Objective To develop a high-resolution melting ( HRM ) assay for rapidly screening Gilbert syndrome ( GS) and Crigler-Najjar syndrome ( CNS) associated with UGT1A1 defects.Method Methodology was developed .Then, we applied the established method to analyze 61 clinical samples from neonatal patients with severe unexplained unconjugated hyperbilirubinemia .Neonates with known risk factors for developing hyperbilirubinemia , such as ABO hemolysis, G6PD deficiency, sepsis, hypoxic ischemic encephalopathy were excluded .Five pairs of PCR primers were designed to detect the five common mutations (G211A, C686A, C1091T, C1352T and T1456G) in Asia population.PCR and HRM Assay conditions were optimized.UGT1A1 genotyping in clinical samples was performed by using the established HRM analysis , and all results were subsequently confirmed by direct DNA sequencing .Results The mutants were readily differentiated by using HRM analysis .In this study, 42 neonates were identified with UGT1A1 mutation, and 4 different known variants were detected .Conclusion HRM analysis in this study was economical, convenient, rapid, effective for screening UGT1A1 gene mutations, which can serve as an reliable method for the clinical diagnosis of GS and CNS and the large-scale molecular epidemiological research of UGT1A1 gene-related diseases.

Clinical medicine and its teaching patterns put forward the new demand to the teaching of laboratory diagnostics. According to the teaching objectives and development tendency of laboratory diagnostics, a novel teaching pattern based on organ-system diseases was established through systematical reform measures. The course content system was reconstructed and focused on the diseases. And the professional teacher group carefully orchestrated, applied new teaching methods,such as case-based learning and problem-based learning. The independent learning on a resource sharing network platform was encouraged, and the evaluation system was innovated. The novel teaching pattern has obtained gratifying achievement, and showed a bright prospect of development.

Objective To study the association between functional genetic polymorphisms of IL-1B(T-31C,C-511T),IL-1RN and the susceptibility to gastric cancers. Methods A case-control study was conducted in 180 gastric cancer cases and 308 age-and sex-matched cancer-free controls.Genotypes were detected by PCR-restriction fragment length polymorphism(PCR-RFLP) assays,and association between genotypes,environmental factors and risk of gastric cancers were determined. Results IL-1B T-31C was in strong linkage disequilibrium with IL-1B C-511T(D'=0.862,R2= 0.721,P=0.000).Multivariate logistic regression analysis revealed that the variant genotypes of IL-1B T-31C and C-511T were not significantly associated with risks for gastric cancers(adjusted OR,0.95 and 95% CI,0.62-1.47 for IL-1B T-31C;and adjusted OR,0.85 and 95% CI,0.55-1.31 for IL-1B C-511T).The variant genotypes(1/2,2/2) in IL-1RN were associated with a non-significantly increased risks for gastric cancers(adjusted OR,1.32 and 95% CI,0.71-2.36) in all subjects and with a significantly increased risks for gastric cancers in subjects with H.pylori infection(adjusted OR,2.03 and 95%CI,1.02-4.80).Conclusion The functional genetic polymorphisms of IL-1RN may contribute to the risks of gastric cancers in high-risk population,particularly in those with H.pylori infection.

Sugarcane stem is an ideal organ for producing foreign pharmaceutical proteins and chemicals by genetic engineering.A perfect promoter driving foreign gene to express strongly and specifically in sugarcane stem is necessary for this purpose.In order to isolate a Sugarcane stem-specific promoter,a fragment of 1968bp nucleotide sequence(Ppst2a)upstream 5' of sugarcane pst2a gene,which was demonstrated to express specifically in sugarcane stem previously was isolated by using chromosomal walking.Bioinformatical analysis of this sequence shows that the sequence contains some typical elements of a promoter.To identify the stem-specific of this promoter,a construct was derived from pCAMBIA1301,which original CaMV 35S promoter was replaced by the 1968bp nucleotide sequence,and named as pCAMBIA1900.Transformations of pCAMBIA1900 and pCAMBIA1301 to leaves and stem pieces of sugarcane were carried out by using particle bombardment.The transient expression of gus showed that the gus expressed specifically in sugarcane with a little higher level compared with CaMV 35S.It is the first report that pst2a promoter is a potential stem-specific promoter which can further be used in transgenes into sugarcane.

OBJECTIVETo discuss the clinical effects of using rotating hinge prosthesis in treating primary malignant tumor of the proximal tibia.

METHODSAfter the malignant tumor of the proximal tibia was extensively resected or radically resected, rotating hinge prosthesis replacement was performed for reconstruction, while neoadjuvant chemotherapy was used in treating the osteosarcoma.

RESULTSThe average follow-up period was 4.2 years. There was no local recurrence and metastasis in 22 patients, though there was recurrence in 8 patients. The MSTS functional scores of all different items 4 years after the operations were above 3, which showed the clinical results were satisfactory.

CONCLUSIONSIt is an efficacious method for the treatment of malignant tumor of the proximal tibia by using rotating hinge prosthesis. It has the advantage of early recovery of joint function.

Adolescent ; Adult ; Arthroplasty, Replacement, Knee ; methods ; Bone Neoplasms ; surgery ; therapy ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Knee Prosthesis ; Male ; Middle Aged ; Tibia ; surgery

OBJECTIVETo develop a method for extracting cytoskeletons and cytoskeleton-associated proteins for proteomic analysis.

METHODSA subcellular sequential proteome extraction method was exploited. The extraction procedure was optimized and controlled according to observed cell morphology changes and one- and two-dimensional electrophoresis images. The extraction efficiency and selectivity were evaluated by Western blotting and mass spectrometry.

RESULTSFour extracted fractions clearly displayed distinct patterns. Western blotting detected the fraction-marker proteins FAK, integrin-β1, histone H1 and cytokeratin 19 only in their expected fractions. About 90% of the protein spots in the cytoskeleton fraction were identified by mass spectrometry as cytoskeleton and/or its associated proteins.

CONCLUSIONThe subcellular proteome sequential fractionation method facilitates the detection of proteins of low abundance and shows a high reproducibility and selectivity, and thus can serve as an ideal pre-fractionation method prior to two-dimensional electrophoresis.

Cytoskeleton ; chemistry ; Electrophoresis, Gel, Two-Dimensional ; methods ; Humans ; Proteome ; analysis ; Proteomics ; methods ; Subcellular Fractions

Objective To analyze the stage characteristics in the exercise behavior improvement of college students and explore the role of Process in the exercise behavior change based on the transtheoretical model, providing basis for the stage-matched intervention for the exercise behavior of college students. Methods There were 932 students who completed the questionnaires, from 5 universities in Shanxi Province were selected by using a stratified random cluster sampling method. Descriptive analysis was used to describe the exercise behavior of college students. Variance analysis and multivariate Logistic regression model were used to analyze the difference of the process of with stage of change among college students’ physical exercise. Multivariate variance analysis was used to analyze how personal characteristics affect process of change. Results Among all participants, 89.4% students knew the importance of physical exercise, and 29.4% students were satisfied with their physical exercise condition. The distribution of students’ physical exercise stage showed an inverted U-shape with left-side peak, and there was a significant difference between gender( 2=54.657, P<0.001). There were significant stage characteristics in the process of students' exercise behavior, gender had a significant main effects on mutual aid relation(F=7.400, P=0.07)and conscious control (F=7.778, P=0.005), gender and grade had interaction effects on social release (F=3.614, P=0.013). Conclusions The college students' exercise behavior showed the characteristics of “knowing but not to do”, which conformed to the Transtheoretical model. It is essential to develop targeted phased exercise intervention strategies according to the relationship between change of phase and change of procedure.

Humans ; Enhanced Recovery After Surgery ; Stomach Neoplasms/surgery* ; Length of Stay ; Laparoscopy ; Postoperative Complications ; Gastrectomy

OBJECTIVETo investigate the effect of simulated microgravity on the proliferation of human monocytic cells THP-1 and the expression of tissue factor (TF) mRNA.

METHODSTHP-1 cells were cultured under a simulated microgravity environment using the rotating cell culture system (RCCS). The changes in the cell proliferation after microgravity culture were assessed by cell counting and cell cycle analysis with flow cytometry. RT-PCR was used to detect the changes in the expression of TF mRNA in THP-1 cells.

RESULTSCulture under simulated microgravity resulted in a significant decrease in the cell number of THP-1 cells in comparison with that of the control cells (P<0.01). After a 24-h culture under microgravity, the G0-Gl phase cells increased from the control level of (46.57∓1.64)% to (67.64∓2.71)% (P<0.05). The cells in both groups showed a low level of TF mRNA expression in the absence of LPS stimulation. A 4-h stimulation with LPS caused up-regulated expression of TF mRNA in both cells, but the microgravity group showed a significantly smaller increase in the expression (2.301∓0.179) than the control group (9.210∓1.328) (P<0.05).

CONCLUSIONMicrogravity can inhibit the proliferation of THP-1 cells and suppress the cellular expression of TF mRNA.

Cell Proliferation ; Cells, Cultured ; Humans ; Monocytes ; cytology ; metabolism ; RNA, Messenger ; genetics ; Thromboplastin ; genetics ; metabolism ; Weightlessness ; Weightlessness Simulation

OBJECTIVETo explore the variations of gene C in hepatitis B viruses between hepatitis B patients and healthy carriers, and provide experimental evidences for analysis of virus gene mutations acting on the virus material science and response of the body to the virus.

METHODSThe virus DNA load in hepatitis B patients and healthy blood donors was investigated by real-time polymerase chain reaction (PCR). Gene sequence analysis was taken to detect gene polymorphism, and all the success samples were compaired with standard strain by DNAstar.

RESULTS(1)G Compared with standard strain, C region in all samples had mutations, there were 31 mutations in at least 2 samples (3 mutations in gene PreC and 28 mutations in gene C), including 9 missense mutations, 1 chain termination mutation and 21 synonymous mutation. Mutations nt 1827 c-->a and nt 2221 c-->t existed in all the samples, and most samples had 6 synonymous mutations. Four hepatitis B patients had mutation nt1896 g-->a, and another 4 patients had 2 mutations, namely, S87G and I97F (or 197L) in HBcAg CTL recognition episome. (2) The success ratio of amplification and sequencing of HBV DNA was closely associated with its copy numbers. In the present study, copy numbers of HBV DNA which were successfully amplified and sequenced were almost more than 40 193/ml.

CONCLUSIONSHBV genome were easily affected by nucleotide mutations, 2 residues had mutations in gene of C region, which is firstly reported, suggesting these mutations may be geographical restricted. Mutations in gene of C region may either change the structure and function of HBeAg and HBcAg, which may further induce the escape of immune clearance for HBV or influence the detection of HBsAg or HBeAg, which may creat new problems for the prevention, diagnosis and treatment of hepatitis B.

Female ; Hepatitis B ; virology ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Mutation ; Polymorphism, Genetic