This paper discussed the application of designing experiments from scientific research in Micro-biology courses and its effects on the teachers and students. The problems of the application of designing experiments in Microbiology courses were analyzed. The practice of the teaching reform showed that it give great advantages for the undergraduates with the enhancement of their ability on theory application and sci-entific innovation. This teaching reform could be widely popularized.

Objective:To explore the prevalence of irritable bowel syndrome (IBS) based on a community population of Shanghai and the treatment of IBS by acupuncture therapy.Methods:A population of 1 685 subjects aged between 18-80 years were randomly selected by clustered sampling from the inhabitants in Changqiao Community of Shanghai to receive a questionnaire,for understanding the symptomatology and prevalence of IBS according to modified Manning and Rome II criteria,and the voluntary IBS patients were treated by acupuncture therapy.Results:The community- based prevalence of IBS was 11.45 % and 5.04% respectively according to modified Manning criteria and Rome II criteria.The ratio of male and female was 0.77:1 in IBS patients and the proportion was 10.62% and 12.19% respectively.But there were no significant difference in prevalence between different age groups (P＞0.05) .IBS is more common in subjects aged between 45-65 years (38.86%).Regarding the 62 IBS patients treated by electroacupuncture (EA),the total effective rate in Tianshu (ST 25) group (n=32) was 84.38%,and it was 56.67% in Daheng (SP 15) group (n=30),which showed a significant difference.(P＜0.05).Conclusion:IBS is a commonly encountered disorder in Changqiao Community of Shanghai and should be taken into consideration for human welfare,disease prevention and further epidemical investigation.Acupuncture treatment is recommended for treating IBS due to its satisfactory therapeutic effect.

DNA, Viral ; chemistry ; Humans ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Torque teno virus ; classification ; genetics

OBJECTIVETo explore therapeutic effects of emergency medial malleolus osteotomy for the treatment of fractures of talar neck and dislocation of talar body.

METHODSFrom 1995. 6 to 2007. 10, among 24 patients with fractures of talar neck and dislocation of talar body, 18 patients were male and 6 patients were female, ranging in age from 28 to 58 years (mean 35.4 years). The duration from injury to the emergency ward ranged from 0.5 to 12 h. All the patients were treated in 5 hours after hospitalization with emergency medial malleolus osteotomy and internal fixation. Firstly, osteotomy was made above the medial malleolus tip; Secondly, the medial malleolus was turned over downward to uncover the talus; Then, the fracture of talus can be reduced in direct visidn.

RESULTSAll the patients were followed up ranged from 6 to 60 months. According to Kenwright evaluation standards, 18 patients obtained an excellent results, 4 good and 2 fair.

CONCLUSIONIt is easy and clearly to perform medial malleolus osteotomy. The blood circulation of talus is preserved. So it is an effective method to treat the fractures of talar neck and dislocation of talar body.

Adult ; Bone Screws ; Female ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Osteotomy ; methods ; Talus ; injuries ; surgery ; Treatment Outcome

To search for potential antitumor drugs with potent efficiency and low toxicity, a novel 1,4,7-triazacyclodecane and its platinum (II) complex were synthesized. These compounds were characterized by elemental analysis, IR, 1H NMR, 13C NMR, MS spectra, thermoanalysis and conductivity measurement. Antitumor activity study indicated these compounds had strong antitumor activity in vitro to some extent. Inhibition of human liver tumor of CA was examined by antitumor rate and growth rate, complex C showed inhibition activity on transplanting-tumor growth of CA, 12 mg x kg(-1) was as potent as cisplatin, its ID50 was 853.6 mg x kg(-1).
Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Screening Assays, Antitumor ; Female ; Humans ; Inhibitory Concentration 50 ; Liver Neoplasms ; pathology ; Male ; Mice ; Neoplasm Transplantation ; Organoplatinum Compounds ; chemical synthesis ; chemistry ; pharmacology ; Random Allocation

OBJECTIVETo explore the molecular mechanisms involved in a pedigree with inherited coagulation factor X (FX) deficiency.

METHODSThe 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).

RESULTSThe 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).

CONCLUSIONThe FX deficiency of the propositus is caused by double heterozygous mutations IVS1 + 1G-->A and Arg347His.

Antigens ; genetics ; Base Sequence ; DNA Mutational Analysis ; Factor X ; genetics ; Factor X Deficiency ; genetics ; Female ; Heterozygote ; Humans ; Male ; Mutation ; Pedigree ; Young Adult

OBJECTIVETo analyze the phenotype and genotype of a family with inherited dysfibrinogenemia.

METHODSLaboratory tests including activated particle thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), and the activity of protein C (PC), protein S(PS) and antithrombin (AT) were conducted in the proband and 4 family members. The activity and antigen of fibrinogen in plasma were measured by functional and immunoturbidimetry assay, respectively. All the exons and exon-intron boundaries of the three fibrinogen genes were analyzed by direct sequencing.

RESULTSThe proband had normal APTT and PT, but prolonged TT. Her plasma fibrinogen levels were extremely reduced, which was also found in her mother. The sequencing results of the proband revealed heterozygous g.5678 G>A in the exon 8 of FGG gene originating from her mother, which caused Arg275His missense mutation.

CONCLUSIONDysfibrinogenemia in the family is caused by Arg275His in the beta chain of fibrinogen and it is the first report on a Chinese family with inherited dysfibrinogenemia.

Adult ; Afibrinogenemia ; blood ; genetics ; Amino Acid Substitution ; Arginine ; genetics ; DNA Mutational Analysis ; Female ; Fibrinogen ; genetics ; metabolism ; Histidine ; genetics ; Humans ; Male ; Pedigree ; Phenotype

OBJECTIVETo get stable cell line expressing B domain-deleted human FVIII (BDDhFVIII) by constructing the eukaryotic expression plasmid.

METHODSEukaryotic expression plasmid containing BDDhFVIII was constructed and transfected into HepG2 cells via electroporation. The expression and purification of the target protein was detected by Western blot.

RESULTSResults of enzyme digestion and sequence analysis demonstrated that the gene of BDDhFVIII was correctly inserted into the eukaryotic expression vector pcDNA4/v5-his. Western blot confirmed the successful expression of BDDhFVIII at the protein levels in HepG2 cells.

CONCLUSIONThe constructed eukaryotic expression vector was able to generate high level expression of human FVIII in HepG2 cells, thus could construct human blood coagulation FVIII stable cell line successfully.

Electroporation ; Factor VIII ; genetics ; Gene Expression ; Genetic Vectors ; biosynthesis ; Hemophilia A ; genetics ; Hep G2 Cells ; Humans ; Plasmids ; biosynthesis ; Recombination, Genetic

BACKGROUNDWe identified the gene mutations in two Chinese pedigree of type I hereditary protein C deficiency and type I hereditary antithrombin deficiency.

METHODSThe 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.

RESULTSThe 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.

CONCLUSIONThe 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.

Adolescent ; Child ; Female ; Fibrin ; deficiency ; Gene Deletion ; Humans ; Male ; Pedigree ; Protein C ; genetics ; Protein C Deficiency ; genetics

Congenital afibrinogenemia is a rare autosomal recessive disorder, characterized by the complete absence or extremely reduced level of fibrinogen. To analyze the phenotype and genotype of a family with inherited afibrinogenemia, laboratory studies including activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT) were tested in the proband and 9 family members. Fibrinogen (Fg) in plasma were measured by both functional and immunoturbidimetry assay. All the exons, exon-intron boundaries and promoter regions of three Fg genes were analyzed by direct sequencing. 102 healthy blood donors were used as normal control. The results showed that phenotype of the proband was diagnosed as afibrinogenemia. Compound heterozygous mutations in Fg FGB gene were detected in the proband. One was a nonsense mutation (Arg17stop) in exon 2, traced back to the proband's mother. The other was a missense mutation (Gly347Arg) in exon 7, which was from the proband' s father. It is concluded that afibrinogenemia is caused by the compound heterozygous mutations Arg17stop and Gly347Arg in the Beta beta-chain of fibrinogen.
Adult ; Afibrinogenemia ; genetics ; Amino Acid Sequence ; Base Sequence ; Child ; Codon, Nonsense ; DNA Mutational Analysis ; Female ; Fibrinogen ; chemistry ; genetics ; Heterozygote ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Pedigree ; Phenotype ; Protein Structure, Secondary ; Sequence Homology, Amino Acid