Epidemiological Survey of Hemoglobinopathies Based on Next-Generation Sequencing Platform in Hunan Province, China / 生物医学与环境科学（英文）

OBJECTIVE@#This study was aimed at investigating the carrier rate of, and molecular variation in, α- and β-globin gene mutations in Hunan Province.@*METHODS@#We recruited 25,946 individuals attending premarital screening from 42 districts and counties in all 14 cities of Hunan Province. Hematological screening was performed, and molecular parameters were assessed.@*RESULTS@#The overall carrier rate of thalassemia was 7.1%, including 4.83% for α-thalassemia, 2.15% for β-thalassemia, and 0.12% for both α- and β-thalassemia. The highest carrier rate of thalassemia was in Yongzhou (14.57%). The most abundant genotype of α-thalassemia and β-thalassemia was -α 3.7/αα (50.23%) and β IVS-II-654/β N (28.23%), respectively. Four α-globin mutations [CD108 (ACC>AAC), CAP +29 (G>C), Hb Agrinio and Hb Cervantes] and six β-globin mutations [CAP +8 (C>T), IVS-II-848 (C>T), -56 (G>C), beta nt-77 (G>C), codon 20/21 (-TGGA) and Hb Knossos] had not previously been identified in China. Furthermore, this study provides the first report of the carrier rates of abnormal hemoglobin variants and α-globin triplication in Hunan Province, which were 0.49% and 1.99%, respectively.@*CONCLUSION@#Our study demonstrates the high complexity and diversity of thalassemia gene mutations in the Hunan population. The results should facilitate genetic counselling and the prevention of severe thalassemia in this region.

Application of Next-Generation Sequencing in Screening of Thalassemia Gene in 11212 Pregnant Women in Suxian and Beihu Districts of Chenzhou City, Hunan Province / 中国实验血液学杂志

OBJECTIVE@#To understand the carrying rate, gene mutation frequency and composition ratio of thalassemia in pregnant women in Suxian and Beihu districts of Chenzhou, Hunan Province.@*METHODS@#Thalassemia gene in 11 212 samples was analyzed by using Next-Generation Sequencing.@*RESULTS@#Among the 11 212 samples, 938 were diagnosed as thalassemia, in which 618 (5.51%) were diagnosed as α-thalassemia, 268 (2.39%) as β-thalassemia, 29（0.26%）as abnormal hemoglobin and 23 (0.21%) as αβ-thalassemia. The gene mutations of --SEA /αα(40.29%) and -α3.7/αα(37.7%) in α-thalassemia were the most common, while for β- thalassemia, the most commonly gene mutation were β41-42M/βN(24.26%) and β654M/βN(23.88%). The detection rate of rare type α,β-thalassemia gene was 0.19%(21/11 212）, 0.53%（59/11 212）, respectively.@*CONCLUSION@#The carrying rate of thalassemia in pregnant women is 8.37% in Suxian and Beihu districts of Chenzhou city, and the genotypes are complex. Next-Generation Sequencing can detect rare thalassemia genes and new gene mutations effectively.

Mechanism of antitumor effect of ursolic acid on T (8;21) leukemia cell kasumi-1 / 中国实验血液学杂志

This study was purposed to investigate the anti-tumor effect of ursolic acid (UA) on t(8;21) leukemia cell line kasumi-1 and its possible mechanisms. The kasumi-1 cells were treated with UA at different concentration for different duration of time. The growth inhibition of kasumi-1 treated with UA was detected by using CCK-8 test, and the morphological changes of kasumi-1 cells were observed by Wright's staining. Furthermore, the apoptosis rate of kasumi-1 was examined by flow cytometry. Lastly, the expression of AML1-ETO, KIT, MYC, CCND1, BCL-2, P53, BAX, MDM2 and protein were detected by using real-time quantitative PCR and Western blot respectively. The results showed that the UA obviously inhibited the growth of kasumi-1 cells in dose- and time-dependent manners. The apoptotic morphological changes of cells were presented when kasumi-1 cells were treated with UA for 48 hours. The apoptotic rate of kasumi-1 cells increased in a dose- and time-dependent ways, and the mRNA levels of AML1-ETO, KIT, MYC, CCND1, BCL2, MDM2 decreased in kasumi-1 cells treated with UA, as well as the protein levels. Meanwhile, UA up-regulated the mRNA and protein levels of P53 in the same manner. It is concluded that UA can exert its anti-tumor effect by inhibiting the proliferation and inducing the apoptosis of kasumi-1 cells in a dose-and time-dependent manners, that may provide the clues for a new targeting therapy to t(8;21) leukemia.

Mechanism concerning antitumor effect of oridonin on multiple myeloma cell line U266 / 中国实验血液学杂志

This study was purposed to investigate the antitumor effect of oridonin on human multiple myeloma cell line U266 and its possible mechanism. The CCK-8 test was used to determine the inhibitory effect of oridonin on proliferation of U266 cells. The morphological changes of U266 cells were observed under optical microscope. The apoptosis rate of U266 cells was detected by flow cytometry. The mRNA levels of FGFR3, BCL2, CCND1 and MYC genes were quantified by using real-time quantitative PCR method, and the protein levels of BCL2, MYC, CCND1, FGFR3 and P53 were detected by Western blot. The results showed that the oridonin obviously inhibited the growth of U266 cell in dose-and time-dependent manners. As for morphological changes, characteristic apoptotic cells presented in U266 cells treated with 10 µmol/L oridonin for 24 hours. The apoptotic rate of U266 cells increased in dose and time dependent manners; after treatment of U266 cells with oridonin the mRNA levels of FGFR3, BCL2, CCND1 and MYC as well as the their protein levels decreased. Occasionally, the oridonin up-regulated the protein levels of P53 in the same manner. It is concluded that the oridonin can exert its anti-tumor effect by inhibiting proliferation and inducing apoptosis of U266 cell in dose dependent and time dependent manners, that maybe give the clues about new program of target therapy for multiple myeloma.

Comparison between application of fecal occult blood quantitive testing instrument and colloidal gold strip method in colorectal cancer screening / 中华预防医学杂志

<p><b>OBJECTIVE</b>To compare the performances of fecal occult blood quantitive testing instrument and colloidal gold strip method in colorectal cancer screening.</p><p><b>METHODS</b>A representative random population of 9000 subjects aging between 40 and 74 years old were selected from Xuxiang, Haining city, Zhejiang province, by random cluster sampling method in year 2011. The fecal samples from each subject were separately detected by the two methods, namely fecal occult blood quantitive testing instrument and colloidal gold strip method. The positive result was standardized by hemoglobin concentration (HGB) ≥ 100 ng/ml under the application of quantitive testing instrument, or color-developing by colloidal gold strip method. The positive subjects from either method would be provided a further colonoscopy examination for pathological diagnosis. The positive rate and consistency of the two methods were compared, as well as the positive predictive value and population detecting rate of the colorectal cancer and adenoma.</p><p><b>RESULTS</b>A total of 6475 (71.9%) subjects submitted their two fecal samples according to our requirement in 9000 subjects. There were separately 319 positive cases (4.9%) and 146 positive cases (2.3%) by the performances of fecal occult blood quantitive testing instrument and colloidal gold strip method, including 45 positive in both tests (Kappa = 0.168, 95%CI:0.119-0.217).184 out of the 319 positive cases (57.7%) in the test by quantitive testing instrument and 89 out of 146 positive cases (61.0%) in the test by colloidal gold strip method received the colonoscopy examination. There were no significant statistical differences between the two methods in the positive predictive value of colorectal cancer (P > 0.05) , developing adenoma and non-developing adenoma.However, the population detecting rate of the colorectal cancer and developing adenoma were higher in the test by quantitive testing instrument (26 cases, 0.402%) than it in the test by colloidal gold strip method (10 cases, 0.154%). The difference showed statistical significance (χ(2) = 7.131, P < 0.01).</p><p><b>CONCLUSION</b>The performances of fecal occult blood quantitive testing instrument might be better than colloidal gold strip method in colorectal cancer screening. However, the results need to be further verified.</p>

Regulating effect and mechanism of electroacupuncture with shuanggu yitong prescription methods on immunosenescence in aging rats with yang deficiency / 中国针灸

<p><b>OBJECTIVE</b>To explore the improvement effect and the regulating mechanism of electroacupuncture with "Shuanggu Yitong" prescription methods on learning memory and immunosenescence in aging rats with yang deficiency.</p><p><b>METHODS</b>Forty female Sprague-Dawley rats, 5-month-old, were randomly divided into a normal control (NC) group, an aging yang deficiency model (M) group, an electroacupuncture with Shuanggu Yitong prescription methods (EA) group and an electroacupuncture control (EAC) group, 10 rats in each group. The aging model rats with yang deficiency were established by hypodermic injection of D-galactose for 40 days and then intramuscular injection of Hydrocortisone for 7 days in the latter three groups except the NC group The EA group was treated with electroacupuncture at "Guanyuan" (CV 4), "Housanli" (ST 36) and "Baihui" (GV 20), and the EAC group was treated with electroacupuncture at "Zhongji" (CV 3), "Yinlingquan" (SP 9) and "Yintang" (EX-HN 3), six times per week for 4 weeks. The escape latency was examined by Morris water maze from the first day of the 4th week. After completion of electroacupuncture treatment, the rate of spleen lymphocyte apoptosis and the serum content of tumor necrosis factor-a (TNF-alpha) were examined by flow cytometry and enzyme linked immunosorbent assay (ELISA) technique respectively, the differences among all the groups above were compared.</p><p><b>RESULTS</b>In comparison with the NC group, the escape latency [(25.4 +/- 3. 6)s vs. (16.23 +/- 2.3)s], the rate of spleen lymphocyte apoptosis [(27.25 +/- 3.3)% vs. (13.2 +/- 3.1)%] and the serum content of TNF-alpha [(15.54 +/- 3.56) pg/mL vs (7.35 +/- 2.89) pg/mL] in the M group those were all significantly increased (all P < 0.01). In the EA group, the escape latency of (17.42 +/- 3.9)s, the rate of spleen lymphocyte apoptosis of (17.2 +/- 3.25)% and the serum content of TNF-alpha of (9.51 +/- 3.53) pg/mL were all significantly lower than those in the M group (all P < 0.01) and the EAC group (all P < 0.05).</p><p><b>CONCLUSION</b>Electroacupuncture with "Shuanggu Yitong" prescription can significantly improve the spatial learning and memory. The protective mechanism is related with the action of electroacupuncture in lowering the rate of spleen lymphocyte apoptosis and the serum content of TNF-alpha, and the therapeutic effect of electroacupuncture at "Guanyuan" (CV 4), "Housanli" (ST 36) and "Baihui" (GV 20) is superior to that of electroacupuncture at "Zhongji" (CV 3), "Yinlingquan" (SP 9) and "Yintang" (EX-HN 3).</p>

Effects of simvastatin on PI3K/AKT signaling pathway in human acute monocytic leukemia cell line SHI-1 / 中国实验血液学杂志

To investigate the effects of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor simvastatin (SV) on proliferation, apoptosis and the PI3K/AKT signaling pathway in human acute monocytic leukemia cell line SHI-1. SHI-1 cells were incubated with different concentrations of SV (5, 10, 15 µmol/L). Otherwise, SHI-1 cells without any treatment were used as control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detection. MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the early stage apoptosis ratio. The human PI3K-AKT Signaling Pathway RT(2) Profiler(TM) PCR Array was used to detect the expression of 84 genes involved in PI3K-AKT signaling. The results indicated that the SV inhibited the proliferation and inducted the apoptosis of SHI-1 cells in time- and dose-dependent manners significantly. The growth inhibition rates of SHI-1 cells treated with 15 µmol/L SV for 24, 48 and 72 h were 26.82, 47.09 and 63.92, respectively; and their early stage apoptosis ratios were 5.75, 13.25 and 15.59, respectively. Compared with the control group, expression levels of 39 genes were changed in the group of 15 µmol/L SV at 48 h, among them 26 genes were down-regulated and 13 genes were up-regulated. It is concluded that the SV can inhibit proliferation and induce apoptosis of SHI-1 cells, and the mechanism may be associated with the changes of gene expression level in PI3K-AKT signaling pathway regulated by SV.

Comparison of allelic frequencies of 15 short tandem repeat loci between chronic myeloid leukemia patients and non-related healthy individuals / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To compare allelic frequencies of 15 short tandem repeat (STR) loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818 and FGA) between chronic myeloid leukemia (CML) patients and non-related healthy individuals from Changzhou region in order to predict genes related with the CML.</p><p><b>METHODS</b>Blood samples were collected from 745 healthy subjects and 132 CML patients with complete remission. Genotypes were determined with gene scan technology and multiplex PCR with fluorescence-labeled primers. Allelic polymorphisms of 15 STR loci were compared between the two groups. Potential genes related with CML were predicted with statistical analysis of differences in allelic frequencies.</p><p><b>RESULTS</b>Allelic frequencies of 3 loci, including CSF1PO, vWA and TPOX, showed a significant difference (P<0.05) between the two groups.</p><p><b>CONCLUSION</b>CSF1PO, vWA and TPOX loci may be related with CML, albeit that the exact biologic mechanisms is unclear.</p>