Hematologic Phenotype and Genotype Analysis of Patients with Hemoglobin Variants / 中国实验血液学杂志

OBJECTIVE@#To study the hematologic and molecular features of 14 patients with hemoglobin (Hb) variants, so as to provide reference data for its laboratory screening.@*METHODS@#A total of 1 029 samples were screened by high performance liquid chromatography (HPLC) on the Bio-Rad VariantⅡHPLC system. GAP-PCR and reverse dot blot (RDB) were used to detect common mutation of α and β globin gene in Chinese. DNA sequencing for α and β globin gene was simultaneously performed in samples with abnormal spectrum peak and negative thalassemia gene.@*RESULTS@#In 1 029 samples, 10 types of structural Hb variants were detected in14 cases (1.36%), including 1 case of Hb E / β- thalassemia, 1 case of Hb E /α- thalassemia (HbH disease), 2 cases of HbG-Taipei, 2 cases of Hb Q-Thailand, 2 cases of Hb Youngstown, 1 case of Hb Guangzhou-Hangzhou, 1 case of Hb M-Boston, 1 case of Hb G-Siriraj, 1 case of Hb J-Baltimore, 1 case of Hb J-Sicilia and 1 case of Hb Tamano.@*CONCLUSION@#The occurrence of abnormal structural Hb variants with many genotypes in Shanghai is unique. Except for Hb E, Hb Youngstown, and Hb M-Boston, other types of heterozygous are normal in phenotypes, and symptoms such as hemolysis and anemia often occur when other diseases are combined.

Therapeutic efficacy of nutritional support by percutaneous endoscopic gastrostomy in critically ill patients: a self-control clinical trial

Background and Objective: Percutaneous endoscopic gastrostomy [PEG] is a procedure to provide enteral nutrition for critically ill patients. It is commonly used in clinical practice; however, the widespread use of PEG is controversial. Our objective was to evaluate the therapeutic effect of nutritional support by PEG in these critically ill patients

Methods: A total of 64 critically ill patients including 41 males and 23 females [aged 23-84] were identified by the Acute Physiology and Chronic Health Evaluation [APACHE] II scoring system during September 2004 to June 2012. The nutritional status before and after PEG was mainly assessed by the tricep skinfold thickness and serum albumin level. The nutritional status and pathological condition were assessed at 4, 8 and 12 weeks before and after PEG feeding. The assessment was according to the classical method of the human nutritional status. Follow-up was performed at one month, three months and 1.5 year after gastrostomy. Statistical analysis was performed by SPSS 11.5 software. The incidence of inhalation pneumonia and gastroesophageal regurgitation was compared by chi square [?[2]] test. P<0.05 were considered statistically significant

Results: Among the 64 patients, 9 patients died of their former diseases or related symptoms. Postoperative follow-up showed that both nutritional status and complications were improved after PEG in 55 patients [P<0.05]. The serum albumin and tricep skinfold thickness levels were significantly increased. The incidence of hypoglycemia, hypocalcemia, hypokalemia and hyponatremia were lower than pre-operation. The frequencies of complications were significantly reduced. No severe complications occurred in any patient

Conclusions: Our study confirmed that PEG was a good long-term route of nutritional supply with no serious complications for critically ill patients

Training model construction and education reform exploration of eight-year medical education / 上海第二医科大学学报

Objective To construct and implement the training model of eight-year medical education with characteristics of Shanghai Jiaotong University. Methods Based on survey,discussion and consultation,the experiences of long schooling medical education in Shanghai Jiaotong University School of Medicine were summarized.Training plan and education reform scheme were established. Results Training objective,guideline and major reform measures had been clarified.The training plan and reform scheme were under process of implementation. Conclusion The training objective of eight-year medical education should be further confirmed.The curriculum should be in accordance with the training objective,and education reform is important and necessary for the eight-year medical education.

Implementation and evaluation of Sino-French cooperation on medical education / 上海第二医科大学学报

Objective To explore the culturing strategies,curiculum provision,courses conferring methods,teaching effects as well as the associated managerial evaluations on the basis of Sino-French cooperation on medical education with the hope of summarizing helpful suggestions to Sino-Foreign cooperation on medical education. Methods The achievements of our Sino-French cooperation on medical education were analyzed and compared in the teaching models,culturing strategies along with courses conferring processes among seven-year medical students from both English-teaching and French-teaching classes. Results Our Sino-French cooperation on medical education was featured in its distinct culturing purposes and effective teaching model.Its scientifically formulated culturing strategy found its full expression in French-teaching atmosphere.The Sino-French cooperation on medical education was consistently welcomed and favorably recommended by both faculties and students. Conclusion The Sino-French cooperation on medical education has not only gained precious experience in culturing the cutting-edge medical talents with the international visions but also conduced to fulfill the goal to establish a modernized and internationalized medical school.

Prospective randomized controlled trial of bilingual education in surgery / 上海第二医科大学学报

Objective To explore the feasibility of "immersion program" in French-taught surgical lessons,as to provide multiple educational methods and practical experiences for the application of bilingual education in clinical medicine.Methods Twenty-nine senior students of French-taught class were randomly divided into group A(n=15) and group B(n=14)."Immersion program" and "transitional bilingual education" were employed for group A and group B,respectively for the first half of teaching session,and "transitional bilingual education" and "immersion program" for the second half,respectively.The differences between the two bilingual education models were compared through quiz.Results In the prior 2 of the 4 quiz,the scores of French quiz and the total scores were much higher in "immersion program" group,and there were significant differences between the two groups(P0.05). Conclusion "Immersion program" helps to improve the ability of presentation,comprehension and application of French in the precondition of equal educational content,and it will be more beneficial when accessing the "immersion program" on the basis of "transitional bilingual education".

Significance of Determination of Argyrophilic Proteins in Nucleolar Organizer Regions of Blood T Lymphocytes before and after Complete Remission in Children with Acute Leukemia / 实用儿科临床杂志

Objective To detect the argyrophilic proteins in nucleolar organizer regions(Ag-NORs) that express rDNA and rRNA proliferation of T lymphocytes before chemotherapy and after complete remission(CR) in children with primary acute leukemia(AL).Methods The argyrophilic granules area of NOR/nuclear area(I.S%) of T lymphocytes was detected by image analysis system in peripheral blood of 42 patients before chemotherapy and after CR and 30 normal children.Results I.S% in the patients before chemotherapy(5.06%?1.36%) were significantly lower than those in the healthy donors(7.51%?1.06%)(t=8.238 P0.05).Conclusion These results suggest that decrease of Ag-NORs expresses the evidence for tumour induced suppression of immune function of T cells in children with AL prior to treatment.

Studies on inherited coagulation factor VII deficiency and tissue factor abnormality in a pedigree / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the mechanism of clinical haemorrhage in an inherited coagulation factor VII (FVII) deficiency and tissue factor abnormality pedigree.</p><p><b>METHODS</b>All exons, exon-intron boundaries and the 3', 5' untranslated sequences of FVII and tissue factor (TF) genes were amplified by PCR and sequenced directly. Any mutation identified by direct sequencing was confirmed by reverse sequencing. FVII cDNA of the proband was synthesized with random primers and amplified by nest PCR.</p><p><b>RESULTS</b>55C-->T heterozygous mutation located in promoter of FVII gene was identified in the proband. The heterozygous mutation was derived from his mother. Tracing the other pedigree members found that his sister had the same heterozygous mutation and the others had wild-type FVII genes. A 9363 C-->T (Arg131Trp) heterozygous polymorphism in TF gene, which was 2.63% frequency of T allele polymorphism, was found in all of the pedigree members.</p><p><b>CONCLUSION</b>It was the first report that the -55C-->T heterozygous mutation in FVII gene and the Arg131Trp heterozygous polymorphism in TF gene explained the clinical symptom of the proband.</p>

A heterozygous point mutation G13328A in antithrombin gene causes thrombosis / 中华血液学杂志

<p><b>OBJECTIVE</b>To identify the phenotype and the gene mutation in a kindred with antithrombin (AT) deficiency.</p><p><b>METHODS</b>Immuno-nephelometry and chromogenic assay were used to detect the plasma level of AT antigen (AT: Ag) and activity (AT: A), respectively. All the seven exons and intron-exon boundaries of AT gene from the propositus were amplified by PCR and direct sequencing of the PCR pro-ducts was performed. Corresponding PCR fragments from the kindred were also sequenced directly. Megaprimer method was used to construct the mutant AT cDNA expressing vector from normal plasmid pCRII AT cDNA. The normal and mutant AT plasmid were transiently transfected into Cos-7 cells and AT: Ag was detected in supernatant and lysate of transfected cell with ELISA.</p><p><b>RESULTS</b>The plasma level of AT: Ag and AT: A for the propositus were 179 mg/L and 42.3%, respectively. A heterozygous G13328A missense mutation in exon 6 was identified, which led to the substitution of Thr (ACC) 404 for Ala (GCC). The sequencing results from the pedigree suggested that three other members also had the mutation. The level of AT:Ag in supernatant and lysate from cells transfected with mutant AT cDNA was 40% and 68% of that of normal AT cDNA transfected cells.</p><p><b>CONCLUSION</b>This is an unreported AT gene mutation in China, which causes type I hereditary antithrombin deficiency and thrombosis in the proposita.</p>

Severe hereditary coagulation factor V deficiency caused by two novel heterozygous mutations / 中华血液学杂志

<p><b>OBJECTIVE</b>To identify gene mutations of a pedigree with inherited factor V (FV) deficiency.</p><p><b>METHODS</b>The activated partial thromboplastin time (APTT), prothrombin time (PT), FV activity (FV:C) and FV antigen (FV:Ag) tests were performed for phenotypic diagnosis. The genomic DNA was extracted from the peripheral blood of the proband and all the 25 exons and their flanks of FV gene were amplified by polymerase chain reaction (PCR). The PCR products were screened by direct sequencing and the mutations were further confirmed by restriction enzyme digestion.</p><p><b>RESULTS</b>APTT, PT, TT, FV:C, FV:Ag of the proband were 249.2 s, 46.6 s, 17.9 s, 0.1% and 1.5%, respectively. FII, FVII, FVIII, FIX, FX activities, vWF and Fg were within normal ranges. Taking the GenBank Z99572 sequence as the reference, four mutations were identified in FV gene of the proband. They were a heterozygous two bases deletion in exon 13 (2238 approximately 2239delAG) introducing a frameshift and a premature stop at codon 689, and a heterozygous missense mutation in exon 23 (G6410T) resulting in the substitution of Gly for Val at codon 2079, respectively. The proband's father and mother were heterozygous for G6410T and for 2238 approximately 2239delAG, respectively.</p><p><b>CONCLUSION</b>The severe FV deficiency of the proband is caused by a frameshift mutation of 2238 approximately 2239delAG and a missense mutation of G6410T, which haven't been identified before.</p>

The inherited coagulation factor XI deficiency caused by intronic mutation IVS J-4delgttg / 中华血液学杂志

<p><b>OBJECTIVE</b>To identify gene defect in a Chinese pedigree of hereditary coagulation factor XI (FXI) deficiency.</p><p><b>METHODS</b>The peripheral blood samples were collected from the proband and her family members. The plasma PT, APTT, FXI:C and FXI:Ag were assayed. The FXI gene exons and exon-intron boundaries of the proband were amplified by PCR and then sequenced directly. The mRNA of FXI in the peripheral blood was analyzed with RT-PCR.</p><p><b>RESULTS</b>The proband and some of her family members had prolonged APTT. The plasma FXI:C and FXI:Ag of the proband, her brother and her parents were lower than 10% and 50% of the normal values, respectively. Nucleotide sequence analysis revealed that the proband and her brother had a homozygous mutation of IVS J-4delgttg in FXI gene. The mutation was inherited from her parents who were heterozygotes. The mutation was not found in 60 normal subjects. No FXI mRNA was detected in peripheral blood sample of the proband.</p><p><b>CONCLUSION</b>The IVS J-4delgttg is a novel mutation causing FXI deficiency, which may interfere with mRNA splicing.</p>

Study on the molecular mechanism of antithrombin gene C2759T (Leu99Phe) mutation causing antithrombin deficiency / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the molecular mechanism of antithrombin (AT) gene C2759T (Leu99Phe) mutation causing AT deficiency.</p><p><b>METHODS</b>A mutated AT cDNA expression plasmid ATM2759 was constructed by mega-primer method. ATM2759 and wild type AT cDNA expression plasmid ATN were transfected into COS7 cells or CHO cells by using Superfect reagent respectively for in vitro expression study and immunofluorescence assay.</p><p><b>RESULTS</b>The antigen levels of AT (AT:Ag) in the cell lysate of ATM2759 transfected COS7 cells and the cell culture supernatant were 174.97% and 35.63% of that of ATN transfected COS7 cells respectively, whereas the AT activity in the cell culture supernatant was 47.73% of the control's. Immunofluorescence analysis showed that the fluorescence intensity was significantly higher in ATM2759 transfected CHO cells than in those transfected with ATN.</p><p><b>CONCLUSIONS</b>Leu99Phe substitution may not affect the binding capacity of AT with heparin. Secretion defect and intracellular accumulation of the mutated AT protein might be the mechanisms of this mutation causing AT deficiency.</p>

FXI gene mutations in two pedigrees of congenital clotting factor XI deficiency / 中华血液学杂志

<p><b>OBJECTIVES</b>To identify the FXI gene mutations in two Chinese pedigrees of congenital factor XI deficiency.</p><p><b>METHODS</b>The peripheral blood samples were collected from the probands and their family members and the plasma FXI:C and FXI:Ag were determined. All the exons and exon-intron boundries of FXI gene were amplified with PCR and sequenced thereafter.</p><p><b>RESULTS</b>A nonsense mutation Trp228stop and two missense mutations Glu323Lys and Leu172Pro were disclosed in the two pedigrees. All mutations existed in a heterozygous state.</p><p><b>CONCLUSION</b>The FXI gene mutations Trp228stop, Glu323Lys and Leu172Pro attribute to the pathogenesis of the congenital factor XI deficiency in Chinese. The Leu172Pro is identified for the first time.</p>

Double heterozygous mutations of non-canonical splice (IVS1a + 5g > a) and His348Gln caused inherited coagulation factor VII deficiency / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the pathogenesis of inherited coagulation factor VII (FVII) deficiency.</p><p><b>METHODS</b>The diagnosis was validated by coagulant parameter assay. FVII gene mutations were analysed in the proband by DNA direct sequencing of PCR products of all exons, exon-intron boundaries and the 3', 5'untranslated sequences. The mutations were confirmed by reverse sequencing. The ectopic transcripts of RT-PCR were used to confirm the characteristics of the mutation in non-canonical splice site (IVS1a + 5g > a).</p><p><b>RESULTS</b>Double heterozygous mutations in the propositus were identified: a T to G mutation at position 10961, resulting in His348Gln substitution, a non-canonical splice site (IVS1a + 5g > a) mutation, causing the new model of splice and frameshift mutation.</p><p><b>CONCLUSION</b>Double heterozygous mutations of His348Gln and IVS1a + 5g > a were identified in a propositus, the splicing pattern of the IVS1a + 5g > a mutation was reported for the first time.</p>

Inherited coagulation factor X deficiency caused by two novel mutations in factor X gene / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the molecular mechanisms involved in a pedigree with inherited coagulation factor X (FX) deficiency.</p><p><b>METHODS</b>The activated partial thromboplastin time (APTT), prothrombin time (PT), FX activity (FX:C) and FX antigen (FX:Ag) test were adopted for phenotype diagnosis. All the 8 exons, intron/exon boundaries and the 5'untranslated regions (UTR) of the FX gene were amplified by polymerase chain reaction (PCR) from the genomic DNA extracted from the peripheral blood of the propositus. The PCR products were screened by direct sequencing. The mutation was confirmed by allele specific PCR (ASPCR).</p><p><b>RESULTS</b>The phenotype of the propositus was identified as FX deficiency (type II). Two novel FX gene mutations were detected in the propositus: one was a donor site splice mutation in intron 1 (IVS1 + 1G-->A), another was a missense mutation 1185G-->A in exon 8 (Arg347His).</p><p><b>CONCLUSION</b>The FX deficiency of the propositus is caused by double heterozygous mutations IVS1 + 1G-->A and Arg347His.</p>

Biochemical activity and gene analysis of inherited protein C and antithrombin deficiency in two Chinese pedigrees / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>We identified the gene mutations in two Chinese pedigree of type I hereditary protein C deficiency and type I hereditary antithrombin deficiency.</p><p><b>METHODS</b>The plasma level of protein C activity (PC:A), protein C antigen (PC:Ag), protein S activity, antithrombin activity (AT:A) and antithrombin antigen (AT:Ag) of propositi and two family members were detected using ELISA and chromogenic assay, respectively. All exons and intron-exon boundaries of protein C gene and antithrombin gene were analyzed by direct sequencing of the corresponding amplified PCR products in DNA from the propositus.</p><p><b>RESULTS</b>The plasma PC:A and PC:Ag of propositus 1 was 26% and 1.43 mg/dl, respectively. The PC:Ag and PC:A of his father were normal. The decreased PC:A level was seen in his mother and 4 of his maternal pedigree. PS:A and AT:A were all normal in pedigree 1 members. A C5498T heterozygous mutation in exon 3 of protein C gene, resulting in the substitution of Arg for Trp at the 15th amino acid, was identified in propositus 1 and 8 of his relatives. The plasma AT:A and AT:Ag of propositus 2 was 48.6% and 10.4 mg/dl, respectively. The reduced AT:A and AT:Ag levels were found in his father and 5 of paternal pedigree. PC:A, PC:Ag and PS:A were all in normal range. A heterozygous 13387-9G deletion in exon 6 of antithrombin gene was identified in propositus 2. This mutation introduced a frameshift and a premature stop at codon 426 and existed in 6 members of pedigree 2.</p><p><b>CONCLUSION</b>The C5498T heterozygous mutation in exon 3 of protein C gene, first reported in China, leads to type I hereditary protein C deficiency. The 13387-9G deletion, a novel mutation, can cause antithrombin deficiency and thrombosis.</p>

Inherited coagulation factor VII deficiency caused by double heterozygotic mutations Arg304Gln and Arg304Trp / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the genotypes of mutations of an inherited coagulation factor VII(F VII) deficiency pedigree.</p><p><b>METHODS</b>The diagnosis was validated by coagulant parameters. F VII gene mutations were analysed in the proband and her family members by DNA direct sequencing. The PCR fragments were cleaved by the Msp I restriction enzyme to confirm the mutations detected by sequencing was performed in this study.</p><p><b>RESULTS</b>Double heterozygous mutations at the same coding site of amino acid were detected in propositus of the pedigree: a C to T mutation at position 11348 resulting in Arg304Trp substitution combined with a G to A mutation at position 11349 resulting in Arg304Gln substitution. Her farther had a G to A mutation at position 11349 and her mother had a C to T mutation at position 11348, respectively. Both were heterozygous mutations. One of her brothers had normal genotype, the other brother and all her three offsprings had heterozygous mutations.</p><p><b>CONCLUSION</b>Double heterozygous mutations coding the same amino acid were found in a pedigree with hereditary coagulation factor VII deficiency.</p>

Identification of two novel factor XI non-sense mutation Trp228stop and Trp383stop in a Chinese pedigree of congenital factor XI deficiency / 中华血液学杂志

<p><b>OBJECTIVE</b>To identify the factor XI gene mutation in a Chinese pedigree of congenital factor XI deficiency.</p><p><b>METHODS</b>The peripheral blood samples were collected from the proband and her family members and the plasma FXI:C and FXI:Ag were assayed. All the exons and their adjacent intron sequences of factor XI were amplified with PCR and sequenced thereafter.</p><p><b>RESULTS</b>Two novel nonsense mutations TGG-->TGA (Trp228stop) and TGG-->TAG (Trp383stop) were identified in the family.</p><p><b>CONCLUSION</b>The compound heterozygous Trp228stop and Trp383stop may attribute to the pathogenesis of the congenital factor deficiency.</p>

Prothrombin deficiency resulted from a homozygous Glu29 to Gly mutation in the prothrombin gene / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the gene mutations in a pedigree with inherited prothrombin (FII) deficiency.</p><p><b>METHODS</b>The activated partial thromboplastin time (APTT), prothrombin time (PT), FII activity (FII:C) and FII antigen (FII:Ag) test were used for phenotype diagnosis. The genomic DNA was extracted from the peripheral blood of the propositus. All the 14 exons, intron/exon boundaries and the 5' and 3' untranslated regions (UTR) of the prothrombin gene were amplified by polymerase chain reaction (PCR). The PCR products were screened by direct sequencing and the mutations detected were further confirmed by restricted enzyme digestion. One hundred and three healthy blood donors were used as controls.</p><p><b>RESULTS</b>The phenotype of the propositus was prothrombin deficiency (type I). With reference to the prothrombin nucleotide sequence published by Degen & Dacie, three variations were found in the FII gene of the propositus. Among them, the novel mutation was a homozygous A601G subtitution in exon 2.</p><p><b>CONCLUSION</b>The prothrombin deficiency of the propositus is caused by a homozygous Glu29 to Gly mutation in the prothrombin gene.</p>