Genotype ; Humans ; beta-Thalassemia

Objective To investigate the correlation between the erythrocyte indices and the genotypes of alpha thalassemia.Methods337 carriers with various genotypes of alpha-thalassaemia ( iron deficiency,alpha-thalassemia double heterozygote and homozygote,α-compounding β-thalassemia and abnormal hemoglobinopathy were excluded) were classified into three groups based on different genotypes of alpha-thalassaemia including silent thalassemia group (ST,83 cases),α-thalassemia trait group (TT,210cases) and intermediate thalassemia group( IT,44 cases),and 154 healthy adults were randomly choosed as normal control The erythrocyte indices involving in RBC,Hb,MCV,MCH,MCHC and RDW-CV were retrospectively analyzed and the difference of which was compared by analysis of variance and SNK test among aboved-mentioned groups.ResultsThere were statistical significance among groups about erythrocyte indices except Hb F.The order of the level of MCV and MCH was NC [( 86.6 ± 5.2) fl,( 29.5 ± 2.1 ) pg] ＞ST[(80.1 ±3.3) fl,(26.7±1.3) pg] ＞TT[(68.5 ±3.4) fl,(22.0 ±1.2) pg] ＞IT[(66.6±7.1)fl,(20.0 ±2.2) pg,F =580.67,761.19,P ＜0.05].And the size of RDW-CV was IT(22.3 ±3.4)％ ＞TT (14.9±1.2) ％ ＞ST(13.8±1.6)％ ＞NC(13.2±1.4)％(F=347.25,P＜0.05).In ST group,the value of MCHC of -α3.7/αα subgroup( 335.6 ± 8.0) g/L was higher than that of -α4.2/αα subgroup( 330 ±7.2) g/L and αTα/αα subgroup (328.4 ±9.5) g/L(F=6.07,P ＜0.05).Meanwhile,in IT group,the value of MCV of αTα/--SEA subgroup( 70.1 ± 7.2 ) fl was higher than that of -α3.7/--SEA subgroup ( 63.4 ±5.9) fl and -α4.2/--SEA subgroup ( 64.1 ± 4.0 ) fl ( F =5.55,P ＜ 0.05 ).However,the value of MCHC of αTα/--SEA subgroup( 289.7 ± 21.2 ) g/L was lower than that of other two subgroups [( 306.3 ± 8.4 ),(306.1 ± 8.7) g/L,F =8.72,P ＜0.05].Except Hb A2 and Hb F,there was positive correlation between the number of deleted α-globin gene and that of RBC and RDW-CV ( r =0.318 and 0.580,P ＜0.01 ).Nevertheless,there was negative correlation between the number of deleted α-globin gene and that of the other erythrocyte indices (r =-0.483,-0.827,-0.744 and -0.684,P all ＜0.01 ).ConclusionsThere is close correlation between the degree of anemia and the number of deleted α-globin gene characterized by Hb reduction and RBC increasing.In addition,the anemia degree of non-deletional Hb H disease is severer than that of deletional Hb H,which of Hb H disease with -α4.2/--SEA is severer than that with -α3.7/--SEA.

ObjectiveTo evaluate the reliability of the probe melting curve analysis (PMCA) based on real-time fluorescent PCR assay for the genetic diagnosis and prenatal diagnosis of β-thalassemia.MethodsA total of 135 cases of peripheral blood samples were collected from Zhuhai Municipal Maternity and Child Healthcare Hospital between 2008 and 2010.All the samples were performed preliminary diagnosis according to the hematological data.Of these,119 cases were diagnosed as β-thalassemia trait,4 cases were diagnosed as severe thalassemia and 12 cases were normal.In addition,44 cases of amniotic fluid and 8 cases of cord blood with high-risk for severe β-thalassemia were also collected.The diagnostic reliability of the PMCA assay and reverse dot blot assay was evaluated on 187 above-mentioned cases by direct DNA sequencing analysis in a double-blind study.ResultsThe genotypes of 185 cases were detected accurately based on the PMCA assay except for two cases:one heterozygote with Ini( ATG ＞ AGG) was omitted and another heterozygote couldn't be distinguished between CD43 ( G ＞ T) and CD37 ( G ＞ A ).For the RDB assay,only one heterozygote with CD71-72 ( + T) was not detected accurately in the above-mentioned cases.Compared with the DNA sequencing analysis,the sensitivity,specificity,negative predictive value,positive predictive value and diagnostic efficiency of the PMCA assay were 98.75％,100.00％,93.10％,100.00％ and 98.93％,respectively.The corresponding value of the RDB assay were 99.38％,100.00％,96.42％,100.00％ and 99.47％,respectively.There were no significant between-group differences in the diagnostic efficiency of the two assays ( P ＞ 0.05 ).The results of prenatal diagnosis were in complete concordance with the follow up results,after the birth or induced labour of the fetuses.ConclusionsThe PMCA assay can be used as an alternative and verified method of RDB assay for the genetic diagnosis and prenatal diagnosis of β-thalassemia.

Objective To explore the feasibility of carrying out the common imprecision range among different biochemical systems in different laboratories.Methods Biochemical professionals visited 18 third-grade class A hospitals in Hebei Province,investigated the internal quality control data of 21 biochemical tests and made classification according to certain parameter criterias.Data were collected from April to September,2010 and calculated for cumulative mean,standard deviation (s) and CV.Data were compared according to 1/3,1/4 of TEa established by CLIA'88 and allowable CVb％ derived from biological variants.Results Among 18 hospitals,5 (27.8％) set their target value by mean value of 6 months,5(27.8％) by continuous 20 days and 8(44.4％) by the given value of the supplier.CVs of 21biochemical tests were quite different among 18 biochemistry laboratories,in which LDH was 6.79 times and CK was 76.79 times different from one another.35.5％-94.1％ biochemical tests met the requirement of CV ＜ 1/3TEa and 0.0％-91.7％ met the requirement of CV ＜ 1/4TEa.0.0％-94.1％ of tests were below allowable CVb％ 16/21 (76.2％) tests could satisfy the requirement of CV ＜ 1/3TEa.The top five tests which didn't meet the requirements were Na,Urea,TBIL,ALT and Glu.Conclusions The internal quality control among biochemistry laboratories in Hebei Province has not been standardized yet.According to the survey data,biochemistry laboratories of third-grade class A hospitals may set 75％ point of imprecision as a reference.After a period of improvement,we will set common imprecision range among biochemistry laborotories in Hebei Province and even in China.

Horizontal gene transfer (HGT), also Lateral gene transfer (LGT), is any process in which an organism transfers genetic material to another species that is not its offspring. With the increase of available genomic data, it has become more convenient to study the way to detect the genes, which are products of horizontal transfers among a given genome. There are few data about known horizontal gene transfers in three bacterium genomes under consideration, so the experiments, which simulated gene transfer by artificially inserting phage genes, were carried out. Combining the feature analysis methods of gene sequences with support vector machine (SVM), a novel method was developed for identifying horizontal gene transfers (HGT) in 3 fully sequenced bacterium genomes (Escherichia coli K12, Borrelia burgdorferi, Bacillus cereus ZK). According to our previous work, codon use frequency (FCU) was selected as the sequence feature, in respect that it is inherently the fusion of both codon usage bias and amino acid composition signals. In addition, another computational method was proposed considering strand asymmetry and predicting horizontal gene transfers of leading strand and lagging strand of genomes under consideration, respectively. To avoid the occasionality of simulating gene transfer through artificially inserting phage genes, 100 times of the transfer-and-recover experiment were repeated and arithmetic average of measurement for each genome being considered were reported to evaluate algorithm's performance. Ten-fold cross-validation was used for both parameter and accuracy estimation. The best results were obtained for C-Support Vector Classification (C-SVC) type by using the radial basis function kernel with ?=100, while for one-class SVM type the best performance was obtained using the polynomial kernel of three degree. The performance of the approach was compared with that of Tsirigos' method ,which is one of the best predictive approachs to date in detecting of horizontal transfer genes. Firstly, for the original method that did not consider the strand asymmetry, the C-SVC type has a high relative improvement(RI) of 31.47% on hit ratio for Escherichia coli K12, while the one-class SVM type has RI of 11.61% for Borrelia burgdorferi. Moreover, as theoretically expected, the method considering the strand asymmetry resulted in higher RI than the original method. In order to examine the approach's performance in detecting factual gene transfer events, the approach was applied in genome of Enterococcus faecalis V583. It is not only succeed in recovering all the seven factual horizontally transferred genes, also found that the whole segment from 7 kb upstream of gene EF2293 to 38 kb downstream of gene EF2299 was probably transferred into E. faecalis V583 genome simultaneously with the above seven genes.

Objective To investigate the clinical value of the probe melting curve analysis‐based PCR assay for the detection of three types of αT .Methods A total of 149 blood and prenatal archival DNA samples (6 of which were amniotic fluid samples)with three knownαT genes ,which included 63 carriers with Hb CS ,22 cases with Hb QS ,43 individuals with Hb WS and 1 double heter‐ozygote with Hb CS and Hb WS) as well as 20 samples with normalα‐globin gene sequence that had been confirmed by RBD com‐bined with DNA sequencing were selected to test the specificity of probe melting curve analysis by blind analysis .Results The probe melting curve analysis accurately detected 100 of the DNA samples previously characterized by S RBD combined with DNA sequencing .Conclusion Probe melting curve analysis‐based PCR assay for the detection ofαT is featured with rapidity and accuracy and can be applied to clinical and prenatal diagnosis .

Aim To investigate the apoptotic mechanism and polyamine transporter recognition of 3-nitro-naphthalimide norspermine conjugate (NNINspm),a novel naphthalimide-polyamine conjugate, in HepG2 cells.Methods The cytotoxicity of NNINspm was assessed by MTT assay.Cell cycle distribution and apoptosis were measured by flow cytometry.The protein expression of cytochrome C,14-3-3,Bad,Bcl-xL,mTOR,p70S6K,Cdk4,p27~(kip1),Akt,Caspase-3,Caspase-9 was evaluated by Western blot.The translocation of Akt was detected by high content screening (HCS) analysis.Results NNINspm induced HepG2 cells apoptosis via Akt dephosphorylation and then triggered a series of signal events, such as Bad dephosphorylation, dissociation of 14-3-3 and Bad, and then binding to Bcl-xL,which finally resulted in mitochondrial disruption,cytochrome c release and caspase cascade activation.Furthermore,the NNINspm-mediated cell cycle arrest was due to mTOR and p70S6K dephosphorylation,Cdk4 down-regulation and p27~(kip1) up-regulation.Conclusion NNINspm induces HepG2 cell apoptosis via PI_3K/Akt signal pathway.

ObjectiveTo study the curative effect and safety of low dose of urokinase (UK) combined with low molecular heparin calcium.MethodsSixty-four cases of sudden cardiac arrest patients were divided into treatment group and control group by random digits table with 32 cases each.Two groups were given cardiopulmonary resuscitation according to the 2005 international guide for cardiopulmonary resuscitation and emergency cardiovascular care.Early in the recovery,the patients in treatment group were pumped in vein with low dose of UK(200 000 U) and injected subcutaneous with low molecular heparin calcium (4100 U ) in 30 minutes.The rate of return of spontaneous circulation (ROSC),survival rate longer than 24 hours and 30 days in two groups and patients dying of bleeding or bleeding conditions in treatment group were observed.ResultsThere were 11 cases (34.4％) of ROSC in control group,and compared with 20 cases (62.5％) in treatment group,there was significant difference (P＜0.05).There were 5 cases (15.6％) of survival longer than 24 hours in control group,and compared with 13 cases(40.6％) in treatment group,there was significant difference (P ＜ 0.05 ).There were 2 cases (6.2％) of survival 30 days in control group,and compared with 8 cases (25.0％) in treatment group,there was significant difference (P＜ 0.01 ).Nobody had subcutaneous bleeding or other organ hemorrhage in control group.But there was I patient who had subcutaneous limited ecchymosis in the injection site in treatment group.The difference of fibrinogen before and after treatment in treatment group was statistically significant(P ＜ 0.01 ),but there was no significant difference in prothrombin time and platelet count before and after treatment in treatment group (P ＞ 0.05).ConclusionsIt is safe and effective in cardiopulmonary resuscitation with low dose of UK combined with low molecular heparin calcium.

Objective To evaluate the effect of neoadjuvant chemotherapy (NAC) by investigating the expression of cyclin D1 in human breast cancer before and after NAC.Methods Eighty-four cases of breast cancer were diagnosed by core biopsies.The expression level of cyclin D1 in cancer tissues was measured by immunohistochemical envision two-step method before and after NAC (pirarubicin and docetaxel regimen for 3-4 cycles).Results Complete remission (CR) occurred in 4 cases of 84 patients (4.76 ％) with 2 pathological complete response cases,partial response (PR) in 54 cases (64.29 ％),stability (SD) in 26 cases (30.95 ％) and no disease progression (PD) patients.The positive rate of cyclin D1 in cancer tissues [65.48 ％ (55/84)] was significantly decreased after NAC [39.29 ％ (33/84)] (x2 =11.55,P =0.001).In clinical level,the ease rate was significantly improved in patients whose cyclin D1 expression switched from positive [86.36 ％ (19/22)] to negative [45.45 ％ (15/33)] after NAC treatment (x2 =9.359,P =0.002).Conclusion NAC significantly decreases the expression of cyclin D1 in breast cancer tissues.Meanwhile,the ease rate is improved when cyclin D1 expression switched from positive to negative after NAC.Therefore,cyclin D1 expression can be used as an evaluation index for the efficiency of NAC.

In the present study, the apoptotic mechanism and polyamine transporter recognition of WJH-6, a novel polyamine conjugate, were investigated in K562 and HL-60 cells. The cytotoxicity of WJH-6 was assessed by MTT assay; cell cycle distribution and apoptosis were measured by flow cytometry; the protein expression of Caspase-3, Caspase-8, Caspase-9, Bid and mitochondrial membrane potential (MMP) were evaluated by high content screening (HCS) analysis; the protein expression of cytochrome c was measured by Western blotting. The results showed that WJH-6 could be recognized and transported by polyamine transporter (PAT). Furthermore, WJH-6 was able to inhibit K562 and HL-60 cells proliferation and induce apoptosis. This apoptotic effect was relative to MMP loss, cytochrome c release from mitochondria to cytoplasm and the activation of Caspase-8, Caspase-9, Caspase-3 and Bid. These results suggested that WJH-6-induced K562 and HL-60 cells apoptosis was related with mitochondrial damage.