Objective To investigate the effects of trichostatin A (TSA) on Th1 and Th17 cells in the mice model of collagen induced arthritis (CIA).Methods Mice model of rheumatoid arthritis (RA) was induced in DBA/1 mice with type Ⅱ collagen.Paws were scored for histological severity of arthritis.The severity of inflammation of mice joints was evaluated by histological examination.Real time polymerase chain reaction (PCR) was used to determine mRNA of cytokines and transcriptional factors.Serum cytokine production was measured by enzyme linked immunosorbent assay (ELISA).T cell proliferation was examined by MTT method.One-way ANOVA and Student-Newman-Keuls were conducted in this study.Results The expressions of IFN-γand IL-17 mRNA of the CIA group were higher than that of the control group (8.27±0.64 vs 2.97±0.25,5.80±0.23 vs 0.70±0.26,all P＜0.01),but were inhibited significantly by TSA introduced at the onset of arthritis(6.60±0.52,2.50±0.41,all P＜0.01).Collagen specific T cell proliferation was significantly suppressed by the introduction of TSA.Increased level of IL-4 was observed in TSA treated group compared to that of CIA group(2.10±0.17 vs 1.01±0.08,P＜0.01).Conclusion Th1 and Th17 cells play crucial roles in the lesions of RA.TSA can suppress the progress of CIA by decreasing the percentage of Th1 and Th17.

Objective: To study the factors effecting the expression of the reporter green fluorescent protein (GFP) gene in the mouse embryo stem cell line R1. Methods: Three different kinds of GFP euko-expression vectors were constructed, and the expression efficiency was contrasted both at mRNA and protein levels after they were integrated into the chromosomes of host cells. Results: At protein level, the GFP expression level of the colonies transfected by the expression vector-pEF-GFP with the promoter of the peptide elongation factor (EF) were significantly higher than that of the colonies transfected by pCMV-GFP with CMV promoter and by pdCMV-GFP with double copies of CMV-GFP expression unit. There was no significant difference between the colonies transfected by pCMV-GFP and pdCMV-GFP. The detection results on mRNA level of GFP had the same tendency as that at protein level. Conclusion: (1) GFP gene expression efficiency controlled by EF promoter is distinctly higher than that by CMV promoter in NIH3T3 and R1 ES cell line.(2) A slight increase of the copy number of the foreign gene expression units in the host chromosome can not make obvious increase of its expression efficiency. (3) The vector express GFP in R1 ES cell line efficiently and stablely is obtained.

Objective: To study the factors effecting the expression of the reporter green fluorescent protein (GFP) gene in the mouse embryo stem cell line R1. Methods: Three different kinds of GFP euko-expression vectors were constructed, and the expression efficiency was contrasted both at mRNA and protein levels after they were integrated into the chromosomes of host cells. Results: At protein level, the GFP expression level of the colonies transfected by the expression vector-pEF-GFP with the promoter of the peptide elongation factor (EF) were significantly higher than that of the colonies transfected by pCMV-GFP with CMV promoter and by pdCMV-GFP with double copies of CMV-GFP expression unit. There was no significant difference between the colonies transfected by pCMV-GFP and pdCMV-GFP. The detection results on mRNA level of GFP had the same tendency as that at protein level. Conclusion: (1) GFP gene expression efficiency controlled by EF promoter is distinctly higher than that by CMV promoter in NIH3T3 and R1 ES cell line.(2) A slight increase of the copy number of the foreign gene expression units in the host chromosome can not make obvious increase of its expression efficiency. (3) The vector express GFP in R1 ES cell line efficiently and stablely is obtained.

OBJECTIVE: To explore the value of estimating chronologic age based on the grades of mandibular third molar development. To evaluate whether mandibular third molar could be used as an indicator for estimating the age under or over 18 years. METHODS: The mineralization status of mandibular third molar of 1 845 individuals aged 10 - 30 was graded and marked based on Demirjian's classification of grades reformed by Orhan. Gender difference was examined by t-test. A cubic regression model was established to analyze the correlation between third molar and chronologic age. Each grade of age cumulative distribution diagram and ROC curve was respectively performed to evaluate the relationship between third molar and the age of 18. Using Bayes discriminant analysis, an equation was established for estimating the age of 18. RESULTS: The inner-rater reliability was 0.903. Statistical analysis showed a moderate correlation between age and grade. Significant differences of both genders were found only in grade D and H (P < 0.05). Males at the grades from 1 to D and females at the grades from 1 to C were under 18 years old, and both males and females at grade H were over 18 years old. The area under the ROC curve was 0.797 (P < 0.05). CONCLUSION Third molar development shows a high correlation with age, and combined with other indicators, it can be used to estimate the age of 18.
Age Determination by Teeth/methods* ; Asian People ; Bayes Theorem ; China ; Female ; Forensic Dentistry ; Humans ; Male ; Molar, Third/diagnostic imaging* ; Radiography, Panoramic ; Reproducibility of Results ; Sex Characteristics

Objective To develop a candidate reference method for the measurement of progesterone in human serum.Methods The serum sample is mixed with the internal standard [3,4-~(13)C_2] progesterone.After extraction with n-hexane and purified by a aqueous solution of 2-Hydroxypropyl-?- cyclodextrin (HP-?-CD),the serum progesterone and labeled progesterone are converted to the 3-enol heptafluorobutyrate and analyzed by gas chromatography mass spectrometry (GC/MS) with selected ion monitoring.The concentration of serum progesterone is calculated by bracketing method.Results The results gave coefficients of variation (CVs) of 0.69% to 2.12%.The analytical recoveries ranged from 98.3% to 100.1%.The results of measuring certified reference materials of serum progesterone are agree with the target value.Conclusion The procedure for measuring progesterone in serum is a highly accurate and precise method and may be used as a candidate reference method for serum progesterone assays.

AIMTo establish a method for determinate of the inhibitory activity of angiotensin-converting enzyme inhibitor captopril by high performance capillary electrophoresis.

METHODSThe characteristic absorptive wavelength of hippuric acid determined by ultraviolet spectrophotometer is 228 nm. The method employed a melted capillary column, 50 mmol.L-1 phosphoric acid (pH 8.3) buffer solution, inject pressure 4.8 kPa, inject time 3 s, separation voltage 20 kV and detection wavelength 228 nm.

RESULTSThe reactant and resultant was separated completed within 7 min. IC50 of captopril was 0.019 mumol.L-1. Captopril is a competitive inhibitor, which was proved by enzyme reaction dynamics.

CONCLUSIONThe method was shown to be accurate, simple and rapid and can be used for determination of the inhibitory activity of captopril.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Captopril ; pharmacology ; Electrophoresis, Capillary ; methods ; Hippurates ; analysis ; Peptidyl-Dipeptidase A ; metabolism

Central randomized system (CRS) is actually a management system for actualizing process of clinical trial, which can not only reduce the factitious jamming during randomization to avoid bias, but also play an important role in drug supervision during long-term and large-scale clinical trials in which a large amount of drugs are applied. This article introduced the two main functions of the CRS, i. e. the randomization of testees and the management of drugs, as well as the key points and instructions for applying clinical research interactive voice responding system (CRIVRS) in operating the project of clinical trial for secondary prevention of myocardial infarction by Qishen Yiqi Drop Pill.
Automatic Data Processing ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Multicenter Studies as Topic ; Myocardial Infarction ; prevention & control ; Phytotherapy ; Randomized Controlled Trials as Topic ; Research Design

OBJECTIVETo study the occurrence, management and prognosis of fatal pulmonary embolism in patients underwent coronary intervention in our department.

METHODSeven patients had fatal pulmonary embolism after coronary intervention in six years, we analysis each patient by the occurrence, prognosis, management of the disease.

RESULTSDuring the last 6 years, 7 [five males, mean age (55.9 +/- 11.7) years, 5 after coronary angiography and 2 after percutaneous coronary intervention] patients developed fatal pulmonary embolism after PCI. All 7 patients presented respiratory and cardiac arrest within 24 hours post coronary intervention. Three patients died, one patient experienced brain death and another three patients survived and are alive without complication till now.

CONCLUSIONThe fatal pulmonary embolism is a scarce complication after coronary intervention with high acute mortality and satisfactory outcome for survivors.

Adult ; Aged ; Angioplasty, Balloon, Coronary ; adverse effects ; Humans ; Male ; Middle Aged ; Prognosis ; Pulmonary Embolism ; diagnosis ; etiology ; Treatment Outcome

This study was aimed to establish the approach of quantitative PCR (q-PCR) for diagnosis of invasive fungal infections (IFI) in patients with hematologic malignancies. Specimens from 40 patients with hematologic malignancies were chosen for q-PCR and galactomannan (GM) test. The 28S rRNA, a real high consensus sequence of fungi, was selected as target gene to design primer and probe. The DNA of fungal species was extracted from serum specimens. The results showed that q-PCR sensitivity, specificity, positive and negative predictive values were 0.89, 0.85, 0.89, 0.85 respectively; GM test sensitivity, specificity, positive and negative predictive values were 0.83, 0.80, 0.88, 0.73 respectively; as combined q-PCR with GM test, these values were 0.94, 0.85, 0.89, 0.92 respectively. It is concluded that the q-PCR assay can be used for early diagnosis for IFI in patients with hematologic malignancies, q-PCR combined with GM test can enhance the diagnosis sensitivity for IFI.
Adolescent ; Adult ; Aged ; Early Diagnosis ; Female ; Fungi ; Hematologic Neoplasms ; microbiology ; Humans ; Male ; Middle Aged ; Mycoses ; diagnosis ; Polymerase Chain Reaction ; methods ; Predictive Value of Tests ; Sensitivity and Specificity ; Young Adult

BACKGROUNDThe Fc receptor associated pathway might improve the immune responses against hepatitis B virus (HBV) as previously described by us. In addition, the Flt3 ligand (FL) has been reported to potentiate antigen presenting cells in vivo and may act as a potential adjuvant to boost antigen-specific immune responses. In this study, the immune efficacies of a set of fusion proteins of HBsAg and Fc and/or FL were evaluated in HBsAg transgenic mice.

METHODSThe fusion proteins composed of HBsAg and the Fc domain of murine IgG1 (HBsAg-Fc) and/or the Flt3 ligand, and yeast-derived recombinant HBsAg were used as immunogen to immunize HBsAg transgenic mice, respectively. Serum and liver HBsAg levels, serum anti-HBsAg and cytokine profile, and the activities of alanine aminotransferase (ALT)/AST were investigated after immunization.

RESULTSAfter six injections, the most pronounced decrease in serum and liver HBsAg levels was observed in the HBsAg-Fc immunized group. In addition, serum Th1 cytokines and ALT/AST activities were highest in this group, indicating an effective induction of a favorable cellular immune response. Interestingly, the fusion protein containing HBsAg-Fc and the Flt3 ligand stimulated an alternative Th1-type immune response featured with high level productions of tumor necrosis factor α (TNF-α) and monocyte chemoabstractant protein 1 (MCP-1), causing a more severe cytotoxicity in hepatocytes while showed less effective in reducing serum HBsAg level.

CONCLUSIONHBsAg-Fc is effective in eliciting both the humoral and cellular immune responses against HBsAg in HBsAg transgenic mice, which makes it a potential immunogen for the immunotherapy of chronic hepatitis B.

Animals ; Chemokine CCL2 ; metabolism ; Cytokines ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Hepatitis B Surface Antigens ; genetics ; immunology ; metabolism ; Immunity, Cellular ; immunology ; Immunity, Humoral ; immunology ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Receptors, Fc ; genetics ; immunology ; metabolism ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism