion group, a 3-cone direct moxibustion group, a 1-cone herbal cake-partitioned moxibustion group, a 2-cone herbal cake-partitioned moxibustion group, and a 3-cone herbal cake-partitioned moxibustion group, with 7 rabbits in each group. Shenque (CV 8), Shenshu (BL 23) and Zusanli (ST 36) were used in each group, but the moxibustion methods, cone numbers and durations of moxibustion were different. Rabbits in each group received moxibustion once every other day for 5 times in total. During the intervention, a thermoelectricity coupled probe and a temperature recorder were used to record the real-time acupoint skin temperature and the temperature at different time points, so as to observe, analyze and process the real-time changes in the temperature difference between the surface and inside of acupoint skin. Results: For herbal cake-partitioned moxibustion, the best temperature for cone changing was (46.38±0.51) ℃ when the highest surface temperature was (49.20±0.52) ℃; the multi-factor comparison of acupoint × cone number × time and acupoint × moxibustion method × time showed that time × acupoint, time × moxibustion method and cone number × acupoint had interactive effects (all P<0.05). Comparing skin temperature differences between different cone numbers at the same acupoint, Shenque (CV 8) on the 1st and the 5th days, Shenshu (BL 23) on the 3rd and the 7th days, Zusanli (ST 36) on the 1st and the 9th days of experiment showed statistically significant differences (all P<0.05). The skin temperature comparison of different moxibustion methods at the same acupoint all had statistical differences (all P<0.05), except for Shenque (CV 8) before moxibustion, Shenshu (BL 23) before moxibustion and on the 5th day; Zusanli (ST 36) only showed statistical differences on the 5th and 7th days (both P<0.05). The skin temperature differences of different acupoints after moxibustion in the 1-cone, 2-cone and 3-cone groups were statistically different (all P<0.05); direct moxibustion and herbal cake-partitioned moxibustion at different acupoints were all statistically different (all P<0.05). Conclusion: Cone changing temperature under the same specifications of herbal cake-partitioned moxibustion was confirmed. Temperature difference between surface and inside of different acupoint skin at the same maximum temperature was significantly different due to the cone numbers and moxibustion methods, which showed the highest at Shenshu (BL 23), the second at Shenque (CV 8), and the lowest at Zusanli (ST 36). The influence of acupoint factor should be considered to determine the quantitative indicators of moxibustion.

Bone marrow mesenchymal stem cells/biphasic calcium phosphate ceramics for cartilage repair in juvenile rats / 中国组织工程研究

BACKGROUND: The pathogenesis of pediatric femoral head necrosis is associated with cartilage injury of the hip joint induced by stress and inflammation. OBJECTIVE: To observe the effect of bone marrow mesenchymal stem cells (BMSCs)/biphasic calcium phosphate ceramics (B-CPC) on cartilage repair in juvenile rats. METHODS: Thirty male Sprague-Dawley rats, aged 1 week, were randomized into three groups. No intervention was done in blank group. A juvenile rat model of articular cartilage injury was made using improved Hulth's method in control and observational groups, followed by implantation of BMSCs/hydroxyapatite and BMSCs/B-CPC,respectively. Four weeks later, the rat articular cartilage was observed pathologically, and MTT and flow cytometry were employed to detect chondrocyte proliferation and apoptosis, respectively. RESULTS AND CONCLUSION: The articular cartilage of the rats in the blank group was smooth and complete. In the control group, articular cartilage damage was obvious, presenting with rupture, defect and irregularity of the articular cartilage surface, as well as unclear four-layer structure of the cartilage. In the observational group, articular cartilage injury was repaired to some extent. At the same observation time, the cell viability was significantly increased in the observational group compared with the control group (all P < 0.05), and the proportion of apoptotic cells was significantly decreased (all P < 0.05). To conclude, BMSCs/B-CPC composite can promote the cartilage repair in juvenile rats.

Bone marrow mesenchymal stem cells/biphasic calcium phosphate ceramics for cartilage repair in juvenile rats / 中国组织工程研究

BACKGROUND: The pathogenesis of pediatric femoral head necrosis is associated with cartilage injury of the hip joint induced by stress and inflammation. OBJECTIVE: To observe the effect of bone marrow mesenchymal stem cells (BMSCs)/biphasic calcium phosphate ceramics (B-CPC) on cartilage repair in juvenile rats. METHODS: Thirty male Sprague-Dawley rats, aged 1 week, were randomized into three groups. No intervention was done in blank group. A juvenile rat model of articular cartilage injury was made using improved Hulth's method in control and observational groups, followed by implantation of BMSCs/hydroxyapatite and BMSCs/B-CPC,respectively. Four weeks later, the rat articular cartilage was observed pathologically, and MTT and flow cytometry were employed to detect chondrocyte proliferation and apoptosis, respectively. RESULTS AND CONCLUSION: The articular cartilage of the rats in the blank group was smooth and complete. In the control group, articular cartilage damage was obvious, presenting with rupture, defect and irregularity of the articular cartilage surface, as well as unclear four-layer structure of the cartilage. In the observational group, articular cartilage injury was repaired to some extent. At the same observation time, the cell viability was significantly increased in the observational group compared with the control group (all P < 0.05), and the proportion of apoptotic cells was significantly decreased (all P < 0.05). To conclude, BMSCs/B-CPC composite can promote the cartilage repair in juvenile rats.

Internal and external fixation materials for the repair of pelvic fracture:reasonable choice and biocompatibility / 中国组织工程研究

BACKGROUND:Pelvic fractures are mostly caused by high energy trauma. With the development of imaging techniques and in-depth study of the anatomical structure of the pelvis and biomechanics, internal fixation and external fixation materials are gradual y being used in the repair of pelvic fracture. OBJECTIVE:To summarize features and applications of external fixation stent material, percutaneous screw fixation, percutaneous sacral iliac screw material for internal fixation and intramedul ary tensile screw material for internal fixation after pelvic fracture. METHODS:We retrieved Wanfang Database and PubMed for studies on the application of internal fixation material and external fixation material in pelvic fracture from 1994 to 2015. Al data were analyzed and summarized. RESULTS AND CONCLUSION:Application of pelvic external fixation materials contributed to the stability of early pelvic fractures, showed smal injury, could increase the reliability of fixation. However, the biomechanical stability of external fixation materials was lower than other internal fixation, could only be used for the early temporary fixation of unstable pelvic fractures in particular cases. Internal fixation materials can achieve anatomical reduction, accorded with the requirements of the physical mechanics of the pelvis, improve the stability of the pelvis, and have become the first choice for repair of unstable pelvic fractures. Currently used methods are percutaneous hol ow screw fixation, percutaneous fixation of the sacral iliac screw, and intramedul ary lag screw fixation. The combination of external fixation and internal fixation can effectively restore the stability of the pelvic cavity. Therefore, we should consider the location, type and stability of the fracture to select the appropriate internal fixation and external fixation materials.

Analysis of Human Platelet Antigen-1 System Alloantibodies Using Recombinant GPIIIa Fragments Coupled to Luminex Beads / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To detect platelet anti-HPA-1a and -1b antibodies using recombinant GPIIIa fragments coupled to Luminex beads.</p><p><b>METHODS</b>The sensitivity of 2 techniques, monoclonal antibody specific immobilization of platelet antigen (MAIPA) and Luminex bead assay, was compared using 12 twofold-serial dilutions (from neat to 1 in 2048) of an anti-HPA-1a WHO international standard. The specificity of Luminex assay to identify anti-HPA-1a and -1b antibodies was assessed using 8 negative or positive controls and 36 blinded samples provided by WHO Platelet Workshop.</p><p><b>RESULTS</b>The sensitivity of MAIPA and Luminex bead assay to detect anti-HPA-1a was dilution 1/64 (i.e. 1.56 IU/ml) and far more than dilution 1/2048 (i.e. 0.049 IU/mL), respectively. The Luminex bead assay could specifically identify negative and positive controls of anti-HPA-1a and -1b. All results of 36 blinded samples by Luminex assay were accordant to reference results except one sample which contained high concentration antithetical antibody and resulted in false positive of anti-HPA-1b. Cross-reactivity was also not observed with the samples containing HLA, ABO or other platelet antibodies.</p><p><b>CONCLUSION</b>The Luminex beads coupled with recombinant GPIIIa fragments can be used to detect HPA-1 system antibodies with sufficient sensitivity and specificity, that is suitable for the detection of platelet alloantibodies in clinical alloimmune thrombocytopenia.</p>

Acute effects of anaerobic exercises by a bicycle ergometer on elastic modulus and local hemodynamics in common carotid arteries / 医用生物力学

Objective To investigate acute effects of anaerobic exercises by a bicycle ergometer on arterial elastic modulus and local hemodynamics in human common carotid arteries with different genders. Methods Nine male and eight female healthy young volunteers at the age of 20-30 year-old successively underwent four groups of exercise trainings with the same workload by an anaerobic bicycle ergometer. The waveforms of arterial diameter and center-line blood velocity were measured in the right common carotid artery using a color Ultrasonic Doppler for each group when at rest and right after exercise training. The heart rate, systolic and diastolic blood pressures were simultaneously measured in brachial artery using an automatic electronic sphygmomanometer. All the measured data were analyzed based upon the principle of classic hemodynamics. The arterial elastic modulus and local hemodynamic parameters, including pressure-strain elastic modulus, flow rate, circumferential strain, wall shear stress and oscillatory shear index (OSI), were then calculated. Results The heart rate and arterial elastic modulus increased after exercises; with the accumulative exercises, in one cardiac cycle, the maximum and mean center-line velocity and flow rate increased while the minimum velocity and flow rate decreased; the systolic and mean blood pressure increased while diastolic blood pressure exhibited no significant change; no significant change could be found in the circumferential strain; the maximum and mean shear stress increased significantly while the minimum shear stress reduced; the oscillatory shear index also increased. Conclusions The anaerobic exercises by a bicycle ergometer may increase the arterial elastic modulus and induce significant effects on local hemodynamic parameters in common carotid arteries for young volunteers with different genders at the age of 20 30 year old. The results in this study could provide useful hemodynamic information for regulation of cerebrovascular function by anaerobic exercises.

Establishment of method detecting CD36 expression on human platelet and its application / 中国实验血液学杂志

The individual with the deficiency of CD36 antigen on platelet displayed the risk of anti-CD36 immune reaction induced by transfusion, which is one of the reasons for platelet transfusion refractoriness (PTR). This study was purposed to detect the expression level of CD36 antigen on platelet by flow cytometry among apheresis platelet donors of Hangzhou area, and the frequency of CD36 deficiency was analyzed. Platelet-rich plasma (PRP) was separated from fresh anticoagulant whole blood by centrifugation, then the platelets were washed and adjusted to 1×10(6). The platelets were incubated with FITC-labeled CD36 and PE-labeled CD41 monoclonal antibodies, then the expression level of CD36 was detected by flow cytometry. The CD36 expression on monocytes for the samples of CD36-deficiency on the platelets was further analyzed. The results showed that 7 samples with CD36 antigen deficiency were found in 192 apheresis platelet donors. The frequency of CD36 deficiency was 3.6% and all of them were typeII deficiency. The significant difference of CD36 antigen expression was observed in the platelet donors of Hangzhou population, among them 59 individuals with low expressed CD36 antigen and 126 individuals with highly expressed CD36 antigen were found according to the geometric mean fluorescence intensity. It is concluded that the CD36 antigen deficient phenotype existed in the population, these data will provide the information for research of the CD36 antigen distribution and help to solve the platelet transfusion refractoriness.

Surgical treatment of isolated fractures of the sustentaculum tali of calcaneus / 中华骨科杂志

Objective To evaluate the clinical results of surgical treatment of isolated fractures of the sustentaculum tali of calcaneus via medial approach.Methods The data of 23 patients with isolated fractures of the sustentaculum tali of calcaneus was retrospectively analyzed who were treated with open reduction and internal fixation with cannulated screw or Kirschner wire via medial approach from September 2006 to March 2011.There were 19 males and 4 females,with an average age of 26.3 years (range,17-41 years).Associated injuries included 4 cases of talus fracture,4 of metatarsal fracture,and 3 of cuboid fracture.The functions of hiudfoot were assessed by American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot scores pre-operation and post-operation respectively.Results Fifteen patients got followed up with an average 20.5 months (10-56 months).Thirteen patients were rated as good,2 as excellent,and the excellent and good rate was 100％(15/15).All the fractures were stabilized reliably,and got clinical union with no obvious complications occurred.Time of fracture union was 8-10 weeks,with an average of 8.5 weeks.Three patients felt mild transient pain during the recovery of walking,but their pain disappeared quickly after physical therapy.No patients developed wound infection,nonunion and other complications.Conclusion For isolated fractures of the sustentaculum tali of caleaneus with articular surface displaced greater than 1 mm or involving the articular surface of middle subtalar joint,open reduction and internal fixation operation via medial approach under direct visualization is recommendable.

Rare blood group screening by serological and molecular methods in Zhejiang Han population / 中国实验血液学杂志

This study was aimed to investigate the distribution of rare blood group in Zhejiang Han population. The H(-) (H system), GPA(-) and s(-) (MNS), Rhnull, Rhmod, D--, CCDEE, CCdEE (variations of Rh), GPC(-) (Gerbich), i(+) (I), Lu(b-) (Lutheran), Js(b-) and k(-) (Kell), Fy(a-) (Duffy), Ok(a-) (Ok), Di(b-) (Diego) phenotypes were screened by serological or molecular methods. Jk (a-b-) phenotype was detected by urea hemolytic test. The results showed that one Di (a+b-) individual was found in 1618 blood donors, three Fy (a-b+) individuals in 1007 donors and one CCdEE individual in 633 Rh negative donors. No Jk (a-b-), H(-), GPA(-), s(-), GPC(-), i(+) (adult), Lu(b-), k(-), Js(b-), Lu(b-) and Ok(a-) phenotypes were found in this large scale survey. It is concluded that Di (a+b-), Fy (a-b+), CCdEE phenotypes are confirmed in the blood donors and this study provides the distribution data of erythrocyte rare blood group in Zhejiang Han population.

Progress on the quantitative modulation of intracellular calcium signals induced by shear flow / 医用生物力学

The intracellular calcium signaling, which is modulated by the microenvironment of cells, is closely related to the cell's self renewal, differentiation, proliferation, and its apoptosis. The study on the quantitative modulation of the intracellular calcium signals could not only help to understand the dynamic behavior of such kind of signaling, but also play a significant role in the control of cell fate, simulation of cell behavior and bionics of cellular biological systems. This paper briefly reviewed the progress on the quantitative modulation of intracellular calcium signals induced by shear flow, including (1) experimental phenomena and the associated mechanisms of shear flow activated intracellular calcium response; (2) mathematical modeling and simulation of the intracellular calcium response induced by shear flow; (3) feedback control of the intracellular calcium signals by shear flow.

C742T mutation of α1, 3 N-acetyl-D-galactosaminyltransferase gene is responsible for A2 subgroup / 中国实验血液学杂志

The objective of this study was to analyze the molecular genetic basis for 2 individuals with A2B phenotype of ABO subtype. The ABO group antigens on red blood cells were identified by monoclonal antibodies and the ABO antibodies in serum were detected by the standard A, B, O cells. The exon 5 to exon 7 coding region of ABO gene was amplified by polymerase chain reaction (PCR) and the PCR product was sequenced directly after the enzymes digested. The amplified product was also cloned by TOPO TA cloning sequencing kit to split the 2 alleles apart and chosen colonies were sequencing bidirectionally for exon 6 to 7 of ABO gene. The results showed that both A and B antigen were identified on red blood cells of the individuals and there was anti-A1 antibody in their serum. There was no 261G deletion and showed 297A/G, 467C/T, 526C/G, 657C/T, 703G/A, 742C/T, 796C/A, 803G/C, 930G/A heterozygotes by direct DNA sequencing. After cloning and sequencing, it was obtained the B101 and one novel A allele. The novel allele has one nucleotide change at 742 position C to T compared with A102, which results in an amino acid Arg to Cys at 248 position and was nominated as A213. It is concluded that C742T mutation of the α1, 3 N-acetyl-D-galactosaminyl-transferase gene can lead to A2 phenotype and with anti-A1 antibody in serum.

Identification of an ABx09 phenotype of ABO subtype / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the molecular basis for an individual with ABx09 phenotype of ABO subtype.</p><p><b>METHODS</b>The ABO group antigens on red blood cells of the proband were identified by monoclonal antibodies, and the ABO antibody in serum was detected by standard A, B, O cells. The exons 1 to 7 of ABO gene were amplified by polymerase chain reaction (PCR) respectively and the PCR products were sequenced directly. The amplified products for exons 5 to 7 were also cloned by TOPO TA cloning sequencing kit to split the two alleles apart, selected colonies were sequenced bidirectionally for exons 5 to 7 of the ABO gene. The samples of the proband's parents were collected, then serological test of the blood group and sequence analysis for exons 6 and 7 of ABO gene were preformed.</p><p><b>RESULTS</b>Both A and B antigens were detected on red blood cells of the proband and there was anti-B antibody in the serum. There was no G deletion at position 261, while 297AG in exon 6, 467CT, 526CG, 657CT, 703GA, 796CA, 803GC, 889GA and 930GA heterozygote in exon 7 were detected by direct DNA sequencing, which can be assigned for A102Bx09 genotype. After cloning and sequencing, two alleles A102 and Bx09 were obtained. The sequence of Bx09 had one nucleotide changes (G to A) at position 889 compared with that of B101, which resulted in an amino acid change of Glu to Lys at 297 position. The Bx09 in the proband was inherited from her mother by family investigation.</p><p><b>CONCLUSION</b>G to A at nt889 of alpha-1,3 galactosyltransferasegene can result in Bx09 phenotype, with the presence of anti-B antibody in serum.</p>

Cloning and sequence analysis of 5'untranslated region of human ABO gene / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To clone and investigate the polymorphism of the 5'-untranslated region (5'-UTR) of the human ABO gene, in order to provide the basis for exploring the transcriptional regulation of the human ABO histo-blood group genes.</p><p><b>METHODS</b>ABO phenotypes of 30 unrelated healthy blood donors were determined by serological technique, their genotypes were analyzed by sequencing the exons 6 and 7 of the ABO gene. Nearly 5 kb of the 5'-UTR of ABO gene was obtained by PCR amplification and sequencing was performed bidirectionally. Haplotypes of samples with heterozygous sites in the 5'-UTR of ABO gene were separated and analyzed after cloning.</p><p><b>RESULTS</b>Twenty polymorphic sites were identified in these samples where ABO genotypes were consistent with serological phenotypes. It included sixteen nucleotide sequence variations, one 8 bp deletion, one 6 bp deletion/insertion, one 36 bp insertion and one 43 bp repeats. Among them, 11 were novel polymorphic sites. Seven different haplotypes of 5'-UTR of ABO gene were defined to the cis/trans linkage of those mutations.</p><p><b>CONCLUSION</b>There were polymorphisms in the 5'-UTR of ABO gene and the nucleotide sequences near the proximal promoter were conservative.</p>

In vitro expression activity of α-(1,2) fucosyltransferase with 35C > T and 682A > G mutations / 中国实验血液学杂志

In order to explore the effects of 35C > T and 682A > G mutations on the activity of alpha-(1,2) fucosyltransferase, the coding region of fut1 gene was amplified by polymerase chain reaction (PCR) from genomic DNA. PCR product was ligated into expression vector using TOPO TA cloning kit to obtain the recombinant plasmids. The recombinant plasmids were transfected into COS-7 cells by liposome method. After screening by using G418, H antigen expression on the COS-7 was tested by flow cytometry and fut1 mRNA was detected by real-time PCR. The results indicated that three kinds of recombinant plasmids pcDNA3.1/V5-His-wild (35C + 682A), pcDNA3.1/V5-His-35T and pcDNA3.1/V5-His-35T-682G were successfully constructed. After transfection, the H antigen expressed on membrane of COS-7 cells at the second day, with the maximum level of expression at the fourth day. When compared with pcDNA3.1/V5-His-wild transfected cells, the H antigen expression level of the 35T and 682G + 35T recombinant plasmids in the transfected cells was 52.7% and 13.3% on the fourth day, respectively. Although the level of fut1 mRNA decreased with prolonging of time, the mRNA expressed on the pcDNA3.1/V5-His-35T-682G transfected cells reached to 14% of the wild plasmids on the first day. It is concluded that 682A > G mutation obviously reduces the activity of alpha-(1,2) fucosyltransferase, while 35C > T mutation leads to partial reduction of H antigen in vitro expression.

Identification of the hemolytic transfusion reaction caused by lewis antibody using serological and molecular biological methods / 中国实验血液学杂志

To analyse the reason for one case of hemolytic transfusion reaction, antibodies in a patient's serum were identified using panel cells and Le (a-b-) phenotype cells, patient phenotype was identified by using anti-Le(a) and anti-Le(b) blood grouping reagents and the entire coding region of FUT3 gene was amplified by PCR and sequenced directly. The results showed that both IgM anti-Le(a) and anti-Le(b) antibodies were detected in patient's serum. Red cells was typed as Le (a-b-) phenotype and the FUT3 genotype was homozygote for non-functional le(59, 508) alleles. In conclusion, anti-Le(b) antibody can result in hemolytic transfusion reaction, FUT3 gene is homozygous for le(59, 508) allele resulting in Le (a-b-) phenotype.

Influencing factors of pulmonary dysfunction in coal worker's pneumoconiosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the possible influencing factors of pulmonary dysfunction in coal worker's pneumoconiosis (CWP).</p><p><b>METHODS</b>A total of 141 patients with CWP and 200 control miners with similar exposure histories but without apparent pulmonary disease or inflammation were interviewed with the detailed questionnaires (including histories of coal dust-exposure, smoking habits, alcohol consumption, protective mask uses, et al). Lung function examinations were performed at the same time. Predicted formula of lung function index were established by the local healthy residents characters and the pulmonary dysfunction was classified by the ratios between tested and predicted values.</p><p><b>RESULTS</b>All parameters of lung function were significantly lower in CWP cases when compared with that of control miners and the healthy controls (P < 0.05). The main types of pulmonary dysfunction were restrictive and mixed ventilation disorders in CWP patients. The factors such as the category of CWP, the mask worn, the smoking quantity and exposure to coal mine dust were included in the unconditional logistic regression model.</p><p><b>CONCLUSIONS</b>The category of CWP, the usage of mask, the smoking and long duration exposure to coal mine dust may be the main possible influencing factors of pulmonary dysfunction of CWP. Influencing factor analyses were given to inform choice of pertinence preventive measures.</p>

278C > T variant of the alpha-1, 3-galactosyltransferase allele responsible for Bw subgroup / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the molecular genetic basis of the Bw variant and identify novel alleles at ABO locus in Chinese Han population.</p><p><b>METHODS</b>Serological techniques were performed to characterize erythrocyte phenotype of a proband. Mutations of the ABO gene were screened by polymerase chain reaction, reverse transcription-polymerase chain reaction and DNA sequencing.</p><p><b>RESULTS</b>The proband was identified as Bw phenotype by serological technology and family study. A novel Bw variant allele was identified in the gDNA and cDNA. The novel allele was observed a missense mutation (278 C to T) at the exon 6 which resulted in an amino acid substitution (P93L) compared with B101 allele. The 278 C to T was the first report mutation position in exon 6 among Bw alleles, so the P93L amino acid substitution was different from others Bw variants which had amino acid substitutions in a conserved functional domain reported previously.</p><p><b>CONCLUSION</b>A novel Bw allele (278 C to T) responsible for Bw variant is reported in Chinese population.</p>

Establishment of genotyping method for DEL phenotype in Zhejiang Han population / 中国实验血液学杂志

In order to establish a genotyping method for DEL phenotype in Zhejiang Han population, an AS-PCR method was developed according to the RHD 1227A allele sequence. Its specificity and sensitivity were assessed in two Rh negative populations whose RHD 1227A or DEL phenotype status was known. The results showed that in evaluation of the method by detecting 50 RHD 1227A positive and 50 RHD 1227A negative individuals, the genotyping method displayed a sensitivity of 100% and a specificity of 100%; in evaluation of the method by detecting 33 DEL positive and 89 DEL negative individuals, the sensitivity was 100%, however, there were two serologically negative samples which were confirmed as positive using genotyping method. After re-testing these two samples with serological method and sequence analysis, it was found that original serological method gave false negative results and genotyping method still showed 100% specificity. The minimal target DNA concentration of this genotyping method is 8.13 ng/microl. In conclusion, designed genotyping method can be used to identify DEL phenotype efficiently in Zhejiang Han Rh negative population.

RHD 1227A allele frequency among Rh negative population and random population / 中国实验血液学杂志

To investigate the frequency of RHD 1227A allele in Rh negative population and random population, an AS-PCR (allele specific-polymerase chain reaction) method was employed to detect RHD 1227A allele. RHD gene copy was determined by D zygosity test and RHD exon 9 nucleotide sequence analysis. The results showed that among 143 Rh negative donors, forty-one RHD 1227A allele carriers were detected, and 8 (19.51%) out of which were RhCCdee, 32 (78.05%) were RhCcdee, and 1 (2.44%) was RhCcdEe. Thirty-five Rh negative RHD 1227A carriers had RHD gene deletion, and the remaining carriers were RHD 1227A homozygous. Seven (1.43%) individuals were detected with RHD 1227A allele among 489 random donors. They were all G/A heterozygous at RHD 1227 site. Serological test indicated that they were normal Rh positive phenotype. It is concluded that the frequency of RHD 1227A allele is 16.43% among Rh negative population and 0.72% among the random population.

FUT3 gene polymorphism associated with Lewis blood group in Chinese Zhejiang population / 中国实验血液学杂志

To investigate the alpha-1, 3/4-fucosyltransferase gene (FUT3) polymorphism associated with Lewis blood group in Zhejiang population, the Lewis phenotypes of 183 random samples from Chinese blood donors in Zhejiang province were identified by standard serological techniques. The entire coding region of FUT3 gene were amplified by PCR from genomic DNA of 39 Lewis negative and 9 Lewis positive phenotype samples and sequenced directly. The haplotypes of FUT3 allele were identified by TOPO cloning sequencing method. The results showed that the frequency of true Le (a-b-) phenotype in Zhejiang population was 10.4% according to serological and molecular biological methods. Five nucleotide acid variant sites (59T > G, 202T > C, 314C > T, 508G > A and 1067T > A) were detected in all 48 sequencing samples. Besides the wild type Le allele, 2 common (le(59, 1067) and le(59, 508) and 3 rare non-functional le alleles (le(59), le(1067) and le(202, 314) were found in this population. In conclusion, the polymorphism of non-functional FUT3 allele was found to be relatively variable in Chinese Zhejiang population.