Terrestrial gamma radiation level around Shidaowan nuclear power plant, China and influencing factors / 中国辐射卫生

@#<b>Objective</b> To monitor the cumulative terrestrial γ radiation dose around Shidaowan nuclear power plant, Shandong, China before operation, to analyze the dose levels and influencing factors, and to estimate the annual effective dose to local residents. <b>Methods</b> Fifty-six monitoring sites were selected within 30 km around the nuclear power plant. The environmental γ radiation dose was measured by the thermoluminescence dosimeter monitoring method. The γ radiation dose levels were investigated for 369 days in four monitoring periods (January 16 to April 14, April 15 to July 20, July 21 to October 21, 2021, and October 22, 2021 to January 20, 2022 for periods I to IV, respectively). Relations between γ radiation and monitoring time, altitude, distance from the nuclear power plant were analyzed, and the annual effective dose of terrestrial γ radiation to residents was estimated to reflect the background terrestrial γ radiation level in the area. <b>Results</b> The average values of terrestrial γ radiation dose rate in the four monitoring periods in the area were (76.196 ± 3.366), (81.773 ± 6.144), (93.554 ± 7.449), and (97.604 ± 9.396) nGy/h, respectively, and the terrestrial γ radiation dose rate in the whole year was (87.282 ± 6.589) nGy/h. The effective dose to residents was 0.428 mSv. The terrestrial γ radiation level was high from July 2021 to January 2022. There was no significant difference in the γ radiation dose rate at the monitoring sites with different distance from the nuclear power plant. No impact upon the terrestrial γ radiation dose by the altitude was observed in this study. <b>Conclusion</b> The terrestrial γ radiation level around Shidaowan nuclear power plant in 2021 was at the background level.

Serological and molecular studies of a rare A subgroup / 中华医学遗传学杂志

OBJECTIVE@#To determine the genotype of an individual suspected for Aw through DNA sequencing.@*METHODS@#Serologic testing was carried out with standard methods. Exons 6 and 7 of the ABO genes were amplified by PCR and subjected to direct sequencing or sequenced after gene cloning.@*RESULTS@#Serological testing showed that the forward typing and reverse typing were Aw and A, respectively. DNA sequencing revealed that the individual has carried an Aw allele and an O allele. Haplotype sequencing of each allele has revealed a nt543 variant (543G>C) in the Aw allele.@*CONCLUSION@#The individual was verified as a rare A subtype, which was previously unreported in mainland China.

Serological and molecular study of a novel B(A) allele with multiple missense mutations / 中华医学遗传学杂志

OBJECTIVE@#To explore the molecular basis for an individual suspected as AwB subtype through DNA sequencing.@*METHODS@#ABO serology was carried out with the standard tube method. To identify the ABO gene haplotype, the amplicons of exon 7 were cloned and sequenced.@*RESULTS@#Serological results showed that the forward typing was AwB and the reverse typing was B. Sequencing analysis revealed that the sample has contained an O01 allele in addition with c.297A>G, c.657C>T, c.796C>A, c.803G>C, c.930G>A variants as compared with the A101 allele.@*CONCLUSION@#Through sequencing analysis, the sample with an AwB subtype by serological testing was identified as a novel B(A) phenotype, which was unreported previously.

Characterization of a pair of twins as blood group chimeras / 中华医学遗传学杂志

OBJECTIVE@#To delineate the blood group for a pair of twins with inconclusive ABO blood typing result.@*METHODS@#Serological test for blood group was carried out by using ABO and Rh Blood Grouping Cards (Microcolumn Gel Immunoassay). Sequence specific primer-PCR (PCR-SSP), direct sequencing and TA clone sequencing were used to analyze the ABO gene. Genetic status was analyzed by using 16 short tandem repeat (STR) markers.@*RESULTS@#Red blood cells of the twins displayed 2+ mixed agglutination phenomenon with anti-A, anti-A1 and anti-E. PCR-SSP and DNA sequencing of exons 6 to 7 revealed that they have an ABO*O.01.01/ABO*O.01.02 genotype. DNA sequencing of microsatellite enhancer region revealed presence of A gene. STR analysis revealed more than two haplotypes for 9 loci between the twins. After clustered by anti-A, the red blood cells were divided into two groups: A, CcDEe and O, CcDee, respectively.@*CONCLUSION@#Serological and molecular techniques have characterized the twins as blood group chimeras.

Construction and validation of eukaryotic expression vectors for human ABO subgroup genes / 中华医学遗传学杂志

OBJECTIVE To construct eukaryotic expression vectors for human ABO and subgroup genes A101, B101, CisAB01, Ael05, B(A)04 and Bw03, and validate their expression in vitro. METHODS Total RNA was isolated from individuals with the A101 and B101 subgroups. cDNA of A101 and B101 was synthesized by reverse transcription and amplified with specific primers. Subgroup genes CisAB01, Ael05, B(A)04 and Bw03 were then amplified with PCR for site-directed mutagenesis. Fragments of the ABO genes were directionally linked to pcDNA3.1 positive-eukaryotie expression vectors. After antibiotic screening, the sequences were analyzed. The vectors were transfected into Hela cells, and the expression of target proteins was detected by Western blotting and immunofluorescence assay. RESULTS Sanger sequencing has confirmed that pDNA3.1-A101, pDNA3.1-CisAB01, pDNA3.1-Ael05, pDNA3.1-B101, pDNA3.1- B(A)04, pDNA3.1-Bw03 positive-eukaryotic expression vector were successfully constructed. The results of Western blotting and immunofluorescence revealed clear presence of the expressed proteins. CONCLUSION Eukaryotic expression vectors for ABO subgroup genes were successfully constructed and worked well in Hela cells in vitro, which can facilitate further study of the ABO blood group proteins.

Polymorphism of RHD gene among RhD negative and D variant blood donors from Qingdao region / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the molecular mechanism and polymorphism of D gene of RhD negative and D variants among voluntary blood donors from Qingdao region.</p><p><b>METHODS</b>For 220 D-negative phenotype cases and 5 D variant cases confirmed by serological test, exons 1 to 10 of the RHD gene were detected by a PCR-SSP method. The samples which contain all or part of the exons were sequenced.</p><p><b>RESULTS</b>Among the 220 cases, 166 (75.45%) had complete absence of the RHD gene, while 54 (24.55%) had retained some or all of the 10 exons. Eight genotypes were identified, which included RHD 1227G>A in 28 cases (12.73%), RHD-CE- (2-9) -D in 19 cases (8.64%), RHD-CE- (3-7)-D in 1 case (0.45%), RHD 3G>A in 1 case (0.45%), RHD 711delC in 2 cases (0.91%), RHD 845G>A in 1 case (0.45%), RHD 1013T>C in 1 case (0.45%), and RHD 1227A/G in 1 case (0.45%). No mutation was found in all of the 10 exons. Two alleles were identified in the 5 cases of D variants, which included RHD 845G>A (4 cases) and RHD 697G>A (1 case).</p><p><b>CONCLUSION</b>Absence of the whole RHD gene is common among RhD negative blood donors from Qingdao region, and there are rich genetic polymorphisms for this locus.</p>

Serological and molecular analysis of a case with para-Bombay phenotype caused by a h(nt328G to A) mutation / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the serological characteristics and molecular basis for an individual with para-Bombay phenotype.</p><p><b>METHODS</b>Blood type of the proband was determined with routine serological methods. Exons 6 and 7 of the ABO gene and coding regions of the FUT1 and FUT2 genes were amplified by PCR and sequenced.</p><p><b>RESULTS</b>The para-Bombay phenotype was confirmed to be of Ah-secretion type. The genotype of the individual was determined as A102/O01. Position 328 of the FUT1 gene was mutated from A to G, resulting in replacement of Alanine (Ala) at position 110 by Threonine (Thr).</p><p><b>CONCLUSION</b>The G to A mutation of nt328 of the FUT1 gene probably underlies the para-Bombay phenotype in this individual.</p>

Serological and molecular study of three cases with a rare Bx02 blood group / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To determine the genotypes of three blood samples suspected as B subtype through DNA sequencing.</p><p><b>METHODS</b>The samples were first genotyped with PCR-SSP. Exons 6 and 7 of the ABO genes were subjected to PCR, direct sequencing, and cloned sequencing to determine the genotypes.</p><p><b>RESULTS</b>Serological results of the three samples were similar, with red cells being weakly agglutinatable by anti-B and serum containing anti-B. The samples were preliminarily genotyped as B/O1. Sequencing analysis showed that all three samples contained an O allele and a 905A>G mutation of the B gene, which was previously defined as Bx02.</p><p><b>CONCLUSION</b>Through sequencing analysis, the three samples typed as B subtype with serological testing were identified as Bx phenotype. The genotype of samples 1 and 2 was Bx02/O101, and that of sample 3 was Bx02/O102.</p>

Effect of vaIproic acid on radiosensitivity to breast cancer ceIIs / 中国药理学与毒理学杂志

OBJECTIVE To study the effect of valproic acid ( VPA) on radiosensitivity to MCF7 breast cancer cells. METHODS MCF7 cells were pretreated with VPA 0.5 and 1 mmol.L-1 for 0, 24, 48 and 72 h respectively, irradiated with 8 Gy lR, and at 6 h post-lR, the γ-H2 AX foci formation in MCF7 cells was tested by immunofluorescence assay. MCF7 cells were pretreated with VPA 0.5 and 1 mmol.L-1 for 72 h, irradiated with 4 Gy lR, and at 48 h post-lR, the cell survival rate was detected by MTT assay. MCF7 cells were pretreated with VPA 0.5 mmol.L-1 for 24 h, and then irradiated according to the amount of cells: 2 Gy (500 and 1000 cells per plate), 4 Gy (2000 and 4000 cells per plate), 6 Gy (8000 and 16000 cells per plate), and the cloning efficiency was calculated. MCF7 cells were pretreated with VPA 0.5 and 1 mmol.L-1 for 0, 24, 48 and 72 h respectively and the cell cycle profile was analyzed via flow cytometry. RESULTS After treatment with VPA alone for 24 h, MCF7 cells showed a significant increase in the amount of γ-H2 AX foci formation ( P < 0. 01). lt was also found that VPA increased lR-induced γ-H2 AX foci formation, which obviously prolonged the pretreatment time of VPA(P<0.01) in a time-dependent manner(r=0.98, P<0.05). VPA 0.5 and 1 mmol.L-1 had the same effect on γ-H2 AX foci formation. Furthermore, VPA was able to cause a significant decrease in lR-induced clonogenic survival but an increase in lR-induced cytotoxicity by MTT assay. Also, VPA alone decreased the plating efficiency of MCF7 cells. However, the cycle profile of MCF7 cells treated with both VPA 0.5 and 1 mmol.L-1 was not changed. CONCLUSION Without affecting the cell cycle profile, both the safe and critical dose of VPA used in clinical epilepsy treatment can significantly increase the accumulation of DNA double strand breaks in the cells and sensitize the cells to lR treatment, suggesting that VPA can induce radio-sensitization of breast cancer cells.

Comparative study on functional characters of MCF7 and HCC1937 cell lines in response to DNA damage / 中华预防医学杂志

<p><b>OBJECTIVE</b>The functional characters of MCF7 and HCC1937 cell lines were compared through the activity of BRCA1 and p53 following DNA damage in order to provide more research evidence for the related studies in both breast cancer cell lines.</p><p><b>METHODS</b>The protein level of BRCA1 and p53 in two breast cancer cell lines and the protein level of BRCA1 in MCF7, HCC1937 and HCC1937 wtBRCA1 breast cancer cell lines treated with 10Gy after 1 h, 4 h or 8 h were detected by western blotting analysis. The distribution and foci formation of BRCA1 in the cells were observed through immunostaining assay and the percentage of BRCA1 or Rad51 foci formation after ionizing radiation was calculated. Cell cycle profiling was analyzed using flow cytometry.</p><p><b>RESULTS</b>Most of BRCA1 and p53 localized in nucleus, and both proteins responded to DNA damage in MCF7 cells. In MCF7 cells,BRCA1 and Rad51 foci formation respectively increased to (59.40 ± 3.66)% from (11.80 ± 3.51)% (t = 16.26, P < 0.05) and (73.90 ± 8.66) % from (16.70 ± 3.76) % (t = 10.49, P < 0.05) after 10 Gy 8 h ; p53 and p21 protein level was further separately induced and enhanced to (82.54 ± 1.04) from (23.75 ± 0.51) (t = 87.90, P < 0.05) and (90.95 ± 1.13) from (50.19 ± 0.89) ( t = 49.11, P < 0.05) after 10 Gy 8 h; and the cells were accumulated in G1 phase. In contrast to MCF7, in HCC1937 cell line, both of BRCA1 and p53 were defective in nucleus since both proteins were mutated; in response to DNA damage, BRCA1 foci formation was not found, p53 and p21 was not induced; there was no cell accumulation in both of G1-S and G2-M phases. However, after complementation of wild-type BRCA1 in HCC1937 cells, DNA damage-induced Rad51 foci formation increased to (61.70 ± 4.03) % from (6.22 ± 2.27) % (t = 20.78, P < 0.05) and accumulation of cells in G2-M phase was also restored after 10 Gy 8h , which was similar to that of in MCF7 cells.</p><p><b>CONCLUSIONS</b>We have identified that BRCA1 and p53 have dramatically different functions in MCF7 and HCC1937 cell lines in response to DNA damage.</p>

Progress in role of breast cancer susceptibility gene 1 in DNA damage repair / 中国药理学与毒理学杂志

Breast cancer susceptibiIity gene 1( BRCA1)is a tumor suppressor,but mutated get BRCA1 is cIoseIy reIated to the deveIopment of breast cancer. Current study has reveaIed that BRCA1 can get invoIved in the DNA damage repair process as a key mediator. DNA doubIe-stranded break (DSB)is the most serious form in DNA Iesions,and BRCA1 pIays an important roIe in repairing DSB through reguIation of homoIogous recombination( HR). In this articIe,the moIecuIar mechanism of BRCA1 in reguIating HR is reviewed with reference to the activity of functionaI domains in BRCA1 struc-ture,the mutations occurring in main domains of BRCA1,the reIationship of BRCA1 with BRCA2, Rad51 or CtIP,and phosphoryIation of BRCA1. In addition,the association of the defective BRCA1-mediated HR repair mechanism with the sensitivity to different DNA damaging agents and synthetic IethaIity in tumor ceIIs is aIso addressed.

Intracellular Ca2+is involved in survival, proliferation and differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes / 中国组织工程研究

BACKGROUND:The mechanism of differentiation and proliferation of bone marrow-derived mesenchymal stem cells remains unclear. In addition, issues such as how signal pathways such as Ca2+and bone marrow-derived mesenchymal stem cellproliferation and differentiation signals form complex signal network remain poorly understood. OBJECTIVE:To investigate the effect of Ca2+in the induced differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes. METHODS:Bone marrow-derived mesenchymal stem cells were isolated from rat bone marrow using whone bone marrow adherence method, purified, amplified, and induced with hepatocyte growth factor. [Ca2+]i in the directional differentiated bone marrow-derived mesenchymal stem cells and control bone marrow-derived mesenchymal stem cells were detected with flow cytometry. Bone marrow-derived mesenchymal stem cells induced with hepatocyte growth factor were mixed with nimodipine of different concentration, and cells were divided into three groups:hepatocyte growth factor+nimodipine 10 mg/L, 50 or 100 mg/L groups. cellgrowth was observed with inverted phase contrast microscope and alpha 1-antitrypsin expression of the cells was confirmed by immunocytochemistry. The calcineurin M and the activation of extracellular signal regulated kinase pathway was detected by reverse transcription-PCR and western blotting, respectively. RESULTS AND CONCLUSION:[Ca2+]i in the directional differentiated bone marrow-derived mesenchymal stem cells was higher than in the control group (P0.05). These findings indicate that Ca2+could participate in the differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes incuded with cytokines, and also maintain the survival and proliferation of bone marrow-derived mesenchymal stem cells.

Effect of taurine on changes of nuclear and cytosolic ［Ca2+］ elicited by H2O2 in rat vascular smooth muscle cells / 中国药理学通报

AIM　As one of the reactive oxygen species, toxic dose of H2O2 leads to the necrosis or apoptosis of many kinds of cells. It is nuclear whether these processes are related with the changes of nuclear Ca2+ elicited by H2O2. This study on the effect of taurine on changes of nuclear and cytosolic (Ca2+) elicited by H2O2 in rat vascular smooth muscle cells. METHODS　The techniques of Fluo-3 and laser scanning confocal microscopy were used. RESULTS　Rapid and sustaining rise of nuclear ［Ca2+］ (［Ca2+］n) and cytosolic ［Ca2+］(［Ca2+］c) were found in cultured rat vascular smooth muscle cells upon adding of 0.5% H2O2. Pretreatment of the cells with 20 mmol*L-1 taurine, a regulator of cellular Ca2+ homestasis, significantly decreased the rise amplitude of ［Ca2+］n (P<0.05), but failed to affect that of ［Ca2+］c (P>0.05) elicited by H2O2. It was suggested that the changes of ［Ca2+］n and ［Ca2+］c have different respond to taurine. CONCLUSION　It was concluded that there exist the independent regulating mechanisms of Ca2+ in the nucleus from differences between the nucleus and cytosol Ca2+ in sensitivity to action of H2O2 and taurine.

Effect of taurine on changes of nuclear and cytosolic [Ca~(2+) ] elicited by H_2O_2 in rat vascular smooth muscle cells / 中国药理学通报

AIM As one of the reactive oxygen species, toxic dose of H 2O 2 leads to the necrosis or apoptosis of many kinds of cells. It is nuclear whether these processes are related with the changes of nuclear Ca 2+ elicited by H 2O 2. This study on the effect of taurine on changes of nuclear and cytosolic (Ca 2+ ) elicited by H 2O 2 in rat vascular smooth muscle cells. METHODS The techniques of Fluo 3 and laser scanning confocal microscopy were used. RESULTS Rapid and sustaining rise of nuclear [Ca 2+ ] ([Ca 2+ ] n) and cytosolic [Ca 2+ ]([Ca 2+ ] c) were found in cultured rat vascular smooth muscle cells upon adding of 0 5% H 2O 2. Pretreatment of the cells with 20 mmol?L -1 taurine, a regulator of cellular Ca 2+ homestasis, significantly decreased the rise amplitude of [Ca 2+ ] n ( P 0 05) elicited by H 2O 2. It was suggested that the changes of [Ca 2+ ] n and [Ca 2+ ] c have different respond to taurine. CONCLUSION It was concluded that there exist the independent regulating mechanisms of Ca 2+ in the nucleus from differences between the nucleus and cytosol Ca 2+ in sensitivity to action of H 2O 2 and taurine.